Title of Invention	A NOVEL ENZYME PREPARATION
Abstract	"A NOVEL ENZYME PREPARATION" An enzyme preparation exhibiting endoglucanase activity, which enzyme is a) encoded by a DNA construct comprising the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, or b) encoded by a DNA construct comprising an analogue of the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, which DNA sequence has at least 75% identity with the DNA sequence shown in SEQ ID No. 8 or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSN 10081, or c) a polypeptide which has at least 70% identity with the amino acid sequence shown in SEQ ID No. 9.

Title of Invention

A NOVEL ENZYME PREPARATION

Abstract

"A NOVEL ENZYME PREPARATION" An enzyme preparation exhibiting endoglucanase activity, which enzyme is a) encoded by a DNA construct comprising the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, or b) encoded by a DNA construct comprising an analogue of the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, which DNA sequence has at least 75% identity with the DNA sequence shown in SEQ ID No. 8 or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSN 10081, or c) a polypeptide which has at least 70% identity with the amino acid sequence shown in SEQ ID No. 9.

Full Text	The present invention relates to novel enzyme preparations cotnprising an enzyme exhibiting endoglucanase activity which performs very good in indus¬trial applications such as laundry compositions, for biopolishing of newly manufactured textiles, for provid¬ing an abraded look of cellulosic fabric or garment, and for treatment of paper pulp. Further, the invention relates to DNA constructs encoding such enzymes, a method for providing a gene encoding for such enzymes, a method of producing the enzyme preparations containing such enzymes, and the use of these enzymes for a number of industrial applications. BACKGROUND OF THE INVENTION Cellulases or cellulytic enzymes are enzymes involved in hydrolyses of cellulose. In the hydrolysis of native cel¬lulose, it is known that there are three major types of cellulase enzymes involved, namely cellobiohydrol'ase (l,4-j3-D-glucan cellobiohydrolase, EC 3.2.1.91), endo-jS-1,4-glucanase (endo-1,4-/3-D-glucan 4-glucanohydrolase, EC 3.2.1.4) and i3-glucosidase (EC 3.2.1.21). Cellulases are synthesized by a large number of microor¬ganisms which include fungi, actinomycetes, myxobacteria and true bacteria but also by plants. Especially endoglucanases of a wide variety of specificities have been identified. 'A very important industrial use of cellulytic enzymes is the use for treatment of cellulosic textile or fabric, e.g. as ingredients in detergent compositions or fabric softener compositions, for bio-polishing of new fabric (garment finishing), and for obtaining a "stone-washed" look of cellulose-containing fabric, especially denim, and several methods for such treatment have been sug¬gested, e.g. in GB-A-1 368 599, EP-A-0 307 564 and EP-A-0 435 876, WO 91/17243, WO 91/10732, WO 91/17244, PCT/DK95/000108 and PCT/DK95/00132. Another important industrial use of cellulytic enzymes is the use for treatment of paper pulp, e.g. for improving the drainage or for deinking of recycled paper. Especially the endoglucanases _(EC No. 3.2.1.4) constitute an interesting group of hydrolases for the mentioned industrial uses. Endoglucanases catalyses endo hydrolysis of l,4-i3-D-glycosidic linkages in cellulose, cellulose derivatives (such as carboxy methyl cellulose and hydroxy' ethyl cellulose), lichenin, /3-l,4 bonds in mixed (3-1,3 glucans such as cereal /3-D—glucans or xyloglucans and other plant material containing cellulosic parts. The authorised name is endo-l,4-^-D-glucan 4-glucano hydrolase, but the abbreviated term endoglucanase is used in the present specification. Reference can be made to T.-M. Enveri, "Microbial Cellulases" in W.M. Fogarty, Microbial Enzymes and Biotechnology, Applied Science Pub¬lishers, p. 183-224 (1983) ; Methods in Enzymology, (1988) Vol. 160, p. 200-391 (edited by Wood, W.A. and Kellogg, S.T.); Beguin, P., "Molecular Biology of Cellulose Degra¬dation", Annu. Rev. Microbiol. (1S90), Vol. 44, pp. 219-24S; Beguin, P. and Aubert, J-P,, "The biological degra¬dation of cellulose", FEMS Microbiology Reviews H (1994) p.25-58; Henrissat, B., "Cellulases and their interaction with cellulose". Cellulose (1994), Vol. 1, pp. 169-196. Fungal endoglucanases have been described in numerous publications, especially those derived from species as e.g. Fusarium oxyspoxrum, Trlchoderma reesei, Trichoderma longibrachiatum, Aspergillus aculeatus, Neocallimastix patriciarum, and e.g. from species of the genera Piromyces, Humicola, Myceliophthora, Geotricum, Penicillium, Irpex, Coprinus. For example, fungal endqgluc_anase_s; have been described by Sheppard, P.O., et al., "The use of conserved cellulase family-specific sequences to clone Cellulase homologue cDNAs from Fusarium oxysporum, Gene, (1994), Vol. 15, pp. 163-167, Saloheimo, A., et al., "A novel, small endoglucanase gene, eglSj from Trichoderma reesei iso¬lated by expression in yeast". Molecular Microbiology (1994), Vol. 13(2), pp. 219-228; van Arsdell, J.N. et al, (1937), Cloning, characterization, and expression in Saccharomyces cerevisiae of endoglucanase I from Trichoderma reesei, Bio/Technology 5: 60-64; Penttila, M. et al., (1986), "Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene". Gene 45:253-263; Saloheimo, M. et al, (1988) , •VEGIII, a new endoglucanase from Trichoderma reeseiz the chaxacterization of both gene and enzyme". Gene 63:11-21; Gonzales, R., et al., "Cloning, sequence analysis and yeast expression of the egll gene fr-ccm Tricho'derma longibrachiatum", Appl. Microbiol. Biotechnol., (1992), Vol. 38, pp. 370-375; Ooi, T. et al. "Cloning and sequence analysis of a cDNA for cellulase (FI-CMCase) from Aspergillus aculeatus", Curr. Genet., (1990), Vol. 18, pp. 217-222; Ooi, T. et al, "Complete nucleotide sequence of a gene coding for Aspergillus aculeatus cellulase (FI-CMCase)", Nucleic Acids Research, (1990), Vol. 18, No. 19, p. 5884; Xue, G. et al., "Clon¬ing and expression of multiple cellulase cDNAs from the anaerobic rumen fungus Neocallimastix patriciairum in E. coli", J. Gen. Microbiol., (1992), Vol. 138, pp. 1413- 1420; -Xue, C. et al., "A novel polysaccharide hydrolase cDNA (celD) from Neocallimastix patriciarum encoding three multi-functional catalytical domains with high endoglucanase, cellobiohydrolase and xylanase activ¬ities", J. Gen. Microbiol., (1992), Vol. 138, pp. 2397-2403; Zhou, L. et al., "Intronless celB from the anaerobic fungus Neocallimastix patriciarum encodes a modular family A endoglucanase", Biochem. J., (1994), Vol. 297, pp. 359-364; Dalboge, H. and Heldt-Hansen, H.P., "A novel method for efficient expression cloning of fungal enzyme genes", Mol. Gen. Genet., (1994), Vol. 243, pp. 253-260; Ali, B.R.S. et al., "Cellulases and hemicellulases of the anaerobic fungus Piromyces consti¬tute a multiprotein cellulose-binding complex and are encoded by multigene families", FEMS Microbiol. Lett., (1995), Vol. 125, No. 1, pp. 15-21. Further, the DNA Data Bank of Japan (DDBJ database publicly available at Internet) comprises two DNA sequences cloned from Penicillium janthinellum encoding endoglucanases (cloned by A. Koch and G. Mernitz, respectively) and a DNA sequence cloned from Humicola qrisea var. thermoidea encoding an endoglucanase (cloned by T. Uozumi). Two endoglucanases from Macrophomina phaseolina have been cloned and sequenced, see Wang, H.Y- and Jones, R.W.: "Cloning, characterization and functional expression of an endoglucanase-encoding gene from the phytopathogenic fungus Wacropho/nina phaseolina" in Gene, 158:125-128, 1995, and Wang, H.Y. and Jones, R.W.: "A unique endoglucanase-encoding gene cloned from the phytopathogenic fungus Macrophomina phaseolina" in Applied And Environmental Microbiology, 61:2004-2006, 199 5. One of these endoglucanases shows high homology to the egl3 endoglucanase from the fungus Trichoderma reesei, the other shows homology to the egll from the microbial phytopathogen Pseudomonas solanacearum indicat¬ing that both endoglucanases belong to family 5 of glycosyl hydrolases (B. Henrissat, Biochem J 280:309-316 (ig^l))"^ Filament-specific expression of a cellulase gene-in the dimorphic fungus Ustilago maydis is disclosed in Schauwecker, F. et al. (1995). wo 91/17243 (Novo Nordisk A/S) discloses a cellulase pre¬paration consisting of a homogenous endoglucanase compo¬nent immunoreactive with an antibody raised against a highly purified 43 kDa endoglucanase derived from Humicola insolens, DSM 1800; WO 91/17244 (Novo Nordisk A/S) discloses a new (hemi)cellulose degrading enzyme, such as an endoglucanase, a cellobiohydrolase or a /3-glucosidase, which may be derived from fungi other than Trichoderma and Phanerochaete; WO 93/20193 discloses an endoglucanase derivable from Aspergillus aculeatus; WO 94/21801 (Genencor Inc.) concerns a cellulase system iso¬lated from Trichoderma longibrachiatum exhibiting endoglucanase activity; WO 94/26880 (Gist Brocades N.V.) discloses an isolated mix±ure of cellulose degrading enzymes, which preferable are obtained from Trichoderma, Aspergillus or Disporotrichum, comprising endoglucanase, cellobiohydrolase, and xyloglucanase activity; and WO 95/02043 (Novo Nordisk A/S) describes an enzyme wirh endoglucanase activity derived from Trichoderma harzianum, which can be used for a number of purposes including e.g. degradation or modification of plant cell walls. It is also known that cellulases may or may not have a cellulose binding domain (a C^BD) . The CBD enhances the binding of the enzyme to a cellulose-containing fiber and increases the efficacy of the catalytic active part of the enzyme. There is an ever existing need for providing novel cellulase enzyme preparations which may be used for applications where cellulase, preferably an endoglucanase, activity is desirable. The object of the present invention is to provide novel enzyme preparations Jiaving substantial cellulytic activ¬ity at acid, neutral or alkaline conditions and improved performance in paper pulp processing, textile treatment, laundry processes or in animal feed; preferably novel cellulases, more preferably well-performing endoglucana-ses, which are contemplated to be producible or produced by recombinant techniques. SUMMARY OF THE INVENTION Surprisingly, it has been found that a group of endoglucanases having certain unique characteristics per¬form very good in those industrial applications for which endoglucanases are conventionally used. These unique characteristics can be described in terms of conserved regions of the amino acid sequence of the enzyme protein and the inventors have found that cellulytic enzymes, i.e. enzymes exhibiting cellulytic activity, having cer¬tain conserved regions arTe very effective e.g. in the treatment of laundry, in the treatment of newly manufac¬tured textile, in the treatment of papermaking pulp. Accordingly, in its first aspect the preseait invention relates to an enzyme preparation consisting essentially of an enzyme having cellulytic activity and comprising a first amino acid sequence consisting of 14 amino acid residues having the following sequence Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp Xaa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 and a second amino acid sequence consisting of 5 amino acid residues having the following sequence Trp Cys Cys Xaa Cys 1 2 ■ 3 4 "3 wherein. in position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 8 of the first sequence, the amino acid is Arg, Lys or His; in position 9, 10, 12 and 14, respectively, of the first sequence, and in position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue in position 12 is Ser, then the amino acid residue in position 14 is not Ser, and (ii) when the amino residue in position 12 is Gly, then the amino acid residue in position 14 is not Ala-. This surprising finding of clearly recognisable conserved regions, in spite of rather prominent variations found within well-performing endoglucanase enzymes, is a result of studies of a number of fungal DNA sequences encoding for specific a^nino acid sequences of enzymes having sig¬nificant cellulytlc, especially endoglucauiase, activ¬ities. Based on this finding, a novel molecular method taylored to screen specifically for genomic DNA or cDNA characterised by encoding the enzymes of the invention has been developed. As tools for this three sets of degenerated primers were constructed. Accordingly, in its second aspect, the invention relates to a method for providing a gene encoding for cellulytic enzymes having the above conserved regions. By using this method, i.e. the set of primers for a PCR screening on genomic DNA, it was surprisingly found that DNA encoding for said enzymes can be found from a broad range of fungi, belonging to taxonomically very different organisms and inhabiting ecologically very different niches. Further, by using this method it has been possible to find DNA sequences encoding for the core regions (cata-lytically active regions or domains) of said enzymes without any attached cellulose binding domain (CBD) which core regions of enzymes would not have been selected by using conventional performance based screening approaches. The inventors have verified experimentally that the linking of a CBD region to a core region enzyme (comprising the catalytically active region or domain of the enzyme) of the present invention results in a sig¬nificantly improved performance, e.g. a fifty times higher performance, of the multiple domain enzyme. Accordingly, the present invention provides novel cellulases, especially endoglucanases, having improved performance in industial applications, either in their native form, or homo- or heterologously produced. Ln further aspects, the present invention relates to novel cellulytic enzyme preparations which are derivable from tax'onomically specific phyli, classes, orders, families, genera, and species; e.g. from Basidiomycotous Hymenomycetes, Zygomycota, Chytridiomycota; or from the classes Discomycstes, Loculoascomycetes, Plectomycetes; Archaeascomycetes, Hernias cornycetes or from the orders Di-aportales, Xylariales, Trichosphaeriales, Phyllachorales; or from the families Nectriaeae, Sordariaceae, Chaetomiaceae, Ceratostomaceae, Lasiosphaeriaceae; or from the genera Cylindrocarpon, Gliocladium, Volutella, Scytalidium, Acremonium, ot from the species Fusarium lycopersici, Fusarium passiflora, Fusarium solani, Fusarium anquioides, Fusarium poae, Humicola nigrescens', Humicola grisea, especially such consisting essentially of an enzyme comprising an amino acid sequence selected from the group consisting of the sequences Xaa Thr Arg Xaa Phe Asp Xaa 12 3 4 5 6 7; Xaa Thr Arg Xaa Tyr Asp Xaa 12 3 4 5 6 7; and Xaa Thr Arg Xaa Trp Asp Xaa 12 3 4 5 6 7 wherein, in position 4, Xaa is Trp, Tyr or Phe; and in position 1 and 7, Xaa is any of the 20 naturally occurring ainino acid residues. More specifically, the enzyme preparation of the invention is preferably obtainable from the taxonoraically specific phyli, classes, orders, families, genera, and" species mentioned above which all produce endoglucanases comprising a first peptide consisting of 13 amino acid residues having trhe following sequence Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp 1 2 3 4 5 6 7 8 9 10 11 12 13 and a second peptide consisting of 5 amino acid residues having the fo11 awing sequence Trp Cys Cys Xaa Cys 12 3 4 5 wherein, in position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; in position S of the first sequence, the amino acid is Arg, Lys or His; in position 9, 10, and 12, respectively, of the first sequence, and in position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues. In yet further aspects, the present invention provides DNA constructs comprising a DNA sequence encoding an en¬zyme exhibiting endoglucanase activity, which DNA sequence comprises the DNA sequence shown in SEQ ID Nos. 1, 4, 6, 8, 10, 12, 16, and 19, respectively, or analogues thereof. The present invention also relates to a recombinant expression vector comprising a DNA construct of the invention; to a cell comprising a DNA construct or a recombinant expression vector of the invention; to a method of producing an enzyme, e.g a recombinant enzyme, of the invention; to a method of providing colour clari¬fication of laundry by using the enzyme of the invention; to a laundry composition comprising the enzyme of the invention; to uses of the enzyme of the invention for degradation or modification of plant material, e.g. cell walls, for treatment of fabric, textile or garment, for treatment of paper pulp; and to an enzyme preparation which is enriched in an enz^Tiie of the present invention. THE DRAVTENGS Figure l is an alignment, of the deduced encoded amino acid sequences of Arrrp-anniixa sp. (I), Volutella colletotrichoides, Cx'inipellis scabella, Acremonium sp. (II), Myceliophthora thermophila, Thielavia terrestris, Macrophomina phaseolina. The Pileup program (Feng and Doolittle, 1987) {GCG package, version 8.0) was used to create the best alignment. Identical residues in at least four sequences (boxed) are indicated around the corre¬sponding amino acids. Figure 2 Figure 2a,b,c illustrates the taxonomic classification within the Fungal Kingdom of all the microorganisms dis¬closed herein as being capable of producing said enzyme preparations and- enaymes of the invention. The taxonomic classification used herein builds primarily on the system used in the :NIH Data Base (Entrez, version spring 1996) available on World Wide Web: (http: //www3 .ncbi.nliti.nih.gov/htbin/ef/entrezTAX) . Regarding classification of organisms which are not included in the Entrez data base the following generally available and world wide accepted reference books have been used: For Ascomycetes: Eriksson, O.E. & Hawksworth, D,L.: Systema Ascomycetum vol 12 (1993). For Basidiomycetes: Jiilich, W. : Higher Taxa of Basidiomycetes, Bibliotheca Mycologia 85, 485pp (1981). For Zygomycetes: O'Donnell, K.: Zygomycetes in culture, University of Georgia, US, 257pp (1979) . General mycological reference books: Hawksworth, D.L., Kirk, P.M., Sutton, B.C. and Pegler, D.N.: Dictionary of the fungi. International Mycological Institute, 616pp (1995); Von Arx, J.A. : The genera of fungi spomlccting in culture, 424pp (1981). The taxonomic implacement of the genus Humicola has untill recently remained unclear- However, studies of 18SRNA of a wide selection of Sordariales has given strong indications of referring Humicola to the order Sordariales (Taylor, Clausen & Oxenboll, unpublished). Further these data suggests Humicola along with Scytalid-ium to be only rather distantly related to the families Sordariaceae, Chaetomiaceae, Ceratostomataceae, and Lasiosphaeriaceae. In accordance with the above Humicola and Scytalidium are here placed within the order Sordariales, with unclassified Family. Figure 3 is an alignment of the deduced partial amino acid sequences derived from a selection of 26 of the 46 microorganisms described in Example—5T—' DETAILED DESCRIPTION OF THE INVENTION In the present context, the term "the 20 naturally occuring amino acid residues" denotes the 20 amino acid residues usually found in proteins and conventionally known as alanine (Ala or A), valine (Val or V), leucine (Leu or L), isoleucine (lie or I), proline (Pro or P), phenylalanine (Phe or F), tryptophan (Trp or W), methionine (Met or M), glycine (Gly or G), serine (Ser or S), threonine (Thr or T), cysteine (Cys or C), tyrosine (Tyr or Y), asparagine (Asn or N), glutamine (Gin or Q), aspartic acid (Asp or D), glutamic acid (Glu or E), lysine (Lys or K), arginine (Arg or R), and histidine (His or H). According to the present invention there is provided novel well-performing endoglucanases comprising consei-ved amino acid sequence regions, especially a first amino acid sequence consisting of 14 amino acid residues having the following sequence Thr Arg Xaa Xaa Asp Cys Cys Xaa Xara Xaa Cys Xaa Trp Xaa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 and a second amino acid sequence consisting of 5 amino acid residues having the following sequence Trp Cys Cys Xaa Cys 12 3 4 5 wherein, in position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; m position 8 of the first sequence, the ammo acid is Arg, Lys or His; in position 9, 10, 12 and 14, respectively, of the first sequence, and in position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue in position 12 is Ser, then the amino acid residue in position 14 is not Ser, and (ii) when the amino residue in position 12 is Gly, then the amino acid residue in position 14 is not Ala. Preferably, the enzyme of the invention is of microbial origin, i.e. obtainable from a microorganism such as a fungus. In a preferred embodiment, the amino acid residue in position 9 of the first sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cysteine, asparaglne, glutamine, tyrosine, serine, methionine and tryptophan, preferably from the group consisting of proline and threonine. In another preferred embodiment, the amino acid r^idne in position 10 of the first sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cysteine, asparagine, glutamine, tyrosine, serine, methionine and tryptophan, preferably serine. In yet another preferred embodiment, the amino acid resi¬ due in position 12 of the first sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cy¬ steine, asparagine, glutamine, tyrosine, serine, methionine and tryptophan, preferably from the group con¬ sisting of alanine and glycine. ' In yet another preferred embodiment, the amino acid resi- due in position 14 of the first sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cy¬steine, asparagine, glutamine, tyrosine, serine, methionine, tryptophan, glutamic acid and aspartic acid, preferably from the group consisting of proline, threonine, serine, alanine, glutamic acid and aspartic acid. Preferably, the amino acid residue in position 4 of the second sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cysteine, asparagine, glutamine, tyrosine, serine, methionine, tryptophan, glutamic acid and aspartic acid, more preferably from the group con¬sisting of alanine, glycine, and glutcunine. Examples of more preferred embodiments are such wherein, in the first sequence, the amino acid residue in position 3 is tyrosine; or the amino acid residue in position 4 is tryptophan; or the amino acid residue in position 8 is lysine. In an especially preferred embodiment,' the enzyme of the invention has a first sequence comprising the amino acid sequence Thr Arg Tyr Trp Asp Cys Cys Lys Pro Ser Cys Ala Trp 1 2 3 4 5 6 7 8 9 10 11 12 13 , or the amino acid sequence Thr Arg Tyr Trp Asp Cys Cys Lys Thr Ser Cys Ala Trp 1 2 3 4 5 6 7 8 9 10 11 12 13 , or the amino acid sequence Thr Arg Tyr Trp Asp Cys Cys Lys Pro Ser Cys Gly Trp 1 2 3 4 5 6 7 8 9 10 11 12 13 . In a second aspect, the present invention provides a method for providing a microbial strain comprising a gene encoding such an enzyme which method comprises hybridization, e.g. PCR amplification, under standard conditions with an oligonucleotide derived from any of the conserved regions, illustrated in Fig.l. A useful oligonucleotide comprises a nucleotide sequence encoding at least a pentapeptide comprised in a peptide selected from the group consisting of a. Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp Xaa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 the amino acid in position 3 or 4 being Trp, Tyr or Phe; the amino acid in postion 8 being Arg, Lys or His; he amino acid in position 9, 10, 12 and 14, respectively, being any of the 20 naturally occurring amino acid resi¬dues ; and b. Trp Cys Cys Xaa Cys Tyr 12 3 4 5 6 the amino acid in position 4 being any of the 2 0 natural¬ly occurring amino acid residues ; and c. Xaa Pro Gly Gly Gly Xaa Gly Xaa Phe 123456789 the amino acid in position 1 being Met or lie; the amino acid in position 6 and 8, respectively, being Leu, lie or Val; and d. Gly Cys Xaa Xaa Arg Xaa Asp Trp Xaa 123456789 the amino acid in position 3 being any of the 20 natural¬ly occurring amino acid residues; the amino acid in position 4 and 6, respectively, being Trp, Tyr or Phe; and the amino acid in position 9 being Phe or Met; The useful oligonucleotides also comprises nucleotide sequences complementary to the sequences mentioned. In a preferred embodiment of the method of the invention, the oligonucleotide corresponds to a PCR primer selected from the PCR primers sense: 5 ' -CCCCAAGCTTACIVCGITA^/TTGGGA^VTTG^/TTG^/TAA^/G^/CC-S ' antisense 1: 5'- CTAGTCTAGATA'^/GCAIGC'^/GCA^/GCACC -3 ' ; antisense 2: CTAGTCTAGAAAIA'^/G/'^ICCIA'^/^/°ICCICCICCIGG -3 ' ; and antisense 3: 5 ' - CTAGTCTAGAIAACCA^/GTCA'^/G^/TAIC°/TCC-3 . In a third aspect, the present invention provides an enzyme preparation which essentially consists of an enzyme having cellulytic activity and having the conserved regions found by the inventors, i.e. which com¬prises a peptide consisting of 7 amino acid residues ha¬ving the following sequence Xaa Thr Arg Xaa Phe Asp Xaa 12 3 4 5 6 7; Xaa Thr Arg Xaa Tyr Asp Xaa 1 2 3 4 ' 5 6 7 ; and Xaa Thr Arg Xaa Trp Asp Xaa 12 3 4 5 6 7 wherein, in position 4, Xaa is Trp, Tyr or Phe; and in position 1 and 7, Xaa is any of the 20 naturally occurring amino acid residues. This enzyme is obtainable from a strain belonging to Basidiomycotous Hymenomycetes (see Fig.2 ), more preferably to the group consisting of the orders Agarica-les, Auriculariales, and Aphyllophorales, even more preferably to the group consisting of the families Exidiaceae, Tricholomataceae, Coprinaceae, Schizophyllaceae, Bjerkanderaceae and Polyporaceae, especially to the group consisting of the genera Exidia, Crinipellis, Fames, Panaeolus, Trametes, Schizophyllum, and Spongipellis. Specific examples are endoglucanases obtainable from a strain belonging to the group consisting of the species Exidia glandulosa, Crinipellis scabella, Fomes fomentarius, and Spongipellis sp., more specific examples being Exidia glandulosa, CBS 277.96, Crinipellis scabella, CBS 280.96, Fames fomentarius, CBS 276.96, and Spongipellis sp., CBS 2 83.96. Exidia glandulosa was deposited at Centraalbureau voor Schimmelcultures, Oosterstraat 1, Postbus 273, NL-3740 AG Baarn, the Netherlands, on 12 March, 1996, under thfe deposition number CBS 277.96; Crinipellis scabella was deposited at Centraalbureau voor Schimmelcultures on 12 March, 19S-6, under the deposition number CBS 280.96, Fames fomentarius was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 276.96, and Spongipellis sp. was deposited at CentraaJLbareau voor Schimmelcultures on 12 Maxcfa, 1996, under the deposition number CBS 283.96; all deposited tinder the Budapest Treaty. The enzyme preparation of the invention is also obtain¬able from a strain belonging to Chytridiomycota, preferably from a strain belonging to the class of Chytridiomycetes, more preferably belonging to the group consisting of the order Spizellomycetales , even more preferably to the family Spizellomycetaceae, especially belonging to the genus Rhizophlyctis. A specific example is a strain belonging to the species Rhizophlyctis rosea, more specifically to Rhizophlyctis rosea, CBS 282.96. Rhizophlyctis rosea was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 28 2.96; under the Budapest Treaty. The enzyme preparation of the invention is also obtain¬able from a strain belonging to Zygomycota, preferably belonging to the class Zygomycetes, more preferably to the order Mucorales, even more preferably to the group of families consisting of Mucoraceae and Thamnidiaceae, especially belonging to the group consisting of the ge¬nera Rhizomucor, Phycomyces and Chaetostylum. Specific examples are strains belonging to the genera Rhizomucor pusillus, Phycomyces nitens, and Chaetostylum fresenii more specifically to Rhizomucor pusillus, IFO 4578, and Phycomyces nitens, IFO 4814 and Chaetostylum fresenii, NRRL 2305. Further, the enzyme preparation of the invention is also obtainable from a strain belonging to the group consist¬ing of Archaeas^cumycetes, Discomycetes, Hemiascomycetes, Loculoascomycetes, and Plectomycetes, preferably belonging to the group consisting of the orders Pezizales, Rhytismatales, Dothideales, and Eurotiales. Especially, the enzyme is obtainaJale from a strain belonging the the group consisting of the families Cucurbitariaceae, Ascobolaceae, Rhytismataceae, and Trichocomaceae, preferably belonging the the group con¬sisting of the genera Diplodia, Microsphaeropsis, Ulospora, Macrophomina, Ascoholus, Saccobolus, Penicil-lium, and Thermomyces. Specific examples are enzymes obtainable from a strain belonging the the group consist¬ing of the species Diplodia gossypina, Microsphaeropsis sp., Ulospora bilgramii, Aureobasidium sp., Macrophomlna phaseolina, Ascobolus stictoides, Saccobolus dilutellus, Peziza, Penicillium verruculosum, Penicillium chrysogenum, and Thermomyces verrucosus; more specifically Diprlodia gossypina, CBS 274.96, Ulospora bilgramii, NKBC 1444, Macrophomina phaseolina, CBS 281.96, Saccobolus dilutellus, CBS 275.96, Penicillium verruculosum, ATCC 62396, Penicillium chrysogenum, ATCC 9480, and Therwomyces verrucosus, CBS 285.96. Diplodia gossypina was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 274.96, Macrophomina phaseolina was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 281.96, Saccoholus dilutellus was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 275.96; Thermomyces verrucosus was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 285.96; all under the Budapest Treaty. Yet fujrther, the enzyme is obtainable from a strain belonging to the group consisting of the orders Dia-portales, Xylariales, Tirlchosphaeriales and Phyllachorales, preferably from a strain belonging to the group consistixjg of the families Xylariaceae, Valsaceae, and PhyllacborBLCBae, more preferably belonging to the ge¬nera Diaporthe, Colletotrichum, Nigrospora, Xylaria, NodulisporuM and Poronia. Specific examples are the specie's Diaporthe syngenesia, Colletotrichum lagenarium, Xylaria hypoxylon, Nigrospora sp., Nodulisporum sp., and Poronia punctata, more specifically Diaporthe syngenesia, CBS 278.96, Colletotrichum lagenarium, ATCC 52 609, Nigrospora sp., CBS 272.96, Xylaria hypoxylon, CBS 284.96. Diaporthe syngenesia was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 278.96, Nigrospora sp. was deposited at Centraalbureau voor Schimmelcultures on 12 March, 199 6, under^he—deposition number CBS 272.96, Xylaria hypoxylon was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 284.96; all under the Budapest Treaty. The enzyme is also obtainable from the unidentified fungal, mitosporic, coleomycetous deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition numbers CBS 270.96, CBS 271.96 and CBS 273.96, respectively, under the Budapest Treaty. The enzyme is also obtainable from a strain belonging to the group consisting of the genera Cylindrocarpon, Gliocladium, Nectria, Volutella, Sordaria, Scytalidium, Thielavia, Syspastospora, Cladorrhinum, Chaetomium, Myce-liphthora and Acremonium, especially from a strain belon¬ging to the group consisting of the species Cylin¬drocarpon sp., Nectria pinea, Volutella colletotrichoides, Sordaria fimicola, Sordaria macrospora, Thielavia terrestris, Thielavia thermophila, Syspastospora boninensis, Cla^dorrhinxrm foecundissimum, Chaetomium muronrm, Chaetomium virescens, Chaetomium brasiliensis, Chaetomium cunicolojrum, Myceliophthora thermophila, Gliocladium catenulatum, Scytalidium thermophila, and Ai:ri^nir>nJmn sp., more specifically from Nectria pinea, CBS 273.96, Volutella colletotrichoides, CBS 400.58, Sordaria fimicola, ATCC 52644, Sordaria macrospora, ATCC 60255, Thielavia terrestris, NRRL 8126, Thielavia thermophila, CCBS 174.70, Chaetomium murorum, CBS 163.52, Chaetomium virescens, CBS 547.75, Chaetomium brasiliensis, CBS 122.65, Chaetomium cunicolorum, CBS 799.83, Syspastospora boninensis, NKBC 1515, Cladorrhinum foecundissimum, ATCC 62373, Myceliophthora thermophila, CBS 117.65, Scytalidiim thermophila, ATCC 28085, Gliocladium catenulatum, ATCC 10523, and Acremonium sp., CBS 478.94. Nectria pinea was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 279.96, and Acremonium sp. was deposited on 28 September 1994 under the deposition number CBS 478.94, both according to the Budapest Treaty. The enzyme is also obtainable from a strain belonging to the group consisting of the species Fusarium solani, Fusarium anguioides, Fusarium poae, Fusarium oxysporum ssp. lycopersici, Fusarium oxysporum ssp. passiflora, Humicola nigrescens and Humicola grisea, especially Fusarium oxysporum ssp lycopersici, CBS 645.78, Fusarium oxysporum ssp passiflora, CBS 744.79, Fusarium solani, IMI 107.511, Fusarium anguioides, IFO 44 67, Fusarium poae, ATCC 60883, Humicola nigrescens, CBS 819.73 and Humicola grisea, ATCC 22726. It is to be noted that Humi¬cola grisea is different from Humicola grisea var. thermoidea. In a preferred embodiment, the enzviae preparation of the invention is derived from the disclosed classes, orders, families, genera and species and essentially consists of an enzyme comprising a first peptide consisting of 13 amino acid residues having the following sequence Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp 1 2 3 4 5 67 8 9 10 11 12 13 and a second peptide consisting of 5 amino acid residues having the following sequence Trp Cys Cys Xaa Cys 12 3 4 5 wherein, in position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 8 of the first sequence-;—Lhi=i 'amino acid is Arg, Lys or His; in position 9, 10, and 12, respectively, of the first sequence, and in position 4 of the second sequence. coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively; or b) an analogue of the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, which i) is homologous with the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, ii) hybridizes with the same oligonucleotide probe as the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequ¬ence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10O31, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, iii) encodes a polypeptide which is homologous with the polypeptide encoded by a DNA sequence comprising the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequ¬ence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, iv) encodes a polypeptide which is immxmologically re¬active with an antibody raised against the purified endoglucanase encoded by the DNA sequence shown in SEQ ID No 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequence obtainable, from the plasmid in Saccharomyces cerevisiae DSM 977 0, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively. Escherichia coli DSM 10512 was deposited under the Buda¬pest Treaty on 2 February, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10511 was deposited under the Buda¬pest Treaty on 2 February, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10571 was deposited under the Buda¬pest Treaty on 6 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10576 was deposited under the Buda¬pest Treaty on 12 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braxonschweig, Germany) . Escherichia coli DSM 10583 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10584 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10585 was deposited under the Buda-pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlufvj-von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10586 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10587 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Escherichia coli DSM 10588 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Saccrharomyces cerevisiae DSM 9770 was deposited under the Budapest Treaty on 24 February, 1995, at DSM (Deutsche ScTncralung von Mikrrrorg^nismeTi und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). SacL.hc±iomyces cerevisiae DSM 100B2 was deposited under the. Budapest Treaty on 3 0 June, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Saccharomyces cerevisiae DSM 10080 was deposited under the Budapest Treaty on 30 June, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). Saccharomyces cerevisiae DSM 10081 was deposited under the Budapest Treaty on 30 June, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany). The DNA construct of■the invention relating to SEQ ID No. 1 can be isolated from or produced on the basis of a DNA library of a strain of Myceliophthora, in particular a strain of M. thermophila, especially M. thermophila, CBS 117.65. The DNA constructs of the invention relating to SEQ ID Nos. 4 and 6 can be isolated from or produced on the basis of a DNA library of a strain of Acremonium, especially Acremonium sp., CBS 478,94. The DNA construct of the invention relating to SEQ ID No. 8 can be isolated from or produced on the basis of a DNA library of a strain of Thielavia in particular a strain of Thielavia terrestris, especially Thielavia terrestris, NRRL 8126. The DNA construct of the invention relating to SEQ ID No. 10 can be isolated from or produced on the basis of a DNA library of a strain of Macrcp2io2±na, in particular a strain of M. phaseolina, especially M.phaseolina, CBS 281.96. The DNA construct of the invention relating to SEQ ID No. 12 can be isolated from or produced on the basis of a DNA library of a strain of .Crinipellis, in particular a strain of C. scaJbeila, especially C.scabella, CBS 280.96. The DNA construct of the invention relating to SEQ ID No. 19 can be isolated from or produced on the basis of a DNA library of a strain of Sordaria, in particular a strain of Sordaria fimicola. In the present context, the "analogue" of the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, is intended to indicate any DNA sequence encoding an enzyme exhibiting endoglucanase activity, which has any or all of the properties i)-iv). The analogous DNA sequence a) may be isolated from another or related (e.g. the same) organism producing the enzyme with endoglucanase activity on the basis of the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, e.g. using the procedures described herein; the homologue may be an allelic variant of the DNA sequence comprising the DNA sequences shown herein, i.e. an alternative form of a gene that arises through mutation; mutations can be silent (no change in the encoded enzyme) or may encode enzymes having altered amino acid sequence; the homologue of the present DNA sequence may also be a genus or spe¬cies homologue, i.e. encoding an enzyme with a similar activity derived from another species, b) may be constructed on the basis of the DNA sequences shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, e.g. by introduction of nucleotide substitutions which do not give rise to another amino acid sequence of the endoglucanase encoded by the DNA sequence, but which correspond to the codon usage of the host organism intfrnded for production of the enzyme, or by introduction of nucleotide substitutions which may give rise to a different amino acid sequence. However, in the latter case amino acid changes are preferably of a minor nature, that is conservative amino acid substitu¬tions that do not significantly affect the folding or activity of the protein, small deletions, typically of one to about 30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue, a small linker peptide of up to about 20-25 residues, or a small extension that facilitates purifica¬tion, such as a poly-histidine tract, an antigenic epitope or a bind-ing domain. See in general Ford et al., Protein Expression and Purification. 2: 95-107, 1991. Examples of conservative substitutions are within the group of basic amino acids (such as arginine, lysine, histidine), acidic amino acids (such as glutamic acid and aspartic acid), polar amino acids (such as glutamine and asparagine), hydrophobic amino acids (such as leucine, isoleucine, valine), aromatic amino acids (such as phenylalanine, tryptophan, tyrosine) and small amino acids (such as glycine, alanine, serine, threonine, methionine). It will be apparent to persons skilled in the art that such substitutions can be made outside the regions criti¬cal to the function of the molecule and still result in an active polypeptide. Amino acids essential to the activity of the polypeptide encoded by the DNA construct of the invention, and therefore preferably not subject to substitution, may be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutage-nersls (Cunninghara and Wells, Science 244. 1081-1085, 1989). In the latter technique mutations are introduced at every residue in the mol¬ecule, and the resultamrt mutant molecules are tested for biological (i.e. endoglucanasa) activity to identify amino acid residues that are critical to the activity of the molecule. Sites of substrate-enzyme interaction can also be determined by analysis of crystal structure as determined by such techniques as nuclear magnetic reson¬ance, crystallography or photoaffinity labeling. See, for example, de Vos et al.. Science 255: 306-312, 1992; Smith et al., J. Mol. Biol. 224: 899-904, 1992; Wlodaver et al., FEBS Lett. 3 09: 59-64, 1992. The endoglucanase encoded by the DNA sequence of the DNA construct of the invention may comprise a cellulose bind¬ing domain (CBD) existing as an integral part of the encoded enzyme,"or a CBD from another origin may be introduced into the endoglucanase enzyme thus creating an enzyme hybride. In this context, the term "cellulose- binding domain" is intended to be understood as defined by Peter Tomme et al. "Cellulose-Binding Domains: Classi¬fication and Properties" in "Enzymatic Degradation of Insoluble Carbohydrates", John N. Saddler and Michael H. Penner (Eds.), ACS Symposium Series, No. 618, 1996. This definition classifies more than 120 cellulose-binding domains (CBDs) into 10 families (I-X), and it demon¬strates that CBDs are found in various enzymes such as cellulases, xylanases, mannanases, arabinofuranosidases, acetyl esterases and chitinases. CBDs have also been found in algae, e.g., the red alga Porphyra purpurea as a non-hydrolytic polysaccharide-binding protein, for refer¬ence see Peter Tomme et al., supra. However, most of the CBDs are from cellulases and xylanases- CBDs are found at the N or C termini of proteins or are internal. Enzyme hybrids are known in the art, see e.g. WO SO/00609 and WO 95/167 82, and may be prepared by transfoi"iaing into a host cell a DNA construct comprising at. least: a fragment of DNA encoding the cellulose-binding domain ligated, with or without a linker, to a DNA sequence encoding the enzyme of interest and growing tlie hosrt cell to express the fused gene. Enzyme hybrids may be described by the following formula: CBD - MR - X, wherein CBD is the N-terminal or the C-terminal region of an amino acid sequence corresponding to at least the cellulose-binding domain; MR is the middle region (the linker), and may be a bond, or a short linking group preferably of from about 2 to about 100 carbon atoms, more preferably of from 2 to 4 0 carbon atoms; or is preferably from about 2 to about 100 amino acids, more preferably of from 2 to 40 amino acids; and X is an N-terminal or C-terminal region of a po^lypeptide encoded by the DNA sequence of the invention. The homology referred to in i) above is determined as the degree of identity between the two sequences indicating a derivation of the first sequence from the second. The homology may suitably be determined by means of computer programs known in the art such as GAP provided in the GCG program package (Needleman, S.B. and Wunsch, CD., Jour¬nal of Molecular Biology, 48_: 443-453, 1970). Using GAP with the following settings for DNA sequence comparison: GAP creation penalty of 5.0 and GAP extension penalty of 0.3, the coding region of the DNA sequence exhibits a degree of identity preferably of at least 60%, more preferably at least 65%, more preferably at least 70%, even more preferably at least 80%, especially at least 90%, with the coding region of the DNA sequence shown in SEQ ID No.l, 4, 6, 8, 10, 12-, or 16, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae, DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherdchia coli, DSH 10512, DSM 10511, DSM 10571, or DSM 10576, respectively. The hybridization referred to in ii) above is intended to i nd i crat.e that the analogous UNA sequence hybridizes to the same probe as the DNA sequence encoding the endoglucanase enzyme under certain specified conditions which are described in detail in the Materials and Methods section hereinafter. The oligonucleotide probe to be used is the DNA sequence corresponding to the endoglucanase encoding part of the DNA sequence shown in SEQ ID NO 1, 4, 6, 8, 10, 12, or 16 respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae, DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively. —5^e homology referred to in iii) above is determined as the degree of identity between the two sequences indicat¬ing a derivation of the first sequence from the second. :he homology may suitably be determined by means of com-5uter programs known in the art such as GAP provided in :he GCG program package (Needleman, S.B. and Wunsch, :.D., Journal of Molecular Biology, AB_: 443-453, 1970). Jsing GAP with the following settings for polypeptide sequence comparison: GAP creation penalty of 3.0 and GAP extension penalty of 0.1, the polypeptide encoded by an analogous DNA sequence exhibits a degree of identity pre¬ferably of at least 55%, more preferably at least 60%, nore preferably at least 65%, even more preferably at least 70%, more preferably at least 80%, especially at least 90%, with the enzyme encoded by a DNA construct :::omprising the DNA sequence shown in SEQ ID No.l, 4, 6, 3, 10, 12, 16 or 19, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae, DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively. In connection with property iv) above it is intended to indicate an endoglucanase encoded by a DNA sequence iso¬lated txom strain Saccharomyces cerevisiae, DSM 9T70, DSM 100B2, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively, arid produced in a host organism transformed with said DNA sequence or the corresponding endoglucanase naturally produced by Myceliophthora thermophila, Acremonium sp., Thielavia terrestris, Macrophomina phaseolina, Crinipellis scabella, Volutella colletotrichoides, or Sordaria fimicola, respectively. The immunological reac¬tivity may be determined by the method described in the Materials and Methods section below. In further aspects the invention relates to an expression vector harbouring a DNA construct of the invention,—a cell comprising the DNA construct or expression vector and a method of producing an enzyme exhibiting vative amino acid substitutions that do not significantly affect the folding or activity of the protein, small deletions, typically of one to about 30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue, a small linker peptide of up to about 20-25 residues, or a small extension that facilitates purification, such as a poly-histidine tract, an antigenic epitope or a binding domain. See in general Ford et al. , Protein Expression and Purification 2: 95-107, 1991. Examples of conservative substitutions are within the group of basic amino acids (such as arginine, lysine, histidine), acidic amino acids (such as glutamic acid and aspartic acid), polar amino acids (such as glutamine and asparagine), hydrophobic amino acids (such as leucine, isoleucine, valine), aromatic amino acids (such as phenylalanine, tryptophan, tyrosine) and small amino acids (such as glycine, alanine, serine, threonine, methionine). It will be apparent to persons skilled in the art that such substitutions can be made outside the regions criti¬cal to the function of tiie laolecule and still result in an active enzyme. Amino acids essential to the activity of the enzyme of the invention, and therefore preferably not subject to substitution, may be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham, 1989). In the latter technique mutations are introduced at every residue in the molecule, and the resultant mutant molecules are tested for cellulytic activity to identify amino acid residues that are critical to the activity of the molecule. Sites of ligand-receptor interaction can also be determined by analysis of crystal structure as determined by such techniques as nuclear magnetic resonancBr;—Crystallography or photoaffinity labelling. See, for example, de Vos et al., 1992; Smith et al., 1992. Wlodaver et al., 1992. The homologue may be an allelic variant, i.e. an alterna¬tive form of a gene that arises through mutation, or an altered enzyme encoded by the mutated gene, but having substantially the same activity as the enzyme of the invention Hence mutations can be silent (no change in the encoded enzyme) or may encode enzymes having altered amino acid sequence. The homologue of the present enzyme may also be a genus or species homologue, i.e. an enzyme with a similar activity derived from another species. A homologue of the enzyme may be isolated by using the procedures described herein. Molecular screening and cloning bv polymerase chain reaction fact) Molecular screening for DNA sequences of the invention may be carried out by polymerase chain reaction (PCR) using genomic DNA or double—strained cDNA isolated from a sraitable source, such as any of the herein mentioned organisms, and synthetic oligonucleotide primers prepared on the basis of the DNA sequences or the amino acid sequences disclosed herein. For instance, suitable oligonucleotide primers may be the primers described in the Materials and Methods section. In accordance with well-known procedures, the PCR fragment generated in the molecular screening may be isolated and suctioned into a suitable vector. The PCR fragment may be used for screening DNA libraries by e.g. colony or plaque hybridization. Expression cloning in yeast The DNA sequence of the invention encoding an enzyme exhibiting endoglucanase activity may be isolated by a general method involving cloning, in suitable vectors, a DNA library from a suitable source, such as any of the herein mentioned organisms transforming suitable yeast host cells with said vectors, culturing the host cells under suitable conditions to express any enzyme of interest encoded by a clone in the DNA library, screening for positive clones by determining any endoglucanase activity of the enzyme produced by such clones, and isolating the enzyme encoding DNA from such clones. The general method is further disclosed in WO 94/14953 the contents of which are hereby incorporated by rejference. A acre detailed description of the screening method is given in Example 1 belov. The DNA sequence coding for the enzyme may for instance be isolated by screening a cDNA library of Macrophomina phaseolina, Crinipellis scabella, Sordaria flmicola or Volutella colletotricboides, and selecting for clones ex¬pressing the appropriate enzyme activity (i.e. endoglucanase activity) or from Escherichia coli DSM 10512 deposited under the Budapest Treaty on 2 February, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany), or- from Escherichia coli DSM lOSl-l—dapcsited under the Budapest Treaty on 2 February, 1996, at DSK, or from Escherichia coli DSM 10576, deposited under the Budapest Treaty on 12 March, 1996, at DSM; or from Escherichia -coli DSM 10571 deposited under the Budapest Treaty on 6 March, 19S6, at DSM; or by screening a cDNA library of Myceliphthora thermophila, CBS 117.55, Acremonium sp., CBS 473.94, or Thielavia terrestris, NRRL 8126, and selecting for clones expres¬sing the appropriate enzyme activity (i.e. endoglucanase activity) or from Saccharomyces cerevisiae DSM 9770 deposited under the Budapest Treaty on 24 February, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany), or from Saccharomyces cerevisiae DSM 10032 deposited under the Budapest Treaty on 30 June, 1995, at DSM, from Saccharomyces cerevisiae DSM lOOSO deposited under the Budapest Treaty on 30 June, 1995, or from Saccharomyces cerevisiae DSM 10081 deposited under the Budapest Treaty on 30 June, 19S5, at DSM. The appropriate DNA sequence may then be isolated from the clone by s~ar.dari proced"ai"es, e-g- as aescrj-sed m Example 1. WTJcIeic acid construct As used herein the term '"nucleic acid construct" is intended to indicate any nucleic acid molecule of cDNA, genomic DNA, syn-hetic DNA or RNA origin. The term "construct" is intended to indicate a nucleic acid segment which may be single- or double-stranded, and which may be based on a complete or partial naturally occurring nucleotide sequence encoding an enzyme of interest. The construct may optionally contain other nucleic acid segments. The nucleic acid construct encoding the enzyme of the invention may suitably be_Qi_genomic orcDNA origin, for instance obtained by preparing a genomic or cDNA library and screening for DNA sequences coding for all or part of the enzyme by hybridization using synthetic oligonucleotide probes in accordance with standard techniques (cf. Sambrook et al., 1989). The nucleic acid construct encoding the enzyme may also be prepared synthetically by established standard methods, e.g. the phosphoamidite method described by Beaucage and Caruthers, (1981), or the method described by Matthes et al., (1984). According to the phospho¬amidite method, oligonucleotides are synthesized, e.g. in an automatic DNA synthesizer, purified, annealed, ligated and cloned in suitable vectors. Furthermore, the nucleic acid construct may be of mixed synthetic and genomic, mixed synthetic and cDNA or mixed genomic and cDNA origin prepared by ligating fragments of s%Tithetic, genomic or cDNA origin (as appropriate), the fragmentLS corresponding to various parts cf rhe entire nucleic acid construct, in accordance with standard tech¬niques. The nncleic acid construct nay also be prepared by polymerase chain reaction using specific priaers, for instance as described in US 4,683,202 or Saiki et al., (1988) . The nucleic acid construct is preferably a DNA construct which term will be used exclusively in this specification and claims. Recombinant vector A recombinant vector comprising a DNA construct encoding the enzyme of the invention may be any -vector which s.^y conveniently be subjected to recombinant DNA procedures, and the choice of vector will often depend on the host cell into which it is to be introduced. Thus, the vector may be an autonomously replicating vector, i.e. a vector which exists as an extrachromosomal entity, the replication of which is independent of chroinosomal replication, e.g. a plasmid. Alternatively, the vector may be one which, when introduced into a host cell, is integrated into the host cell genome and replicated together with the chromosome(s) into which it has been integrated. The vector is preferably an expression vector in which the DNA seguence encoding the enzyme of the invention is operably linked to additional segments required for transcription of the DNA. In general, the expression vector is derived from plasmid or viral DNA, or may contain elements of both. The term, "operably linked" indicates that the segments are arranged so that they function in concert for their intended purposes, e.g. transcription initiates in a promoter and proceeds through the DNA sequence coding for the enzyme. The prnmnter may be any DNA sequence which shows transcriptional activity in the host cell of choice and may be derived from genes encoding proteins either homologous or heterologous to the host cell. Examples of suitable promoters for use in yeast host cells include promoters from yeast glycolytic genes (Hitzeman et al., J. Biol. Chem. 255 (1980), 12073 -12080; Alber and Kawasaki, J. Mol. APPI. Gen. 1 (1982), 419 - 434) or alcohol dehydrogenase genes (Young et al., in Genetic Engineering of Microorganisms for Chemicals (Hollaender et al, eds.), Plenum Press, New York, 1982), or the TPIl (US 4,599,311) or ADH2-4C (Russell et al.. Nature 3 0^ rT98 3). 652 - 654) promoters. Examples of suitable promoters for use in filamentous fungus host cells are, for instance, the ADH3 promoter (McKnight et al.. The EMBO J. i (1985), 2093 - 2099) or the tpJA promoter. Examples of other useful promoters are those derived from the gene encoding A. oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, A. niger neutral a-amylase, A. niger acid stable a-amylase, A. niger or A. avamori glucoamylase (gluA), Rhizomucor miehei lipase, A. oryzae alkaline protease, A. oryzae triose phosphate isomerase or A. nidulans acetamidase. Preferred are the TAKA-amylase and gluA promoters. Examples of suitable promoters for use in bacterial host cells include the promoter of the Bacillus stearothermophilus maltogenic amylase gene, the Bacillus licheniformis alpha-amylase. gene, the Bacillus . : . , ajuyloliquefaciens BAN amylase gene, the Bacillus subtilis alkaline protease gen, or the Bacillus pumilus xylcsidase gene, cr by the phage Lambda PR or P^ promoters or rhe E. coli lac, trp or tac promoters. Th-e DNA sequence encoding th-e enzyme of the invention may also. i:f necessary, be operably connected to a suitable terminator. The recombinant vector of the invention may further comprise a DNA sequence enabling the vector to replicate in the host cell in question. The vector may also comprise a selectable marker, e.g. a gene the product of which complements a defect in the host cell, such as the gene coding for dihydrofolate reductase (DHFR) or the Schizosaccharomyces pombe TPI gene (described by P.R. Russell, Gene 40, 19S5, pp. 125-130). For filamentous fungi, selectable markers include amdS7~PVrG. argB, niaP, sC. To direct an enzyme of the present invention into the secretory pathway of the host cells, a secretory signal sequence (also known as a leader sequence, prepro sequence or pre sequence) rr.ay be provided in the recombinant vector. The secretory signal sequence is joined to the DNA sequence encoding the enzyme in the correct reading frame. Secretory signal sequences are commonly positioned 5' to rhe DliA sequence encoding the enzyme. The secretory signal sequence may be that normally associated with the enzyme or may be from a gene encoding another secreted protein. For secretion from yeast cells, the secretory signal sequence may encode any signal peptide which ensures efficient direction of the expressed enzyme into the secretory pathway of the cell. The signal peptide may be naturally occurring signal peptide, or a functional part thereof, or it may be a syntheric peptide. Suitable signal peptides have been fO'jnd to be che a-facror signal peptide (cf. US 4,370,COS), the signal peptide of mouse salivary amylase (of. O. Kagencuchle et al., Nature 289, 1981, pp. 643-646), a modified carbcxypeptidase signal peptide (cf. L.A. Vails er al.. Cell 48, 19S7, pp. 887-897), the yeast BARl signal peptide (cf. WO 87/02670), or the yeast aspartic protease 3 (YAP3) signal peptide (cf. M. Egel-Mitani et al., Yeast 6, 1990, pp. 127-137). For efficient secretion in yeasr, a sequence encoding a leader peptide may also be inserted downstream of the signal sequence and upstream of the DNA sequence encoding the enzyme. The function of the leader peptide is to allow the expressed enzyme to be directed from the endoplasmic reticulum to the Golgi apparatus and further to a secretory vesicle for secrerion into the culture medium, (i.e. exportation of the enzyme across the cell wail or at least through the cellxrXar m.embrane into the periplasmic space of the yeast cell). The leader peptide may be the yeast a-factor leader (the use of which is described in e.g. US 4,546,082, EP 16 201, EP 123 294, EP 123 544 and EP 163 529). Alternatively, the leader peptide may be a synthetic leader peptide, which is to say a leader peptide not found in nature. Synthetic leader peptides may, for instance, be constructed as described in WO S9/02463 or WO 92/11378. For use in filamentous fungi, the signal peptide may conveniently be derived from a gene encoding an Aspergillus sp. amylase or glucoaraylase, a gene encoding a Rhizomucor miehei lipase or protease, a Humicola lanuginosa lipase. The signal peptide is preferably derived from, a gene encoding .4. oryzae TAKA amylase, A. niger neutral a-amylase, .-.. niger acid-stable amylase, or A. niger glucoam.ylase. ^ - The procedures used to ligate the DN'A sequences coding for the present enzyme, the promoter and optionally the terminator and/or secretory signal sequence, respectively, and to insert them into suitable vectors containing the information necessary for replication, are well known to persons skilled in the art (cf., for instance, Sambrook et al., op.cit.). Host cells The DNA sequence encoding the present enzyme introduced into the host cell may be either homologous or heterologous to the host in question. If homologous to the host cell, i.e. produced by the host cell in nature, it will typically be operably connected to another promoter sequence or, if applicable, anoTiher secretory signal sequence and/or terminator sequence than in its natural environment. The term "homologous" is intended to include a cDNA sequence encoding an enzyme native to the host organism in question. The term "heterologous" is intended to include a DNA sequence not expressed by the host cell in nature. Thus, the DNA sequence may he from another organism, or it may be a synthetic sequence. The host cell into which the DNA construct or the recombinant vector of the invention is introduced may be any cell which is capable of producing the present enzyme and includes bacteria, yeast, fungi and higher eukaryotic cells. Examples of bacterial host cells which, on cultivation, are capable of producing the enzyme of the invention are gram-positive bacteria such as strains of Bacillus, such as strains of B. subtilis, B. licheniformis, 5. ler.tus, B. brevis, B. stearothermophilus, B. alkalophilus, 3. amyloliquefacier.s, B. coagulans, B. circvlans, B. lautus, B. megatherium or 3. tburingier.sis, or strair.s cf Szrepzoi?.yces, such as S. livida::s or S. jnurinus, or gram-negative bacteria such as Echerichia coli. The transformation of the bacteria may be effected by protoplast transformation or by using comperent cells in a manner known per se (cf. Sambrook et al., suora). When expressing the enzyme in bacteria' such as E. coli, the enzyme may be retained in the cytoplasm, typically as insoluble granules (known as inclusion bodies), or may be directed to the periplasmic space by a bacterial secretion sequence. In the for»-er case, the cells are lysed and the granules are recovered and denatured after which the enzyme is refolded by diluting the denaturing agent. In the latter case, the enzyme may be recovered from the periplasmic space by disrupting the cells, e.g. by sonication or osmotic shock, to release the contents of the periplasmic space and recovering the enzyme. Examples of suitable yeasts cells include cells of Saccharomyces spp. or Schizosaccharomyces spp., in pa:.-ticular strains of Saccharomyces cerevisiae or Saccharomyces kluyveri. Methods for transforming yeast cells with heterologous DNA and producing heterologous enzymes therefrom are described, e.g. in US 4,599,311, US 4,931,373, US 4,870,008, 5,037,743, and US 4,845,075, all of which are hereby incorporated by reference. Transformed cells are selected by a phenotype determined by a selectable marker, commonly drug resistance or the ability to grow in the absence of a particular nutrient, e.g. leucine. A preferred vector for use in yeast is the POTl vector disclosed in US 4,931,373. The DNA sequence encoding the enzyme of the invention may be preceded by a signal sequence and optionally a leader sequence , e.g. as described above. Further examples of suitable yeasr cells are strains of Kluyveromyces, such as .^'. lactis, Hansenula, e.g. H. polymorpha, or Pichia, e.g. P. pastoris (cf. Gleescn er al., J. Gen. Microbiol. 132, 1986, pp. 3455-3465; US 4,882,275). Examples cf other fungal cells are cells of filamentcus fungi, e.g. Aspergillus spp., Neurospora spp., Fusarium spp. cr Trichoderma spp., in particular strains of A. oryzae, A. nldulans, A. niger, or Fusarium graminearum. The use of Aspergillus spp. for the expression of pro¬teins is described in, e.g., EP 272 277, EP 230 023. The transformation of F. oxysporuw may, for instance, be carried out as described by Malardier et al., 1989, Gene 73: 147-156. When a filamentous fungus is used as the host cell, it may be transformed with the DNA construct of the invention, conveniently by integrating the DNA construct in the host chromosome to obtain a recombinant host cell. This integration is- generally considered to be an advantage as the DNA sequence is more likely to be stably maintained in the cell. Integration of the DNA constructs into the host chromosome may be performed according to conventional methods, e.g. by homologous or heterologous recombination. The transformed or transfected host cell described above is then cultured in a suitable nutrient medium under conditions permitting the expression of the present enzyme, after which the resulting enzyme is recovered from the culture. The medium used to culture the cells may be any conventional medium suitable for growing the host cells, such as minimal or complex media containing appropriate supplements. Suitable media are available from commercial suppliers or may be prepared according to published recipes (e.g. in catalogues of the American Type Culture Collection) . The enz^ine produced by the cells may then be recovered from the culture medium by conventional procedures including separating the host cells from z'r.e. medium by csnrrifugarion or fiitra-icn. precipiraning zr.e proteinaceous components of the supernatant or filtrate by means of a salt, e.g. ammonium sulphate, purification by a variety of chromarographic procedures, e.g. ion exchange chromatography, gel filtration chromiatography, affinity chromatography, or the like, dependent on the type of enzyme in question. In a still further aspect, the present invention relares to a method of producing an enzyme according to the invention, wherein a suitable host cell transformed with a DNA sequence encoding the enzyme is cultured under conditions permitting the production of the enzyme, and the resulting enzyme is recovered from the culture. Enzyme Screening driven by taxonomy as well as ecology: A powerful tool like the molecular screening disclosed herein, designed to detect and select said type of interesting enzymes, can still not stand on its own. In order to maximize the chances of making interesting discoveries the molecular screening approach was in the present investigation combined with careful selection of which fungi to screen. The selection was done through a thorough insight in the identification of fungi, in taxonom.ical classification and in phylogenetic relationships. A taxonomic hot spot for production of cellulytic enzymes can further only be fully explored if also the ecological approach is included. Thorough knowledge about the adaptation to various substrates (especially saprotrophic, necrotrophic or biotrophic degradation of •plant m-aterials) are prerequisites for designing an intelligent screening and for managing a successful selection of strains and ecological niches to be searc¬hed . Both the taxonomy and the ecological approach disclosed herein aim at maximizing discovery" of said enzymes in the molecular screening program. However, still several hundreds (or if all preliminary work is included) several thousand fungi have been brought in culture in order to detect the 53 hits of said type of cellulytic enzy^m.e here reported. The screening and cloning may be carried out using the following: MATERIALS AND METHODS List of organisms: Saccharomyces cerevisiaB, -USiL. 917 0, DSM 10082, DSM 10080, DSM 10081, or Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, containing the plasmid comprising the full length DNA sequence, coding for the endoglucanase of the invention, in the shuttle, vector pYES 2.0. Escherichia coli DSM 10583, 10584, 10585, 10586, 10587, and 10588. Diplodia gossypir.a Cooke Deposit of Strain, Ace No: CBS 274.96 Classification: Ascomycota , Loculoascorr.ycetes , Dothideales, Cucurbitariaceae Ulospora bilgramii {Hawksw. et al.) Hawksw. et al . Ace No of strain: NK3C 1444, Nippon University, (Prof. 'Tubaki collection) Classification: Ascorriycota , Loculoascomycetes, Dothideales, (family unclassified) Microsphaeropsis sp. Isolated from: Leaf of Camellia japonica (Theaceae, Guttifsrales), grown in Kunming Botanical garden, Yunnan Province, China Classification: Ascomycota, Loculoascomycetes, Dothideales, (family unclassified) Macrophomina phaseolina (Tassi) Goidannich Syn: Rhizoctonia baraticola Deposit of Strain, Ace No.:CBS 281.96 Isolated from seed of Glycine max (Leguminosa), cv CMM 60, grown in Thailand, 1990 Classification: Ascomycota, Discomycetes, Rhytism.atales, Rhytismataceae Ascojbolus stictoideus Speg. Isolated from goose dung, Svalbard, "TTorway Classification: Ascomycota, Discomycetes, Pezizales, Ascobolaceae Saccobolus dilutellus (Fuck.) Sacc. Deposit of strain: Ace No CBS 275.96 Classification: Ascomycota, Discomycetes, Pezizales, Ascobolaceae Penicillium verruculosum Peyronel Ex on Ace No of species: ATCC 62396 Classification: Ascomycota, Plectomycetes, Eurotiales, Trichocomaceae Penicillium ckrysogenum Thorn Ace No of Strain: ATCC 9480 Classification: Ascomycota, Plectomycetes, Eurotiales, Trichocomaceae Thermomyces verrucosus Pugh et ai Deposit of Strain, Ace No.: CES 235.96 Classification: Aseor.yecra, PlectcmyceTies, Eurcriales, (family unclassified; affiliation based on ISS RNA, sequencing and homologies) Xylaria hypcxylcn L. ex Greville Deposit of srrain, Ace No: CBS 2S4.96 Classification: Ascomycota, Pyrenomycetes, Xylariales, Xylariaceae Poronia punctata (Fr.ex L.} Fr. Classification: Ascomycota, Pyrenomycetes, Xylariales, Xylariaceae Nodulisporum sp Isolated from leaf of Camellia reticulata (Theaeeae, Guttiferales), grown in Kunming Botanical Garden, Yunnan Province, China Classification: Ascomycota-,—Pyrenomycetes, Xylariales, Xylariaceae Cylindrocarpon sp Isolated from marine sample, the Bahamas Classification: Ascomycota, Pyrenomycetes, Hypocreales (unclassified) Acremonium so Deposit of Strain, Ace. No.: CBS 478.94 Classification: Ascomycota, Pyrenomycetes, Hypocreales, Hypocreaceae Fusarium anguioides Sherbakoff Ace No of strain: IFO 4467 Classification: Ascomycota, Pyrenomycetes, Hypocreales, H>'pocr eaceae Fusarium poae (Peck) Wr. Ex on Ace No of species: ATCC 6OS S3 i_iassir ica~ ion: Ascomyccta , Pyrenomyce'ces , nypocreaJ.es, Hypocreaceae Fusariun solani (Mart.)Sacc.emnd,Snyd 5 Hans. Ace No of strain: IMI 107.511 Classification: Ascomycota, Pyrenomycetes, Hypocreales, Hypocreaceae Fusariuui oxysporum ssp lycopersici (Sacc .) Sr.yd . & Har.s . Ace No of strain: CBS 64 5.7 3 Classification: Ascomycota, Pyrenomyeeres, Hypocreales, Hypocreaceae Fusarium oxysporum ssp passiflora hoc No of strain: CBS 7 4 4.79 ::lassif ication: Ascomycota, Pyrenomycetes, Hypocreales, -lypocreaceae Gliocladium catenulatum Gillman & Abbott Ace. No. of strain: CBS 227.48 classification: Ascoinycota, Pyrenomycetes, Hypocreales, Hypocreaceae Nectria pinea Dingley Deposit of Strain, Ace. No. CBS 279.96 Classification: Ascomycota, Pyrenomycetes, Hypocreales, Nectriaceae Volutella colletotrichoides Ace No of Strain: CBS 400.53 Classification: Ascoinycota, Pyrenomycetes, Hypocreales (unclassified) Sordaria macrospcra Auerswala Ex on Ace No of species: ATCC 60255 Classification: Ascomycota, Pyrenomycetes, Sordariales, Sordariaceae Sordaria fimicola (Roberge) Cesati ez De '^otaris Ex on Ace. No. for the species: ATCC 52644 Isolat.ed from dung by H.Dissi.ng, ISP, KU, Denmark Classification: Ascomycota, Py^enonycetes, Sordariales, Sordariaceae Humicola grisea Traeen ex on Ace No for the species: ATCC 2 2726 Source: Hatfield Polytechnic Classification: Ascomycota, Pyrenomycetes, Sordariales, (fam. unclassified) Humicola nigrescens Omvlk Ace No of strain: CBS S19.73 Classification: Ascomycota, Pyrenomycetes, Sordariales, (fam. unclassified) Scytalidium thermophilum (Cooney et Emerson) Austwick Ace No of strain: ATCC 28085 Classification: Ascomycota, PyrenomycGtes, Sordariales, (fam. unclassified) Thielavia thermophila Fergus et Sinden I (syn Corynascus thermophilus) Ace No of strain: CBS 174.70, IMI 145.136 Classification: Ascomycota, Pyrenomycetes, Sordariales, Chaetomiaceae Isolated from Mushroom compost Thielavia terrestris (Appinis) Malloch et Cain Ace No of strain: NRRL3126 Classification: Ascomycota, Pyrenomycetes, Sordariales, Chaetomiaceae Cladorrhinum foecundissimum Saccardo et Marchal Ex on Ace No of species: ATCC 62 27 3 Ciassif icatio.n: A.sccr.ycora , ?'j"rencmyce"ces , Sordariales, Lasiosphaeriaceae Isolared from leaf of Selandin sp. (Ccmpositaceae, A.sterales) , Dallas .Mountain, Jamaica Syspastospora boninensis Ace No of strain: NKBC 1515 (Nippon University, profe Tubaki Collection) Classification: Ascomycota, Pyrenomycetes, Sordariales, Cerastomataceae Chaetomium cuniculorum Fuckel Ace. No. of strain: CBS 799.83 Classification: Ascomycota, Pyrenomycetes, Sordariales, Chaetomiaceae Chaetomium brasiliense Batista et Potual Ace No of strain; CBS -L22^ 65 Classification: Ascomycota, Pyrenomycetes, Sordariales, Chaetomiaceae Chaetojp.ium murorum Corda Ace No of strain: CES 163.52 Classification: Ascomycota, Pyrenomycetes, Sordariales, Chaetomiaceae Chaetowium virescens (von Arx) Udagawa Ace.No. of strain: CBS 5 4 7.75 Classification: Ascomycota, Pyrenomycetes, Sordariales, Chaetomiaceae Myceliophthora tbermophlla (Apinis) Oorschot Deposit of Strain, Ace No:CBS 117.65 Classification: Ascomycota, Pyrenomycetes, Sordariales, Chaetomiaceae Nigrospora sp Deposit of strain. Ace No: CBS 272.96 Isolated from leaf of Artocarp'uS aitiiis, Moraceae, Urticales qrowv, in Christiana, Jamaica Classification: Ascomycota, pyrenomycetes, Trichcsphaeriales, (family unclassified) Nigrospora sp Isolated from leaf of Pinus yuannanensis, Botanical Garden, Kuning, Yunnan. Classification: A.scomycota, Pyrenomycetes, Triehosphaeriales, Abietaceae, Pinales. Diaporthe syngenesia Deposit of strain, .Ace No: CBS 278.96 Classification: .Ascomycota, Pyrenomycetes, Diaporthales, Valsaceae Colletotrichum lagenarium (Passerini) Ellis at Halsted syn Glomerella cinguiata var orbiculare Jenkins et Winstead Ex on ace No of species: ATCC 52609 Classification: Ascomycota, Pyrenomycetes, Phyllachorales Exidia glandulosa Fr. Deposit of Strain, Ace No: CBS 277.96 Classification: Basidiomycota, Hymenoiriycetes, Auriculariales, Exidiaceae Crinipellis scabella (Alb.&Schw.:Fr.)Murr Deposit of strain: Ace No CBS 280.96 Classification: Basidiomycota, Hymenomycetes, Agaricales, Panaeolus retivugis (Fr.) Gill. Ace.No. of strain: CBS 275.47 Classification: Basidiomycota, Kynenc-ycetes, Agaricales, Coprinaceae Fojnes fomentarius (L.) Fr. Deposit of strain: Ace No. CBS 2 7 6.96 Classification: Basidiorr.ycora, Hymenomycetes, Aphyllophorales, Fomitaceae Spongipellis sp. Deposit of Strain: Ace No CBS 283.96 Classification: Basidiomycota, Hymenomycetes, Aphyllophorales, Bjerkanderaceae (identified and affiliated taxonomieally by ISS sequence and homology) Trametes sanguinea (Fr.) Lloyd syn: Polyporus sanguineus; Pycnoporus sanguineus (L.:Fr.) Murrill Ace No of strain: AKU 5062 (Kyoto University Culture Collection) Classification: Basidiomycota, Aphy-Llophorales, Polyporaceae Schizophyllum commune Fr Ace. No. of species: ATCC 38548 Classification: Basidiomycota, Aphyllophorales, Schizophyllaceae Rhizophlyctis rosea (de Bary & Wor) Fischer Deposit of Strain: Ace No.: CBS 282.96 Classification: Chytridiomycota, Chytridiomycetes, Spizellomycetales, Spizellomycetaceae Rhizomucor pusillus (Lindt) Schipper syn: Mucor pusillus Ace No of strain: IFO 4 578 Ex en Ace No of species: ATCC 468S3 Classification: Zygomycota, Zygomycetes, Mucorales, Mucoraceae Phycomyces nitens (Kur.ze) van Tieghem & Le Monnier Ace No of strain: IFO.4314 Ex en Kco No of species: ATCC 16327 Classification: Zygomycota, Zygomycetes, Mucorales, Mucoraceae Chaetostylum fresenii van Tieghem & Le Monnier syn. Helicostylum fresenii Ace No of strain NRRL 2 3 05 Classification: Zygomycota, Zygomyceres, Mucorales, Thamnidiaceae Unclassified: Trichothecium roseum Ace No of strain: IFO 5372 Coniothecium sp Endophyte, isolated from leaf of unidentifed higher plant, growing in Kunming, Yunnan, China Unclassified and Un-identified: Deposit of strain, Ace No.: CBS 271.96 Isolated from leaf of Artocarpus altilis (Moraceae, Urticales), grown in Christiana, Jamaica Deposit of strain. Ace No.: CBS 273.96 Isolated from leaf of Pimenta dioica (Myrtaceae, Myrtales) grown in Dallas Mountain, Jamaica Deposit of strain: CBS 270.96 Isolated from leaf of PseudocalyroiBa alliaceum - -. ..- (Bignoniaceae, Solanales) growing in Dallas Mountain, u s m a 1 c a Other strains: Escherichia coli MC1061 and DKIOB. Yeast strain: The Saccharomyces csrsvisiae strain used was W3124 (MATa; ura 3-52; leu 2-3, 112; his 3-D200; pep 4-1137; prcl::HIS3; prbl:: LEU2; cir+). Plasmids: The Aspergillus expression vector pHD414 is a derivative of the plasmid p775 (described in EP 238 023). The construction of pHD414 is further described in WO 93/11249. pYES 2.C (Invitrogen) pA2C477, PA2C193, pA2C357, pA2C371, pA2C385, pA2C475, pA2C438, p/v2C502 (See example 1, 2, 2 and 4). Isolation of the DNA sequence shown in SEQ ID No. l, 4, 6, 8, 10, 12, 16, or 19 respectively: The full length DNA sequence, comprising the cDNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, coding for the endoglucanase of the invention, can be obtained from the deposited organism S. cerevisiae, DSM 9770, DSM 100S2, DSM 10080, DSM 10081, E. coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively, by extraction of plasmid DNA by methods known in the art (Sambrook et al. (1989) Molecular cloning: A laboratory manual. Cold Spring Harbor lab.. Cold Spring Harbor, NY). PCR primers for molecular screening of cellulases of the present invention: The four degenerate, deoxyinosine-conraining cligcnucieotide primers (sense; s and antisense; asl, as2 and as3) corresponding to four highly conserved amino acid regions fou.nd in the deduced amino acid sequences of Thielavia terrestris cellulase, Myceliophthora thermophllum cellulase, and two cellulases frcs Acremonlum sp. The residues are numbered according to the Myceliophthora thermophilum sequence. The deoxyinosines are depicted by an I in the primer sequences, and the restriction sites are underlined. (klippe-klistre fra SKa Molecular screening by polymerase chain reaction (PCR): In vitro amplification of genomic DNA and double-stranded cDNA. j Directional, double-stranded cDNA was synthesized from 5 /ig of poly(A)RNA as described below. Genomic DNA was isolated according to Yelton et al. Approximately 10 to 20 ng of double-stranded, cellulase-induced cDNA or 100 to 200 ng of genomic DNA from a selection of fungal strains was PCR amplified in PCR buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 1.5 mM MgClj, 0.01 % (w/v) gelatin) containing 200 ^M of each dNTF and 100 pmoi of each degenerate primer in three combinations: 1) sense, D — V-'v_'^ _ .-..^. vj c i lA'-—! /(-oiiA' / JTGLJGA /-[TG /[-i'lj /-[/"..-i /,; /(-C—-i antisense 1, 5'- CTAGTCTAGATA\',;CAIGC\/J-;C.VV,;CACC -3'; or 2) sense, 5 ' - CCCCAAGCTTAClVrGITA'-7TTGGGA'7TTG^/TTG'7T^-A'^/n^/cC-3 ' antisense 2, CTAGTCTAGA.-J'.IAV.;/'^ICCIAV'7"ICCICCICCIGG -3 ■ ; or 3) sense, 5' - CCCCAAGCTTACIVCC-ITA^7TTGGGA'7TTG'7TTG'7T^"V\/,/'/,.C-3 ■ antisense 3, 5 ■ - CTAGTCTAGAIA;^.CCA\/nTCA'\/n-7'TAIC a DNA thermal cycler (Landgraf, Germany) and 2.5 units of Taq polymerase (Perkln-Elmer, Cetus, USA). Thirty cycles of PCR were performed using a cycle profile of denaturation at 94 °C for 1 min, annealing at 64 °C for 2 min, and extension at 72 °C for 3 min. Ten-/xl aliquots of the amplification products were analyzed by electrophoresis in 3 % agarose gels (NuSieve, FMC) with Haelll-digested ^XIV^ RF DNA as a size marker. Direct sequencing of the PCR products. Eighty-/il aliquots of the PCR products were purified using the QIAquick PCR purification kit (Qiagen, USA) according to the manufacturer's instructions. The nucleotide sequences of I the amplified PCR fragments were determined directly on the purified PCR products by the dideoxy chain-termination method, using 50-150 ng template, the Taq deoxy-terminal cycle sequencing kit (Perkin-Elmer, US.A) , fluorescent labeled terminators and 5 pmol of the i sense primer: 5'- CCCCAAGCTTACI"/(-GITA'\'TTGGGA'~/-T- G'^/T-TG'/-AA'^/G'/(-C-3 ' . Analysis of the sequence data were perforned according to Devereux et al. Cloning by polymerase chain reaction (PCR): Su±)cloning of PCR fragments. Twentyfive-^i aliquots of the PCR products generared as described above were electrophoresed in 0.8 % low gelling temperature agarose (SeaPlaque GTG, FMC) gels, the relevant fragments were excised from the gels, and recovered by agarase treatment by adding 0.1 vol of 10 x agarase buffer (New England Biolabs) and 2 units per 100 jj.1 molten agarose to the sample, followed by incubation at 45 °C for 1.5 h. The sample was phenol and chloroform extracted, and precipitated by addition of 2 vols of 96 % EtOH and 0.1 of 3 M NaAc, pH 5.2. The PCR fragments were recovered by centrifugarion, washed in 70 % EtOH, dried and resuspended in 20 fxl of restriction enzyme buffer (10 mM Tris-HCl, 10 mM MgCl2, 50 mM NaCl, 1 mM DTT) . The fragments were digested witrh—^indlll and Xbal, phenol and chloroform extracted, recovered by precipitation with 2 vols of 96 % EtOH and 0.1 of 3 M NaAc, pH 5.2, and subcloned into HindIII/Xi>aI-cleaved pYES 2.0 vector. Screening of cDNA libraries and characterization of the positive clones. cDNA libraries in S. cerevisiae or E. coli, constructed as described below, were screened by colony hybridization (Sambrook, 1989) using the corresponding randoiTi-primed (Feinberg and Vogelstein) "p-labeled (>1 x lo"cpm/^q) PCR products as probes. The hybridizations were carried out in 2 x SSC (Sambrook, 1989), 5 X Denhardfs solution (Sambrook, 1989), 0.5 % (w/v) SDS, 100 fig/ml denatured salmon sperm DNA for 20 h at 65^C followed by washes in 5 x SSC at 25°C (2 x 15 min), 2 X SSC, 0.5 % SDS at 65°C (30 min), 0.2 x SSC, 0.5 % SDS at- 65°C (30 min) and finally in 5 x SSC ( 2 x 15 ' ' min) at 25=C. The positive cDNA clones were characterized by sequencing rhe ends of the cD.N'.A inserts with pYES 2.0 polylinker primers (Invitrogen, USA.), and by derermining the nucleotide seuence of the longest cDNA from both strands by the dideoxy chain termination method (Sanger et al.) using fluorescent labeled terminators. Qiagen purified plasmid D.^^'.-. (Qiagen, USA) was sequenced with the Taq deoxy terminal cycle sequencing kit (Perkin Elmer, USA) and either pYES 2.0 polylinker primers (Invitrogen, US.A) or synthetic oligonucleotide primers using an Applied Biosystems 373A automated sequencer according to the ma.nufacturers instructions. Analysis of the sequence data was performed according to Devereux et al. Extraction of total RNA was performed with guanidinium thiocya.nate followed by ultracentrifugation through a 5.7 M CsCl cushion, and isolation of poly(A)"RNA was carried out by oligo(dT)-cellulose affinity chromatography using the procedures described in WO 94/14953. cDNA synthesis: Double-stranded cDNA was synthesized from 5 /ig poly (A)* RNA by the RNase H method (Gubler and Hoffman (1983) Gene 25:263-269, Sambrook et al. (1989) Molecular cloning: A laboratory manual, Cold Spring Harbor lab.. Cold Spring Harbor, NY) using the hair-pin modification developed by F. S. Hagen (pers. comm.). The poly(A)' RNA (5 Mg in 5 ^1 of DEPC-treated water) was heated at 70°C for 8 min. in a pre-siliconized, RNase-free Eppendorph tube, quenched on ice and combined in a final volume of 50 IJ.1 with reverse transcriptase buffer (50 mM Tris-Cl, pH 8.3, 75 mM KCl, 3 mM MgCl,, 10 mM DTT, Bethesda Research Laboratories) containing 1 mM of dATP, dGTP and dTTP and 0.5 mM 5-methyl-dCTP (Pharmacia), 40 units human placental ribonuclease inhibitor (RNasin, Promega) , 1.45 ug cf cligc (dT) ,,-Not I prim.er (Pharmacia) and 1000 units Superscript II RNase H reverse - trans-criptase (Bethesda Research Laboratories) --First- '• . , strand cDNA was synthesized by incubating the reacrion mixture at 4 5"C fcr 1 hour. After synthesis, rhe mirture was gelfij.'craTec ~hrougn a MicroSpin S-4 00 HR (Pharmacia) spin column according ro the manufacturer's instructions. After rhe gelfiltration, the hybrids were diluted in 2 50 /il second strand buffer (20 mM Tris-Cl, pH 7.4, 90 miM KCl, 4.6 mM'MgCl,, 10 mM (NH4)2S04, 0.16 mM iSViAD^) containing 200 ^M of each dNTP, 60 units I. coli DNA polymerase I (Pharmacia), 5.25 units RNase K (Promega) and 15 units E. coli DNA ligase (Boehringer Mannheim). Second strand cDNA synTihesis was performied by incubating the reaction tube at 16°C for 2 hours and additional 15 min. at 25°C. The reaction was stopped by addition of EDTA to a final concentration of 2 0 inM followed by phenol and chloroform extractions. Mung bean nuclease treatment: The double-stranded cDNA was precipitateti—at -20°C for 12 hours by addition of 2 vols 96% EtOH, 0.2 vol 10 M NH4AC, recovered by centrifugation, washed in 70% EtOH, dried and resuspended in 30 Ml Mung bean nuclease buffer (30 mM NaAc, pH 4.6, 300 mM NaCl, 1 rc.M ZnSO,, 0.35 mM DTT, 2% glycerol) containing 25 units Mung bean nuclease (Pharmacia). The single-stranded hair-pin DNA was clipped by incubating the reaction at 30^C for 30 min., followed by addition of 70 /il 10 mM Tris-Cl, pH 7.5, 1 mM EDTA, phenol extraction and precipitation with 2 vols of 96% EtOH and 0.1 vol 3 M NaAc, pH 5.2 on ice for 30 min. Blunt-ending with T4 DNA polymerase: The double-stranded cDNAs were recovered by centrifugation and blunt-ended in 30 fil T4 DNA polymerase buffer (20 mM Tr is-acetate, pH 7.9, 10 mM MgAc. 50 mM KAc, 1 mM DTT) containing 0.5 ir.M of each dNTP and 5 units T4 DNA polymerase (N'ew England Biolabs) by incubating, the reaction mixture at 16°C for 1 hour. The reaction was stopped by addition of EDTA to a final coficentrarion of 20 mM, followed by phenol and chlcrcfcr:?. e>rcrac-icns, and precipitarion for 12 hours at -20-C by adding 2 vols 96% EtOH and 0.1 vol 3 M NaAc pH Adapter ligation/ Not I digestion and size selection: After the fill-in reaction the cDNAs were recovered by centrifugation, washed in 70% EtOH and dried. The cDNA pellet was resuspended in 25 /il ligation buffer (30 mM Tris-Cl, pH 7.S, 10 -uM MgCl,, 10 mM DTT, 0. 5 mM ATP) containing 2.5 ^g non-palindromic BstXI adaptors (Invitrogen) and 30 units T4 ligase (Promega) and incubated at 16=C for 12 hours. The reaction was stopped by heating at 65°C for 20 min. and then cooling on ice for 5 min. The adapted cDNA was digested with Not I restriction enzvTue by addition of 2 0 ul water, 5 ul ICx Not I restriction enzyme buffer (New England Biolabs) and 50 units Not I (New England Biolabs), followed by incubation for 2.5 hours' at 37='C. The reaction was stopped by heating at 65°C for 10 min. The cDNAs were size-fractionated by gel electrophoresis on a 0.8% SeaPlaque GTG low melting temperature agarose gel (FMC) in Ix TBE to separate unligated adaptors and small cDNAs. The cDNA was size-selected with a cut-off at 0.7 kb and rescued from the gel by use of /J-Agarase (New England Biolabs) according to the manufacturer's instructions and precipitated for 12 hours at -20°C by adding 2 vols 96% EtOH and 0.1 vol 3 M NaAc pH 5.2. Construction of libraries: The directional, size-selected cDNA was recovered by centrifugation, washed in 70% EtOH, dried and resuspended in 30 ^1 10 mM Tris-Cl, pH 7.5, 1 mM EDTA. The cDNAs were desalted by gelfiltration through a MicroSpin S-300 HR (Pharmacia) spin coluinn according to the manufacturer's instructions. Three test ligations were carried out in 10 /il.ligation buffer (30 mM Tris-Cl, pH 7.8, 10 mM MgCl., 10 mM DTT, 0.5 nsM ATP) containing 5 til double-stranded cDNA (reacricn rubes -1 and #'2), 15 units T4 iigase (Fromega) and 50 ng (zube fl), 40 ng (tube #2) and 40 ng (tube #3, the vector background control) of BstXI-NctI cleaved pYES 2.0 vector. The ligation reactions were perfcruied by incubation at 16'C for 12 hours, heating at 70'C for 20 ir.in. and addition of 10 fil water to each tube. 1 ^1 of each ligation mixture was electroporated into 40'^1 electrocompetent E. coli DHIOB cells (Bethesda research Laboratories) as described (Sambrook et al. (1939) Molecular cloning: A laboratory manual, Cold Spring Harbor lab., Cold Spring Harbor, NY). Using the optimal conditions a library was established in E. coli consisting of pools. Each pool was made by spreading transformed E. coli on LB+ampicillin agar plates giving 15.000-30.000 colonies/plate after incubation at 37°C for 24 hours. 20 ml LB+ampicillin was added to the plate and the cells were suspended herein. The cell suspension was shaked in a 50 mi tube for 1 hour at 37°C. Plasmid DNA was isolated from the cells according to the manufacturer's instructions using QIAGEN plasmid kit and stored at -20°C. 1 ^1 aliquots of purified plasmid DNA (100 ng//il) from individual pools were transformed into S. cerevisiae W3124 by electroporation (Becker and Guarante (1991) Methods Enzymol. 194:182-137) and the transformants were plated on SC agar containing 2% glucose and incubated at 30°C. Identification of positive colonies: After 3-5 days of growth, the agar plates were replica plated onto a set of SC + galactose-uracil agar plates containing 0.1% AZCL HE cellulose. These plates were incubated for 3-7 days at 30°C. Endoglucanase positive colonies were identified as colonies surrounded by a blue halo. Cells from enz-v'-rrie-positive colonies were spread for single colony isolation on agar, and an enzy-e-producing single colony was selected for each of the endoglucanase- Characterization of positive clones: The positive clones were obtained as single colonies, the cDNA inserts were aaplified direcrly from the yeast colony using biotinylated polylinker primers, purified by magnetic beads (Dynabead M-280, Dynal) system and characterized individually by sequencing the 5"-end of each cDNA clone using the chain-termination method (Sanger et al. (1977) Proc. Natl. Acad. Sci. U.S.A. 74:54 63-5467) and the Soquenase system (United States Biochemical). The nucleotide sequence was determined of the longest cDNA from, both strands by the dideoxy chain termination -cethod (Sanger et al.) using fluorescent labeled terminators. Plasmid DNA was rescued by transformation into E. coll as described below. Qiagen purified plasmid DNA (Qiagen, USA) was sequenced with the Tag deoxy terminal cycle sequencing kit (Perkin Elmer, USA) and either pYES 2.0 polylinker primers (Invitrogen, USA) or synthetic oligonucleotide primers using an Applied Biosystems 373A automated sequencer according to the manufacturers instructions. Analysis or the sequence data was performed according to Devereux et al . '■) Isolation of a cDNA gene for expression in Aspergillus: An endoglucanase-producing yeast colony was inoculated into 20 ml YPD broth in a 50 ml glass test tube. The tube was shaken for 2 days at 30°C. The cells were harvested I by centrifugation for 10 min. at 3000 rpm. DNA was isolated according to WO 94/14953 and dissolved in 50 /il water. The DNA was transformed into E. coli by standard procedures. Flasmid DNA was isolated from E. coli using standard procedures, and analyzed by restriction enzyme analysis. The cDNA insert was excised using appropriate restriction enzymes and ligated into an Aspergillus expression vector. Transformation of Aspergillus oryzae or Aspergillus niger Protoplasts may be prepared as described in WO 95/0204 2.. p. 16, line 21 - page 17, line 12, which is hereby incorporated by reference. 100 ^1 of protoplast suspension is mixed with 5-25 ug of the appropriate DNA in 10 ,ul of STC (1.2 M sorbitol, 10 my. Tris-HCl, pK = 7.5, 10 mM CaCl-,) . Protoplasts are mixed with p3SR2 (an A. nidulans amdS gene carrying plasmid). The mixture is left at room temperature for 25 minutes. 0.2 ml of 60% PEG 4000 (BDH 29576), 10 ir.M CaCl, and 10 muM Tris-HCl, pH 7.5 is added and carefully mixed (twice) and. finally 0.85 ml of the same solution is added and carefully mixed. The mixture is left at room temperature . f-cy^ 2 5 minutes, spun at 2 500 g for 15 minutes and the pellet is resuspended in 2 ml of 1.2 M sorbitol. After one more sedimentation the protoplasts are spread on mininial plates (Cove, Biochem. Biophys. Acta 113 (1966) 51-56) containing 1.0 M sucrose, pH 7.0, 10 mM acetamide as nitrogen source and 20 mM CsCl to inhibit background growth. After incubation for 4-7 days at 37=C spores are picked and spread for single colonies. This procedure is repeated and spores of a single colony after the second reisolation is stored as a defined transformant. Test of A. oryzae transformants Each of the transformants were inoculated in 10 ml YPM and propagated. After 2-5 days of incubation at 37°C, 10 ml supernatant was removed. The endoglucanase activity was identified by AZCL HE cellulose as described above. Hybridization conditions (to be used in evaluating property ii) of the DN'A construct of the invention): Suirable ccnditicns for determining hybridizarion between a nucleotide probe and a homologous DNA or RNA sequence involves presoaki.ng of the filter containing the DN.-. fragments or RN.A. to hybridize in 5 x SSC (standard saline citrate) for 10 min, and prehybridization of the filter in a solution of 5 x SSC (Sambrook et al. 1939), 5 x Denhardt's solution (Sambrook et al. 1989), 0:5 % SDS and 100 ^g/ml of denatured sonicated salmon sperm DNA (Sambrook et al. 19S9), followed by hybridization in the same solution containing a random-primed (Feinberg, A. ?. and Vogelstein, B. (19o3) Anal. Biochem. 132:6-13), 'P-dCTP-labeled (specific activity > 1 x lO' cpm/^g ) probe for 12 hours at ca. 45»C. The filter is then washed two times for 30 minutes in 2 x SSC, 0.5 % SDS at preferably not higher than 50°C, more preferably not higher than 55°C, more preferably not higher than 60°C, more preferably not higher than 65°C, even more preferably not "higher than 70°C, especially not higher than 75s€-,— The nucleotide probe to be used in the hybridization is the DNA sequence corresponding to the endoglucanase encoding part of the DNA sequence shown in SEQ ID No. l, 4, 6, 8, 10, 12, or 16, resepctively, and/or the DNA sequence obtainable from the plasmid in S. cerevisiae, DSM 9770, DSM 10082, DSM 10030, DSM 10081, E. coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively. Immunological cross-reactivity: Antibodies tc be used in determining imrriunological cross-reactivity may be prepared by use of a purified cellulase. More specifically, antiserum against the cellulase of the invention may be raised by immunizing rabbits (or other rodents) according to the procedure described by N. Axelsen ejt a 1. in: A Manual of Quantitative Immunoelec¬trophoresis , Blackvell Scientific Publications, 1973, Chapter 2 3, or A. Johnstone and R. Thorpe, Immunochemistrv in Practice. Blackwell Scientific Publications, 1932 fr.cre specifically pp. 27-21). Purified im-unociooulins may be obtained from the anti-sera, for example by salt precipitation ( (NH/) , SOj) , followed by dialysis and ion exchange c.hromiatography, e.g. on DEAE-Sephadex. IiruEunochemicai characterization cf proteins may be done either by Gutcherlony double-diffusion analysis (O. Cuchterlony in: Handbook of Experimental I~munologv (D.M. Weir, Ed.), Blackwell Scientific Publications, 1967, pp. 655-706), by crossed immunoelectrophoresis (N. .Axelsen et al. , supra, Chapters 3 and 4), or by rocket immiunoelectrophoresis (N. Axelsen et al., Chapter 2). Media YPD: 10 g yeast extract, 20 g peptone, H^O to 900 m^l. Autoclaved, 100 ml 20% glucose (sterile filtered) added. YPM: 10 g yeast extract, 20 g peptone, H^O to 900 ml. Autoclaved-,—;-ao ml 20% maltodextrin (sterile filtered) added. 10 X Basal salt: /5 g yeast nitrogen base, 113 g succinic acid, 63 g NaOH, H,0 ad 1000 ml, sterile filtered. SC-\JPJ\: 100 ml 10 X Basal salt, 28 ml 20% casamino acids 'j without vitamins, 10 ml 1% tryptophan, H^O ad 900 ml, autoclaved, 3.6 mi 5% threonine and 100 ml 20% glucose or 20% galactose added. SC-URA agar: SC-URA, 20 g/1 agar added. PD agar: 39g potato dextrose agar, DIFCO 0013; add deionized water up to 1000ml; autoclave (12l"C for 15-20 min) . PC agar: Potatoes and carrots (grinded, 20 g of each) and water, added up to IGOGm.l, are boiled for 1 hr; agar (20g/l of Merck 16:4); autoclave (12l"C for 20 m.in) PC liquid broth: as PC agar but without the Agar ?D liquid bro~h: 24g po~ax.o dextrose broth, Difco 0545, deionized water up to lOOOml; autoclave (121'~C for 15-20 min) PC and PD liquid broth with cellulose: add 30 g Solcafloc (Dicacel available from, Dicalite-Eurcpe-Nord, 9000 Gent, Belgium) per lOOOmI PB-9 liquid broth: 12 g Rofec (Roquette 101-0441) and 24 g glucose are added to lOOOmI water; pH is adjusted to 5.5; 5ml mineral oil and 5 g CaCo, are added per lOOOr.l. autoclave (121'C for 40 min) YPG liquid broth: 4g yeast extract (Difco 0127), Ig KK^PO, (Merck4873), 0.5g MgS04.7H20 Merck 5886, 15g Dextrose, Roquette 101-0441, 0.1ml Pluronic (101-3088); deionized water up to lOOOml; autoclave (20min at 121°C) Dilute salt solution (DS): Make up two stock solutions: P-stock: 13.61g KH.POj,- 13.21g (NHJ2P0j, 17.4 2g KH,POj; deionized water up to 100ml Ca/Mg stock: 7.35g CaCl,, 2H2O, 10.17g MgCl,, 6H_,0, deionized water up to lOOml; pH adjusted to 7.0; autoclaving (121'C; 2 0min) Mix O..51T1I P-stock with 0.1ml Ca/Mg stock add deionized water up to 1000ml AZCL HE cellulose (Megazyme, Australia). Uses During washing and wearing, dyestuff from dyed fabrics cr garment will conventionally bleed from the fabric which then looks faded and worn. Removal of surface fibers from the fabric will partly restore the original colours and looks cf the fabric. By the term "colour clarification", as used herein, is meant the partly restoration cf the initial colours of fabric cr garment throughout m.ultiple washing cycles. The term "de-pilling" denotes removing of pills fro- the fabric surface. The term "soaking liquor" denotes an aqueous liquor in which laundry may be immersed prior to being subjected to a conventional washing proccoc. The soaking liquor may contain one or m.ore ingredients conventionally used in a washing or laundering process. The term, "washing liquor" denotes an aqueous liquor in which laundry is subjected to a washi.ng process, i.e. usually a combined chemical and mechanical action either manually or in a washing machine. Conventionally, the washing liquor is an aqueous solution of a powder or liquid detergent composition. The term "rinsing liquor" denotes an aqueous liquor in which laundry is immersed and treated, conventionally immediately after being subjected to a washing process, in order to rinse the laundry, i.e. essentially remove the detergent solution from the laundry. The rinsing liquor may contain a fabric conditioning or softening composition. The laundry subjected to the method of the present inven¬tion may be conventional washable laundry. Preferably, the major part of the laundry is sewn or unsewn fabrics, including knits, wover.s,_ denims, yarns, and toweling,' made from cotton, cotton blends or natural or manmade cellulosics (e.g. originating frr- xylan-containing cel¬lulose fibers s_ch as from wood pulp) or blends rhereoi. Examples of blends are blends of cotton or rayon/viscose with one or -ore companion material such as wool, syn¬thetic fibers (e.g. pclyamide fibers, acrylic fibers, polyester fibers, polyvinyl alcohol fibers, polyvinyl chloride fibers, pclyvinylidene chloride fibers, polyurethane fibers, polyurea fibers, aramid fibers), and cellulose-containing fibers (e.g. rayon/viscose, ramie, flax/linen, jute, cellulose acetate fibers, lyocell). Detergent Compositions According to one aspect of the present invention, the present enaoglucanases m.ay typically be componenrs of a detergent composition. As such, rhey may be included in the detergent comiposition in the form of a non-dusting granulate, a stabilized JJ-quid, or protected enzymes. Non-dusting granulates may be produced, e.g., as dis¬closed in US 4,106,991 and 4,661,452 (both to Novo Industri A/S) and may optionally be coated by methods known in the art. Examples of waxy coating materials are poly(ethylene oxide) products (polyethyleneglycol, PEG) with mean molecular weights of 1000 to 20000; ethoxylated nonylphenols having from 16 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon aroms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglycerides of fatty acids. Examples of film-forming coating materials suitable for application by fluid bed techniques are given in patent GB 1483591. Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Other enzyme stabilizers are well known in the art. Protected enzymes may be prepared according to the method disclosed in EP The detergent composition of the invention may be in any convenient form, e.g. as powder, granules, paste or liquid. .-. liquid detergent may be aqueous, typically con¬taining up to 70% water and 0-30% organic solvent, or nonaqueous. The detergent composition comprises one or m.ore surf¬actants, each of which may be anionic, nonionic, cationic, or zwittericnic. The detergent will usually contain 0-50% of anionic surfactant such as linear alkylbenzenesulfonate (LAS), alpha-olefinsulfonate (AOS), alkyl sulfate (fatry alcohol sulfate) (AS), alcohol ethoxysulfate (.AEOS or AES) , secondary alkanesulfonates (SAS), alpha-sulfo fatty acid methyl esters, alkyl- or alkenylsuccinic acid, or soap. It may also contain 0-40% of nonionic surfaxrtant such as alcohol ethoxylate (AEO or AE), carboxylated alcohol ethoxylates, nonyiphenol ethoxylate, alkylpolyglycoside, alkyldimethylamine oxide. ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, or polyhydroxy alkyl fatty acid amide (e.g. as described in WO 92/06154). The detergent composition may additionally comprise one or more other enzymes such as amylase, lipase, cutinase, protease, peroxidase, and oxidase, e.g. laccase. The detergent may contain 1-65% of a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, citrate, nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTMPA), alkyl- or =Ikenylsuccinic acid, soluble silicates or layered silicates (e.g. SKS-6 from.. Hoechst) . The detergent may also be unbuilt, i.e. essentially free of detergent JDUiider. The detergent may comprise one or more polymers. Examiples are carboxymethylcellulose (CMC), poly(vinylpyrrolidone) (?'/?) , polyethyleneglycol (PEG; , pcly(vir.yl alcohol) (PVA), polycarboxylates such as pclyacrylates, maleic/acrylic acid copolymers and lauryl methacrylate/acrylic acid 'copolymers. The detergent m.ay contain a bleaching system which may comprise a H;0; source such as perborate or percarbonare which -ay be combined with a peracid-forming bleach activator such as tetraacetylethylenediamine (TAED) or nonajnoyloxybenzenesulfonate (NOBS) . Alternatively, the bleaching system may comprise peroxyacids of, e.g., the amide, imide, or sulfone type. The enzymes of the detergent composition of the invention may be stabilized using convfiai_i.onal stabilizing agents, e.g. a polyol such as propylene glycol or glycerol, a sugar or sugar alcohol, lactic acid, boric acid, or a boric acid derivative such as, e.g., an aromatic borate ester, and the composition may be formulated as described in, e.g., WO 92/19709 and WO 92/19708. The detergent may also contain other conventional deter¬gent ingredients such as, e.g., fabric conditioners in¬cluding clays, foam boosters, suds suppressors, anti-corrosion agents, soil-suspending agents, anti-soil-redeposit ion agents, dyes, bactericides, optical brighteners, or perfume. The pH (measured in aqueous solution at use concentra¬tion) will usually be neutral or alkaline, e.g. in the ranee of 7—li. Particular forms of detergent compositions within the scope of -he invention include: 1} .A deterge.nt composition formulated as a granulate having a bulk density of at least 600 g/l comprising 00 01 - 0.1" 2) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising 3) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising detergent liquid comprising a liquid nonionic surfactant such as, e.g., linear alkoxylated primary alcohol, a builder system (e.g. phosphate), enzyme and alkali The detergent may also corr.prise anionic surfactant and/or a bleach system. The endoglucanase may be incorporated in concentrations conventionally employed in detergents. It is at present contemplated that, in the laundry composition of the in¬vention, the cellulase may be added in an amount corre¬sponding to 0.0001-10 mg (calculated as pure enzyme protein) of cellulase per liter of wash liquor. According to yet another aspect of the present invention, e.ndoglucanase may typically be a component of a conditioning or softener composition. Examples of softener co-positions are disclosed in e.g. EP 0 textile applications In another embodiment, the present invention relates to use of the endoglucanase of the invention in the bio-polishing process. Bio-Polishing is a specific treatment of the yarn surface which improves fabric quality with respect to handle and appearance without loss of fabric wet ability. The most important effects of Bio-Polishing can be characterized by less fuzz and pilling, increased gloss/luster, improved fabric handle, increased durable softness and altered water absorbency. Bio-Polishing usually takes place in the wet processing of the manufacture of knitted and woven fabrics. Wet processing comprises such steps as e.g. desisting, scouring, bleaching, washing, dying/printing and finishing. During each of these steps, the fabric is more or less subjected to mechanical action. In general, after the textiles have been knitted or woven, the fabric proceeds to a desizing stage, followed by a scouring stage, etc. Desizing is the act of removing size from textiles. Prior to weaving on mechanical looms, warp yarns are often coated with size starch or starch derivatives in order to increase their tensile strength. After weaving, the size coating must be removed before further processing the fabric in order to ensure a homogeneous and wash-proof result. It is known that in order to achieve the effects of Bio-Polishing, a combination of cellulytic and mechanical action is required. It is also known that "super-softness" is achievable when the treatment with a cellulase is combined with a conventional treatment with softening agents. It is contemplated that use of the endoglucanase of . the invention for bio-polishing of -cellulosic fabrics is advantageous, e.g. a more thorough polishing can be achieved. Bio-polishing may be obtained cy applying zhe -ernod described e.g. in WO S3/'2027£. Stone-washing It is known to provide a "stone-washed" look (localized abrasion of zhe colour) in dyed fabric, especially in denim fabric or jeans, either by washing the denim or jeans made from such fabric in the presence of pumice stones to provide the desired localized lightening of the colour of the fabric or by treating the fabric enzymatic ally, in particular with cellulytic enzymes. The treatment with an endoglucanase of the present invention may be carried out either alone such as disclosed in US 4,832,864, together with a smaller amount of pumice than required in the traditional process, or together with prelate such as disclosed in WO 95/09225. Pulp and paper applications In the papermaking pulp industry, the endoglucanase of the present invention may be applied advantageously e.g. as follows: - For debarking: pretreatraent with the endoglucanase may degrade the cambium layer prior to debarking in mechanical drums resulting in advantageous energy savings. - For defibration: treatment of a material containing cellulosic fibers with the endoglucanase prior to refin¬ing or beating may result in reduction of the energy consumption due to the hydrolysing effect of the cellulase on the interfere surfaces. Use of the endoglucanase may result in improved energy savings as compared to the use of known enzymes, since i~ is believed that the enzyme composition of the invention may possess a higher ability to penetrate fibre walls. - For fibre modification, i.e. improvemenr of fibre prop¬erties where partial hydrolysis across the fibre wall is needed which requires deeper penetrating enzymes (e.g. in order to make coarse fibers more flexible). Deep treatment of fibers has so far not been possible for high yield pulps e.g. mechanical pulps or mixtures of recycled pulps. This has been ascribed to the nature of the fibre wall structure that prevents the passage of enzyme molecules due to physical restriction of the pore matrix of the fibre wall. It is contemplated that the present endoglucanase is capable of penetrating into the fibre wall. - For drainage improvement. The — papermaking pulps may be improved by treatment of the pulp with hydrolysing enzymes, e.g. cellulose. Use of the present endoglucanase may be more effective, e.g. result in a higher degree of loosening bundles of strongly hydrated micro-fibrils in the fines fraction (consisting of fibre debris) that limits the rate of drainage by blocking hollow spaces between fibers and in the wire mesh of the paper machine. The Canadian standard freeness (CSF) increases and the Schopper-Riegler drainage index decreases when pulp in subjected to cellulase treatment, see e.g. US patent 4,923,565; TAPPI T227, SCAN 019:65.ence. - For inter fibre bonding. Hydrolytic enzymes are applied in the manufacture of papermaking pulps for improving the inter fibre bending. The enzymes rinse the fibre surfaces for impurities e.g. cellulosic debris, thus enhancing the area of exposed cellulose with attachment to the fibre wall, thus improving the fibre-to-fibre hydrogen binding capaci'cv. This -recess is also referred tr as dehcrnification. Paper and board produced wirh a cellulase containing enzyme preparation r.ay have an improved strength or a reduced grammage,, a smoother surface and an improved printability. - For enzymatic deinking. Partial hydrolysis of recycled paper during or upon pulping by use of hydrolysing enzymes such as cellulases are known to facilitate the removal and agglomeration of ink particles. Use of the present endoglucanse may give a more effective loosening of ink from the surface structure due to a better pen¬ etration of the enzyme molecules into the fibrillar matrix of the fibre wall, thus softening the surface whereby ink particles are effectively loosened. The agglomeration of loosened ink particles are also improved, due to a more efficient hydrolysis of cellulo- sic fragments found attached to ink particles or-i-ginating from the fibres. The treatment of lignocellulosic pulp may, e.g., be performed as described in WO 91/14819, WO 91/14822, WO 92/17573 and WO 92/18688. Degradation of plant material In yet another embodiment, the present invention relates to use of the endoglucanase and/or enzyme preparation according to the invention for degradation of plant material e.g. cell walls. It is contemplated that the novel endoglucanase and/or enzyme preparation of the invention is useful in the pre¬paration of wine, fruit or vegetable juice in order to increase yield. Endoglucanases according to the invention may also be applied for enzymatic hydrolysis of various plant cell-wall derived materials cr waste materials, e.g. agricultural residues such as wheat-straw, corn cobs, whole corn plants, nut shells, grass, vegetable hulls, bean .hulls, spent grains, sugar beet pulp, and the like. The plant marerial may be degraded in order to improve different kinds of processing, facilitate purification or extraction of other components like purification of beta-glucan or beta-glucan oligomers from cereals, improve the feed value, decrease the water binding capacity, improve the degradability in waste v.'ater plants, improve the conversion of e.g. grass and corn to ensilage, etc. The following examples illustrate the invention EXAMPLE 1 Cellulytic enzymes from 4 fungi, belonging to 3 families under two orders within the Ascomycetes were detected by expression cloning; corresponding DNA sequences were determined; the enzymes heterologously expressed, and produced by liquid fermentation, characterized and demonstrated to give good performance in colour clarifi¬cation assays. Isolate CBS 117.65, CBS 478.94, NRRL 8126, and ATCC 10523 were grown in shake flask cultures on cellulose enriched potato dextrose broth, incubated for 5 days at 2 6"c (shaking conditions, 150 rpm). A. Cloning and expression of an endoglucanase from Myceliophthora tbermophila, Acremonlum sp.. and Thielavia terrssTiris and Volutella collstotrichcldes mRNA was isolated from Myceliophthora thermophila, Acremcnium sp., Thielavia terrestris and"'/olu tell a colletotrichoides, respectively, crcwTi in a cellulose-containing fermentation medium with agitation to ensure sufficient aeration. Mycelia were harvested after 3-5 days' growth, immediately frozen in liquid nitrogen and stored at -80°C. Libraries from y.yceliophthora thermophila, Acreiuohium sp., Thielavia terrestris and Volutella colletotrichoides, respectively, each consisting of approx. 10* individual clones were constructed i.n E. coli as described with a vector background of 1%. Plasmid DNA from some of the pools from each library was transformed into yeast, and 50-100 plates containing 250-400 yeast colonies were obtained—^om each pool. Endoglucanase-positive colonies were identified and isolated on SC-agar plates with the AZCL HE cellulose assay. cDNA inserts were amplified directly from the yeast colonies and characterized as described in the Materials and Methods section above. The DNA sequence of the cDNA encoding the endoglucanase from Myceliophthora thermophila is shown in SEQ ID No. l and the corresponding amino acid sequence is also shown in SEQ ID No. 1. The cDNA is obtainable from the plasmid in DSM 9770. The DNA sequence of the cDNA encoding the endoglucanase from Acremonium sp. is shown in SEQ ID No. 4 and the corresponding amino acid sequence is shown in SEQ ID No. 5. The cDN.A is obtainable from^ the pl3sm,id in DSM 10082. The DNA sequence of the cDKA encoding the endoglucanase rrorT; __\nej.5V'i3 ^err^s^rxs is snovn m ^r-v XLJ HO. O anc the corresponding amino acid sequence is shown in SEQ ID No. 9. The cDNA is obtainable from the plasmid in DSM 10031.' The DNA sequence of the cDNA encoding the endoglucanase from Volutella colletotrichoides is shown in SEQ ID No. 16 and the corresponding amino acid sequence is shown in SEQ ID No. 17. The cDNA is obtainable from the plasmid in DSM 10571. Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanases in Aspergillus, the D.N'A was digested with appropriate restriction enzymes, size fractionated on gel, and a fragment corresponding to the endoglucanase gene from Hyceliophthora thermophila, Acrewdnium sp., ThTWlavia terrestris and Volutella colletotrichoides, respectively, was purified. The genes were subsequently ligated to pHD414, digested with appropriate restriction enzymes, resulting in the plasmids pA2Cl93, pA2C357, pA2C385 and pA2C488, respectively. After amplification of the DNA in E. coli the plasmids were transformed into Aspergillus oryzae as described above. Test of A. oryzae transformants Each of the transformants were tested for endoglucanase activity as described above. Some of the transformants had endoglucanase activity which was significantly larger than the Aspergillus oryzae background. This demonstrates efficient expression of the eridoglucanases in Aspergillus oryzae. The transformants with the highest endoglucanase activity were selected and inoculated in a 500 ml shake flask vith YPM r.edia. .After 3-5 days of fer-entation with sufficient agira~io- to ensure good aerarion, "he culture broth was centrifuged for 10 minutes at 20C0 g and the suoernatant recovered. B. Determination of endoglucanase activity The cellulytic activity of the endoglucanase may be determined relative to an analytical standard and expressed in the unit S-CEVU. Cellulytic enzymes hydrolyse CMC, thereby decreasing the viscosity of the incubation mixture. The resulting reduction in viscosity may be determined by a vibration viscosimeter (e.g. MIVI 3000 from Sofraser, France). Determination of the cellulytic activity, measured in terms of S-CEVU, may be determined according to the anal^Lsis method AF 301.1 which is available fro- the-Applicant upon request. The S-CEV^U assay quantifies the amount of catalytic activity present in the sample by measuring the ability of the sample to reduce the viscosity of a solution of carboxy-methylcellulose (CMC). The assay is carried out at 40°C, pH 7.5 using a relative enzyme standard for reducing the viscosity of the CMC substrate. Assay for determination of endoglucanase activity in terms of SAVI units using phosphoric-acid swollen cellu¬lose (PASC): Definition: 1 SAVI-U is the amount of enzyme which form.s an amount of reducing carbohydrates equivalent to 1 umol of glucose per minute. Assay condition: Enzyme solution: 0,5 ml 4 g/1 PASC in 0,1 M Buffer: 2.0 ml 2 0 m,i.n, 40 ■ °C Sensitivity: Max 0.1 5AVIU/ml = approx. 1 S-CE\nj/mi (CMC viscosity) Min 0.01 SAVIU/ml = approx. 0.1 S-CE'vTO/ml Determination of for.~ation of reducing sugars: The reducing groups assay was performed according to Lever, M. A new reaction for colormetric determination of carbohydrates. Anal. Biochem, 1972. Vol 47 (273-279). Reagent mixture was prepared by mixing 1,5 gram, p-hydroxybenzoic-acide hydracide (PHBAH) with 5 gram sodium tartrate in IGO ml 2 % sodium hydroxide. Substrate: PASC stock solution was prepared the following way using ice cold acetone and phosphoric acid. 5 gram of cellulose (Avicel) was moistered with water, and 150 ml ice cold 85% ortho-phosphoric acid was added. The mixture was placed in ice bath under slow stirring for 1 hr. Then 100 ml ice cold acetone was added with stirring. The slurry was transferred to a Buchner filter with pyrex sintered disc number 3 and then washed three times with 100 ml ice cold acetone, and sucked as dry as possible after each wash. Finally, the filter cake was washed twice with 500 ml water, sucked as dry as possible after each wash. The PASC was mixed with deionized water to a total volume of 300 ml, blended to homogeneity (using the Ultra Turrax Homogenizer) and stored in refrigerator (up to one month). Substrate equilibration with buffer: 20 gram phosphoric acid swollen cellulose PASC stock solution was centrifu-ged for 20 r.in ar 5CC0 rpm. , rhe superna-an- was poured of; the sediment was resuspended in 30 ml of buffer and centrifuged for 20 -in. at 5000 rpm., the supernatant was poured of, and the sediment was resu^spended i.n buffer io a total of 60 g corresponding to a subsrrate concentra¬tion of 5 g cellulose/litre. Buffer for pH S,5 determination: 0.1 M Barbital. Buffer for pH 10 determination: 0.1 M Glycine. 1. Dilution of enzyme samples The enzyme solution is diluted in the same buffer as the substrate. 2. Enzyme reaction The substrate in buffer solution is preheated for 5 min. at 40C (2 ml). Then the enzyme solution (diluted to between 0.2 and 1 S-CEVU/ml) 0,5 ml is added and mixed for 5 sec. Enzymes blanks are obtained by adding the stop reagent before enzyme solution. Incubate for 20 min. at 40 'c. The reaction is stopped by adding 0.5 ml 2% NaOH solution and mixing for 5 sec. The samples are centrifuged for 20 min. at 5000 rpm. 1 ml supernatant is mixed with 0.5 ml PHBAH reagent and boiled for 10 min. The test tubes are cooled in a ice water bath. 3. Determination of reducing end groups: The absorbancy at 410 nm is measured usi.ng a spectrophotom.eter. Blanks are prepared by adding sodium hydroxide before adding enzyme solution. k standard glucose curve was obtained by using glucose ccncentraticns of 5, 10, 15 and 25 mg/1 in the sane buffer and addin^ ?HH.-vH reacent before boilinc. The release of reducing glucose equivalent is calculated using this standard curve. 4. Calculation cf catalytic activity: Measure absorbance at 410 nm 1) Standard curve (Glucose) - (HpO) vs concentration of glucose 2) Enzyme sample (Sample) - (Blank) Calculate glucose concentration according to a standard Activity (SAVIU/ml): X (mg glucose/1) * Dilution ISO.16 (MW of glucose) * 20 (min) C. Purification and characterisation of the endoglucanase from M. thermophila Aspergillus oryzae transformed with pA2C193 was grown on YPM medium for 4 days. The liquid was then centrifuged and sterile filtered. -The.sample was.concentrated by ultrafilrration -on AMICON cells using a DOW membrane GR61P? with cut-off 20 kD. The Uf-concentrate was analyzed for S-CE'v'U/ml and SaviL'/mi '—■ ^ ^ '^- V* X ; UF-concentrate \ S- i ; CE'.'TF/ml Saviu/ml 9.25 ml 1 570 1 41 Purification: 2 ml of the UF-concentrate was diluted 5 times to lower the ionic strength and filtered through 0.2 2 um disk filter. This sample was applied to a Mono Q* HR5/5 Phar¬macia column, equilibrated with 50 mM Tris/HCl buffer. pH 7.5, (buffer A) and a flow of 1 ml/min. After wash to baseline, with buffer h, the colum.n was eluted with a Tris/HCl buffer, pK 7.5, containing 1 M IJaCl (buffer B) , the elation gradient was 0-50% buffer B in 1 hour. After 36 min. a peak complex showed up, 1 ml fractions were picked up and the first 10 fractions showed cellulase activity on CMC/Agarose/congo-red plates. These fractions were pooled and concentrated, by ultrafiltration on AMICON cells using a DOW membrane GR61PP with cut-off 20 kD, to 3 ml. This sample was applied to a HiLoad 26/60 Superdex 75"" prep grade Pharmacia column, equilibrated with 100 mM Na-Acetate buffer, pH 6.35, and a 1 ml/min flow. After 82 min. a peak showed up, 1 ml fractions were picked up and.the first 10 fractions showed cellulase activity on CMC/Agarose/congo-red plates. These fractions were pooled and the following results were obtained: A;so='^- 15, A2so/A26o=l-62 Mw(5DS:=2 2 kD pl=3.5-5 Purity on SDS-PAGE =100% S-CEVU/ml=28.5 S-CEVU/A:so=lS3 S-CEV~u/mg=4 3 6 Extinction ccefficient=54880 (calculated) •Mw(calculated)=22 kD The Extinction coefficient is based on the content of tyrosine, tryptophane and cystein calculated fro- the sequence cf the enclcsad SEQ ID No. 1 (the amino acid sequence). SDS-Page was performed on NOVEX Pre-Cast Gels 4-20% Tris-Glycine Gel 1.0 mm x 10 Well lEF was performed on Pharmacia FAGplate pH 3.5 - 9.5, the activity was visualized by CMC-Congored overlaying. DPtPrmination of K^, & k^..,: k„ and k„, was determined in the same manner as the determination of SAVI Units at pH 8.5 with a substrate concentration up to 8 g/1. The following results were obtained: k^.i 3 8 per sec. >v„, 5 g/1, phosporic acid swollen cellulose, pH S Specific activity on CMC at pH 7.5: 435 S-CEVU per mg protein. D. Determination of pH and temperature profile of the endoglucanase from M. thermophila The pK profile was determined at the following conditions: mixing G. IM Tri-sodium phosphate with O.IM cimc acid. Purified -endoglucanase was diluted to ensure rhe assay response to be within the linear range of the assay. The substrate was a 0.4% suspension of AZCL-HZ-cellulcse (MegaZyme) mixed 1:1 with the citrate/phosphate buffer to a final substrate concentration of 0.2% AZCL-HE-cellulose. 1 ml substrate in Eppendorf® 1.5-i polypropylene tubes were added 10 p.1 of enzyme solution and incubated for 15 minutes in Eppendorf^' temperature controlled Thermomixers before heat-inactivation of enzymes for 2U minutes at 95°C in a separate Thermomixer. The tubes were centrirugea and 200 /il of eacn supernatant was transferred to a well in a 96 well micrctiter plate and OD was measured at 620nm in an ELISA reader (Labsystems Multiskan® MCC/340). For the pH optiraumi incubations took place an 30°C. For each pH value, three tubes were added enzyme and incubated before heat-inactivation, whereas one tube (the blank) was added enzyme and heat-inactivated immediately. The mean value of the three incubated samples was calculated and the blank value was substracted. The following pH profile was determined: pH Relative Activity 2.5 :10% ■'10% 3 . 5 4 . 5 5 100% 1 S 6.5 S6% 7 7S% 7.5 8 68% S. 5 = ■ 3-_/ -t o 9 31% 10 lO-S It is seen that the endoglucanase has more than 60% activity between pH 4.0 and 8.0 and optimal activity at pH 5.0-6.0. ~ Temperature profile: The temperature optimum was determined in the same manner at pH 5.5. The temperatures ranged from 30^0 to 80°C. For each temperature three incubations were carried out and the mean calculated. Three blanks were produced by immediate heat-inactivation of enzyme and the mean was subtracted from the incubated sample values. It is seen that the endoglucanase has optimal activity at 50-70°C. Temp. {°C) 30 40 50 60 70 80 Relative Activity 7 4^0 77% 99% 100% 93% 62% The temperature stability was determined in the same manner at pH 5.5 and 3 0°C, and, further, the enzyme solu¬tions were preneate'.i _or 1 hour at une accuaj. cemperatur^ and cooled on ice. The residual activity is shown below in % of the activitv of a non-oreheated enzvme samnle: 40 50 60 70 80 Relative Activity 95% 84% 92% 86% 24% E. Color clarification of Myceliophthora cellulase (SEQ ID No. 1) measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers Apparatus Liquid volume Agitation stirrer Rinse time Washing temp Washing liqour pH Washing time Repetitions Enzymes Dosage Texrile Drying Evaluation Terg-o-tometer 100 rol 150 movements/min with vertical 5 min in tapwater 40° 0.05 M phosphate buffer 7.0 3 0 nin 2 Myceliophthora SEQ ID N'o. IB 500 and 2500 S-CEVU/1 2 swatches cf aged clack 100% ccrrcn 5x6 cm (0.9 gram) Tumble dry The light re-issicn is -easured by a Datacolcr Elrepho Remission spectrophotometer. Rem.issicn is calculated as delta L (Hunter Lab-values) . When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker, and lower L values are obtained. The sample is compared with a blind sample, i.e. washed without enzyme: No cellulase 500 ECU/1 2500 ECU/i 0.00 -1.41 -1.3-3.— Delta L-values compared to blind sample. The data shows that Mvceliophthora cellulase without CBD gives good color clarification under the conditions tested. F. Construction of the gene fusions between the endoglucanase from Myceliophthora thermophila and the 43k.D endoglucanase from Humicola insolens The purpose of the two constructions was to make derivatives of the endoglucanase from M. thermophila with the linker and CBD from the 43kD endoglucanase from H. insolens (disclosed in WO 91/17243). The native endoglucanase from M. thermophila do not have a linker and/or a cellulose binding domain, CBD. CMl: Construction 1 consist of the endoglucanase from .M. thermophila (225 amino acids) and the 72 C-ter~inai amiinc acids from the H. insolens 43kD endoglucanase. CM2: Construction 2 consist of the endoglucanase from .M. thermophila (225 amino acids) and the S3 C-terminal amino acids from the H. insolens 43kD endoglucanase. The 43kD endoglucanase cDNA from H. insolens was cloned into prtD414 in such a way that the endoglucanase gene was transcribed from the Taka-promoter. The resulting plasm.id v;as nam.ed pCaHj4 1S. In a similar way the cDNA encoding the endoglucanase from M. thermophila was cloned into pHD414 and the resulting plasm.id was named pA2C193. Primers: primer 1: 5'- CGGAGCTCACGTCCAAGAGCGGCTGCTCCCGTCCCTCCAGCAGCACCAGCTCTCCGG -3' primer 2: 5' CCGGAGAGCTGGTGCTGCTGGAGGGACGGGAGCAGCCGCTCTTGGACGTGAGCTCCG -3 ' primer 3: 5'- CGGAGCTCACGTCCAAGAGCGGCTGCTCCCGTAACGACGACGGCAACTTCCCTGCCG -3' primer 4: 5'- CGGCAGGGiiJvGTTGCCGTCGTCGTTACGGGAGCAGCCGCTCTTGC-ACGTGAGCTCCG -3 ' Taka-pro. primer: 5' CAACATCACATCAAGCTCTCC -2' AMG-term. primer: 5' CCCCATCCTTTAACTATAGCG -3' The endoglucanase fusions were constructed by the PCR overlap-extension method as described by Higuchi et al. 1988. Construction 1: Reaction A: The Polymerase Chain Reaction (PCR) was used to amplify the fragment of pCaKj418 between primer 1 and ;i>IG-term. primer (the linker and CBD from the 4 3kD endoglucanase from H.insolens). Reaction B: PCR amplification of the fragment between Taka-pro. primer and primer 2 in pA2C193, the endoglucanase gene from M.thermophila. Reaction C: The two purified fragments were used in a third PCR in the presence of the primers flanking the total region, i.e. Taka-pro. primer and AMG-term. primer. Cons^ifttction 2 : The same procedure was used where primer 3 and primer 4 had replaced respectively primer 1 and primer 2. The fragment amplified in reaction C was purified, digested with restriction enzymes Xba I and BsstE II. The purified digested fragment was ligated into pA2C193 digested with restriction enzymes Xba I and BsstE II. Competent cells from E. coli strain DH5aF' (New England Biolabs.) were transformed with the ligated plasmid and colonies containing the gene fusion were isolated. The sequence of the cloned part was verified by DNA sequencing. The sequence of the gene in the two constructs are shown in SEQ ID No. 2A and SEQ ID No. 3A. Polymerase Chain Reactions were carried out under standard conditiicns, as reccmmended by Perkin-Elmer. Reaction A and B started with 2 min. at 94°C followed by 20 cycles of {30 sec. at 94-C, 30 sec. at 50°C and 1 min. at 72°C) and end with 4 min. at 72 -C. Reaction C started with (2 min. at 94=C, 1 min. at 52°C and 2 min. at 72°C), followed by 15 cycles of (30 sec. at 94°C, 30 sec. at 52°C and 90 sec. at 72°C) and end with 4 min. at 72°C. The two constructs were transformed into Aspergillus ory-zae as described above. G. Purification and characterisation of cloned cellulases with cellulose binding domains: The cloned product is recovered after fermentation by separation of the extracellular fluid from the production organism. About one gram of cellulase is then highly purified by affinity chromatography using 150 gram of Avicel in a slurry with 20 mm Sodium- phosphate pH 7.5. The Avicel is mixed with the crude fermentation broth which contain total about 1 gram of cellulase. .After mixing at 4 C for 20 rain the Avicel enzyme is packed into a column with a dimension of 50 times 200 mm about 400 ml total. The column is washed with the 200 ml buffer, then washed with 0.5 M NaCl in the same buffer until no more protein elutes. Then washed with 500 ml 20 mir. Tris pH 8.5. Finally the pure full length enzyme is eluted with 1% triethylainine pH 11.8. The eiured enzy~e solution is adjusted ro pH S and concentrated using a Amicon cell unit with a membrane DOW GR61?? (polypropylene with a cut off of 20 KD) to above 5 The purified cellulases were characterised as follow: Mw pi Molar E.280 S-CEVU per SDS-PAGE A.280 Myceliophthora /4.950 13 5 Acremonium (SEQ ID No.5) 40 kD 5 68.020 185 Thielavia (SEC ID No.S) 3 5 kD 4.3 52.4 70 ~5 instructions. Two of the cellulases were blocked, this is due to the N-terminal glutamine which form a pyroglutamate which can not be detected and which block for further sequencing. DSC Differential scanning calometry was done at neutral pH (7.0) using a MicroCalc Inc. MC calorimeter with a constant scan rate and raising the temperature from 20 to 90° at a rate of 90° per hour. Raising antibody. The cellulases from Myceliophthora, Acremonium and Thielavia were used for raising antibody in rabits. 0.1 mg of the purified cellulase in 0.9 % NaCl solution mixed with Freunds adjuvant immediately prior to injection. The rabits were immunized 10 times with one week interval. The immunoglobulin G fraction (IgG) was purified by a-r.cnium sulfate precipitation (25% saturation), the precipitate was solubiiized in water and then dialyzed extensively against sodium acetate buffer (pH 5.0, 50 m-M) altering with deionized water. .A.fter filtration, the IgG fraction was stabilized with sodium" azide (0.01%) . Using immunodiffusion in agar plates all three cellulases form a single immunoprecipitate with its homologous antiserum and no precipitate was seen between the 3 cloned cellulases and the sera raised against the other two cellulases. H-I. Performance of endoglucanase of construction 1 (SEQ ID No. 2) measured in buffer as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers Apparatus Liquid volume Agitation Rinse time Washing temp Water Hardness Washing liquor PH Washing rime Repetitions Textile Drying Terg-o-tometer 100 ml 150 movements/min (rpm) 5 min in tap water 40°C 1 mM CaCl, 0.05 M phosphate buffer 7 .0 30 min 2 2 swatches of aged black, 100% cotton 5x6 c- Tumfcle dry Evaluation: The light "remission was measured by a Macberh Color Eye 7000 Remission spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn were removed by the cellulase, the surface appeared more bright, and lower L values were obtained. Results: 2 50 1 1000 Inventive enzyme 0 -1.4 -1. 6 , The data show that the enzyme of the invention gives very good'color clarification under the conditions tested a-II. Performance of cloned endoglucanase from Thielavia terrestris (SEQ ID No.9) in buffer measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers Apparatus Liquid voluine Agitation Rinse time Washing temp Washing liqour pH Washing time Repetitions Textile Terg-o-tometer 100 ml 150 movements/min with vertical stirrer 10 min in tapwater 40° 0.05 M phosphate buffer. 7.0 30 min 2 2 swatches of aged black cotton 5x6 cm (app. 150 g/m2) :Lva j.uation Tumble dry ~oior Elrepho Remission- spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker and nicer, and lower L values are obtained. Results S-CEVU/1 0 50 2 00 Inventive enzyme 0 -0.56±0.10 -1.3210.06 The data show that the cellulase gives good color cLaxification under the conditions tested. H-III. Performance of endoglucanase of Volutella colletrichoides (SEQ ID No. 17) measured in buffer as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers Apparatus Liquid volume Agitation Rinse time Washing temp Washing liqour pH Washing time Repetitions Dosage Textile Terg-o-tometer 100 ml 150 movements/min with vertical stirrer 5 min in tapwater 40° 0.05 M phosphate buffer 7.0 30 min 2 2.5 S-CEVU/ml 2 swatches of aged black 100% 5x5 cm (0.5 gra.~) Drying : Tumble dry Evaluation: The light remission is measured by a Datacoior Eirephc Remission spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker, and lower L values are obtained. The sample is compared with a blind sample, i.e. washed without enzyme: No cellulase With cellulase O.CO -0.57 Delta L remission values compared to blind sample. The data~"STTows that the Volutella colletrichoides cellulase gives good color clarification under the conditions tested. H-IV. Performance of cloned cellulases from Thielavia terrestris and Acremonium sp. CBS 4 7 8.94 in high pH heavy duty detergent measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers Apparatus Liquid volume Agitation Rinse time Washing temp Washing liqour Terg-o-toineter 150 ml 150 movements/min with vertical stirrer 10 min in tapwater 35'C l.D g/1 US type KDG (zeolite/soda built, anionic/ncnionic weight ratio > 2.5) PH Hardness Washing time Repetitions Textile 10. 0 1.0 sM CaCl2 ■-'.—' T iil^ - jn -. >:6 2 swatches of aged black co: Cm (app. 150 g/rri2) 2 swatches of heavy knitted co~zor. 5x6 cm (app. 600 g/in2) Drying : Tumble dry Evaluation : The light remission is measured by a Datacolor Elrepho Remission spectrophotometer. Remission is caiculared as delta L (Hunter Lab-values). When the surface fibrils and fibcLti pruLruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker and nicer, and lower L values are obtained. Different dosages of cloned cellulases from Thielavia terrestris (SEQ ID No. 9) and Acremonium sp. CBS 478.94 (SEQ ID No.5), respectively, (denoted A and B, respectively) were tested. Jesuits: S-CEVU/1 0 500 2000 A 0 -2.09 ±0.22 -2 .36 ±0 . 19 B 0 -0. 60 ±0.36 -1.96 ±0. 23 The data show that both cellulases gives good color clarification under the conditions tested. K-V. Performance of cellulases cloned from Tbielavia terrestris and Acremonium sp. CBS 47S.94, and construction 1 (SEQ ID No.2) measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers .ppc Liauid volume Rinse time Washing temp Hardness Washing liqour Washing time Repetitions Textile Terg-o-tometer 150 ml 150 mcvemencs/ir.in virh vertical stirrer 10 min in tapwater 35°C 1.0 mM CaCl2 0.3 4 mM MgC12 2.0 g/1 HDL {neutral, citrate built HDL, with nonionic/anionic weight ra¬tion > 0.5) 7 . 5 3 0 min 2 2 swatches of aged black cotton 5x6 cm (app. I5a g/m2} , 2 swatches of heavy knitted cotton 4x7 cm (app. 600 g/m2) Drying : Tumble dry- Evaluation : The light remission is measured by a Datacolor Elrepho Remission spectrophotometer. Remission is calculated as delta L (CIE Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker and nicer, and lower L values are obtained. Three different dosages Different dosages of cloned cellulases from Thielavia terrestris (SEQ ID No. 9) and Acremonium sp. CBS 478.94 (SEQ ID No.5) and the construction 1 (SEQ ID No.2), respectively, (denoted A and B and C, respectively) were tested. Results: s- CEVU/1 0 100 200 400 ^ '^ -3 . 06 ±0. 24 -3. 15 ±0. 27 -3.92 ±0.26 B c -1.75 ±0.27 -3 . OS ±0.32 -3.51 ±0.44 C 0 -1.84 ±0.39 -1.70 ±0.47 -2.30 ±0.61 The data show that all cellulases gives very good color clarification under the conditions tested. I. Application of endoglucanases from Thielavia terre¬stris, Acremonium sp. and construction 1 (SEQ ID No. 2) in denim finishing Experimental Apparatus: Liquid volume: Fabric: Washing machine Wascator FL 120 20 L 1.1 kg denim fabric, 14% oz 100 cotton Desizing: 10 min, 55°C, pH 7 50 ml Aquazyme 120L 2.5 g/1 Phosphate buffer Abrasion: 2 hours pH and temperature varied according to the following table: Enzyme Activity pH/temp Buffer system SEQ ID No. 2 1400 S-CEVU/g 6/55°C 2.5 g/1 phosphate buffer No. 9 292 S-CEVU/g 5/65"C 1 g/1 citrate buffer No. 5 782 S-CEVU/g 7/45'C 2.5 g/1 phosphate buffer Inactivaricn: 15 -in, SO'C 1 g/ sodium carbonate Rinses: Three rinse cycles of 5 min in cold tap water Evaluation: Abrasion: The remission' from the fabric was determined at 420 nm using a Ticklish 2000 as a measure of the abrasion level. The results from the treatment of the denim fabric with different endoglucanases of the invention is shown in the following table: Enzyme Dosage Trial conditions Abrasion 42 0 mn Blank 0 S-CEVU/g textile pH 6, 55'C 9 . 96 SEQ ID No. 2 10 S-CEVU/g textile pH 6, 55°C 14 . 37 Blank 0 S-CEVU/g textile pH 5, 65C 9. 26 SEQ ID No. 9 10 S-CEVU/g textile pH 5, 65°C 16.86 Blank 0 S-CZ\Ti/g textile pH 7, 45C 9 . 47 SEQ ID No. 5 1 10 S-CEVU/g pH 7, 45C 14 . 03 1 i All rested cellulases show excellent performance in denim finishing, although each enz>Tne is unique in its own way. When applying the enzyme corresponding ro SEQ ID No. 2 for denim finishing it is possible to reach a high abrasion level with a minimum of strength loss. When treating denim with the enzyme corresponding to SEQ ilD No. 9, a very high wash down can be reached which leaves the fabric with an almost bleached appearance. Denim finishing with the enzyme corresponding to SEQ ID No. 5 gives a high abrasion level at a low temperature optimum which makes it possible to reduce the processing tempera¬ture and save energy. Lyocell fibers which are sold under the trade name Te' are spun from wood pulp cellulose in a more environmentally friendly waterbased solvent than is the case for normal viscose production). However, the fibers have a tendency to fibrillate when they are processed into textiles which is seen on the surface and denoted "fuzz". By using cellulases it is possible to permanently remove the exposed and fuzzy fibers and significantly improve the look of the finished fabric, the treatment generally known as Biopolishing. The endoglucanases of the present invention are especially suited for the removal of Lyocell surface fibers. MATERIALS AND METHODS The textile substrate was either 100 % woven or different kinds of jersey knitted dark blue Tencel. The dark -colour and jersey knit was preferred in order to enhance the visual effects which simplifed the evaluation. A woven 70/3C Tencel/Kayon blend was also used to a lesser extent. The assays were either performed in 200 ml scale using a Launder-c-meter or in the 20 1 scale using a Wascator. The treatment time was 6Cmin at 55' C in Wascator and SO-SO min in LOM. The buffer was 2 g/i sodium acetate adjusted to pH 5 with acetic acid. The fabric to liquid ratio was 1:10 but in the Launder-o-meter 20 steel balls with a diameter of 14 mm (11 g each) was used to obtain sufficient mechanical abrasion. The biopolishing was immediately followed by inactivation using 2 g/lit sodium carbonate at 80° C for 15 min followed by rinsing in cold water. The results were evaluated using a fuzz note scale from 1 - 5 were 1 is the fibrillated look of the starting material and 5 is a high quality look with no visible fibers on the surface. Since the performance of endocellulases is specific towards a'^snxface treatment the weightloss is below 2 % and is therefore not included in the evaluation. Two cellulases were evaluated: the cellulases cloned from Acremonium sp. (SEQ ID No. 5) and from Thielavia terrestris (SEQ ID No. 9). The two cellulases are able to defibrillate both Tencel and Tencel blended fabrics. By using an endoglucanase of the invention, only small fibrils are removed rather than whole fibers such as is the case when using acid cellulase mixtures from Trichoderma. The strength loss of the treated fabric is threrefore kept at a minimum when using endoglucanases of the present invention. The following dosages gave a superior defibrillation, i.e. fuzz note 4 or above: 15 S-CEVU/g fabric of cellulase from Acrerao.iium sp (SEQ ID No. 5); and 10 S-CEVU/g fabric of cellulase from Thelavia terrestris (SEQ ID No.S). EXAMPLE 2 A new cellulytic enzyme was by expression cloning as well as by PCR cloning detected to be produced by a plant pat¬hogen, isolated from soy bean seeds and identified as Ma-crophomina phaseolina. Production of biomass for PCR and expression cloning procedures: Isolate CBS 281.96 was grown in shake flask cultures on I cellulose enriched potato dextrose broth, incubated for 5 days at 260C (shaking conditions: 150 rpm). A. Cloning and expression of an endoglucanase from Macrophomina phaseolina mRNA was isolated from Macrophomina phaseolina, grown in a cellulose-containing fermentation medium with agitation to ensure sufficient aeration. Mycelia were harvested after 3-5 days' growth, immediately frozen in liquid nitrogen and stored at -30°C. A library from Macrophomina phaseolina, consisting of approx. 10" individual clones was constructed in E. coli as described with a vector background of 1%. Plasmid DNA from some of the pools was transformed into yeast, and 50-100 plates containing 250-400 yeast colonies were obtained from each pool. Endoglucanase-positive colonies were identified and isolated on SC-agar plates with the AZCL HE cellulose assay. cDNA inserts were amplified directly from the yeast colonies and characterized as described in the Materials and Methods section above. The D^ The cDNA' is obtainable from rhe plasmid in DSM 10512. Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanse in Aspergillus, the DNA was digested with appropriate restriction enzymes, size fractionated on gel, and a fragment corresponding to the endoglucanase gene was purified. The gene was subsequently ligated to pHD414, digested with appropriate restriction enzymes, resulting in the plasmid pA2C477. After amplification of the DNA in E. coli the plas-.id was transformed into Aspergillus oryzaa as described above. Screening of the cDNA library by hybridization and characterization of the positive clones. Approximately 6000 colony forming units (c.f.u.) from the Macrophomina phaseolina cDNA library in E. coli was screened by colony hybridization using a random-primed "P-labeled PCR product from M. phaseolina as probe. The PCR product was generated as described in the Materials and methods section. The positive cDNA clones were characterized by sequencing the ends of the cDNA inserts, and by determining the nucleotide seuence of the longest cDNA from both strands. The DNA sequence of the cDNA encoding the endoglucanase is shown in SEQ ID No. 10 and the corresponding amino acid sequence is shown in SEQ ID No. 11. B. Construction of gene fusion between the endoglucanase from Macrophomina phaseolina and the 4 3 k.D endoglucanase from Humicola insolens One construction was prepared in order to make a derivative of the endoglucanase from M. phaseolina with the linker and C5D from the 4 3 kD endoglucanase from H. insolens (disclosed in WO 91/17243). The native endoglucanase from M. phaseolina does not have a linker and/or a cellulose binding domain, CBD. The construction consists of the endoglucanase from, M. phaseolina (223 anino acids) and the 72 C-terminal amino acids from the H. insolens 4 3 kD endoglucanase. The 43 kD endoglucanase cDNA from H. insolens is cloned into pHD414 in such a way that the endoglucanase gene is transcribed from the Taka-promoter. The resulting plasmid is named pCaHj4lS. The'cDNA encodirrg"the endoglucanase from M. phaseolina is cloned into pYES2.0 as a BstX I/Not I fragment and the resulting plasmid is named pClC477. Primers: primer 1: 5'- GGTCGCCCGGACTGGCTGTTCCCGTACCCCCTCCAGCAGCACCAGCTCTCCGG -3' primer 2: 5' CCGGAGAGCTGGTGCTGCTGGAGGGGGTACGGGAACAGCCAGTCCGGGCGACC -3' pYES2.0 F.HT primer: 5' CGGACTACTAGCAGCTGTAATACG -3' AMG-term. primer: 5' CCCCATCCTTTAACTATAGCG -3' The endoglucanase fusion is constructed by the PCR overlap-extension method as described by Higuchi et al. 1988. Reaction A: The Pclymerase Chain Reaction (PCR) is used to'amplify the fragment of pCaHj418 between printer 1 and AMG-term. primer (the linker and CBD from the 4 3 kD endoglucanase from H.insolens). Reaction B: PC?, amplification of the fragmen" berwier. pYES2.0 F.HT primer and primer 2 in pClC477, the endoglucanase gene from M. p'naseolina. Reaction C: The two purified fragments are used in a third PCR in the presence of the primers flanking rhe total region, i.e. pYES2.0 F.HT primer and AMG-term. primer. The fragment amplified in reaction C is purified, digested with restriction enzymes, e.g. Xba I and BamH I. The purified digested fragment is ligated into pHD4 14 digested with restriction enzymes, e.g. Xba I and BamH I. Competent cells from E. coli strain DHSaF' (New England Biolabs) are transformed with the ligated plasmid and colonies containing the gene fusion are isolated. The sequence of the cloned part was verified by DNA sequen¬ cing. Polymerase Chain Reactions are carried out under standard conditions, as recommended by Perkin-Elmer. Reaction A and B start with 2 min. at 94°C followed by 20 cycles of (30 sec. at 94°C, 30 sec. at 52°C and 1 min. at 72°C) and ends with 4 min. at 72 =C. Reaction C starts with (2 min. at 94°C, 1 min. at 52°C and 2 min. at 72°C), followed by 20 cycles of (30 sec. at 94°C, 30 sec. at 52°C and 90 sec. at 72°C) and ends with 4 min. at 72°C. The construct may be transformed into Aspergillus oryzae as described above. EXAMPLE 3 Cloning and expression of an endoglucanase froE Acremonlum sp. and Sordaria fimicola Production of biomass for expression cloning procedures: Isolates CBS 473.94 and ATCC 52644, respec~iveiy, were grown in shake flask cultures on cellulose enriched pota¬to dextrose broth, incubated for 5 days at 260C (shaking conditions: 150 rpm). mRNA was isolated from Acremonium sp., C5S 478.94, and Scrdaria flrrdcola, ATCC 52644, respectively, grown in a cellulose-containing fermentation medium with agitation to ensure sufficient aeration. Mycelia were harvested after 3-5 days' growth, immediately frozen in liquid nitrogen and stored at -80°C. Libraries from Acrer-:onlum sp., and Sordaria fimicola, respectively, each consisting of approx. lo' individual clones were constructed in E. jzpli as described with a vector background ofTT. Plasmid DNA from some of the pools from each library was transformed into yeast, and 50-100 plates containing 250-400 yeast colonies were obtained from each pool. Endoglucanase-positive colonies were identified and isolated on SC-agar plates with the AZCL HE cellulose assay. cDNA inserts were amplified directly from the yeast colonies and characterized as described in the Materials and Methods section above. The DNA sequence of the cDNA encoding the endoglucanase from Acremonium sp. is shown in SEQ ID No. 6 and the corresponding amino acid sequence is shown in SEQ ID No.7. The cDNA is obtainable from the plasmid in DSM 1003 0. The partial DNA sequence of the cDNA encoding the endoglucanase from Sordaria fi~dccla is shown in SEQ ID No. 19 (Nucleotide sequence of rhe 5'-e.nd of the cDNA) and the corresponding amino acid sequence is shown in SEQ ID No. 20. The cDNA is obtainable from the plasmid in DSM Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanase in Aspergrillus, the Dl^k was digested with appropriate restriction enzymes, size fractionated on gel, and a fragm.ent corresponding to the endoglucanase gene from Acremonium sp. and Sordaria fimicola, respectively, was purified. The genes were subsequently ligated to pHD414, digested with appropriate restriction enzymes, resulting in the plasmids pA2C371 and pA2C502, respectively. After amplification of the DNA in E. coli the plasmids were trarrsformed into Aspergillus oryzae as described above. EXAMPLE 4 A. Cloning by PCR an endoglucanase from Crinipellis sca-bella, CBS 280.96 Isolate CBS 280.96 was grown in static flask cultures, holding wheat bran medium (per flask: 300g wheat bran added 450 ml salt solution), incubated for 6 days at 26C. After incubation the wheat bran was extracted with destilled water (300ml per flask) and the extract tested for endoglucanase activity (0.1% AZCL-HE-Cellu]ose (megazyme) in 1% agarose (Litex agarose, Medinova). Activity was observed on the plates holding pH of 3.0, 7.0 and S.5. mRN-ii was isolated from Crinipellis scabella grown as describe above. Mycelia were harvested after 3-5 days' growxh, luiTTiediatiely froze.n m liquid nitrogen and stored at -60'C. A library from Crinipellis scabella, consisting of apprcx. iO' individual clones was constructed in I. coli as described with a vector background of 1%. Approximately 10 000 colony forming units (c.f.u.) from the Crinipellis scabella cDNA library in E. coli was screened by colony hybridization using a random-primed "P-labeled PCR product from C. scabella as probe. The PCR product was generated as described in the Materials and methods section. The positive cDNA clones were characterized by sequencing the ends of the cDNA inserts, and by determining the nucleotide seuence of the longest cDNA from both strands. The DNA sequence of the cDNA encoding the endoglucanase is shown in SEQ ID No. 12 and the corresponding amino acid-ssqiience is shown in SEQ ID No. 13. The cDNA is obtainable from the plasmid in DSM 10511. Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanse in Aspergillus, the DNA was digested with appropriate restriction enzymes, size fractionated on gel, and a fragment corresponding to the endoglucanase gene was purified. The gene was subsequently ligated to pHD414, digested with appropriate restriction enzymes, resulting in the plasmid pA2C475. After amplification of the DNA in E. coli the plasmid was transformed into Aspergillus oryzae as described above. Construction of two gene fusions between the endoglucanase from Crinipellis scahella and the linker/CBD region of the 43 kDa endoglucanase from cusj.ccxa x~i.sox^ns. The native endoglucanase from Crinipellis scabella neit¬her has a linker nor a cellulose binding domain (CBD). In addition, the full-length cDNA contains an ATG start codon upstream from the endoglucanase encoding open reading frame (ORF), presumably resulting in scrambled translation initiation upon heterologous expression of the cDNA, such as in the yeast Saccharomyces cerevisiae and the filamentous fungus Aspergillus oryzae. Thus, two gene fusions between the endoglucanase from Crinipellis scabella and the linker/CBD region of the 43kD endoglucanase from Humicola insolens (disclosed in WO 91/17243) has been constructed using splicing by overlap extension (SOE) (Horton et al, 1S39). Construction 1 consists of the cDNA encoding the 226-re-sidue fendoglucanase from C. scabella fused by PCR with the 3'-end cDNA of H. insolens coding for the linker and CBD region (72 amino acids) at the COOH-terminus of the H. insolens 43kD endoglucanase. The second hybrid construct is identical to the abovementioned gene fusion, except that the first five residues from the putative signal peptide have been deleted by PCR resulting in a shorter signal, which starts with the second in-frame ATG start codon. Plasmid constructs The plasmid pClC475 contains the full-length cDNA from C. scabella, cloned into BstXI/Notl-cut yeast expression vector pYES 2.0, the plasmid pClC144 contains the full-length cDNA from H. insoiens, cloned into the BstXI site of pYES 2.0. ■■ Splicing by overlap extension Two PCR fragments encoding the core region of the endog¬lucanase from C. scahella were generated in PCR buffer (10 mM Tris-HCl, pH 8.3, 50 inM"KCl, 1.5 r^M MgCl-,, 0.01 % gelatin; containing 200 .LLM each dNTP), usi.ng 50-100 ng of pClC475 as template, and 250 pmol of the reverse primer (5'-GACCGGAGAGCTGGTGCTGCTGGAGGGTTTACGAACACAGCCCGAGATATTAG TG- 3) in two combinations with 300-350 pmol of each forward primer (forward no. 1 5'- CCCCA„AGCTTGACTTGGAACCAJ^TGGTCCATCC-3 " , forward no. 2 5"-CCCCAAGCTTCCATCCAAACATGCTTAAAACGCTCG- 3"), a DNA thermal cycler (Landgraf, Germany) and 2.5 units of Tag polymerase (Perkin-Elmer, Cetus, USA). Thirty cycles of PCR were performed using a cycle profile of denaturation at 94 °C for 1 min, annealing at 55 °C for 2 m.in, and extension at 72 °C for 3 m.in. The PCR' fragffient coding for the linker and CBD of the endoglucanase of H. insolens was generated in PCR- buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 1.5 mM MgCl,, 0.01 % gelatin; containing 200 ^M each dNTP) using 100 ng of the pClC144 template, 250 pmol forward primer (5'-CACTAATATCTCGGGC- TGTGTTCGTAAACCCTCCAGCAGCACCA-GCTCTCCGGTC-3') , 250 pmol of the pYES 2.0 reverse primer (5'-GGGCGTGAATGTAAGCGTGACATA-3■) a DNA thermal cycler (Landgraf, Germany) and 2.5 units of Tag polymerase (Perkin-Elmer, USA). Thirty cycles of PCR were performed as above. The PCR products were electrophoresed in 0.7 % low gelling temperature agarose gels (SeaPlaque, FMC), the fragments of interest were excised from the gel and recovered by treatment with agarase (New England Biolabs, USA) according to the manufacturer's instructions, followed by phenol extraction and ethanol precipitation at - 20 °C for 12 h by adding 2 vols of 96 % EtOH and 0. 1 vol of 3.M NaAc. The recombinant hybrid genes between the endoglucanase from Crinipallis scabella and the linker/CBD region of the 4 3 kD endoglucanase from Humicola insolsr.s were generated by combining the overlapping PCR fragments from above (ca. 50 ng of each template) in two combinations in PCR buffer (10 mM Tris-HCl, pH S.3, 50 ir^M KCl, 1.5 mM MgCl-,, C.Ol % gelatin; containing 200 uM each dNT?) . The SOE reaction was carried out using the DNA thermal cycler (Landgraf, Germany) and 2.5 units of Taq polymerase (Perkin-Elmer, Cetus, USA). Two cycles of PCR were performed using a cycle profile of denaturation at S4 °C for 1 min, annealing at 55 °C for 2 min, and extension at 72 °C for 3 min, the reaction was stopped, 2 50 pmol of each end-primer (forward no. 1 5'-CCCCAAGCTTGACTTGGAACCAATGGTCCATCC-3', forward no. 2 5 ' -CCCCAAGCTTCCATCC.ILAJ\CATGCTTAAAACGCTCG-3 ' , reverse primer 5 '-GGGCGTGAATGT.AAGCGTGACATA-3 ' ) was added to rhe reaction mixture, and an additional 30 cycles of PCR were performed using a cycle profile of denaturation at 94 °C for 1 min, annealing at 55 'tnor 2 min, and extension at 72 °C for 3 min. Construction of the expression cassettes for heterologous expression in Aspergillus oryzae The PCR-generated, recombinant fragments were electrophoresed in a 0.7 % low gelling temperature agarose gel (SeaPlaque, FMC), the fragments of interest were excised from the gel and recovered by treatment with agarase (New England Biolabs, USA) according to the manufacturer's instructions, followed by phenol extraction and ethanol precipitation at - 20 °C for 12 h. The DNA fragments were digested to completion with Hindlll and Xbal, and ligated into Hi/idlll/A'bal-cleaved pHD414 vector followed by electroporation of the constructs into £. coli DHIOB cells according to the manufacturer's instructions (Life Technologies, -USA). - The nucleotide sequence of the resulting gene fusions were cererTrined froir, born strands as described in rne Materials and methods section, SEQ ID no. 14A and 15A. The constructs may be transformed into A. oryzae as described. EXAMPLE 5 PCR facilitated detection of said type of cellulytic en¬zyme from 46 filamentous and monocentric fungi, representing 32 genera, from 23 families, belonging to 15 orders of 7 classes, covering all in all all four groups of the true Fungi: Ascomycetous, Basidiomycetous, Chytri-diomycetous and Zygomycetous fungi 5.1 Materials l.Diplodia gossypina Cooke Deposit of Strain, Ace No: CBS 274.S6 2.Ulospora bilgramii (Hawksw. et al.) Hawksw. et al. Ace No of strain: NKBC 1444, 3. Microsphaeropsis sp 4. Ascobolus stictoideus Speg. Ace No of strain: Q026 (Novo Nordisk collection) Isolated from goose dung, Svalbard, Norway 5. Saccobolus dilutellus (Fuck) Sacc. Deposit of strain: Ace No CBS 275.96 6. Penicillium verruculosum Peyronel Ex on Ace No of species: ATCC 62396 7. Penicillium chrysogenum Thorn Ace No of Strain: ATCC 9480 8. Thermomyces verrucosus Pugh et al Deposit of Strain, Ace No.: CES 2 3 5.56 9. Xylaria hypoxylon L. ex Greville Deposit of Strain, Ace No: CBS 2 3 4.96 10. Poronia punctata (Fr.ex L.) Fr. Ref:A.Munk: Danish Pyrenomyeetes, Dansk Botanisk Arkiv, Voll7,1 1957 11. Nodulisporum sp Isolated from leaf of Camellia reticulata (Theaceae, Guttiferales), Kunming Botanical Garden, Yunnan Province, China 12. Cylindrocarpon sp Isolated from marine sample, the Bahamas 13. Fusarium anguioides Sherbakoff Ace No of strain: IFO 4467 14. Fusarium poae (Peck) Wr. Ex on Ace No of species: ATCC 50883 15. Fusarium solani (Mart.)Sacc.emnd.Snyd & Hans. Ace No of strain: IMI 107.511 16. Fusarium oxysporum ssp lycopersici (Sacc.)Snyd. & Hans. Ace No of strain: CBS 645.78 17. Fusarium oxysporum ssp passiflora Ace No of strain: CBS 744.79 IS. Glioeladium catenulatum Gillman & Abbott Ace. No. of strain: ATCC 10523 19. Nectria pinea Dingley Deposit of Surain, Ace. No. CBS 279.95 20. Sordaria macrospora .a.uersvald Ex on Ace No of species: .a.TCC 602 55 21. Humicola grisea Traeen ex on Ace No for the species: ATCC 22726 22. Humicola nigrescens Omvik Ace No of strain: CBS 819.73 23. Scytalidium thermophilum (Cooney et Emerson) Austwick Ace No of strain: ATCC 28085 24. T.hielavia thermophila Fergus et Sinden (syn Corynascus thermophiius) Ace No of strain: CBS 174.70, IMI 145.136 25. Cladorrhinum foecundissimum Saccardo et Marchal Ex on Ace No of species: ATCC 62373 26. Syspastospora boninensis Ace No of strain: NKBC 1515 (Nippon University, profe Tubaki Collection) 27. Chaetoir.ium cuniculorum Fuckel Ace. No. of strain: CBS 799.83 28. Chaetomium brasiliense Batista et Potual Ace No of strain: CBS 122.65 29. Chaetomium murorum Corda Aec No of strain: CBS 163.52 30. Chaetomium virescens (von Arx) Udagawa Ace.No. of strain: C5S 547.75 31. Nigrcspora sp Deposit of strain, Ace No: CBS 2"2.96 32. Nigrospora sp Isolated from: 33. Diaport-he syngenesia Deposit of strain, Aec No: CBS 278.96 34. Colletotrichum lagenarium (Passerini) Ellis et Kalsted syn Glomerella cingulata var orbiculare Jenkins et Winstead Ex on a^^c No u£ species: ATCC 52609 35. Exidia glandulosa Fr. Deposit of Strain, Aec No: CBS 277.96 36. Pomes fomentarius (L.) Fr. Deposit of strain: Aec No. CBS 276.96 37. Spongipellis (?) Deposit of Strain: Ace No CBS 233.96 38. Rhizophlyctis rosea (de Bary & Wor) Fischer Deposit of Strain: Ace No.: CBS 282.96 39. Rhizomucor pusillus (Lindt) Schipper syn: Mucor pusillus Ace No of strain: IFO 4578 40. Phycomyces nitens (Kunze) van Tieghem & Le Monnier Ace No of strain: IFO 4814 41 Chaetostylum fresenii van Tieghem & Le Monnier syn. Helicostylum fresenii Ace No of strain NRRL 2 3 05 D'.neciuiTi rcseum, Ace No or s-crain: xru o j, /: 43. Coniothecium sp Endophyte, isolated from leaf of flevering plant, Kunming , Yunnan, China 44. Deposit of strain, Ace No.: CBS 271.96 Coelomycete, Isolated from leaf of Artocarpus altilis (Moraceae, Urticales), Christiana, Jamaica 45. Deposit of strain. Ace No.: CBS 273.96 Coelomycete, isolated from leaf of Pimenta dioica (Myrtaceae, Myrtales), Dallas Mountain, Jamaica 45. Deposit of strain: CBS 270.96 Coelomycete, isolated from leaf of Pseudocalymma alliaceum (Bignoniaceae, Solanales) growing in Dallas Mountain, Jamaica " JO 5.2 Procedure Maintenance of strains and production of biomass: The strains were maintained on agar in petrie dishes (9cm) or on slants (see list of Media: PCA and PDA). 44 of the strains were grown in shake flasks under the following growth conditions: general fungal media as PC, PD and PB 9 or YPG (see list of media); incubation time from 3 to 9 days; temperature 26"C; rpm between 150 and 175 . Strain No 14 (F.poae) was grown on wheat bran for 15 days (26"C; static). Strain No 38 was grown in dilute salt solution (DS/2), added 1 cm- pieces of autoclaved filter paper. Activity test: Activity was tested on 0.1% AZCL-HE-Cellulose (Megazyme) plates (14 era Petrie dishes), made up in 1% agarose (HSE, Lirex Agarose, Medmova). .Ail tests were done in triplicate, viz. A2CL-HE-Cellulose dissolved in three buffers, adjusted ro pH 3, 7 or S.5 (using various proportions of t:he following two ingredients Citric acid sonohydrat, Merck art. No 100244 (21.0 g) dissolved in water, making a total of 1000 ml; 0.IM tri-Sodium dodeca-brohydrate, Merck art.no. 6578 (38 g), dissolved in wa¬ter, making a total of 1000 ml. The mixing is done imrnidiately before use. Harvesting of Biomass: The biomass was harvested by filtering (mesh adjusted to the growth of the fungus, the finest used for fungi which have highly sporulating mycelium as e.g. Fusarium spp.). The biomass on the filter was scraped into a sterile plastic bag and imrnidiately frozen (by submerging into liquid nitrogen). 5.3 Results I. Using the PCR screening and amplification techniques described in Materials and Methods the following partial cDNA sequences were obtained: Saccobolus dilutellus (Fuck) Sacc, CBS 275.96: SEQ ID No. 21 (and the deduced amino acid sequence in SEQ ID No. 22) ; Thermomyces verrucosus, CBS 285.96: SEQ ID No. 23 (and the deduced amino acid sequence in SEQ ID No. 24); Xylaria hypoxylon, CBS 284.96: SEQ ID No. 25 (and the deduced amino acid sequence in SEQ ID No. 26); Fusarium oxysporum ssp lycopersici, CBS 645.78: SEQ ID No. 27 (and the deduced amino acid sequence in SEQ ID No. 28); Nectria pinea, CBS 279.96: SEQ ID No. 29 (and the deduced amino acid sequence in SEQ ID No. 30); Humicola grisea, ATCC 2272 6: SEQ ID No. 31 (and rhe deduced amino acid sequence in SEQ ID No. 32); Humicola nigrescens, C3S 819.73: SEQ ID No. 33 (and the deduced amino acid sequence in SEQ" ID No. 34); Cladorrhinum foecundissimum, ATCC 62373: SEQ ID No. 35 (and the deduced amino acid sequence in SEQ ID No. 36); Syspastospora boninensis, NKBC 1515: SEQ ID No. 37 (and the deduced amino acid sequence in SEQ ID No. 38); Nigrospora sp., CBS 272.96: SEQ ID No. 39 (and the deduced amino acid sequence in SEQ ID No. 40); Chaetostylum fresenii: SEQ ID No. 41 (and the deduced amino acid sequence in SEQ ID No. 42); Exidia glandulosa, CBS 277.96: SEQ ID No. 43 (and the deduced amino acid sequence in SEQ ID No. 44); Coniothecium sp.: SEQ ID No. 45 (and the deduced amino acid sequence in SEQ ID No. 46); Deposition No. CBS 271.96: SEQ ID No. 47 (and the deduced amino acid sequence in S£i2—LD No. 48); Deposition No. CBS 270.96: SEQ ID No. 49 (and the deduced amino acid sequence in SEQ ID No. 50); Diplodia gossypina, CBS 274.96: SEQ ID No. 51 (and the deduced amino acid sequence in SEQ ID No. 52); Ulospora bilgramii, NKBC 1444: SEQ ID No. 53 (and the deduced amino acid sequence in SEQ ID No. 54); Penicillium verruculosum, ATCC 62396: SEQ ID No. 55 (and the deduced amino acid sequence in SEQ ID No. 56); Poronia punctata: SEQ ID No. 57 (and the deduced amino acid sequence in SEQ ID No. 58); Fusarium anguioides, IFO 4467: SEQ ID No. 59 (and the deduced amino acid sequence in SEQ ID No. 60); Thielavia thermophila, CBS 174.70: SEQ ID No. 61 (and the deduced amino acid sequence in SEQ ID No. 62); Chaetomium cuniculorum, CBS 799.83: SEQ ID No. 63 (and the deduced amino acid sequence in SEQ ID No. 64); Chaetomium virescens: SEQ ID No. 65 (and the deduced amino acid sequence in SEQ ID No. 66); Colletctrichum lagenarium: SEQ ID No. 67 (and the deduced amino acid sequence in SEQ ID No. 65); Phycomyces nitens: SEQ ID No. 69 (and the deduced amino acid sequence in SEQ ID No. 70); and Trichothecium roseum: SEQ ID No. 71 (and the deduced amino acid sequence in SEQ ID No. 72); II. Using the PCR screening and amplification techniques described in Materials and Methods partial cDNA encoding partially for the enzyme of the invention was obtained and the plasmid was deposited according to the Budapest Treaty: Escherichia coli, DSM 10583, deposition date 13 March, 1996; cDNA from Trichothecium roseum; Escherichia coli, DSM 10584, deposition date 13 March, 1996; cDNA from Syspastospora boninensis; Escherichia coli, DSM 10585, depbsitrTon date 13 March, 1996; cDNA from Cheatomium murorum Escherichia coli, DSM 10587, deposition date 13 March, 1996; cDNA from Sordaria fimicola; Escherichia coli, DSM 10588, deposition date 13 March, 1996; cDNA from the unidentified strain CBS 273.96; Escherichia coli, DSM 10586, deposition date 13 March, 1996; cDNA from Spongipellis sp. Color clarification of crude supernatants During normal wash the fabric will often fade. However, the fabric appearance is improved and the original colours are much better preserved or maintained if the fabric is washed with a cellulase giving color clarificarion. Color clarification is measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers. Apparatus Liquid volume Agitation Rinse time Washing temp Washing liqour PH Washing time Repetitions Enzymes Dosage Textile cotton Terg-o-tometer 100 ml 150 movements/min with vertical stirrer 5 min in tapwater 40° 0.05 M phosphate buffer 7.0 30 min 2 Crude supernatants from the strains shown below. Two dosages from : 200, 500, 1000 or 2500—S=CEVU/1 2 swatches of aged black 100% 5x6 cm (0.9 gram) Drying : Tumble dry- Evaluation: The light remission is measured by a Datacolor Elrepho Remission spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker, and lower L values are obtained. The samples are compared with a blind sample, i.e. washed without enzyme. Below is shown the delta L remission values compared to a blind sample. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT (A) NAME: Novo Nordisk A/S (B) STREET: Novo Alle (C) CITY: DK-2880 Bagsvaerd (E) COUNTRY: Denmark (F) POSTAL CODE (ZIP): DK-2880 (G) TELEPHONE: +45 44 88 88 (H) TELEFAX: +45 44 49 32 56 (ii) TITLE OF INVENTION: NOVEL ENDOGLUCANASES (iii) NU'MBER OF SEQUENCES: 72 (iv) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.30 (EPO) (2) INFORMATION FOR SEQ ID NO: lA: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 891 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Saccharomyces cerevisiae (B) STRAIN: DSM 9770 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: lA: AAAGAAAGGC TCTCTGCTGT CGTCGCTCTC GTCGCTCTCG TCGGCATCCT CCATCCGTCC 60 GCCTTTGATA ACCCGCTCCC CGACTCAGTC AAGACGACGC ATACTTGGCA CCATGCATCT 12 0 CTCCGCCACC ACCGGGTTCC TCGCCCTCCC GGTCCTGGCC CTGGACCAGC TCTCGGGCAT 180 CGGCCAGACG ACCCGGTACT GGGACTGCTG CAAGCCGAGC TGCGCCTGGC CCGGCAAGGG 2 4 0 CCCCTCGTCT CCGGTGCAGG CCTGCGACAA GAACGACAAC CCGCTCAACG ACGGCGGCTC 3 00 CACCCGGTCC GGCTGCGACG CGGGCGGCAG CGCCTACATG TGCTCCTCCC AGAGCCCCTG 3 60 GGCCGTCAGC GACGAGCTGT CGTACGGCTG GGCGGCCGTC AAGCTCGCCG GCAGCTCCGA 4 2 0 GTCGCAGTGG TGCTGCGCCT GCTACGAGCT GACCTTCACC AGCGGGCCGG TCGCGGGCAA 480 GAAGATGATT GTGCAGGCGA CCAACACCGG TGGCGACCTG GGCGACAACC ACTTTGACCT 540 GGCCATCCCC GGTGGCGGTG TCGGTATTTT CAACGCCTGC ACCGACCAGT ACGGCGCTCC 600 CCCGAACGGC TGGGGCGACC GCTACGGCGG CATCCATTCC AAGGAAGAGT GCGAATCCTT 660 CCCGGAGGCC CTCAAGCCCG GCTGCAACTG GCGCTTCGAC TGGTTCCAAA ACGCCGACAA 72 0 CCCGTCGGTC ACCTTCCAGG AGGTGGCCTG CCCGTCGGAG CTCACGTCCA AGAGCGGCTG 780 CTCCCGTTAA GAGGGAAGAG AGGGGGCTGG 7AGGACCGAA AGATTCAACC TCTGCTCCTG 84 0 CTGGGGAAGC TCGGGCGCGA GTGTGAAACT GGTGTAAATA TTGTGGCACA CACAAGCTAC 900 TACAGTCCGT CTCGCCGTCC GGCTAACTAG CCTTGCTGCG GATCTGTCCA AAAAAAAAAA 960 (2) INFORMATION FOR SEQ ID NO: IB: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 225 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: IB: Met His Leu Ser Ala Thr Thr Gly Phe Leu Ala Leu Pro Val Leu Ala 15 10 15 Leu Asp Gin Leu Ser Gly lie Gly Gin Thr Thr Arg Tyr Trp Asp Cys 20 25 30 Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Gly Pro Ser Ser Pro Val 35 40 45 Gin Ala Cys Asp Lys Asn Asp Asn Pro Leu Asn Asp Gly Gly Ser Thr 50 55 60 Arg Ser Gly Cys Asp Ala Gly Gly Ser Ala Tyr Met Cys Ser Ser Gin 65 70 75 80 Ser Pro Trp Ala Val Ser Asp Glu Leu Ser Tyr Gly Trp Ala Ala Val 85 90 95 Lys Leu Ala Gly Ser Ser Glu Ser Gin Trp Cys Cys Ala Cys Tyr Glu 100 105 110 Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met lie Val Gin 115 120 125 Ala Thr Asn Thr Gly Gly Asp Leu Gly Asp Asn His Phe Asp Leu Ala 130 135 140 lie Pro Gly Gly Gly Val Gly lie Phe Asn Ala Cys Thr Asp Gin Tyr 145 150 155 160 Gly Ala Pro Pro Asn Gly Trp Gly Asp Arg Tyr Gly Gly lie His Ser 165 170 175 Lys Glu Glu Cys Glu Ser Phe Pro Glu Ala Leu Lys Pro Gly Cys Asn 180 185 190 Trp Arg Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Ser Val Thr Phe 195 200 205 Gin Glu Val Ala Cys Pro Ser Glu Leu Thr Ser Lys Ser Gly Cys Ser 210 215 220 Arg 225 2) INFORMATION FOR SEQ ID NO: 2A: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 894 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: "Construction 1" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2A: \TGCATCTCT CCGCCACCAC CGGGTTCCTC GCCCTCCCGG TCCTGGCCCT GGACCAGCTC 60 rCGGGCATCG GCCAGACGAC CCGGTACTGG GACTGCTGCA AGCCGAGCTG CGCCTGGCCC 120 GGCAAGGGCC CCTCGTCTCC GGTGCAGGCC TGCGACAAGA ACGACAACCC GCTCAACGAC 180 GGCGGCTCCA CCCGGTCCGG CTGCGACGCG GGCGGCAGCG CCTACATGTG CTCCTCCCAG 240 AGCCCCTGGG CCGTCAGCGA CGAGCTGTCG TACGGCTGGG CGGCCGTCAA GCTCGCCGGC 300 AGCTCCGAGT CGCAGTGGTG CTGCGCCTGC TACGAGCTGA CCTTCACCAG CGGGCCGGTC 360 GCGGGCAAGA AGATGATTGT GCAGGCGACC AACACCGGTG GCGACCTGGG CGACAACCAC 420 TTTGACCTGG CCATCCCCGG TGGCGGTGTC GGTATTTTCA ACGCCTGCAC CGACCAGTAC 480 GGCGCTCCCC CGAACGGCTG GGGCGACCGC TACGGCGGCA TCCATTCCAA GGAAGAGTGC 540 GAATCCTTCC CGGAGGCCCT CAAGCCCGGC TGCAACTGGC GCTTCGACTG GTTCCAAAAC 600 GCCGACAACC CGTCGGTCAC CTTCCAGGAG GTGGCCTGCC CGTCGGAGCT CACGTCCAAG 660 AGCGGCTGCT CCCGTCCCTC CAGCAGCACC AGCTCTCCGG TCAACCAGCC TACCAGCACC 720 AGCACCACGT CCACCTCCAC CACCTCGAGC CCGCCAGTCC AGCCTACGAC TCCCAGCGGC 780 TGCACTGCTG AGAGGTGGGC TCAGTGCGGC GGCAATGGCT GGAGCGGCTG CACCACCTGC 840 GTCGCTGGCA GCACTTGCAC GAAGATTAAT GACTGGTACC ATCAGTGCCT GTAG 894 INFORMATION FOR SEQ ID NO: 2B: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 297 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2B: Met His Leu Ser Ala Thr Thr Gly Phe Leu Ala Leu Pro Val Leu Ala 15 10 15 Leu Asp Gin Leu Ser Gly lie Gly Gin Thr Thr Arg Tyr Trp Asp Cys 20 25 30 Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Gly Pro Ser Ser Pro Val 35 40 45 Gin Ala Cys Asp Lys Asn Asp Asn Pro Leu Asn Asp Gly Gly Ser Thr 50 55 60 Arg Ser Gly Cys Asp Ala Gly Gly Ser Ala Tyr Met Cys Ser Ser Gin 65 70 75 80 Ser Pro Trp Ala Val Ser Asp Glu Leu Ser Tyr Gly Trp Ala Ala Val 85 90 95 Lys Leu Ala Gly Ser Ser Glu Ser Gin Trp Cys Cys Ala Cys Tyr Glu 100 105 110 Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met lie Val Gin 115 120 125 Ala Thr Asn Thr Gly Gly Asp Leu Gly Asp Asn His Phe Asp Leu Ala 130 135 140 lie Pro Gly Gly Gly Val Gly lie Phe Asn Ala Cys Thr Asp Gin Tyr 145 150 155 160 Gly Ala Pro Pro Asn Gly Trp Gly Asp Arg Tyr Gly Gly He His Ser 165 170 175 Lys Glu Glu Cys Glu Ser Phe Pro Glu Ala Leu Lys Pro Gly Cys Asn 180 185 190 Trp Arg Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Ser Val Thr Phe 195 200 205 Gin Glu Val Ala Cys Pro Ser Glu Leu Thr Ser Lys Ser Gly Cys Ser 210 215 220 Arg Pro Ser Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr 225 230 235 240 Ser Thr Thr Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr 245 250 255 Thr Pro Ser Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn 260 265 270 Gly Trp Ser Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys 275 280 285 lie Asn Asp Trp Tyr His Gin Cys Leu 290 295 (2) INFORMATION FOR SEQ ID NO: 3A: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 927 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: "Construction 2" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3A: ATGCATCTCT CCGCCACCAC CGGGTTCCTC GCCCTCCCGG TCCTGGCCCT GGACCAGCTC 60 TCGGGCATCG GCCAGACGAC CCGGTACTGG GACTGCTGCA AGCCGAGCTG CGCCTGGCCC 120 GGCAAGGGCC CCTCGTCTCC GGTGCAGGCC TGCGACAAGA ACGACAACCC GCTCAACGAC 180 GGCGGCTCCA CCCGGTCCGG CTGCGACGCG GGCGGCAGCG CCTACATGTG CTCCTCCCAG 240 hGCCCCTGGG CCGTCAGCGA CGAGCTGTCG TACGGCTGGG CGGCCGTCAA GCTCGCCGGC 300 AGCTCCGAGT CGCAGTGGTG CTGCGCCTGC TACGAGCTGA CCTTCACCAG CGGGCCGGTC 360 GCGGGCAAGA AGATGATTGT GCAGGCGACC AACACCGGTG GCGACCTGGG CGACAACCAC 420 TTTGACCTGG CCATCCCCGG TGGCGGTGTC GGTATTTTCA ACGCCTGCAC CGACCAGTAC 480 GGCGCTCCCC CGAACGGCTG GGGCGACCGC TACGGCGGCA TCCATTCCAA GGAAGAGTGC 540 GAATCCTTCC CGGAGGCCCT CAAGCCCGGC TGCAACTGGC GCTTCGACTG GTTCCAAAAC 600 GCCGACAACC CGTCGGTCAC CTTCCAGGAG GTGGCCTGCC CGTCGGAGCT CACGTCCAAG 660 AGCGGCTGCT CCCGTAACGA CGACGGCAAC TTCCCTGCCG TCCAGATCCC CTCCAGCAGC 720 ACCAGCTCTC CGGTCAACCA GCCTACCAGC ACCAGCACCA CGTCCACCTC CACCACCTCG 780 AGCCCGCCAG TCCAGCCTAC GACTCCCAGC GGCTGCACTG CTGAGAGGTG GGCTCAGTGC 840 GGCGGCAATG GCTGGAGCGG CTGCACCACC TGCGTCGCTG GCAGCACTTG CACGAAGATT 900 AATGACTGGT ACCATCAGTG CCTGTAG 927 ) INFORMATION FOR SEQ ID NO: 3B: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3 08 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3B: Mer His Leu Ser .Ala Thr,Thr,Gly P.he Leu Ala Leu Pro Val Leu Ala 15 10 15 Leu Asp Gin Leu Ser Gly lie Gly Gin Thr Thr Arg Tyr Trp Asp Cys 20 25 30 Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Gly Pro Ser Ser Pro Val 35 40 45 Gin Ala Cys Asp Lys Asn Asp Asn Pro Leu Asn Asp Gly Gly Ser Thr 50 55 50 Arg Ser Gly Cys Asp Ala Gly Gly Ser Ala Tyr Met Cys Ser Ser Gin 65 70 75 80 Ser Pro Trp Ala Val Ser Asp Glu Leu Ser Tyr Gly Trp Ala Ala Val 85 90 95 Lys Leu Ala Gly Ser Ser Glu Ser Gin Trp Cys Cys Ala Cys Tyr Glu 100 105 110 Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met lie Val Gin 115 120 12 5 Ala Thr Asn Thr Gly Gly Asp Leu Gly Asp Asn His Phe Asp Leu Ala 130 135 140 lie Pro Gly Gly Gly Val Gly lie Phe Asn Ala Cys Thr Asp Gin Tyr 145 150 155 160 Gly Ala Pro Pro Asn Gly Trp Gly Asp Arg Tyr Gly Gly lie His Ser 165 170 175 Lys Glu Glu Cys Glu Ser Phe Pro Glu Ala Leu Lys Pro Gly Cys Asn 180 185 190 Trp Arg Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Ser Val Thr Phe 195 200 205 Gin Glu Val Ala Cys Pro Ser Glu Leu Thr Ser Lys Ser Gly Cys Ser 210 215 220 Arg Asn Asp Asp Gly Asn Phe Pro Ala Val Gin lie Pro Ser Ser Ser 225 230 235 240 Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr Ser Thr Thr Ser Thr 245 250 255 Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser Gly Cys 260 265 270 Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser Gly Cys 275 2S0 285 Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys lie Asn Asp Trp Tyr 290 255 300 His Gin Cys Leu 305 (2) INFORMATION FOR SEQ ID NO: 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 888 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Saccharoinyces cerevisiae, DSM 10082 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: CCAGTGTGCT GGAAAGCCTT CGTGCTGTCC CCGACGTATC CCTGACCGCC ATGCGTTCCA 60 CCAGCATCTT GATCGGCCTT GTTGCCGGCG TCGCTGCTCA GAGCTCTGGC TCTGGCCATA 120 CAACCAGGTA CTGGGACTGC TGCAAGCCCT CATGCGCCTG GGATGAGAAG GCGGCTGTCA 180 GCCGGCCGGT CACAACATGC GACAGGAACA ACAGCCCCCT TTCGCCCGGC GCTGTGAGCG 240 GCTGCGACCC CAACGGCGTT GCATTCACCT GCAACGACAA CCAGCCTTGG GCCGTAAACA 300 ACAATGTCGC CTACGGTTTT GCGGCTACCG CCTTCCCTGG TGGCAATGAG GCGTCGTGGT 360 GCTGTGCCTG CTATGCTCTT CAATTCACAT CCGGCCCCGT TGCTGGCAAG ACGATGGTTG 420 TGCAATCCAC CAACACTGGC GGAGATCTCA GCGGCACTCA CTTCGATATC CAGATGCCCG 480 GTGGAGGTCT CGGCATCTTC GACGGCTGCA CCCCGCAGTT CGGCTTCACG TTCCCCGGCA 540 ACCGCTACGG CGGTACCACG AGCCGCAGCC AGTGCGCCGA GCTGCCCTCC GTCCTCCGTG 600 ACGGCTGCCA CTGGCGTTAC GACTGGTTCA ACGATGCCGA CAACCCCAAC GTCAACTGGC 660 GCCGCGTCCG ATGCCCGGCG GCCCTCACGA ACCGCTCCGG CTGCGTCCGC AACGACGACA 720 ACAGCTACCC CGTCTTCGAG CCCGGCACGG GCACCCCGCC GACCCCCACG ACCACGACTA 780 CCAGCTCCCC TCCTCAGCCC ACCAACGGCG GAGGCGGCGG CACTTCTCCT CACTGGGGCC 840 AGTGCGGCGG CCAGGGCTGG TCTGGCCCGA CGGCCTGTGC CGGTGGGTCG ACCTGCAACC 900 TGATCAACCC GTGGTACTCC CAGTGCATTC CCAACTAAGT GATCCGGGCA TTGCGGTCGA 960 AAGGGGACCG TTAGTCGACA AGGCCCAGCC AGACCTCAGG CAGGTGGCTG CCATGGCAGA 1020 TTGTATATAG TCTTCCGAGT ACATACTATT GAATGAAAAT AAGAGCGGCT CGGACCATGA 1080 GCAGATGCCA TTTGATAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 1140 AAAAAAAAAA AAAA 1154 (2) INFORMATION FOR SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 295 amino acids (B) TYPE: amino acid (C) STRANDEDNESS; single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: Met Arg Ser Thr Ser lie Leu lie Gly Leu Val Ala Gly Val Ala Ala 15 10 15 Gin Ser Ser Gly Ser Gly His Thr Thr Arg Tyr Trp Asp Cys Cys Lys 20 25 30 Pro Ser Cys Ala Trp Asp Glu Lys Ala Ala Val Ser Arg Pro Val Thr 35 40 45 Thr Cys Asp Arg Asn Asn Ser Pro Leu Ser Pro Gly Ala Val Ser Gly 50 55 60 Cys Asp Pro Asn Gly Val Ala Phe Thr Cys Asn Asp Asn Gin Pro Trp 65 70 75 80 Ala Val Asn Asn Asn Val Ala Tyr Gly Phe Ala Ala Thr Ala Phe Pro 85 90 95 Gly Gly Asn Glu Ala Ser Trp Cys Cys Ala Cys Tyr Ala Leu Gin Phe 100 105 110 Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val Gin Ser Thr Asn 115 120 125 Thr Gly Gly Asp Leu Ser Gly Thr His Phe Asp lie Gin Met Pro Gly 130 135 140 Gly Gly Leu Gly lie Phe Asp Gly Cys Thr Pro Gin Phe Gly Phe Thr 145 150 155 160 Phe Pro Gly Asn Arg Tyr Gly Gly Thr Thr Ser Arg Ser Gin Cys Ala 165 170 175 Glu Leu Pro Ser Val Leu Arg Asp Gly Cys His Tjrp Arg Tyr Asp Trp 180 185 190 Phe Asn Asp Ala Asp Asn Pro Asn Val Asn Trp Arg Arg Val Arg Cys 195 200 205 Pro Ala Ala Leu Thr Asn Arg Ser Gly Cys Val Arg Asn Asp Asp Asn 210 215 220 Ser Tyr Pro Val Phe Glu Pro Gly Thr Gly Thr Pro Pro Thr Pro Thr 225 230 235 240 Thr Thr Thr Thr Ser Ser Pro Pro Gin Pro Thr Asn Gly Gly Gly Gly 245 250 255 Gly Thr Ser Pro His Trp Gly Gin Cys Gly Gly Gin Gly Trp Ser Gly 260 265 270 Pro Thr Ala Cys Ala Gly Gly Ser Thr Cys Asn Leu lie Asn Pro Trp 275 280 285 Tyr Ser Gin Cys lie Pro Asn 290 295 (2) INFORMATION FOR SEQ ID NO: 6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1423 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (Vi) ORIGINAL SOURCE: (A) ORGANISM: Saccharomyces cerevisiae, DSM 10080 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: AAAGTTCTGG CCGGAACAGA TCTCCGTTGT CGATCTTCGA TTTTCCAGAC TCAGTCTGTG 60 ACACTCCTTC AATCCACATT CCTTTACTTC TTCGTCACTC ATTCACATCA TGATTTCAGC 120 TTGGATTCTC CTGGGGCTGG TAGGCGCCGT GCCCTCCTCC GTCATGGCCG CCTCGGGCAA 180 AGGCCACACC ACCCGCTACT GGGATTGCTG CAAGACTTCT TGCGCATGGG AGGGCAAGGC 240 ATCCGTCTCC GAGCCTGTCC TGACCTGTAA CAAGCAGGAC AACCCCATCG TCGATGCCAA 300 CGCCAGAAGC GGCTGCGACG GCGGCGGGGC ATTTGCCTGT ACCAACAATT CCCCTTGGGC 360 CGTGAGCGAG GACCTGGCCT ACGGATTTGC TGCCACAGCC CTCAGCGGCG GCACTGAGGG 420 CAGCTGGTGC TGCGCGTGTT ACGCCATCAC ATTCACGAGT GGCCCTGTGG CTGGCAAGAA 480 GATGGTCGTC CAGTCCACGA ACACGGGAGG CGACCTGTCC AACAACCACT TTGACCTGAT 540 GATTCCCGGT GGAGGCCTCG GCATCTTTGA CGGTTGCTCG GCTCAGTTCG GACAACTTCT 600 TCCCGGCGAG CGTTACGGAG GTGTTTCGTC CCGCTCTCAA TGCGATGGCA TGCCCGAGCT 660 CTTGAAAGAC GGTTGCCAGT GGCGCTTCGA CTGGTTCAAG AACTCAGACA ACCCTGACAT 720 CGAGTTCGAG CAGGTCCAGT GTCCCAAAGA GCTCATTGCG GTCTCTGGGT GCGTCCGTGA 780 CGACGATAGC AGCTTTCCCG TCTTCCAAGG TTCGGGCTCA GGAGATGTCA ACCCACCTCC 840 CAAGCCGACT ACGACTACGA CCTCGTCAAA GCCGAAAACA ACCTCTGCAC CATCCACTCT 900 CTCGAACCCA TCCGCCCCTC AACAGCCAGG GAACACTGAT AGACCTGCCG AGACAACCAC 960 TACCAAGCTG CCTGCCCTGC CGGCCACGAC GAGCAGCCCT GCTGTCTCAG TTCCTTCGTC 1020 CAGCGCTCGC GTGCCTTTGT GGGGGCAATG CGACTCGGAA GCTTCATGGG ACGCACCTAA 1080 GAAGTGTGCA AAGGGCACCA AGTGTGTCTA CGTCAACGAC TGGTACTCTC AATGCCAGCC 1140 GAAGAACTCT TGTGCTTGAG AAGCAATGCT CACAGCATGT CCTCTTGTCA CCCCTTCTTT 1200 TCATTCCCAA ACATACTTAC TGTATTATTA TTTCCGATGC TTCATTTCTT GCTTGTTTCT 1260 GTCTTTCCTG CACGCAGCTT TCAACGATAC CCTTCATGCG ATTGCCCTAC GATCAGATGA 1320 TGGGCACGAC ATGGAGGATG GTTTGGGCAC TCACGCGTTC AGGACGGGAA AATTTATTAG 1380 GGCTGAGATC CGTGAATTGA CTTCATTTCG GCGGAATGTC TGC 1423 INFORMATION FOR SEQ ID NO: 7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 349 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: Met lie Ser Ala Trp lie Leu Leu Gly Leu Val Gly Ala Val Pro Ser 15 10 15 Ser Val Met Ala Ala Ser Gly Lys Gly'His Thr Thr Arg Tyr Trp Asp 20 25 30 Cys Cys Lye Thr Ser Cys Ala Trp Glu Gly Lys Ala Ser Val Ser Glu 35 40 45 Pro Val Leu Thr Cys Asn Lys Gin Asp Asn Pro lie Val Asp Ala Asn 50 55 60 Ala Arg Ser Gly Cys Asp Gly Gly Gly Ala Phe Ala Cys Thr Asn Asn 65 70 75 80 Ser Pro Trp Ala Val Ser Glu Asp Leu Ala Tyr Gly Phe Ala Ala Thr 85 90 95 Ala Leu Ser Gly Gly Thr Glu Gly Ser Trp Cys Cys Ala Cys Tyr Ala 100 105 110 He Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met Val Val Gin 115 120 125 Ser Thr Asn Thr Gly Gly Asp Leu Ser Asn Asn His Phe Asp Leu Met 130 135 140 He Pro Gly Gly Gly Leu Gly He Phe Asp Gly Cys Ser Ala Gin Phe 145 150 155 160 Gly Gin Leu Leu Pro Gly Glu Arg Tyr Gly Gly Val Ser Ser Arg Ser 165 170 175 Gin Cys Asp Gly Met Pro Glu Leu Leu Lys Asp Gly Cys Gin Trp Arg 180 185 190 Phe Asp Trp Phe Lys Asn Ser Asp Asn Pro Asp He Glu Phe Glu Gin 195 200 205 Val Gin Cys Pro Lys Glu Leu He Ala Val Ser Gly Cys Val Arg Asp 210 215 220 Asp Asp Ser Ser Phe Pro Val Phe Gin Gly Ser Gly Ser Gly Asp Val 225 230 235 240 Asn Pro Pro Pro Lys Pro Thr Thr Thr Thr Thr Ser Ser Lys Pro Lys 245 250 255 Thr Thr Ser Ala Pro Ser Thr Leu Ser Asn Pro Ser Ala Pro Gin Gin 250 265 270 Pro Gly Asn Thr Asp Arg Pro Ala Glu Thr Thr Thr Thr Lys Leu Pro 275 280 285 Ala Leu Pro Ala Thr Thr Ser Ser Pro Ala Val Ser Val Pro Ser Ser 290 295 300 Ser Ala Arg Val Pro Leu Trp Gly Gin Cys Asp Ser Glu Ala Ser Trp 305 310 315 320 Asp Ala Pro Lys Lys Cys Ala Lys Gly Thr Lys Cys Val Tyr Val Asn 325 330 335 Asp Trp Tyr Ser Gin Cys Gin Pro Lys Asn Ser Cys Ala 340 345 (2) INFORMATION FOR SEQ ID NO: 8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1174 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Saccharomyces cerevisiae, DSM ; 10081 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8 I GAGCAGCACC CCTCAAGCTG TACAGTTTCC ACCCCGCTCT CTTTTCTTCG GCCCCCAGGA 60 TGCGCTCTAC TCCCGTTCTT CGCACAACCC TGGCOGCTGC ACTTCCTCTG GTCGCCTCCG 120 CGGCCAGTGG CAGTGGCCAG TCCACXJAGAT ACTGGGACTG CTGC3AGCCG TCGTGCGCTT 180 GGCCCGGGAA GGCCGCCGTC AGCCAACCGG TCTACGCGTG CGATGCCAAC TTCCAGCGCC 240 TGTCCGACTT CAATGTCCAG TCGGGCTGCA ACGGCGGCTC GGCCTACTCC TGCGCCGACC 300 AGACTCCCTG GGCGGTGAAC GACAATCTCG CCTACGGCTT CGCCGCGACG AGCATCGCCG 360 GCGGGTCCGA ATCCTCGTGG TGCTGCGCCT GCTACGCGCT CACCTTCACT TCCGGTCCCG 420 TCGCCGGCAA GACAATGGTG GTGCAGTCAA CGAGCACTGG CGGCGACCTG GGAAGTAACC 480 AGTTCGATAT CGCCATGCCC GGCGGCGGCG TGGGCATCTT CAACGGCTGC AGCTCGCAGT 540 TCGGCGGCCT CCCCGGCGCT CAATACGGCG GCATTTCGTC GCGCGACCAG TGCGATTCCT 600 TCCCCGCGCC GCTCAAGCCC GGCTGCCAGT GGCGGTTTGA CTGGTTCCAG AACGCCGACA 660 ACCCGACGTT CACGTTCCAG CAGGTGCAGT GCCCCGCCGA GATCGTTGCC CGCTCCGGCT 720 GCAAGCGCAA CGACGACTCC AGCTTCCCCG TCTTCACCCC CCCAAGCGGT GGCAACGGTG 780 GCACCGGGAC GCCCACGTCG ACTGCGCCTG GGTCGGGCCA GACGTCTCCC GGCGGCGGCA 840 GTGGCTGCAC GTCTCAGAAG TGGGCTCAGT GCGGTGGCAT CGGCTTCAGC GGATGCACCA 900 CCTGTGTCTC TGGCACCACC TGCCAGAAGT TGAACGACTA CTACTCGCAG TGCCTCTAAA 960 CAGCTTTTCG CACGAGGTGG CGGGAC»GAG CAAGGAGACC GTCAACTTCG TCATGCATAT 1020 TTTTTGAGCG CTCAATACAT ACATAACCTT CGATTCTTGT ACATAGCACG CCGGTACACA 1080 TCTCACACCG ACTTTGGGGG CGGAATCAGG CCCGTTTTAA AAAAAAAAAA AAAAAAAAAA 1140 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAA 1174 (2) INFORMATION FOR SEQ ID NO: 9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 299 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9: Met Arg Ser Thr Pro Val Leu Arg Thr Thr Leu Ala Ala Ala Leu Pro 15 10 15 Leu Val Ala Ser Ala Ala Ser Gly Ser Gly Gin Ser Thr Arg Tyr Trp 20 25 30 Asp Cys Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Ala Ala Val Ser 35 40 45 Gin Pro Val Tyr Ala Cys Asp Ala Asn Phe Gin Arg Leu Ser Asp Phe 50 55 60 Asn Val Gin Ser Gly Cys Asn Gly Gly Ser Ala Tyr Ser Cys Ala Asp 65 70 75 80 Gin Thr Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala 85 90 95 Thr Ser lie Ala Gly Gly Ser Glu Ser Ser Trp Cys Cys Ala Cys Tyr 100 105 110 Ala Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val 115 120 125 Gin Ser Thr Ser Thr Gly Gly Asp Leu Gly Ser Asn Gin Phe Asp lie 130 135 140 Ala Met Pro Gly Gly Gly Val Gly lie Phe Asn Gly Cys Ser Ser Gin 145 150 155 160 Phe Gly Gly Leu Pro Gly Ala Gin Tyr Gly Gly lie Ser Ser Arg Asp 165 170 175 Gin Cys Asp Ser Phe Pro Ala Pro Leu Lys Pro Gly Cys Gin Trp Arg 180 185 190 Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Thr Phe Thr Phe Gin Gin 195 200 205 Val Gin Cys Pro Ala Glu lie Val Ala Arg Ser Gly Cya Lys Arg Asn 210 215 220 Asp Asp Ser Ser Phe Pro Val Phe Thr Pro Pro Ser Gly Gly Asn Gly 225 230 235 240 Gly Thr Gly Thr Pro Thr Ser Thr Ala Pro Gly Ser Gly Gin Thr Ser 245 250 255 Pro Gly Gly Gly Ser Gly Cys Thr Ser Gin Lys Trp Ala Gin Cys Gly 260 265 270 Gly lie Gly Phe Ser Gly Cys Thr Thr Cys Val Ser Gly Thr Thr Cys 275 280 285 Gin Lys Leu Asn Asp Tyr Tyr Ser Gin Cys Leu 290 295 (2) INFORMATION FOR SEQ ID NO: 10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 913 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli, DSM 10512 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: GCACTATTCT CAGCTCCATT CTCCCTTGAA GTAATTCACC ATGTTCTCTC CGCTCTGGGC 60 CCTGTCGGCT CTGCTCCTAT TTCCTGCCAC TGAAGCCACT AGCGGCGTGA CAACCAGGTA 120 CTGGGACTGC TGCAAGCCGT CTTGTGCTTG GACGGGCAAA GCATCCGTCT CCAAGCCCGT 180 CGGAACCTGC GACATCAACG ACAACGCCCA GACGCCGAGC GATCTGCTCA AGTCGTCCTG 240 TGATGGCGGC AGCGCCTACT ACTGCAGCAA CCAGGGCCCA TGGGCCGTGA ACGACAGCCT 300 TTCCTACGGC TTCGCTGCCG CCAAGCTGTC CGGAAAGCAG GAGACTGATT GGTGCTGTGG 360 CTGCTACAAG CTCACATTCA CCTCCACCGC CGTTTCCGGC AAGCAAATGA TCGTGCAAAT 420 CACGAACACG GGCGGCGhCC TCGGCAACAA CCACTTCGAC ATCGCCATGC CGGGCGGCGG 480 CGTCGGCATC TTCAACGGGT GCTCCAAGCA ATGGAACGGC ATCAATCTGG GCAACCAGTA 540 TGGCGGCTTC ACTGACCGCT CGCAATGTGC GACGCTCCCG TCCAAGTGGC AGGCCAGCTG 600 CAACTGGCGC TTCGACTGGT TCGAGAATGC CGACAACCCC ACCGTCGATT GGGAGCCTGT 660 CACTTGCCCA CAGGAATTGG TCGCCCGGAC TGGCTGTTCC CGTACCTAAG TGGGGGTGGA 720 ACCTCCATGT GAATTGGTGT ATATAGCTCC TGCCTGAGCA TCCACCAGTT CGCATGTGTT 780 6ATCAGGAGT TGTGTTGCCT TGCTAGGAAA GACTTTGTTG GAAACTTGCG TGTTTATTCC 840 AATTGAATAA CCCTGTATAG ACCGGTCACA TTTTTCTCTG AAAAAAAAAA AAAAAAAAAA 900 AAAAAAAAAA AAA 913 (2) INFORMATION FOR SEQ ID NO: 11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 222 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11: Met Phe Ser Pro Leu Trp Ala Leu Ser Ala Leu Leu Leu Phe Pro Ala 1 5 10 15 Thr Glu Ala Thr Ser Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys 20 25 30 Pro Ser Cys Ala Trp Thr Gly Lys Ala Ser Val Ser Lys Pro Val Gly 35 40 45 Thr Cys Asp lie Asn Asp Asn Ala Gin Thr Pro Ser Asp Leu Leu Lys 50 55 60 Ser Ser Cys Asp Gly Gly Ser Ala Tyr Tyr Cya Ser Asn Gin Gly Pro 65 70 75 80 Trp Ala Val Asn Asp Ser Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu 85 90 95 Ser Gly Lys Gin Glu Thr Asp Trp Cys Cys Gly Cys Tyr Lys Leu Thr 100 105 110 Phe Thr Ser Thr Ala Val Ser Gly Lys Gin Met He Val Gin He Thr 115 120 125 Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp He Ala Met Pro 130 135 140 Gly Gly Gly Val Gly He Phe Asn Gly Cys Ser Lys Gin Trp Asn Gly 145 150 155 160 He Asn Leu Gly Asn Gin Tyr Gly Gly Phe Thr Asp Arg Ser Gin Cys 165 170 175 Ala Thr Leu Pro Ser Lys Trp Gin Ala Ser Cys Asn Trp Arg Phe Asp 180 185 190 Trp Phe Glu Asn Ala Asp Asn Pro Thr Val Asp Trp Glu Pro Val Thr 195 200 205 Cys Pro Gin Glu Leu Val Ala Arg Thr Gly Cys Ser Arg Thr 210 215 220 2) INFORMATION FOR SEQ ID NO: 12: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 808 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli, DSM 10511 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12: CCGCTGCTGG GTATATAATG CTCAGACTTG GAACCAATGG TCCATCCAAA CATGCTTAAA 60 ACGCTCGCTC CATTGATCAT CTTGGCCGCC TCGGTCACAG CGCAAACAGC AGGAGTTACG 120 ACCCGCTACT GGGACTGCTG CAAGCCAAGC TGTGGATGGA GTGGAAAGGC TTCTGTTTCT 180 GCTCCAGTCA GAACTTGCGA TCGTAATGGA AATACACTTG GCCCAGACGT GAAAAGCGGA 240 TGTGATAGCG GTGGAACGTC ATTCACTTGC GCGAACAATG GTCCATTTGC GATTGACAAT 300 AACACTGCAT ATGGTTTTGC TGCAGCCCAC TTAGCGGGCT CTAGCGAAGC A6CCTGGTGT 360 TGCCAGT6CT ACGAATTGAC GTTTACGAGT GGACCCGTAG TTGGGAAGAA ACTGACCGTT 420 CAAGTCACAA ACACGGGAGG TGACCTCGGA AATAATCACT TTGACCTGAT GATCCCCGGT 480 GGAGGTGTTG GCCTCTTCAC ACAAGGATGT CCTGCTCAGT TTGGGAGCTG GAACGGGGGT 540 GCTCAATACG GGGGTGTGTC CAGCCGTGAC CAATGCTCCC AACTTCCAGC AGCTGTGCAA 600 GCTGGATGTC AATTCCGTTT CGACTGGATG GGTGGCGCGG ATAACCCCAA CGTCACCTTC 660 CGACCTGTGA CCTGCCCAGC GCAGCTCACT AATATCTCGG GCTGTGTTCG TAAATGATTC 720 f ACGAATATGT AGTGTCGAAT ATGTACATGT GTATGTACTA TAGCTTCAAA GATGGAGGGT 780 CTGTTTAAAA AAAAAAAAAA AAAAAAAA 808 ) (2) INFORMATION FOR SEQ ID NO: 13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 226 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13: Met Val His Pro Asn Met Leu Lys Thr Leu Ala Pro Leu lie lie Leu 15 10 15 Ala Ala Ser Val Thr Ala Gin Thr Ala Gly Val Thr Thr Arg Tyr Trp 20 25 30 Asp Cys Cys Lys Pro Ser Cys Gly Trp Ser Gly Lys Ala Ser Val Ser 35 40 45 Ala Pro Val Arg Thr Cys Asp Arg Asn Gly Asn Thr Leu Gly Pro Asp 50 55 60 Val Lys Ser Gly Cys Asp Ser Gly Gly Thr Ser Phe Thr Cys Ala Asn 65 70 75 80 Asn Gly Pro Phe Ala lie Asp Asn Asn Thr Ala Tyr Gly Phe Ala Ala 85 90 95 Ala His Leu Ala Gly Ser Ser Glu Ala Ala Trp Cys Cys Gin Cys Tyr 100 105 110 Glu Leu Thr Phe Thr Ser Gly Pro Val Val Gly Lys Lys Leu Thr Val 115 120 125 Gin Val Thr Asn Thr Gly Gly Asp Leu Gly Asn Aen His Phe Asp Leu 130 135 140 Met lie Pro Gly Gly Gly Val Gly Leu Phe Thr Gin Gly Cys Pro Ala 145 150 155 160 Gin Phe Gly Ser Trp Asn Gly Gly Ala Gin Tyr Gly Gly Val Ser Ser 165 170 175 Arg Asp Gin Cys Ser Gin Leu Pro Ala Ala Val Gin Ala Gly Cys Gin 180 185 190 Phe Arg Phe Asp Trp Met Gly Gly Ala Asp Asn Pro Asn Val Thr Phe 195 200 205 Arg Pro Val Thr Cys Pro Ala Gin Leu Thr Asn lie Ser Gly Cys Val 210 215 220 Arg Lys 225 (2) INFORMATION FOR SEQ ID NO: 14-A: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1048 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14-A: GACTTGGAAC CAATGGTCCA TCCAAACATG CTTAAAACGC TCGCTCCATT GATCATCTTG 60 GCCGCCTCGG TCACAGCGCA AACAGCAGGA GTTACGACCC GCTACTGGGA CTGCTGCAAG 120 CCAAGCTGTG GATGGAGTGG AAAGGCTTCT GTTTCTGCTC CAGTCAGAAC TTGCGATCGT 180 AATGGAAATA CACTTGGCCC AGACGTGAAA AGCGGATGTG ATAGCGGTGG AACGTCATTC 240 ACTTGCGCGA ACAATGGTCC ATTTGCGATT GACAATAACA CTGCATATGG TTTTGCTGCA 300 GCCCACTTAG CGGGCTCTAG CGAAGCAGCC TGGTGTTGCC AGTGCTACGA ATTGACGTTT 360 ACGAGTGGAC CCGTAGTTGG GAAGAAACTG ACCGTTCAAG TCACAAACAC GGGAGGTGAC 420 CTCGGAAATA ATCACTTTGA CCTGATGATC CCCGGTGGAG GTGTTGGCCT CTTCACACAA 480 GGATGTCCTG CTCAGTTTGG GAGCTGGAAC GGGGGTGCTC AATACGGGGG TGTGTCCAGC 540 CGTGACCAAT GCTCCCAACT TCCAGCAGCT GTGCAAGCTG GATGTCAATT CCGTTTCGAC 600 TGGATGGGTG GCGCGGATAA CCCCAACGTC ACCTTCCGAC CTGTGACCTG CCCAGCGCAG 660 CTCACTAATA TCTCGGGCTG TGTTCGTAAA CCCTCCAGCA GCACCAGCTC TCCGGTCAAC 720 CAGCCTACCA GCACCAGCAC CACGTCCACC TCCACCACCT CGAGCCCGCC AGTCCAGCCT 780 ACGACTCCCA GCGGCTGCAC TGCTGAGAGG TGGGCTCAGT GCGGCGGCAA TGGCTGGAGC 840 G6CTGCACCA CCTGCGTCGC TGGCAGCACT TGCACGAAGA TTAATGACTG GTACCATCAG 900 TGCCTGTAGA CGCAGGGCAG CTTGAGGGCC TTACTGGTGG CGCAACGAAA TGACACTCCC 960 AATCACTGTA TTAGTTCTTG TACATAATTT CGTCATCCCT CCAGGGATTG TCACATAAAT 1020 GCAATGAGGA ACAATGAGTA CAGAATTC 1048 (2) INFORMATION FOR SEQ ID NO: 14-B: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 298 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14-B: Met Val His Pro Asn Met Leu Lys Thr Leu Ala Pro Leu lie lie Leu 15 10 15 Ala Ala Ser Val Thr Ala Gin Thr Ala Gly Val Thr Thr Arg Tyr Trp 20 25 30 Asp Cys Cys Lys Pro Ser Cys Gly Trp Ser Gly Lys Ala Ser Val Ser 35 40 45 Ala Pro Val Arg Thr Cys Asp Arg Asn Gly Asn Thr Leu Gly Pro Asp 50 55 60 Val Lys Ser Gly Cys Asp Ser Gly Gly Thr Ser Phe Thr Cys Ala Asn 65 70 75 80 Asn Gly Pro Phe Ala lie Asp Asn Asn Thr Ala Tyr Gly Phe Ala Ala 85 90 95 Ala His Leu Ala Gly Ser Ser Glu Ala Ala Trp Cys Cys Gin Cys Tyr 100 105 110 Glu Leu Thr Phe Thr Ser Gly Pro Val Val Gly Lys Lys Leu Thr Val 115 120 125 Gin Val Thr Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp Leu 130 135 140 Met lie Pro Gly Gly Gly Val Gly Leu Phe Thr Gin Gly Cys Pro Ala 145 150 155 160 Gin Phe Gly Ser Trp Asn Gly Gly Ala Gin Tyr Gly Gly Val Ser Ser 165 170 175 Arg Asp Gin Cys Ser Gin Leu Pro Ala Ala Val Gin Ala Gly Cys Gin 180 185 190 Phe Arg Phe Asp Trp Met Gly Gly Ala Asp Asn Pro Asn Val Thr Phe 195 200 205 Arg Pro Val Thr Cys Pro Ala Gin Leu Thr Asn lie Ser Gly Cys Val 210 215 220 Arg Lys Pro Ser Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser 225 230 235 240 Thr Ser Thr Thr Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro 245 250 255 Thr Thr Pro Ser Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly 260 265 270 Asn Gly Trp Ser Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr 275 280 285 Lys lie Asn Asp Trp Tyr His Gin Cys Leu 290 295 (2) INFORMATION FOR SEQ ID NO: 15-A: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1031 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15-A: CCATCCAAAC ATGCTTAAAA CGCTCGCTCC ATTGATCATC TTGGCCGCCT C6GTCACAGC 60 GCAAACAGCA GGAGTTACGA CCCGCTACTG GGACTGCTGC AAGCCAAGCT GTGGATGGAG 120 TGGAAAGGCT TCTGTTTCTG CTCCAGTCAG AACTTGCGAT CGTAATGGAA ATACACTTGG 180 CCCAGACGTG AAAAGCGGAT GTGATAGCGG TGGAACGTCA TTCACTTGCG CGAACAATGG 240 TCCATTTGCG ATTGACAATA ACACTGCATA TGGTTTTGCT GCAGCCCACT TAGCGGGCTC 300 TAGCGAAGCA GCCTGGTGTT GCCAGTGCTA CGAATTGACG TTTACGAGTG GACCCGTAGT 360 TGGGAAGAAA CTGACCGTTC AAGTCACAAA CACGGGAGGT GACCTCGGAA ATAATCACTT 420 TGACCTGATG ATCCCCGGTG GAGGTGTTGG CCTCTTCACA CAAGGATGTC CTGCTCAGTT 480 TGGGAGCTGG AACGGGGGTG CTCAATACGG GGGTGTGTCC AGCCGTGACC AATGCTCCCA 540 ACTTCCAGCA GCTGTGCAAG CTGGATGTCA ATTCCGTTTC GACTGGATGG GTGGCGCGGA 600 TAACCCCAAC GTCACCTTCC GACCTGTGAC CTGCCCAGCG CAGCTCACTA ATATCTCGGG 660 CTGTGTTCGT AAACCCTCCA GCAGCACCAG CTCTCCGGTC AACCAGCCTA CCAGCACCAG 720 CACCACGTCC ACCTCCACCA CCTCGAGCCC GCCAGTCCAG CCTACGACTC CCAGCGGCTG 780 CACTGCTGAG AGGTGGGCTC AGTGCGGCGG CAATGGCTGG AGCGGCTGCA CCACCTGCGT 840 CGCTGGCAGC ACTTGCACGA AGATTAATGA CTGGTACCAT CAGTGCCTGT AGACGCAGGG 900 CAGCTTGAGG GCCTTACTGG TGGCGCAACG AAATGACACT CCCAATCACT GTATTAGTTC 960 TTGTACATAA TTTCGTCATC CCTCCAGGGA TTGTCACATA AATGCAATGA GGAACAATGA 1020 GTACAGAATT C 1031 (2) INFORMATION FOR SEQ ID NO: 15-B: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 293 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15-B: Met Leu Lya Thr Leu Ala Pro Leu lie lie Leu Ala Ala Ser Val Thr 15 10 15 Ala Gin Thr Ala Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys Pro 20 25 30 Ser Cys Gly Trp Ser Gly Lys Ala Ser Val Ser Ala Pro Val Arg Thr 35 40 45 Cys Asp Arg Asn Gly Asn Thr Leu Gly Pro Asp Val Lys Ser Gly Cys 50 55 60 Asp Ser Gly Gly Thr Ser Phe Thr Cys Ala Asn Asn Gly Pro Phe Ala 65 70 75 80 lie Asp Asn Asn Thr Ala Tyr Gly Phe Ala Ala Ala His Leu Ala Gly 85 90 95 Ser Ser Glu Ala Ala Trp Cys Cys Gin Cys Tyr Glu Leu Thr Phe Thr 100 105 110 Ser Gly Pro Val Val Gly Lys Lys Leu Thr Val Gin Val Thr Asn Thr 115 120 125 Gly Gly Asp Leu Gly Asn Asn His Phe Asp Leu Met lie Pro Gly Gly 130 135 140 Gly Val Gly Leu Phe Thr Gin Gly Cys Pro Ala Gin Phe Gly Ser Trp 145 150 155 160 Asn Gly Gly Ala Gin Tyr Gly Gly Val Ser Ser Arg Asp Gin Cys Ser 165 170 175 Gin Leu Pro Ala Ala Val Gin Ala Gly Cys Gin Phe Arg Phe Asp Trp 180 185 190 Met Gly Gly Ala Asp Asn Pro Asn Val Thr Phe Arg Pro Val Thr Cys 195 200 205 Pro Ala Gin Leu Thr Asn He Ser Gly Cys Val Arg Lys Pro Ser Ser 210 215 220 Ser Thr Ser Ser Pro Val Asn Gin Pro Thr. Ser Thr Ser Thr Thr Ser 225 230 235 240 Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser Gly 245 250 255 Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser Gly 260 265 270 Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys He Asn Asp Trp 275 280 285 Tyr His Gin Cys Leu 290 (2) INFORMATION FOR SEQ ID NO: 16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1132 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECtTLE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli, DSM 10571 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16: CAACAGTTCA AACACCTACA AGGTCCCGTG CCCTGTAGAC CATGCGTTCC TCTGCAGTCC 60 TCATCGGCCT CGTGGCCGGT GTGGCCGCCC AGTCCTCTGG CACCGGCCGC ACCACCAGAT 120 ACTGGGACTG CTGCAAGCCG TCCTGCGGGT GGGACGAAAA GGCCTCCGTC AGCCAGCCCG 180 TCAAGACGTG CGATAGGAAC AACAACCCTC TCGCGTCCAC GGCCAGGAGC GGCTGCGATT 240 CCAACGGCGT CGCCTACACG TGCAACGATA ACCAGCCGTG GGCTGTCAAC GATAACCTGG 300 CCTATGGTTT TGCTGCCACG GCTTTCAGTG GTGGATCGGA GGCCAGCTGG TGCTGTGCCT 360 GCTATGCCCT TCAGTTCACC TCCGGCCCTG TTGCGGGAAA GACCATGGTC GTCCAGTCGA 420 CAAACACCGG CGGCGACCTC AGCGGCAACC ACTTTGACAT CCTCATGCCC GGCGGCGGCC 480 TGGGCATCTT CGACGGCTGC ACCCCGCAAT GGGGCGTCAG CTTCCCCGGA AACCGCTACG 540 GCGGCACCAC CAGCCGCAGC CAGTGCTCCC AAATCCCCTC GGCCCTGCAG CCCGGCTGCA 600 ACTGGCGGTA CGACTGGTTC AACGACGCCG ACAACCCCGA CGTCTCGTGG CGCCGCGTCC 660 AGTGCCCCGC CGCACTCACC GACCGCACCG GCTGCCGCCG CTCCGATGAC GGGAACTATC 720 CCGTCTTCCA GCCCGGTCCG CCCCCGGCCA CGACGATCAG GACATCGACT ACCATCACAG 780 CCTCATCGTC GTCTTCGTCT TCGTCGTCGT CGACTACGGC TGGTAGCCCG CCTGTGCCGA 840 CTGGTGGTGG TAGTGGGCCA ACGTCGCCTG TCTGGGGACA GTGCGGCGGT CAGGGATGGA 900 GTGGTCCTAC GCGTTGTGTT GCTGGGTCGA CATGCAGTGT GGTCAACCCG TGGTACTCGC 960 AGTGTTTTCC TTAAGGAGCC TCTGGCTGAG CAGATCCTTT CGAAGAGGAG GGTCTCTCTG 1020 CTCTTTCAGT CTGTTCAGGG AACGGCCGTC TCGGCTACAT TGTACATATC CCACCTCGTA 1080 TATAGCTAGC TCATCTACAC TTGTGATCTC CAAAAAAAAA AAAAAAAAAA AA 1132 (2) INFORMATION FOR SEQ ID NO: 17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 310 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17: Met Arg Ser Ser Ala Val Leu lie Gly Leu Val Ala Gly Val Ala Ala 15 10 15 Gin Ser Ser Gly Thr Gly Arg Thr Thr Arg Tyr Trp Asp Cys Cys Lys 20 25 30 Pro Ser Cys Gly Trp Asp Glu Lys Ala Ser Val Ser Gin Pro Val Lys 35 40 45 Thr Cys Asp Arg Asn Asn Asn Pro Leu Ala Ser Thr Ala Arg Ser Gly 50 55 60 Cys Asp Ser Asn Gly Val Ala Tyr Thr Cys Asn Asp Asn Gin Pro Trp 65 70 75 80 Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Ala Phe Ser 85 90 95 Gly Gly Ser Glu Ala Ser Trp Cys Cys Ala Cys Tyr Ala Leu Gin Phe 100 105 110 Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val Gin Ser Thr Asn 115 120 125 Thr Gly Gly Asp Leu Ser Gly Asn His Phe Asp He Leu Met Pro Gly 130 135 140 Gly Gly Leu Gly He Phe Asp Gly Cys Thr Pro Gin Trp Gly Val Ser 145 150 155 160 Phe Pro Gly Asn Arg Tyr Gly Gly Thr Thr Ser Arg Ser Gin Cys Ser 165 170 175 Gin He Pro Ser Ala Leu Gin Pro Gly Cys Asn Trp Arg Tyr Asp Trp 180 185 190 Phe Asn Asp Ala Asp Asn Pro Asp Val Ser Trp Arg Arg Val Gin Cyj 195 200 205 Pro Ala Ala Leu Thr Asp Arg Thr Gly Cys Arg Arg Ser Asp Asp Gly 210 215 220 Asn Tyr Pro Val Phe Gin Pro Gly Pro Pro Pro Ala Thr Thr lie Arg 225 230 235 240 Thr Ser Thr Thr lie Thr Ala Ser Ser Ser Ser Ser Ser Ser Ser Ser 245 250 255 Ser Thr Thr Ala Gly Ser Pro Pro Val Pro Thr Gly Gly Gly Ser Gly 260 265 270 Pro Thr Ser Pro Val Trp Gly Gin Cys Gly Gly Gin Gly Trp Ser Gly 275 280 285 Pro Thr Arg Cys Val Ala Gly Ser Thr Cys Ser Val Val Asn Pro Trp 290 295 300 Tyr Ser Gin Cys Phe Pro 305 310 (2) INFORMATION FOR SEQ ID NO: 18-A: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 885 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (vi) ORIGINAL SOURCE: "Construction from Macrophomina" (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..885 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18-A: ATG TTC TCT CCG CTC TGG GCC CTG TCG GCT CTG CTC CTA TTT CCT GCC 48 Met Phe Ser Pro Leu Trp Ala Leu Ser Ala Leu Leu Leu Phe Pro Ala 145 150 155 ACT GAA GCC ACT AGC GGC GTG ACA ACC AGG TAC TGG GAC TGC TGC AAG 96 Thr Glu Ala Thr Ser Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys 160 165 170 CCG TCT TGT GCT TGG ACG GGC AAA GCA TCC GTC TCC AAG CCC GTC GGA 144 Pro Ser Cys Ala Trp Thr Gly Lys Ala Ser Val Ser Lys Pro Val Gly 175 180 185 ACC TGC GAC ATC AAC GAC AAC GCC CAG ACG CCG AGC GAT CTG CTC AAG 192 Thr Cyg Asp He Asn Asp Asn Ala Gin Thr Pro Ser Asp Leu Leu Lys 190 195 200 205 TCG TCC TGT GAT GGC GGC AGC GCC TAC TAC TGC AGC AAC CAG GGC CCA 240 Ser Ser Cys Asp Gly Gly Ser Ala Tyr Tyr Cys Ser Asn Gin Gly Pro 210 215 220 TGG GCC GTG AAC GAC AGC CTT TCC TAC GGC TTC GCT GCC GCC AAG CTG 288 Trp Ala Val Asn Asp Ser Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu 225 230 235 TCC GGA AAG CAG GAG ACT GAT TGG TGC TGT GGC TGC TAC AAG CTC ACA 336 Ser Gly Lys Gin Glu Thr Asp Trp Cys Cys Gly Cys Tyr Lys Leu Thr 240 245 250 TTC ACC TCC ACC GCC GTT TCC GGC AAG CAA ATG ATC GTG CAA ATC ACG 384 Phe Thr Ser Thr Ala Val Ser Gly Lys Gin Met lie Val Gin lie Thr 255 260 265 i AAC ACG GGC GGC GAC CTC GGC AAC AAC CAC TTC GAC ATC GCC ATG CCG 432 Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp He Ala Met Pro 270 275 280 285 GGC GGC GGC GTC GGC ATC TTC AAC GGG TGC TCC AAG CAA TGG AAC GGC 480 Gly Gly Gly Val Gly He Phe Asn Gly Cys Ser Lys Gin Trp Asn Gly 290 295 300 I ATC AAT CTG GGC AAC CAG TAT GGC GGC TTC ACT GAC CGC TCG CAA TGT 528 He Asn Leu Gly Asn Gin Tyr Gly Gly Phe Thr Asp Arg Ser Gin Cys 305 310 315 GCG ACG CTC CCG TCC AAG TGG CAG GCC AGC TGC AAC TGG CGC TTC GAC 576 Ala Thr Leu Pro Ser Lys Trp Gin Ala Ser Cys Asn Trp Arg Phe Asp 320 325 330 TGG TTC GAG AAT GCC GAC AAC CCC ACC GTC GAT TGG GAG COT GTC ACT 624 Trp Phe Glu Asn Ala Asp Asn Pro Thr Val Asp Trp Glu Pro Val Thr 335 340 345 TGC CCA CAG GAA TTG GTC GCC CGG ACT GGC TGT TCC CGT ACC CCC TCC 672 Cys Pro Gin Glu Leu Val Ala Arg Thr Gly Cys Ser Arg Thr Pro Ser 350 355 360 365 AGC AGC ACC AGC TCT CCG GTC AAC CAG CCT ACC AGC ACC AGC ACC ACG 720 Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr Ser Thr Thr 370 375 380 TCC ACC TCC ACC ACC TCG AGC CCG CCA GTC CAG CCT ACG ACT CCC AGC 768 Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser 385 390 395 GGC TGC ACT GCT GAG AGG TGG GCT CAG TGC GGC GGC AAT GGC TGG AGC 816 Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser 400 405 410 GGC TGC ACC ACC TGC GTC GCT GGC AGC ACT TGC ACG AAG ATT AAT GAC 864 Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys lie Asn Asp 415 420 425 TGG TAC CAT CAG TGC CTG TAG 885 Trp Tyr His Gin Cys Leu * 430 435 (2) INFORMATION FOR SEQ ID NO: 18-B: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 295 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18-B: Met Phe Ser Pro Leu Trp Ala Leu Ser Ala Leu Leu Leu Phe Pro Ala 15 10 15 Thr Glu Ala Thr Ser Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys 20 25 30 Pro Ser Cys Ala Trp Thr Gly Lys Ala Ser Val Ser Lys Pro Val Gly 35 40 45 Thr Cys Asp lie Asn Asp Asn Ala Gin Thr Pro Ser Asp Leu Leu Lys 50 55 60 Ser Ser Cys Asp Gly Gly Ser Ala Tyr Tyr Cys Ser Asn Gin Gly Pro 65 70 75 80 Trp Ala Val Asn Asp Ser Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu 85 90 95 Ser Gly Lys Gin Glu Thr Asp. .Trp Cys Cys Gly Cys Tyr Lys Leu Thr 100 105 110 Phe Thr Ser Thr Ala Val Ser Gly Lys Gin Met lie Val Gin lie Thr 115 120 125 Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp lie Ala Met Pro 130 135 140 Gly Gly Gly Val Gly lie Phe Asn Gly Cys Ser Lys Gin Trp Asn Gly 145 150 155 160 lie Asn Leu Gly Asn Gin Tyr Gly Gly Phe Thr Asp Arg Ser Gin Cys 165 170 175 Ala Thr Leu Pro Ser Lys Trp Gin Ala Ser Cys Asn Trp Arg Phe Asp 180 185 190 Trp Phe Glu Asn Ala Asp Asn Pro Thr Val Asp Trp Glu Pro Val Thr 195 200 205 Cys Pro Gin Glu Leu Val Ala Arg Thr Gly Cys Ser Arg Thr Pro Ser 210 215 220 Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr Ser Thr Thr 225 230 235 240 Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser 245 250 255 Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser 260 265 270 Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys He Asn Asp 275 280 285 Trp Tyr His Gin Cys Leu * 290 295 (2) INFORMATION FOR SEQ ID NO: 19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 425 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli, DSM 10576 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:l..4 25 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19: CAAGATACAA TATGCGTTCC TCCACTATTT TGCAAACCGG CCTGGTGGCC GTTCTCCCCT 60 TCGCCGTCCA GGCCGCCTCA GGATCCGGCA AGTCCACCAG ATATTGGGAC TGCTGCAAAC 120 CATCTTGTGC CTGGTCCGGC AAGGCTTCTG TCAACCGCCC TGTTCTCGCC TGCAACGCAA 180 ACAACAACCC GCTGAACGAC GCCAACGTCA AGTCAGGATG TGATGGCGGT TCTGCATACA 240 CCTGTGCCAA CAACTCTCCC TGGGCAGTGA ATGACAATCT GGCCTACGGC TTCGCGGCCA 300 CAAAACTCAG CGGGGGGACC GAGTCATCTT GGTGCTGCGC CTGTTATGCC CTCACATTCA 360 CATCGGGTCC TGTTTCTGGC AAAACCTTGG TTGTCCAGTC TACCAGTACC GGTGGTGATC 420 TTGGC 425 (2) INFORMATION FOR SEQ ID NO: 20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 141 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20: Met Arg Ser Ser Thr He Leu Gin Thr Gly Leu Val Ala Val Leu Pro 15 10 15 Phe Ala Val Gin Ala Ala Ser Gly Ser Gly Lys Ser Thr Arg Tyr Trp 20 25 30 Asp Cys Cys Lys Pro Ser Cys Ala Trp Ser Gly Lys Ala Ser Val Asn 35 40 45 Arg Pro Val Leu Ala Cys Asn Ala Asn Asn Asn Pro Leu Asn Asp Ala Asn Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys Ala Asn 65 70 75 80 Asn Ser Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala 85 90 95 Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser Trp Cys Cys Ala Cys Tyr 100 105 110 Ala Leu Thr Phe Thr Ser Gly Pro Val Ser Gly Lys Thr Leu Val Val 115 120 125 Gin Ser Thr Ser Thr Gly Gly Asp Leu Gly 130 135 2) INFORMATION FOR SEQ ID NO: 21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 108 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Saccobolus dilutellus (B) STRAIN: CBS 275.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:!..108 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: TCG GOT TGC GAT AAC GGT GGT GGC ACT GCA TAC ATG TGT GCC AGC CAG Ser Ala Cys Asp Asn Gly Gly Gly Thr Ala Tyr Met Cys Ala Ser Gin is 10 15 GAG CCG TGG GCA GTG AGC TCC AAC GTC GCG TAC GGC TTT GCT GCA GTT Glu Pro Trp Ala Val Ser Ser Asn Val Ala Tyr Gly Phe Ala Ala Val 20 25 30 AGA ATC AGC GGA Arg lie Ser Gly 35 (2) INFORMATION FOR SEQ ID NO: 22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22: Ser Ala Cys Asp Asn Gly Gly Gly Thr Ala Tyr Met Cys Ala Ser Gin 15 10 15 Glu Pro Trp Ala Val Ser Ser Asn Val Ala Tyr Gly Phe Ala Ala Val 20 25 30 Arg He Ser Gly 35 (2) INFORMATION FOR SEQ ID NO: 23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 99 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Thermomyces verrucosus (B) STRAIN: CBS 285.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:!..99 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 23: GCC TGC AAC GCA AAC TTC CAG CGC ATC AGT GAC CCC AAC GCC AAG TCG Ala Cys Asn Ala Asn Phe Gin Arg lie Ser Asp Pro Asn Ala Lys Ser GGC TGC GAT GGT GGC TCG GCC TTC TCT TGC GCC AAA CAA ACC CCT TGG Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Lys Gin Thr Pro Trp GCC Ala (2) INFORMATION FOR SEQ ID NO: 24: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 33 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24: Ala Cys Asn Ala Asn Phe Gin Arg lie Ser Asp Pro Asn Ala Lys Ser 1 5 10 15 Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Lys Gin Thr Pro Trp 20 25 30 Ala (2) INFORMATION FOR SEQ ID NO: 25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 225 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Xylaria hypoxylon (B) STRAIN: CBS 284.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..225 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25: GAC CAG CCG CTC GGC GGA CAA CGG ACG CGA CCA AGG AGC GCG TGC GAC 48 Asp Gin Pro Leu Gly Gly Gin Arg Thr Arg Pro Arg Ser Ala Cys Asp 35 40 45 AAT GGC GGC TCT GCA TAG ATG TGC AGC AAC CAG AGC CCG TGG GCC GTC 96 Asn Gly Gly Ser Ala Tyr Met Cys Ser Asn Gin Ser Pro Trp Ala Val 50 55 60 65 GAC GAT TCT CTC AGT TAG GGA TGG GCT GCC GTT AGG ATC TAT GGA CAT 144 Asp Asp Ser Leu Ser Tyr Gly Trp Ala Ala Val Arg lie Tyr Gly His 70 75 80 ACC GAA ACT ACT TGG TGC TGC GCT TGC TAC GAG TTG ACT TTT ACC AGC 192 Thr Glu Thr Thr Trp Cys Cys Ala Cys Tyr Glu Leu Thr Phe Thr Ser 85 90 95 GGT CCG GTT AGC GGC AAG AAG ATG ATT GTT CAG 225 Gly Pro Val Ser Gly Lys Lys Met lie Val Gin 100 105 (2) INFORMATION FOR SEQ ID NO: 26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 75 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26: Asp Gin Pro Leu Gly Gly Gin Arg Thr Arg Pro Arg Ser Ala Cys Asp 15 10 15 Asn Gly Gly Ser Ala Tyr Met Cys Ser Asn Gin Ser Pro Trp Ala Val 20 25 30 Asp Asp Ser Leu Ser Tyr Gly Trp Ala Ala Val Arg lie Tyr Gly His 35 40 45 Thr Glu Thr Thr Trp Cys Cys Ala Cys Tyr Glu Leu Thr Phe Thr Ser 50 55 60 Gly Pro Val Ser Gly Lys Lys Met lie Val Gin 65 70 75 (2) INFORMATION FOR SEQ ID NO: 27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Fusarium oxysporum ssp lycopersici (B) STRAIN: CBS 645.78 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:!..177 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27: AGA AAC GAC AAC CCC ATC TCC AAC ACC AAC GCT GTC AAC GGT TGT GAG 48 Arg Asn Asp Asn Pro lie Ser Asn Thr Asn Ala Val Asn Gly Cys Glu 30 35 40 45 GGT GGT GGT TCT GCT TAT GCT TGC ACC AAC TAC TCT CCC TGG GCT GTC 96 Gly Gly Gly Ser Ala Tyr Ala Cys Thr Asn Tyr Ser Pro Trp Ala Val 50 55 60 AAC GAT GAG CTT GCC TAC GGT TTC GCT GCT ACC AAG ATC TCC GGT GGC 144 Asn Asp Glu Leu Ala Tyr Gly Phe Ala Ala Thr Lys lie Ser Gly Gly 65 70 75 TCC GAG GCC AGC TGG TGC TGT GCC TGC TAT CTA 177 Ser Glu Ala Ser Trp Cys Cys Ala Cys Tyr Leu 80 85 (2) INFORMATION FOR SEQ ID NO: 28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 59 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28: Arg Asn Asp Asn Pro lie Ser Asn Thr Asn Ala Val Asn Gly Cys Glu 15 10 15 Gly Gly Gly Ser Ala Tyr Ala Cys Thr Asn Tyr Ser Pro Trp Ala Val 20 25 30 Asn Asp Glu Leu Ala Tyr Gly Phe Ala Ala Thr Lys lie Ser Gly Gly 35 40 45 Ser Glu Ala Ser Trp Cys Cys Ala Cys Tyr Leu 50 55 (2) INFORMATION FOR SEQ ID NO: 29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 63 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: LINEAR (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Nectria pinea (B) STRAIN: CBS 279.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..63 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 29: AGC GGC TGT GAC GGT GGT TCT GCC TAC GCC TGT GCA AAC AAC TCC CCT 4 8 Ser Gly Cys Asp Gly Gly Ser Ala Tyr Ala Cys Ala Asn Asn Ser Pro 60 65 70 75 TGG GCT GTC AAC GAT 63 Trp Ala Val Asn Asp 80 (2) INFORMATION FOR SEQ ID NO: 30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 30: Ser Gly Cys Asp Gly Gly Ser Ala Tyr Ala Cys Ala Asn Asn Ser Pro 15 10 15 Trp Ala Val Asn Asp 20 (2) INFORMATION FOR SEQ ID NO: 31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Humicola grisea Traeen (B) STRAIN: ATCC 22726 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..177 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 31: AAC CAG CCT GTC TTC ACT TGC GAC GCC AAA TTC CAG CGC ATC ACC GAC 48 Asn Gin Pro Val Phe Thr Cys Asp Ala Lys Phe Gin Arg lie Thr Asp 25 30 35 CCC AAT ACC AAG TCG GGC TGC GAT GGC GGC TCG GCC TTT TCG TGT GCT 96 Pro Asn Thr Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala 40 45 50 GAC CAA ACC CCC TGG GCT CTG AAC GAC GAT TTC GCC TAT GGC TTC GCT 144 Asp Gin Thr Pro Trp Ala Leu Asn Asp Asp Phe Ala Tyr Gly Phe Ala 55 60 65 GCC ACG GCT ATT TCG GGT GGA TCG GAA GCC TCG 177 Ala Thr Ala lie Ser Gly Gly Ser Glu Ala Ser 70 75 80 (2) INFORMATION FOR SEQ ID NO: 32: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 59 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 32: Asn Gin Pro Val Phe Thr Cys Asp Ala Lys Phe Gin Arg lie Thr Asp 15 10 15 Pro Asn Thr Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala 20 25 30 Asp Gin Thr Pro Trp Ala Leu Asn Asp Asp Phe Ala Tyr Gly Phe Ala 35 40 45 Ala Thr Ala lie Ser Gly Gly Ser Glu Ala Ser 50 55 (2) INFORMATION FOR SEQ ID NO: 33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 153 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Humicola nigrescens Oinvik (B) STRAIN: CBS 819.73 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:!..15 3 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 33: GTC TAC GCC TGC AAC GCA AAC TTC CAG CGC ATC ACC GAC GCC AAC GCC 48 Val Tyr Ala Cys Asn Ala Asn Phe Gin Arg lie Thr Asp Ala Asn Ala 60 65 70 75 AAG TCC GGC TGC GAT GGC GGC TCC GCC TTC TCG TGC GCC AAC CAG ACC 96 Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Asn Gin Thr 80 85 90 CCG TGG GCC GTG AGC GAC GAC TTT GCC TAC GGT TTC GCG GCT ACG GCG 144 Pro Trp Ala Val Ser Asp Asp Phe Ala Tyr Gly Phe Ala Ala Thr Ala 95 100 105 CTC GCC GGC 153 Leu Ala Gly 110 (2) INFORMATION FOR SEQ ID NO: 34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 51 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein . (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 34: Val Tyr Ala Cys Asn Ala Asn Phe Gin Arg lie Thr Asp Ala Asn Ala 15 10 15 Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Asn Gin Thr 20 25 30 Pro Trp Ala Val Ser Asp Asp Phe Ala Tyr Gly Phe Ala Ala Thr Ala 35 40 45 Leu Ala Gly 50 (2) INFORMATION FOR SEQ ID NO: 35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 181 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Cladorrhinum foecundissimum (B) STRAIN: ATCC 62373 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..181 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 35; GTC AAC CGC CCT GTC CTC GCC TGC GAC GCA AAC AAC AAC CCT CTG ACC 48 Val Asn Arg Pro Val Leu Ala Cys Asp Ala Asn Asn Asn Pro Leu Thr 15 10 15 GAC GCC GGC GTC AAG TCC GGA TGT GAT GGC GGT TCT GCA TAC ACC TGT 96 Asp Ala Gly Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys 20 25 30 GCC AAC AAC TCC CCA TGG GCA GTG AAC GAC CAG CTC GCC TAC GGC TTT 144 Ala Asn Asn Ser Pro Trp Ala Val Asn Asp Gin Leu Ala Tyr Gly Phe 35 40 45 GCC GCC ACC AAA CTG AGC GGC GGA ACT GAG TCG TCA 180 Ala Ala Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser 50 55 60 (2) INFORMATION FOR SEQ ID NO: 36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 60 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 36; Val Asn Arg Pro Val Leu Ala Cys Asp Ala Asn Asn Asn Pro Leu Thr 15 10 15 Asp Ala Gly Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys 20 25 30 Ala Asn Asn Ser Pro Trp Ala Val Asn Asp Gin Leu Ala Tyr Gly Phe 35 40 45 Ala Ala Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser 50 55 60 (2) INFORMATION FOR SEQ ID NO: 37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 64 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Syspastospora boninensis (B) STRAIN: NKBC 1515 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:!..64 (xi) SEQUENCE DESCRIPTION; SEQ ID NO: 37: GGC TGC GAC GGC GGC AGC GCC TTC ACC TGC TCC AAC AAC TCT CCA TGG 48 Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser Pro Trp GCT GTG AAC GAA GAT 63 Ala Val Asn Glu Asp (2) INFORMATION FOR SEQ ID NO: 38: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 38: Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser Pro Trp Ala Val Asn Glu Asp (2) INFORMATION FOR SEQ ID NO: 39: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 153 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Nigrospora sp (B) STRAIN: CBS 272.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1.-153 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 39: ACA AGA AAC GAC GGG CCC CTG TCC AGC CCC GAT GCC GCC TCC GGC TGT 48 Thr Arg Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser Gly Cys 25 30 35 GAT GGC GGC GAA GCC TTT GCC TGT TCT AAT ACC TCG CCT TGG GCC GTC 95 Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp Ala Val 40 45 50 AGC GAC GAG CTC GCG TAG GGA TAG GTC GCC ACG TCC ATC TCC GGC GGC 144 Ser Asp Gin Leu Ala Tyr Gly Tyr Val Ala Thr Ser lie Ser Gly Gly 55 60 65 ACC GAG TCA 153 Thr Glu Ser 70 (2) INFORMATION FOR SEQ ID NO: 40: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 51 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 40: Thr Arg Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser Gly Cys 15 10 15 Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp Ala Val 20 25 30 Ser Asp Gin Leu Ala Tyr Gly Tyr Val Ala Thr Ser lie Ser Gly Gly 35 40 45 Thr Glu Ser 50 (2) INFORMATION FOR SEQ ID NO: 41: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 159 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (Vi) ORIGINAL SOURCE: (A) ORGANISM: Chaetostylum fresenii (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..159 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 41: GTC CGA ACG TGT AGT GCC AAC GAC TCG CCC TTG TCC GAC CCA AAT GCC 48 Val Arg Thr Cys Ser Ala Asn Asp Ser Pro Leu Ser Asp Pro Asn Ala 55 60 65 CCA AGT GGG TGT GAC GGT GGT AGC GCC TTC ACT TGT TCC AAC AAC TCC 9 6 Pro Ser Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser 70 75 80 CCG TGG GCA GTC GAT GAC CAG ACA GCT TAT GGC TTT GCG GCA ACA GCC 14 4 Pro Trp Ala Val Asp Asp Gin Thr Ala Tyr Gly Phe Ala Ala Thr Ala 85 90 95 ATC AGT GGC CAG TCC 159 lie Ser Gly Gin Ser 100 (2) INFORMATION FOR SEQ ID NO: 42: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 53 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 42: Val Arg Thr Cys Ser Ala Asn Asp Ser Pro Leu Ser Asp Pro Asn Ala 15 10 15 Pro Ser Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser 20 25 30 Pro Trp Ala Val Asp Asp Gin Thr Ala Tyr Gly Phe Ala Ala Thr Ala 35 40 45 lie Ser Gly Gin Ser 50 (2) INFORMATION FOR SEQ ID NO: 43; (i) SEQUENCE CHARACTERISTICS; (A) LENGTH: 153 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Exidia glandulosa (B) STRAIN: CBS 277.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..153 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 43: TGT GAG AAG AAC GAC AAC CCC TTA GCT GAC TTC AGC ACG AAA TCC GGG 48 Cys Glu Lys Asn Asp Asn Pro Leu Ala Asp Phe Ser Thr Lys Ser Gly 55 60 65 TGT GAA AGC GGA GGT TCG GCT TAT ACG TGT AAC AAC CAA TCA CCA TGG 96 Cys Glu Ser Gly Gly Ser Ala Tyr Thr Cys Asn Asn Gin Ser Pro Trp 70 75 80 85 GCC GTC AAT GAC TTG GTG TCG TAT GGC TTC GCC GCC ACA GCG ATC AAT 144 Ala Val Asn Asp Leu Val Ser Tyr Gly Phe Ala Ala Thr Ala lie Asn 90 95 100 GGT GGC AAT 153 Gly Gly Asn (2) INFORMATION FOR SEQ ID NO: 44: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 51 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 44: Cys Glu Lys Asn Asp Asn Pro Leu Ala Asp Phe Ser Thr Lys Ser Gly 15 10 15 Cys Glu Ser Gly Gly Ser Ala Tyr Thr Cys Asn Asn Gin Ser Pro Trp 20 25 30 Ala Val Asn Asp Leu Val Ser Tyr Gly Phe Ala Ala Thr Ala lie Asn 35 40 45 Gly Gly Asn 50 (2) INFORMATION FOR SEQ ID NO: 45: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 171 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Coniothecium sp (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:l..171 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 45: AGG GGC CCC GTC GGA ACC TGC AAG AGG AAC GAG AAC CCC CTC TCC GAC 48 Ser Arg Pro Val Gly Thr Cys Lys Arg Asn Asp Asn Pro Leu Ser Asp 55 60 65 CCC GAT GCC AAG TCC GGC TGC GAC GGC GGC GGC GCC TTC ATG TGC TCC 96 Pro Asp Ala Lys Ser Gly Cys Asp Gly Gly Gly Ala Phe Met Cys Ser 70 75 80 ACC CAG CAG CCG TGG GCC GTC AAC GAC AAT CTG GCA TAT GGC TTC GCC 144 Thr Gin Gin Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala 85 90 95 GCC ACG GCC ATC AGC GGC GGC AAC GAG 171 5 Ala Thr Ala lie Ser Gly Gly Asn Glu 100 105 (2) INFORMATION FOR SEQ ID NO; 46: ) (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 57 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 46: Ser Arg Pro Val Gly Thr Cys Lys Arg Asn Asp Asn Pro Leu Ser Asp 15 10 15 Pro Asp Ala Lys Ser Gly Cys Asp Gly Gly Gly Ala Phe Met Cys Ser 20 25 30 Thr Gin Gin Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala 35 40 45 Ala Thr Ala lie Ser Gly Gly Asn Glu 50 55 (2) INFORMATION FOR SEQ ID NO: 47: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 159 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: CBS 271.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:!..159 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 47: ACT TGC AAC AAG AAC GAC GGG CCC CTG TCG AGC CCC GAT GCC I GCC TCC 4 8 Thr Cys Asn Lys Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser 60 65 70 GGC TGT GAT GGC GGC GAA GCC TTT GCC TGT TCT AAT AGC TCG CCT TGG 96 Gly Cys Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp 75 80 85 GCC GTC AGC GAC GAG CTC GCG TAG GGA TAG CTC GCC ACG TCC ATC TCC 144 Ala Val Ser Asp Gin Leu Ala Tyr Gly Tyr Leu Ala Thr Ser lie Ser 90 95 100 105 GGC GGC ACC GAG TCG 159 Gly Gly Thr Glu Ser 110 (2) INFORMATION FOR SEQ ID NO: 48: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 53 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 48: Thr Cys Asn Lys Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser 15 10 15 Gly Cys Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp 20 25 30 Ala Val Ser Asp Gin Leu Ala Tyr Gly Tyr Leu Ala Thr Ser He Ser 35 40 45 Gly Gly Thr Glu Ser 50 (2) INFORMATION FOR SEQ ID NO: 49 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 84 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: CBS 270.96 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:1..84 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 49: CCA GTT TTC TCC TGT GAC AAG TAC GAC AAC CCT CTA CCT GAC GCC AAT 4 8 Pro Val Phe Ser Cys Asp Lys Tyr Asp Asn Pro Leu Pro Asp Ala Asn 55 60 65 GCT GTG TCC GGG TGT GAC CCC GGA GGT ACT GCC TTC 84 Ala Val Ser Gly Cys Asp Pro Gly Gly Thr Ala Phe 70 75 80 (2) INFORMATION FOR SEQ ID NO: 50: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 28 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 50: Pro Val Phe Ser Cys Asp Lys Tyr Asp Asn Pro Leu Pro Asp 5 Ala Asn 15 10 15 Ala Val Ser Gly Cys Asp Pro Gly Gly Thr Ala Phe 20 25 (2) INFORMATION FOR SEQ ID NO: 51: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 147 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Diplodia gossypina, CBS 274.96 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 51: ACCTGCGACG CCTGCGACAG CCCCCTCAGC GACTACGACG CCAAGTCCGG CTGCGACGGC 60 GGTAGCGCAT ACACCTGCAC CTACTCTACC CCCTGGGCCG TCGACGACAA CCTCTCCTAC 120 GGTTTCGCCG CCGCCAAGCT GAGCGGA 147 (2) INFORMATION FOR SEQ ID NO: 52: (i) SEQUENCE CPiARACTERISTICS: (A) LENGTH: 49 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 52: Thr Cys Asp Ala Cys Asp Ser Pro Leu Ser Asp Tyr Asp Ala Lys Ser 15 10 15 Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys Thr Tyr Ser Thr Pro Trp 20 25 30 Ala Val Asp Asp Asn Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu Ser 35 40 45 Gly (2) INFORMATION FOR SEQ ID NO: 53: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 13 5 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Ulospora bilgramii, NKBC 1444 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 53: CCACTAGCAG ATTTCACCGG TGGAACCGGC TGTAATGGCG GTTCGACATT CTCATGCTCA 60 AACCAACAAC CATGGGCGGT CAACGACACA TTCTCGTACG GCTTTGCGGG CATCTTTATC 120 ACAGGCCATG TCGAG 135 (2) INFORMATION FOR SEQ ID NO: 54: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 5 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 54: Pro Leu Ala Asp Phe Thr Gly Gly Thr Gly Cys Asn Gly Gly Ser Thr 15 10 15 Phe Ser Cys Ser Asn Gin Gin Pro Trp Ala Val Asn Asp Thr Phe Ser 20 25 30 Tyr Gly Phe Ala Gly He Phe He Thr Gly His Val Glu 35 40 45 (2) INFORMATION FOR SEQ ID NO: 55: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 114 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Penicillium verruculosum, ATCC 62396 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 55: GCCAAATCTG GATGTGATGC TGGTGGAGGT CAAGCCTACA TGTGCTCCAA CCAACAACCT 60 TGGGTAGTCA ACGACAACCT CGCCTACGGT TTCGCCGCAG TCAACATTGC CGGC 114 (2) INFORMATION FOR SEQ ID NO: 56: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 56: Ala Lys Ser Gly Cys Asp Ala Gly Gly Gly Gin Ala Tyr Met Cys Ser 15 10 15 Asn Gin Gin Pro Trp Val Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala 20 25 30 Ala Val Asn lie Ala Gly 35 (2) INFORMATION FOR SEQ ID NO: 57: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 113 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Poronia punctata (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 57: TTCGACGTCC GGGTGCGACA ATGGCGGCAG CGCCTTCATG TGCTCTAACC AAAGCCCCTG 60 GGCCGTCAAC GACGATCTGG CCTACGGCTG GGCCGCCGTC TCAATCGCGG GCC 113 (2) INFORMATION FOR SEQ ID NO: 58: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 37 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 58: Ser Thr Ser Gly Cys Asp Asn Gly Gly Ser Ala Phe Met Cys Ser Asn 15 10 15 Gin Ser Pro Trp Ala Val Asn Asp Asp Leu Ala Tyr Gly Trp Ala Ala 20 25 30 Val Ser lie Ala Gly 35 (2) INFORMATION FOR SEQ ID NO: 59: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Fusariura anguioides, IFO 4467 (XI) SEQUENCE DESCRIPTION: SEQ ID NO: 59: [■CAACACCGG TGCAGACGTG CGACCGCAAC GACAACCCGC TCTACGACGG :GGGTCGACG 6O '.GGTCCGGCT GCGACGCCGG CGGCGGCGCC TACATGTGCT CGTCGCACAG ICCGTGGGCC 120 TCAGCGACA GCCTCTCGTA CGGCTGGGCG GCCGTCCGCA TCGCCGGCCA TCCGAG 177 2) INFORMATION FOR SEQ ID NO: 60: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 59 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 60: Ser Thr Pro Val Gin Thr Cys Asp Arg Asn Asp Asn Pro Leu Tyr Asp' 15 10 15 Gly Gly Ser Thr Arg Ser Gly Cys Asp Ala Gly Gly Gly Ala Tyr Met 20 25 30 Cys Ser Ser His Ser Pro Trp Ala Val Ser Asp Ser Leu Ser Tyr Gly 35 40 45 Trp Ala Ala Val Arg lie Ala Gly Gin Ser Glu 50 55 (2) INFORMATION FOR SEQ ID NO: 61: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 150 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Thielavia thermophila, CBS 174.70 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 61: AACGACAACC CCATCTCCAA CACCAACGCT GTCAACGGTT GTGAGGGTGG TGGTTCTGCT 60 TACGCTTGCT CCAACTACTC TCCCTGGGCT GTCAACGATG ACCTTGCCTA CGGTTTCGCT 12 0 GTTACCAAGA TCTCCGGTGG CTCCGAGGCC 150 (2) INFORMATION FOR SEQ ID NO: 62: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 50 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 62: Asn Asp Asn Pro lie Ser Asn Thr Asn Ala Val Asn Gly Cys Glu Gly 15 10 15 Gly Gly Ser Ala Tyr Ala Cys Ser Asn Tyr Ser Pro Trp Ala Val Asn 20 25 30 Asp Asp Leu Ala Tyr Gly Phe Ala Val Thr Lys lie Ser Gly Gly Ser 35 40 45 Glu Ala 50 (2) INFORMATION FOR SEQ ID NO: 63: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 180 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (A) ORGANISM: Chaetomium cuniculoruin, CBS 799-83 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 63: GTCAATCAGC CCATCCGAAC GTGTAGTGCC AACGACTCGC CCTTGTCCGA CCCAAATACC 60 CCAAGTGGCT GTGACGGTGG TAGCGCCTTC ACTTGTTCCA ACAACTCCCC GTGGGCAGTC 120 GATGACCAGA CAGCTTATGG CTTTGCGGCA ACAGCCATCA GTGGCCAGTC CGAGAGCAGC 180 (2) INFORMATION FOR SEQ ID NO: 64: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 60 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 64: Val Asn Gin Pro lie Arg Thr Cys Ser Ala Asn Asp Ser Pro Leu Ser 15 10 15 Asp Pro Asn Thr Pro Ser Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys 20 25 30 Ser Asn Asn Ser Pro Trp Ala Val Asp Asp Gin Thr Ala Tyr Gly Phe 35 40 45 Ala Ala Thr Ala lie Ser Gly Gin Ser Glu Ser Ser 50 55 60 (2) INFORMATION FOR SEQ ID NO: 65: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 159 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (iv) ORIGINAL SOURCE: Chaetomium virescens, CBS 547.75 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 65: ACCTGCGACA AGAAGGACAA CCCCATCTCT GATGCCAACG CCAAGAGCGG CTGTGATGGC 60 5 GGTTCTGCTT TCGCCTGCAC CAACTACTCT CCCTTCGCCG TCAACGACAA CCTCGCCTAC 120 GGTTTCGCTG CCACCAAGCT TGCTGGAGGC TCCGAGGCT I 159 (2) INFORMATION FOR SEQ ID NO: 66: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 53 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 66: Thr Cys Asp Lys Lys Asp Asn Pro lie Ser Asp Ala Asn Ala Lys Ser 15 10 15 Gly Cys Asp Gly Gly Ser Ala Phe Ala Cys Thr Asn Tyr Ser Pro Phe 20 25 30 Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Lys Leu Ala 35 40 45 Gly Gly Ser Glu Ala 50 (2) INFORMATION FOR SEQ ID NO: 67: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 81 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Colletotrichum lagenarium (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 67: ACCTGCTACG CCAATGACCA GCGCATCGCC GACCGCAGCA CCAAGTCCGG CTGCGACGGC 60 GGCTCGGCCT ACTCCTGTTC T 81 (2) INFORMATION FOR SEQ ID NO: 68: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 27 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 68: Thr Cys Tyr Ala Asn Asp Gin Arg lie Ala Asp Arg Ser Thr Lys Ser 15 10 15 Gly Cys Asp Gly Gly Ser Ala Tyr Ser Cys Ser 20 25 (2) INFORMATION FOR SEQ ID NO: 69: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 160 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Phycomyces nitens (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 69: ACCTGTGACA AGAAGGACAA CCCCATCTCA AACTTGAACG CTGTCAACGG TTGTGAGGGT 60 GGTGGTTCTG CCTTCGCCTG CACCAACTAC TCTCCTTGGG CGGTCAATGA CAACCTTGCC 12 0 TACGGCTTCG CTGCAACCAA GCTTGCCGGT GGCTCCGAGG 160 (2) INFORMATION FOR SEQ ID NO: 70: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 53 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 70: Thr Cys Asp Lys Lys Asp Asn Pro lie Ser Asn Leu Asn Ala Val Asn 15 10 15 Gly Cys Glu Gly Gly Gly Ser Ala Phe Ala Cys Thr Asn Tyr Ser Pro 20 25 30 Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Lys Leu 35 40 45 Ala Gly Gly Ser Glu 50 (2) INFORMATION FOR SEQ ID NO: 71: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 165 base pairs (B) TYPE: nucleic acid 5372 (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (iv) ORIGINAL SOURCE: Trichothecium roseum, IFO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 71: CCAGTAGGCA CCTGCGACGC CGGCAACAGC CCCCTCGGCG ACCCCCTGGC CAAGTCTGGC 60 TGCGAGGGCG GCCCGTCGTA CACGTGCGCC AACTACCAGC CGTGGGCGGT CAACGACCAG 120 CTGGCCTACG GCTTCGCGGC CACGGCCATC AACGGCGGCA CCGAG 165 (2) INFORMATION FOR SEQ ID NO: 72: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 55 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 72: Pro Val Gly Thr Cys Asp Ala Gly Asn Ser Pro Leu Gly Asp Pro Leu 1 5 10 15 Ala Lys Ser Gly Cys Glu Gly Gly Pro Ser Tyr Thr Cys Ala Asn Tyr 20 25 30 Gin Pro Trp Ala Val Asn Asp Gin Leu Ala Tyr Gly Phe Ala Ala Thr 35 40 45 Ala lie Asn Gly Gly Thr Glu 50 55 REFERENCES Background of the invention: GB-A-1368599 EP-A-0 307 564 EP-A-0 435 876 WO 91/17243 WO 91/10732 WO 91/17244 WO 95/24471 WO 95/26398 Methods in Enzymology, 1988, Vol. 160, p. 200-391 (edited by Wood, W.A. and Kellogg, S.T.). Beguin, P., "Molecular Biology of Cellulose Degradation", Annu. Rev. Microbiol. (1990), Vol. 44, pp. 219-248. Henrissat, B., "Cellulases and their interaction with cellulose". Cellulose (1994), Vol. 1, pp. 169-196. T.-M. Enveri, "Microbial Cellulases" in W.M. Fogarty, Microbial Enzymes and Biotechnology, Applied Science Publishers, 183-224 (1983). Beguin, P. and Aubert, J-P., "The biological degradation of cellulose", FEMS Microbiology Reviews IJ. (1994) 25-58. Sheppard, P.O., et al., "The use of conserved cellulase family-specific sequences to clone Cellulase horoologue cDNAs from Fusarium oxysporum, Gene, (1994), Vol. 15, pp. 163-167. Saloheimo, A., et al., "A novel, small endoglucnaase gene, eglS, from Trichoderma reesei isolated by expression in yeast", Molecular Microbiology (1994), Vol. 13(2), pp. 219-228. van Arsdell, J.N. et al, (1987) Cloning, characterization, and expression in Saccharomyces cerevisiae of endoglucanase I from Trichoderma reesei, Bio/Technology 5: 60-64. Penttila, M. et al, (1986) Homology between cellulase gnees of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene. Gene 45:253-263. Saloheimo, M. et al, (1988) EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme. Gene 63:11-21. Gonzales, R., et al., "Cloning, sequence analysis and yeast expression of the egll gene from Trichoderma longibrachiatum", Appl. Microbiol. Biotechnol. (1992), Vol. 38, pp. 370-375. Ooi, T. et al. "Cloning and sequence analysis of a cDNA for cellulase (FI-CMCase) from Aspergillus aculeatus" Curr. Genet. (1990), Vol. 18, pp. 217-222. Ooi, T. et al, "Complete nucleotide sequence of a gene coding for Aspergillus aculeatus cellulase (FI-CMCase)" Nucleic Acids Research (1990), Vol. 18, No. 19, p. 5884. Xue, G. et al., "Cloning and expression of multiple cellulase cDNAs fromthe anaerobic rumen fungus Neocallimastix patriciarum in E. coli, J. Gen. Microbiol. (1992), Vol. 138, pp. 1413-1420. Xue, G. et al., "A novel polysaccharide hydrolase cDNA {celD) from Neocallimastix patriciarum encoding three multi-functional catalytical domains with high endoglucanase, cellobiohydrolase and xylanase activities", J. Gen. Microbiol. (1992), Vol. 138, pp. 2397-2403. Zhou, L. et al., "Intronless celB from the anaerobic fungus Neocallimastix patriciarum encodes a modular family A endoglucanase", Biochem. J. (1994), Vol. 297, pp. 359-364. Dalboge, H. and Heldt-Hansen, H.P., "A novel method for efficient expression cloning of fungal enzyme genes", Mol. Gen. Genet. (1994), Vol. 243, pp. 253-250. Ali, B.R.S. et al., "Cellulases and hemicellulases of the anaerobic fungus Pirojnyces constitute a multiprotein cellulose-binding complex and are encoded by multigene families", FEMS Microbiol. Lett. (1995), Vol. 125, No. 1, pp. 15-21. DNA Data Bank of Japan (DDBJ). Wang, H.Y. and Jones, R.W.: "Cloning, characterization and functional expression of an endoglucanase-encoding gene from the phytopathogenic fungus Macrophomina phaseplina", Gene, 158:125-128, 1995. Wang, H.Y. and Jones, R.W.: "A unique endoglucanase-encoding gene cloned from the phytopathogenic fungus Macrophomina phasedina", Appl. and Environm. Microbiology, 61:2004-2006, 1995. B. Henrissat: Biochem. J., 280:309-316, 1991. Schauwecker, F., Wanner, G., Kahmann, R.: "Filament-specific expression of a cellulase gene in the dimorphic fungus Ustilago maydis", 1995, Biological Chemistry Hoppe-Seyler, 376:617-625. WO 93/20193 WO 94/21801 WO 94/26880 WO 95/02043 The Drawings: Feng and Doolittle, 1987, J. Mol. Evol. 25: 351-3 60. NIH Data Base (Entrez, version spring 1996) available on World Wide Web: (htt- p://www3.ncbi.nlm.nih.gov/htbin/ef/entrezTAX). Eriksson, O.E. & Hawksworth, D.L.: Systema Ascomycetum vol 12 (1993). Jiilich, W. : Higher Taxa of Basidiomycetes, Bibliotheca Mycologia 85, 485pp (1981). O'Donnell, K.: Zygomycetes in culture, University of Georgia, US, 257pp (1979). Hawksworth, D.L., Kirk, P.M., Sutton, B.C. and Pegler, D.N.: Dictionary of the fungi, International Mycological Institute, 616pp (1995); Von Arx, J.A.: The genera of fungi sporulating in culture, 424pp (1981). Detailed Description: Ford, et al., Protein Expression and Purification 2: 95-107, 1991. Cunningham and Wells, Science 244. 1081-1085, 1989. de Vos et al.. Science 255: 306-312, 1992. Smith et al., J. Mol. Biol. 224: 899-904, 1992. Wlodaver et al., FEES Lett. 309: 59-64, 1992. Tomme, P. et al. "Cellulose-Binding Domains: Classi¬fication and Properties" in "Enzymatic Degradation of Insoluble Carbohydrates", John N. Saddler and Michael H. Penner (Eds.), ACS Symposium Series, No. 618, 1996. WO 90/00609 WO 95/16782 Needleman, S.B. and Wunsch, CD., Journal of Molecular Biology, 41- 443-453, 1970. WO 94/14953 Sambrook, J., Fritsch, E. F. & Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Lab., Cold Spring Harbor, NY. > Beaucage and Caruthers, Tetrahedron Letters 22 (1981), 1859 - 1869. Matthes et al., EMBO Journal 3. (1984), 801 - 805. US 4,683,202 Saiki et al.. Science 239 (1988), 487 - 491. Hitzeman et al., J. Biol. Chero. 255 (1980), 12073 -12080. Alber and Kawasaki, J. Mol. Appl. Gen. 2, (1982), 419 -434) . Young et al., in Genetic Engineering of Microorganisms for Chemicals (Hollaender et al, eds.). Plenum Press, New York, 1982). US 4,599,311 Russell et al., -Nature 304 (1983), 652 - 654. McKnight et al., The EMBO J. 4 (1985), 2093 - 2099 P.R. Russell, Gene 40, 1985, pp. 125-130. US 4,870,008 O. Hagenbuchle et al., Nature 289. 1981, pp. 643-646. L.A. Vails et al., Cell 48., 1987, pp. 887-897. WO 87/02670 M. Egel-Mitani et al., Yeast 6. 1990, pp. 127-137. US 4,546,082 EP 16 201 EP 123 294 EP 123 544 EP 163 529 WO 89/02463 WO 92/11378 US 4,599,311 US 4,931,373 US 4,870,008 US 5,037,743 US 4,845,075 US 4,931,373 Gleeson et al., J. Gen. Microbiol. 132. 1986, pp. 3459-3465. US 4,882,279 EP 272 277 EP 230 023 Malardier et al., 1989, Gene 78: 147-156. WO 93/11249. WO 94/14953. wo 95/02043. Horton, R. M., Hunt, H. D., Ho, S. N., Pullen, J. K., and Pease, L. R. (1989) Gene 77, 61-68 i Dalbcsge, H., and Heldt-Hansen, H. (1994) Mol. Gen. Genet. 243, 253-260 Christensen, T., Woldike, H., Boel, E., Mortensen, S. B., ) Hjortshcj, K., Thim, L., and Hansen, M. T. (1988) Bio/Technology 6, 1419-1422 Sanger, F., Nicklen, S., and Coulson, A. R. (1977) Proc. Natl. Acad. Sci. D. S. A. 74, 5463-5467 Devereux, J., Haeberli, P., and Smithies, O. (1984) Nucleic Acids Res. 12, 387-395 Becker, D. M. & Guarante, L. 1991. Methods Enzymol. 194: ) 182-187. Gubler, U. & Hoffman, B. J. 1983. Gene 25: 263-269. R. Higuchi, B. Krummel, and R.K. Saiki (1988). A general > method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucl. Acids Res. 16: 7351-7367. Sanger, F., Nicklen, S. & Coulson, A. R. 1977. Proc. ) Natl. Acad. Sci. U. S. A. 74: 5463-5467. N. Axelsen et al., A Manual of Quantitative Immunoelectrophoresis. Blackwell Scientific Publications, 1973, Chapters 2,3,4 and 23. WE CLAIM; 1. An enzyme preparation exhibiting endoglucanase activity, wherein the enzyme is a) encoded by a DNA construct comprising the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, or b) encoded by a DNA construct comprising an analogue of the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, which DNA sequence has at least 75% identity with the DNA sequence shown in SEQ ID No. 8 or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSN 10081, or c) a polypeptide which has at least 70% identity with the amino acid sequence shown _ in SEQ ID No. 9. 2. The enzyme preparation according to claim 1, wherein it said enzyme. 3. The enzyme preparation according to claim 1 or 2, wherein it additionally comprises one or more enzymes selected from the group consisting of galactanases, xylanases, arabinanases, pectin acetyl esterases, polygalacturonases, rhamnogalacturonases, pectin lyases, pectate lyases, endogucanases, pectin methylesterases, proteases, lipases, amylases, cutinases, peroxidases, laccases, cellobiohydrolases and transglutaminases. 4. A detergent composition comprising the enzyme preparation according to any of the claims 1 to 3, and a compound selected from the group consisting of a surfactant, a builder compound, and a fabric softening agent. 5. The composition according to claim 4, wherein the surfactant is a nonionic, anionic, cationic, zwitterionic, ampholytic or amphoteric surfactant. -270- 6. The composition according to claim 4, wherein the fabric softening agent is a cationic or nonionic softening agent, preferably a quaternary ammonium compound, and which optionally further comprises one or more compounds selected from a surfactant, an electrolyte, a buffer, an antioxidant and a liquid carrier. 7. A method of providing colour clarification of laundry, wherein said method comprises treating the laundry with a soaking, washing or rinsing liquor comprising an enzyme preparation according to any of claims 1 to 3 or with a detergent composition according to any one of the claims 4 to 6. 8. The method according to claim 7, wherein the laundry is treated in a washing machine. 9. The method according to claim 7 or 8, wherein the endoglucanase is present in the soaking, washing, or rinsing liquor in an effective amount of between 1 and 1000 S— CEVU, preferably between 5 and 200 S-CEVU, per liter of liquor during machine cycle use conditions. 10. The method according to any of the claims 7 to 9, wherein the pH of the soaking, washing, or rinsing liquor is between 4 and 11, preferably between 6 and 10.5. 11. The method according to any of the claims 7 to 10, wherein the temperature is between 15°C and 60°C. 12. A method for biopolishing of a cellulosic fabric which method comprises treating the yam surface of said fabric with the enzyme preparation according to any of claims lto3. -271- 13. The method according to claim 12, wherein it takes place in the wet processing of the manufacture of knitted and woven fabrics. 14. A method for "stone-washing" which method comprises treating a dyed cellulosic fabric with the enzyme preparation according to any of claims 1 to 3. 15. An enzyme preparation exhibiting endoglucanase activity substantially as herein described and exemplified. 16. A detergent composition comprising the enzyme preparation substantially as herein described and exemplified. 17. A method of providing colour clarification substantially as herein described and exemplified. 18. A method for biopolishing of a cellulosic fabric substantially as herein described and exemplified. 19. A method for "stone-washing" substantially as herein described and exemplified.

Full Text

The present invention relates to novel enzyme preparations cotnprising an enzyme exhibiting endoglucanase activity which performs very good in indus¬trial applications such as laundry compositions, for biopolishing of newly manufactured textiles, for provid¬ing an abraded look of cellulosic fabric or garment, and for treatment of paper pulp. Further, the invention relates to DNA constructs encoding such enzymes, a method for providing a gene encoding for such enzymes, a method of producing the enzyme preparations containing such enzymes, and the use of these enzymes for a number of industrial applications.
BACKGROUND OF THE INVENTION
Cellulases or cellulytic enzymes are enzymes involved in hydrolyses of cellulose. In the hydrolysis of native cel¬lulose, it is known that there are three major types of cellulase enzymes involved, namely cellobiohydrol'ase (l,4-j3-D-glucan cellobiohydrolase, EC 3.2.1.91), endo-jS-1,4-glucanase (endo-1,4-/3-D-glucan 4-glucanohydrolase, EC 3.2.1.4) and i3-glucosidase (EC 3.2.1.21).
Cellulases are synthesized by a large number of microor¬ganisms which include fungi, actinomycetes, myxobacteria and true bacteria but also by plants. Especially endoglucanases of a wide variety of specificities have been identified.
'A very important industrial use of cellulytic enzymes is the use for treatment of cellulosic textile or fabric, e.g. as ingredients in detergent compositions or fabric

softener compositions, for bio-polishing of new fabric (garment finishing), and for obtaining a "stone-washed" look of cellulose-containing fabric, especially denim, and several methods for such treatment have been sug¬gested, e.g. in GB-A-1 368 599, EP-A-0 307 564 and EP-A-0 435 876, WO 91/17243, WO 91/10732, WO 91/17244, PCT/DK95/000108 and PCT/DK95/00132.
Another important industrial use of cellulytic enzymes is the use for treatment of paper pulp, e.g. for improving the drainage or for deinking of recycled paper.
Especially the endoglucanases _(EC No. 3.2.1.4) constitute an interesting group of hydrolases for the mentioned industrial uses. Endoglucanases catalyses endo hydrolysis of l,4-i3-D-glycosidic linkages in cellulose, cellulose derivatives (such as carboxy methyl cellulose and hydroxy' ethyl cellulose), lichenin, /3-l,4 bonds in mixed (3-1,3 glucans such as cereal /3-D—glucans or xyloglucans and other plant material containing cellulosic parts. The authorised name is endo-l,4-^-D-glucan 4-glucano hydrolase, but the abbreviated term endoglucanase is used in the present specification. Reference can be made to T.-M. Enveri, "Microbial Cellulases" in W.M. Fogarty, Microbial Enzymes and Biotechnology, Applied Science Pub¬lishers, p. 183-224 (1983) ; Methods in Enzymology, (1988) Vol. 160, p. 200-391 (edited by Wood, W.A. and Kellogg, S.T.); Beguin, P., "Molecular Biology of Cellulose Degra¬dation", Annu. Rev. Microbiol. (1S90), Vol. 44, pp. 219-24S; Beguin, P. and Aubert, J-P,, "The biological degra¬dation of cellulose", FEMS Microbiology Reviews H (1994) p.25-58; Henrissat, B., "Cellulases and their interaction with cellulose". Cellulose (1994), Vol. 1, pp. 169-196.
Fungal endoglucanases have been described in numerous publications, especially those derived from species as e.g. Fusarium oxyspoxrum, Trlchoderma reesei, Trichoderma

longibrachiatum, Aspergillus aculeatus, Neocallimastix patriciarum, and e.g. from species of the genera Piromyces, Humicola, Myceliophthora, Geotricum, Penicillium, Irpex, Coprinus.
For example, fungal endqgluc_anase_s; have been described by Sheppard, P.O., et al., "The use of conserved cellulase family-specific sequences to clone Cellulase homologue cDNAs from Fusarium oxysporum, Gene, (1994), Vol. 15, pp. 163-167, Saloheimo, A., et al., "A novel, small endoglucanase gene, eglSj from Trichoderma reesei iso¬lated by expression in yeast". Molecular Microbiology (1994), Vol. 13(2), pp. 219-228; van Arsdell, J.N. et al, (1937), Cloning, characterization, and expression in Saccharomyces cerevisiae of endoglucanase I from Trichoderma reesei, Bio/Technology 5: 60-64; Penttila, M. et al., (1986), "Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene". Gene 45:253-263; Saloheimo, M. et al, (1988) , •VEGIII, a new endoglucanase from Trichoderma reeseiz the chaxacterization of both gene and enzyme". Gene 63:11-21; Gonzales, R., et al., "Cloning, sequence analysis and yeast expression of the egll gene fr-ccm Tricho'derma longibrachiatum", Appl. Microbiol. Biotechnol., (1992), Vol. 38, pp. 370-375; Ooi, T. et al. "Cloning and sequence analysis of a cDNA for cellulase (FI-CMCase) from Aspergillus aculeatus", Curr. Genet., (1990), Vol. 18, pp. 217-222; Ooi, T. et al, "Complete nucleotide sequence of a gene coding for Aspergillus aculeatus cellulase (FI-CMCase)", Nucleic Acids Research, (1990), Vol. 18, No. 19, p. 5884; Xue, G. et al., "Clon¬ing and expression of multiple cellulase cDNAs from the anaerobic rumen fungus Neocallimastix patriciairum in E. coli", J. Gen. Microbiol., (1992), Vol. 138, pp. 1413-
1420; -Xue, C. et al., "A novel polysaccharide hydrolase cDNA (celD) from Neocallimastix patriciarum encoding three multi-functional catalytical domains with high

endoglucanase, cellobiohydrolase and xylanase activ¬ities", J. Gen. Microbiol., (1992), Vol. 138, pp. 2397-2403; Zhou, L. et al., "Intronless celB from the anaerobic fungus Neocallimastix patriciarum encodes a modular family A endoglucanase", Biochem. J., (1994), Vol. 297, pp. 359-364; Dalboge, H. and Heldt-Hansen, H.P., "A novel method for efficient expression cloning of fungal enzyme genes", Mol. Gen. Genet., (1994), Vol. 243, pp. 253-260; Ali, B.R.S. et al., "Cellulases and hemicellulases of the anaerobic fungus Piromyces consti¬tute a multiprotein cellulose-binding complex and are encoded by multigene families", FEMS Microbiol. Lett., (1995), Vol. 125, No. 1, pp. 15-21. Further, the DNA Data Bank of Japan (DDBJ database publicly available at Internet) comprises two DNA sequences cloned from Penicillium janthinellum encoding endoglucanases (cloned by A. Koch and G. Mernitz, respectively) and a DNA sequence cloned from Humicola qrisea var. thermoidea encoding an endoglucanase (cloned by T. Uozumi). Two endoglucanases from Macrophomina phaseolina have been cloned and sequenced, see Wang, H.Y- and Jones, R.W.: "Cloning, characterization and functional expression of an endoglucanase-encoding gene from the phytopathogenic fungus Wacropho/nina phaseolina" in Gene, 158:125-128, 1995, and Wang, H.Y. and Jones, R.W.: "A unique endoglucanase-encoding gene cloned from the phytopathogenic fungus Macrophomina phaseolina" in Applied And Environmental Microbiology, 61:2004-2006, 199 5. One of these endoglucanases shows high homology to the egl3 endoglucanase from the fungus Trichoderma reesei, the other shows homology to the egll from the microbial phytopathogen Pseudomonas solanacearum indicat¬ing that both endoglucanases belong to family 5 of glycosyl hydrolases (B. Henrissat, Biochem J 280:309-316 (ig^l))"^ Filament-specific expression of a cellulase gene-in the dimorphic fungus Ustilago maydis is disclosed in Schauwecker, F. et al. (1995).

wo 91/17243 (Novo Nordisk A/S) discloses a cellulase pre¬paration consisting of a homogenous endoglucanase compo¬nent immunoreactive with an antibody raised against a highly purified 43 kDa endoglucanase derived from Humicola insolens, DSM 1800; WO 91/17244 (Novo Nordisk A/S) discloses a new (hemi)cellulose degrading enzyme, such as an endoglucanase, a cellobiohydrolase or a /3-glucosidase, which may be derived from fungi other than Trichoderma and Phanerochaete; WO 93/20193 discloses an endoglucanase derivable from Aspergillus aculeatus; WO 94/21801 (Genencor Inc.) concerns a cellulase system iso¬lated from Trichoderma longibrachiatum exhibiting endoglucanase activity; WO 94/26880 (Gist Brocades N.V.) discloses an isolated mix±ure of cellulose degrading enzymes, which preferable are obtained from Trichoderma, Aspergillus or Disporotrichum, comprising endoglucanase, cellobiohydrolase, and xyloglucanase activity; and WO 95/02043 (Novo Nordisk A/S) describes an enzyme wirh endoglucanase activity derived from Trichoderma harzianum, which can be used for a number of purposes including e.g. degradation or modification of plant cell walls.
It is also known that cellulases may or may not have a cellulose binding domain (a C^BD) . The CBD enhances the binding of the enzyme to a cellulose-containing fiber and increases the efficacy of the catalytic active part of the enzyme.
There is an ever existing need for providing novel cellulase enzyme preparations which may be used for applications where cellulase, preferably an endoglucanase, activity is desirable.
The object of the present invention is to provide novel enzyme preparations Jiaving substantial cellulytic activ¬ity at acid, neutral or alkaline conditions and improved

performance in paper pulp processing, textile treatment, laundry processes or in animal feed; preferably novel cellulases, more preferably well-performing endoglucana-ses, which are contemplated to be producible or produced
by recombinant techniques.
SUMMARY OF THE INVENTION
Surprisingly, it has been found that a group of endoglucanases having certain unique characteristics per¬form very good in those industrial applications for which endoglucanases are conventionally used. These unique characteristics can be described in terms of conserved regions of the amino acid sequence of the enzyme protein and the inventors have found that cellulytic enzymes, i.e. enzymes exhibiting cellulytic activity, having cer¬tain conserved regions arTe very effective e.g. in the treatment of laundry, in the treatment of newly manufac¬tured textile, in the treatment of papermaking pulp.
Accordingly, in its first aspect the preseait invention relates to an enzyme preparation consisting essentially of an enzyme having cellulytic activity and comprising a first amino acid sequence consisting of 14 amino acid residues having the following sequence
Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp Xaa 1 2 3 4 5 6 7 8 9 10 11 12 13 14
and a second amino acid sequence consisting of 5 amino acid residues having the following sequence
Trp Cys Cys Xaa Cys 1 2 ■ 3 4 "3
wherein.

in position 3 of the first sequence, the amino acid is
Trp, Tyr or Phe;
in position 4 of the first sequence, the amino acid is
Trp, Tyr or Phe;
in position 8 of the first sequence, the amino acid is
Arg, Lys or His;
in position 9, 10, 12 and 14, respectively, of the first
sequence, and in position 4 of the second sequence, the
amino acid is any of the 20 naturally occurring amino
acid residues with the provisos that, in the first amino
acid sequence, (i) when the amino residue in position 12
is Ser, then the amino acid residue in position 14 is not
Ser, and (ii) when the amino residue in position 12 is
Gly, then the amino acid residue in position 14 is not
Ala-.
This surprising finding of clearly recognisable conserved regions, in spite of rather prominent variations found within well-performing endoglucanase enzymes, is a result of studies of a number of fungal DNA sequences encoding for specific a^nino acid sequences of enzymes having sig¬nificant cellulytlc, especially endoglucauiase, activ¬ities.
Based on this finding, a novel molecular method taylored to screen specifically for genomic DNA or cDNA characterised by encoding the enzymes of the invention has been developed. As tools for this three sets of degenerated primers were constructed. Accordingly, in its second aspect, the invention relates to a method for providing a gene encoding for cellulytic enzymes having the above conserved regions.
By using this method, i.e. the set of primers for a PCR screening on genomic DNA, it was surprisingly found that DNA encoding for said enzymes can be found from a broad range of fungi, belonging to taxonomically very different

organisms and inhabiting ecologically very different niches.
Further, by using this method it has been possible to find DNA sequences encoding for the core regions (cata-lytically active regions or domains) of said enzymes without any attached cellulose binding domain (CBD) which core regions of enzymes would not have been selected by using conventional performance based screening approaches. The inventors have verified experimentally that the linking of a CBD region to a core region enzyme (comprising the catalytically active region or domain of the enzyme) of the present invention results in a sig¬nificantly improved performance, e.g. a fifty times higher performance, of the multiple domain enzyme.
Accordingly, the present invention provides novel cellulases, especially endoglucanases, having improved performance in industial applications, either in their native form, or homo- or heterologously produced.
Ln further aspects, the present invention relates to novel cellulytic enzyme preparations which are derivable from tax'onomically specific phyli, classes, orders, families, genera, and species; e.g. from Basidiomycotous Hymenomycetes, Zygomycota, Chytridiomycota; or from the classes Discomycstes, Loculoascomycetes, Plectomycetes; Archaeascomycetes, Hernias cornycetes or from the orders Di-aportales, Xylariales, Trichosphaeriales, Phyllachorales; or from the families Nectriaeae, Sordariaceae, Chaetomiaceae, Ceratostomaceae, Lasiosphaeriaceae; or from the genera Cylindrocarpon, Gliocladium, Volutella, Scytalidium, Acremonium, ot from the species Fusarium lycopersici, Fusarium passiflora, Fusarium solani, Fusarium anquioides, Fusarium poae, Humicola nigrescens', Humicola grisea, especially such consisting essentially of an enzyme comprising an amino acid sequence selected

from the group consisting of the sequences Xaa Thr Arg Xaa Phe Asp Xaa 12 3 4 5 6 7; Xaa Thr Arg Xaa Tyr Asp Xaa 12 3 4 5 6 7; and
Xaa Thr Arg Xaa Trp Asp Xaa
12 3 4 5 6 7
wherein, in position 4, Xaa is Trp, Tyr or Phe; and
in position 1 and 7, Xaa is any of the 20 naturally
occurring ainino acid residues.
More specifically, the enzyme preparation of the
invention is preferably obtainable from the taxonoraically
specific phyli, classes, orders, families, genera, and"
species mentioned above which all produce endoglucanases
comprising a first peptide consisting of 13 amino acid
residues having trhe following sequence
Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp
1 2 3 4 5 6 7 8 9 10 11 12 13
and a second peptide consisting of 5 amino acid residues
having the fo11 awing sequence
Trp Cys Cys Xaa Cys
12 3 4 5
wherein, in position 3 of the first sequence, the amino
acid is Trp, Tyr or Phe; in position 4 of the first
sequence, the amino acid is Trp, Tyr or Phe; in position
S of the first sequence, the amino acid is Arg, Lys or
His; in position 9, 10, and 12, respectively, of the
first sequence, and in position 4 of the second sequence,
the amino acid is any of the 20 naturally occurring amino
acid residues.
In yet further aspects, the present invention provides DNA constructs comprising a DNA sequence encoding an en¬zyme exhibiting endoglucanase activity, which DNA sequence comprises the DNA sequence shown in SEQ ID Nos.

1, 4, 6, 8, 10, 12, 16, and 19, respectively, or analogues thereof.
The present invention also relates to a recombinant expression vector comprising a DNA construct of the invention; to a cell comprising a DNA construct or a recombinant expression vector of the invention; to a method of producing an enzyme, e.g a recombinant enzyme, of the invention; to a method of providing colour clari¬fication of laundry by using the enzyme of the invention; to a laundry composition comprising the enzyme of the invention; to uses of the enzyme of the invention for degradation or modification of plant material, e.g. cell walls, for treatment of fabric, textile or garment, for treatment of paper pulp; and to an enzyme preparation which is enriched in an enz^Tiie of the present invention.
THE DRAVTENGS
Figure l is an alignment, of the deduced encoded amino acid sequences of Arrrp-anniixa sp. (I), Volutella colletotrichoides, Cx'inipellis scabella, Acremonium sp. (II), Myceliophthora thermophila, Thielavia terrestris, Macrophomina phaseolina. The Pileup program (Feng and Doolittle, 1987) {GCG package, version 8.0) was used to create the best alignment. Identical residues in at least four sequences (boxed) are indicated around the corre¬sponding amino acids.
Figure 2
Figure 2a,b,c illustrates the taxonomic classification within the Fungal Kingdom of all the microorganisms dis¬closed herein as being capable of producing said enzyme preparations and- enaymes of the invention.
The taxonomic classification used herein builds primarily

on the system used in the :NIH Data Base (Entrez, version
spring 1996) available on World Wide Web:
(http: //www3 .ncbi.nliti.nih.gov/htbin/ef/entrezTAX) .
Regarding classification of organisms which are not
included in the Entrez data base the following generally
available and world wide accepted reference books have
been used:
For Ascomycetes: Eriksson, O.E. & Hawksworth, D,L.:
Systema Ascomycetum vol 12 (1993).
For Basidiomycetes: Jiilich, W. : Higher Taxa of
Basidiomycetes, Bibliotheca Mycologia 85, 485pp (1981).
For Zygomycetes: O'Donnell, K.: Zygomycetes in culture,
University of Georgia, US, 257pp (1979) .
General mycological reference books:
Hawksworth, D.L., Kirk, P.M., Sutton, B.C. and Pegler,
D.N.: Dictionary of the fungi. International Mycological
Institute, 616pp (1995);
Von Arx, J.A. : The genera of fungi spomlccting in
culture, 424pp (1981).
The taxonomic implacement of the genus Humicola has untill recently remained unclear- However, studies of 18SRNA of a wide selection of Sordariales has given strong indications of referring Humicola to the order Sordariales (Taylor, Clausen & Oxenboll, unpublished). Further these data suggests Humicola along with Scytalid-ium to be only rather distantly related to the families Sordariaceae, Chaetomiaceae, Ceratostomataceae, and Lasiosphaeriaceae. In accordance with the above Humicola and Scytalidium are here placed within the order Sordariales, with unclassified Family.
Figure 3 is an alignment of the deduced partial amino acid sequences derived from a selection of 26 of the 46 microorganisms described in Example—5T—'

DETAILED DESCRIPTION OF THE INVENTION
In the present context, the term "the 20 naturally occuring amino acid residues" denotes the 20 amino acid residues usually found in proteins and conventionally known as alanine (Ala or A), valine (Val or V), leucine (Leu or L), isoleucine (lie or I), proline (Pro or P), phenylalanine (Phe or F), tryptophan (Trp or W), methionine (Met or M), glycine (Gly or G), serine (Ser or S), threonine (Thr or T), cysteine (Cys or C), tyrosine (Tyr or Y), asparagine (Asn or N), glutamine (Gin or Q), aspartic acid (Asp or D), glutamic acid (Glu or E), lysine (Lys or K), arginine (Arg or R), and histidine (His or H).
According to the present invention there is provided novel well-performing endoglucanases comprising consei-ved amino acid sequence regions, especially a first amino acid sequence consisting of 14 amino acid residues having the following sequence
Thr Arg Xaa Xaa Asp Cys Cys Xaa Xara Xaa Cys Xaa Trp Xaa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 and a second amino acid sequence consisting of 5 amino acid residues having the following sequence
Trp Cys Cys Xaa Cys
12 3 4 5
wherein,
in position 3 of the first sequence, the amino acid is
Trp, Tyr or Phe;
in position 4 of the first sequence, the amino acid is
Trp, Tyr or Phe;
m position 8 of the first sequence, the ammo acid is
Arg, Lys or His;
in position 9, 10, 12 and 14, respectively, of the first

sequence, and in position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue in position 12 is Ser, then the amino acid residue in position 14 is not Ser, and (ii) when the amino residue in position 12 is Gly, then the amino acid residue in position 14 is not Ala.
Preferably, the enzyme of the invention is of microbial origin, i.e. obtainable from a microorganism such as a fungus.
In a preferred embodiment, the amino acid residue in position 9 of the first sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cysteine, asparaglne, glutamine, tyrosine, serine, methionine and tryptophan, preferably from the group consisting of proline and threonine.
In another preferred embodiment, the amino acid r^idne in position 10 of the first sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cysteine, asparagine, glutamine, tyrosine, serine, methionine and tryptophan, preferably serine.
In yet another preferred embodiment, the amino acid resi¬
due in position 12 of the first sequence is selected from
the group consisting of proline, threonine, valine,
alanine, leucine, isoleucine, phenylalanine, glycine, cy¬
steine, asparagine, glutamine, tyrosine, serine,
methionine and tryptophan, preferably from the group con¬
sisting of alanine and glycine. '
In yet another preferred embodiment, the amino acid resi-

due in position 14 of the first sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cy¬steine, asparagine, glutamine, tyrosine, serine, methionine, tryptophan, glutamic acid and aspartic acid, preferably from the group consisting of proline, threonine, serine, alanine, glutamic acid and aspartic acid.
Preferably, the amino acid residue in position 4 of the second sequence is selected from the group consisting of proline, threonine, valine, alanine, leucine, isoleucine, phenylalanine, glycine, cysteine, asparagine, glutamine, tyrosine, serine, methionine, tryptophan, glutamic acid and aspartic acid, more preferably from the group con¬sisting of alanine, glycine, and glutcunine.
Examples of more preferred embodiments are such wherein, in the first sequence, the amino acid residue in position 3 is tyrosine; or the amino acid residue in position 4 is tryptophan; or the amino acid residue in position 8 is lysine.
In an especially preferred embodiment,' the enzyme of the
invention has a first sequence comprising the amino acid
sequence
Thr Arg Tyr Trp Asp Cys Cys Lys Pro Ser Cys Ala Trp
1 2 3 4 5 6 7 8 9 10 11 12 13 ,
or the amino acid sequence
Thr Arg Tyr Trp Asp Cys Cys Lys Thr Ser Cys Ala Trp
1 2 3 4 5 6 7 8 9 10 11 12 13 ,
or the amino acid sequence
Thr Arg Tyr Trp Asp Cys Cys Lys Pro Ser Cys Gly Trp
1 2 3 4 5 6 7 8 9 10 11 12 13 .
In a second aspect, the present invention provides a method for providing a microbial strain comprising a gene

encoding such an enzyme which method comprises hybridization, e.g. PCR amplification, under standard conditions with an oligonucleotide derived from any of the conserved regions, illustrated in Fig.l.
A useful oligonucleotide comprises a nucleotide sequence encoding at least a pentapeptide comprised in a peptide selected from the group consisting of a.
Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp Xaa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 the amino acid in position 3 or 4 being Trp, Tyr or Phe; the amino acid in postion 8 being Arg, Lys or His; he amino acid in position 9, 10, 12 and 14, respectively, being any of the 20 naturally occurring amino acid resi¬dues ; and b.
Trp Cys Cys Xaa Cys Tyr 12 3 4 5 6
the amino acid in position 4 being any of the 2 0 natural¬ly occurring amino acid residues ; and c.
Xaa Pro Gly Gly Gly Xaa Gly Xaa Phe 123456789 the amino acid in position 1 being Met or lie; the amino acid in position 6 and 8, respectively, being Leu, lie or Val; and d.
Gly Cys Xaa Xaa Arg Xaa Asp Trp Xaa 123456789
the amino acid in position 3 being any of the 20 natural¬ly occurring amino acid residues;
the amino acid in position 4 and 6, respectively, being Trp, Tyr or Phe; and the amino acid in position 9 being Phe or Met;
The useful oligonucleotides also comprises nucleotide

sequences complementary to the sequences mentioned.
In a preferred embodiment of the method of the invention,
the oligonucleotide corresponds to a PCR primer selected
from the PCR primers
sense:
5 ' -CCCCAAGCTTACIVCGITA^/TTGGGA^VTTG^/TTG^/TAA^/G^/CC-S '
antisense 1:
5'- CTAGTCTAGATA'^/GCAIGC'^/GCA^/GCACC -3 ' ;
antisense 2:
CTAGTCTAGAAAIA'^/G/'^ICCIA'^/^/°ICCICCICCIGG -3 ' ; and
antisense 3:
5 ' - CTAGTCTAGAIAACCA^/GTCA'^/G^/TAIC°/TCC-3 .
In a third aspect, the present invention provides an enzyme preparation which essentially consists of an enzyme having cellulytic activity and having the conserved regions found by the inventors, i.e. which com¬prises a peptide consisting of 7 amino acid residues ha¬ving the following sequence Xaa Thr Arg Xaa Phe Asp Xaa 12 3 4 5 6 7; Xaa Thr Arg Xaa Tyr Asp Xaa 1 2 3 4 ' 5 6 7 ; and Xaa Thr Arg Xaa Trp Asp Xaa 12 3 4 5 6 7
wherein, in position 4, Xaa is Trp, Tyr or Phe; and in position 1 and 7, Xaa is any of the 20 naturally occurring amino acid residues.
This enzyme is obtainable from a strain belonging to Basidiomycotous Hymenomycetes (see Fig.2 ), more preferably to the group consisting of the orders Agarica-les, Auriculariales, and Aphyllophorales, even more preferably to the group consisting of the families Exidiaceae, Tricholomataceae, Coprinaceae, Schizophyllaceae, Bjerkanderaceae and Polyporaceae,

especially to the group consisting of the genera Exidia, Crinipellis, Fames, Panaeolus, Trametes, Schizophyllum, and Spongipellis.
Specific examples are endoglucanases obtainable from a strain belonging to the group consisting of the species Exidia glandulosa, Crinipellis scabella, Fomes fomentarius, and Spongipellis sp., more specific examples being Exidia glandulosa, CBS 277.96, Crinipellis scabella, CBS 280.96, Fames fomentarius, CBS 276.96, and Spongipellis sp., CBS 2 83.96.
Exidia glandulosa was deposited at Centraalbureau voor Schimmelcultures, Oosterstraat 1, Postbus 273, NL-3740 AG Baarn, the Netherlands, on 12 March, 1996, under thfe deposition number CBS 277.96; Crinipellis scabella was deposited at Centraalbureau voor Schimmelcultures on 12 March, 19S-6, under the deposition number CBS 280.96, Fames fomentarius was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 276.96, and Spongipellis sp. was deposited at CentraaJLbareau voor Schimmelcultures on 12 Maxcfa, 1996, under the deposition number CBS 283.96; all deposited tinder the Budapest Treaty.
The enzyme preparation of the invention is also obtain¬able from a strain belonging to Chytridiomycota, preferably from a strain belonging to the class of Chytridiomycetes, more preferably belonging to the group consisting of the order Spizellomycetales , even more preferably to the family Spizellomycetaceae, especially belonging to the genus Rhizophlyctis. A specific example is a strain belonging to the species Rhizophlyctis rosea, more specifically to Rhizophlyctis rosea, CBS 282.96.
Rhizophlyctis rosea was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition

number CBS 28 2.96; under the Budapest Treaty.
The enzyme preparation of the invention is also obtain¬able from a strain belonging to Zygomycota, preferably belonging to the class Zygomycetes, more preferably to the order Mucorales, even more preferably to the group of families consisting of Mucoraceae and Thamnidiaceae, especially belonging to the group consisting of the ge¬nera Rhizomucor, Phycomyces and Chaetostylum. Specific examples are strains belonging to the genera Rhizomucor pusillus, Phycomyces nitens, and Chaetostylum fresenii more specifically to Rhizomucor pusillus, IFO 4578, and Phycomyces nitens, IFO 4814 and Chaetostylum fresenii, NRRL 2305.
Further, the enzyme preparation of the invention is also obtainable from a strain belonging to the group consist¬ing of Archaeas^cumycetes, Discomycetes, Hemiascomycetes, Loculoascomycetes, and Plectomycetes, preferably belonging to the group consisting of the orders Pezizales, Rhytismatales, Dothideales, and Eurotiales. Especially, the enzyme is obtainaJale from a strain belonging the the group consisting of the families Cucurbitariaceae, Ascobolaceae, Rhytismataceae, and Trichocomaceae, preferably belonging the the group con¬sisting of the genera Diplodia, Microsphaeropsis, Ulospora, Macrophomina, Ascoholus, Saccobolus, Penicil-lium, and Thermomyces. Specific examples are enzymes obtainable from a strain belonging the the group consist¬ing of the species Diplodia gossypina, Microsphaeropsis sp., Ulospora bilgramii, Aureobasidium sp., Macrophomlna phaseolina, Ascobolus stictoides, Saccobolus dilutellus, Peziza, Penicillium verruculosum, Penicillium chrysogenum, and Thermomyces verrucosus; more specifically Diprlodia gossypina, CBS 274.96, Ulospora bilgramii, NKBC 1444, Macrophomina phaseolina, CBS 281.96, Saccobolus dilutellus, CBS 275.96, Penicillium

verruculosum, ATCC 62396, Penicillium chrysogenum, ATCC 9480, and Therwomyces verrucosus, CBS 285.96.
Diplodia gossypina was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 274.96, Macrophomina phaseolina was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 281.96, Saccoholus dilutellus was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 275.96; Thermomyces verrucosus was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 285.96; all under the Budapest Treaty.
Yet fujrther, the enzyme is obtainable from a strain belonging to the group consisting of the orders Dia-portales, Xylariales, Tirlchosphaeriales and Phyllachorales, preferably from a strain belonging to the group consistixjg of the families Xylariaceae, Valsaceae, and PhyllacborBLCBae, more preferably belonging to the ge¬nera Diaporthe, Colletotrichum, Nigrospora, Xylaria, NodulisporuM and Poronia. Specific examples are the specie's Diaporthe syngenesia, Colletotrichum lagenarium, Xylaria hypoxylon, Nigrospora sp., Nodulisporum sp., and Poronia punctata, more specifically Diaporthe syngenesia, CBS 278.96, Colletotrichum lagenarium, ATCC 52 609, Nigrospora sp., CBS 272.96, Xylaria hypoxylon, CBS 284.96.
Diaporthe syngenesia was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 278.96, Nigrospora sp. was deposited at Centraalbureau voor Schimmelcultures on 12 March, 199 6, under^he—deposition number CBS 272.96, Xylaria hypoxylon was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 284.96;

all under the Budapest Treaty.
The enzyme is also obtainable from the unidentified fungal, mitosporic, coleomycetous deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition numbers CBS 270.96, CBS 271.96 and CBS 273.96, respectively, under the Budapest Treaty.
The enzyme is also obtainable from a strain belonging to the group consisting of the genera Cylindrocarpon, Gliocladium, Nectria, Volutella, Sordaria, Scytalidium, Thielavia, Syspastospora, Cladorrhinum, Chaetomium, Myce-liphthora and Acremonium, especially from a strain belon¬ging to the group consisting of the species Cylin¬drocarpon sp., Nectria pinea, Volutella colletotrichoides, Sordaria fimicola, Sordaria macrospora, Thielavia terrestris, Thielavia thermophila, Syspastospora boninensis, Cla^dorrhinxrm foecundissimum, Chaetomium muronrm, Chaetomium virescens, Chaetomium brasiliensis, Chaetomium cunicolojrum, Myceliophthora thermophila, Gliocladium catenulatum, Scytalidium thermophila, and Ai:ri^nir>nJmn sp., more specifically from Nectria pinea, CBS 273.96, Volutella colletotrichoides, CBS 400.58, Sordaria fimicola, ATCC 52644, Sordaria macrospora, ATCC 60255, Thielavia terrestris, NRRL 8126, Thielavia thermophila, CCBS 174.70, Chaetomium murorum, CBS 163.52, Chaetomium virescens, CBS 547.75, Chaetomium brasiliensis, CBS 122.65, Chaetomium cunicolorum, CBS 799.83, Syspastospora boninensis, NKBC 1515, Cladorrhinum foecundissimum, ATCC 62373, Myceliophthora thermophila, CBS 117.65, Scytalidiim thermophila, ATCC 28085, Gliocladium catenulatum, ATCC 10523, and Acremonium sp., CBS 478.94.
Nectria pinea was deposited at Centraalbureau voor Schimmelcultures on 12 March, 1996, under the deposition number CBS 279.96, and Acremonium sp. was deposited on

28 September 1994 under the deposition number CBS 478.94, both according to the Budapest Treaty.
The enzyme is also obtainable from a strain belonging to the group consisting of the species Fusarium solani, Fusarium anguioides, Fusarium poae, Fusarium oxysporum ssp. lycopersici, Fusarium oxysporum ssp. passiflora, Humicola nigrescens and Humicola grisea, especially Fusarium oxysporum ssp lycopersici, CBS 645.78, Fusarium oxysporum ssp passiflora, CBS 744.79, Fusarium solani, IMI 107.511, Fusarium anguioides, IFO 44 67, Fusarium poae, ATCC 60883, Humicola nigrescens, CBS 819.73 and Humicola grisea, ATCC 22726. It is to be noted that Humi¬cola grisea is different from Humicola grisea var. thermoidea.
In a preferred embodiment, the enzviae preparation of the invention is derived from the disclosed classes, orders, families, genera and species and essentially consists of an enzyme comprising a first peptide consisting of 13 amino acid residues having the following sequence
Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa Trp 1 2 3 4 5 6*7 8 9 10 11 12 13
and a second peptide consisting of 5 amino acid residues having the following sequence
Trp Cys Cys Xaa Cys 12 3 4 5
wherein, in position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; in position 8 of the first sequence-;—Lhi=i 'amino acid is Arg, Lys or His; in position 9, 10, and 12, respectively, of the first sequence, and in position 4 of the second sequence.

coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively; or
b) an analogue of the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, which
i) is homologous with the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, ii) hybridizes with the same oligonucleotide probe as the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequ¬ence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10O31, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, iii) encodes a polypeptide which is homologous with the polypeptide encoded by a DNA sequence comprising the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequ¬ence obtainable from the plasmid in Saccharomyces cerevisiae DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, iv) encodes a polypeptide which is immxmologically re¬active with an antibody raised against the purified endoglucanase encoded by the DNA sequence shown in SEQ ID No 1, 4, 6, 8, 10, 12, 16, or 19, respectively, or the DNA sequence obtainable, from the plasmid in Saccharomyces cerevisiae DSM 977 0, DSM 10082, DSM 10080, DSM 10081, Escherichia coli,

DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively.
Escherichia coli DSM 10512 was deposited under the Buda¬pest Treaty on 2 February, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Escherichia coli DSM 10511 was deposited under the Buda¬pest Treaty on 2 February, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Escherichia coli DSM 10571 was deposited under the Buda¬pest Treaty on 6 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Escherichia coli DSM 10576 was deposited under the Buda¬pest Treaty on 12 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braxonschweig, Germany) .
Escherichia coli DSM 10583 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Escherichia coli DSM 10584 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Escherichia coli DSM 10585 was deposited under the Buda-pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlufvj-von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).

Escherichia coli DSM 10586 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Escherichia coli DSM 10587 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Escherichia coli DSM 10588 was deposited under the Buda¬pest Treaty on 13 March, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Saccrharomyces cerevisiae DSM 9770 was deposited under the Budapest Treaty on 24 February, 1995, at DSM (Deutsche ScTncralung von Mikrrrorg^nismeTi und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
SacL.hc±iomyces cerevisiae DSM 100B2 was deposited under the. Budapest Treaty on 3 0 June, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Saccharomyces cerevisiae DSM 10080 was deposited under the Budapest Treaty on 30 June, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
Saccharomyces cerevisiae DSM 10081 was deposited under the Budapest Treaty on 30 June, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany).
The DNA construct of■the invention relating to SEQ ID No. 1 can be isolated from or produced on the basis of a DNA

library of a strain of Myceliophthora, in particular a strain of M. thermophila, especially M. thermophila, CBS 117.65.
The DNA constructs of the invention relating to SEQ ID Nos. 4 and 6 can be isolated from or produced on the basis of a DNA library of a strain of Acremonium, especially Acremonium sp., CBS 478,94.
The DNA construct of the invention relating to SEQ ID No. 8 can be isolated from or produced on the basis of a DNA library of a strain of Thielavia in particular a strain of Thielavia terrestris, especially Thielavia terrestris, NRRL 8126.
The DNA construct of the invention relating to SEQ ID No. 10 can be isolated from or produced on the basis of a DNA library of a strain of Macrcp2io2±na, in particular a strain of M. phaseolina, especially M.phaseolina, CBS 281.96.
The DNA construct of the invention relating to SEQ ID No. 12 can be isolated from or produced on the basis of a DNA library of a strain of .Crinipellis, in particular a strain of C. scaJbeila, especially C.scabella, CBS 280.96.
The DNA construct of the invention relating to SEQ ID No. 19 can be isolated from or produced on the basis of a DNA library of a strain of Sordaria, in particular a strain of Sordaria fimicola.
In the present context, the "analogue" of the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, is intended to indicate any DNA sequence encoding an enzyme exhibiting endoglucanase activity, which has any or all of the properties i)-iv).

The analogous DNA sequence
a) may be isolated from another or related (e.g. the same) organism producing the enzyme with endoglucanase activity on the basis of the DNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, e.g. using the procedures described herein; the homologue may be an allelic variant of the DNA sequence comprising the DNA sequences shown herein, i.e. an alternative form of a gene that arises through mutation; mutations can be silent (no change in the encoded enzyme) or may encode enzymes having altered amino acid sequence; the homologue of the present DNA sequence may also be a genus or spe¬cies homologue, i.e. encoding an enzyme with a similar activity derived from another species,
b) may be constructed on the basis of the DNA sequences shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, e.g. by introduction of nucleotide substitutions which do not give rise to another amino acid sequence of the endoglucanase encoded by the DNA sequence, but which correspond to the codon usage of the host organism intfrnded for production of the enzyme, or by introduction of nucleotide substitutions which may give rise to a different amino acid sequence. However, in the latter case amino acid changes are preferably of a minor nature, that is conservative amino acid substitu¬tions that do not significantly affect the folding or activity of the protein, small deletions, typically of one to about 30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue, a small linker peptide of up to about 20-25 residues, or a small extension that facilitates purifica¬tion, such as a poly-histidine tract, an antigenic epitope or a bind-ing domain. See in general Ford et al., Protein Expression and Purification. 2: 95-107, 1991. Examples of conservative substitutions are within the

group of basic amino acids (such as arginine, lysine, histidine), acidic amino acids (such as glutamic acid and aspartic acid), polar amino acids (such as glutamine and asparagine), hydrophobic amino acids (such as leucine, isoleucine, valine), aromatic amino acids (such as phenylalanine, tryptophan, tyrosine) and small amino acids (such as glycine, alanine, serine, threonine, methionine).
It will be apparent to persons skilled in the art that such substitutions can be made outside the regions criti¬cal to the function of the molecule and still result in an active polypeptide. Amino acids essential to the activity of the polypeptide encoded by the DNA construct of the invention, and therefore preferably not subject to substitution, may be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutage-nersls (Cunninghara and Wells, Science 244. 1081-1085, 1989). In the latter technique mutations are introduced at every residue in the mol¬ecule, and the resultamrt mutant molecules are tested for biological (i.e. endoglucanasa) activity to identify amino acid residues that are critical to the activity of the molecule. Sites of substrate-enzyme interaction can also be determined by analysis of crystal structure as determined by such techniques as nuclear magnetic reson¬ance, crystallography or photoaffinity labeling. See, for example, de Vos et al.. Science 255: 306-312, 1992; Smith et al., J. Mol. Biol. 224: 899-904, 1992; Wlodaver et al., FEBS Lett. 3 09: 59-64, 1992.
The endoglucanase encoded by the DNA sequence of the DNA construct of the invention may comprise a cellulose bind¬ing domain (CBD) existing as an integral part of the encoded enzyme,"or a CBD from another origin may be introduced into the endoglucanase enzyme thus creating an enzyme hybride. In this context, the term "cellulose-

binding domain" is intended to be understood as defined by Peter Tomme et al. "Cellulose-Binding Domains: Classi¬fication and Properties" in "Enzymatic Degradation of Insoluble Carbohydrates", John N. Saddler and Michael H. Penner (Eds.), ACS Symposium Series, No. 618, 1996. This definition classifies more than 120 cellulose-binding domains (CBDs) into 10 families (I-X), and it demon¬strates that CBDs are found in various enzymes such as cellulases, xylanases, mannanases, arabinofuranosidases, acetyl esterases and chitinases. CBDs have also been found in algae, e.g., the red alga Porphyra purpurea as a non-hydrolytic polysaccharide-binding protein, for refer¬ence see Peter Tomme et al., supra. However, most of the CBDs are from cellulases and xylanases- CBDs are found at the N or C termini of proteins or are internal. Enzyme hybrids are known in the art, see e.g. WO SO/00609 and WO 95/167 82, and may be prepared by transfoi"iaing into a host cell a DNA construct comprising at. least: a fragment of DNA encoding the cellulose-binding domain ligated, with or without a linker, to a DNA sequence encoding the enzyme of interest and growing tlie hosrt cell to express the fused gene. Enzyme hybrids may be described by the following formula:
CBD - MR - X,
wherein CBD is the N-terminal or the C-terminal region of an amino acid sequence corresponding to at least the cellulose-binding domain; MR is the middle region (the linker), and may be a bond, or a short linking group preferably of from about 2 to about 100 carbon atoms, more preferably of from 2 to 4 0 carbon atoms; or is preferably from about 2 to about 100 amino acids, more preferably of from 2 to 40 amino acids; and X is an N-terminal or C-terminal region of a po^lypeptide encoded by the DNA sequence of the invention.

The homology referred to in i) above is determined as the degree of identity between the two sequences indicating a derivation of the first sequence from the second. The homology may suitably be determined by means of computer programs known in the art such as GAP provided in the GCG program package (Needleman, S.B. and Wunsch, CD., Jour¬nal of Molecular Biology, 48_: 443-453, 1970). Using GAP with the following settings for DNA sequence comparison: GAP creation penalty of 5.0 and GAP extension penalty of 0.3, the coding region of the DNA sequence exhibits a degree of identity preferably of at least 60%, more preferably at least 65%, more preferably at least 70%, even more preferably at least 80%, especially at least 90%, with the coding region of the DNA sequence shown in SEQ ID No.l, 4, 6, 8, 10, 12-, or 16, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae, DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherdchia coli, DSH 10512, DSM 10511, DSM 10571, or DSM 10576, respectively.
The hybridization referred to in ii) above is intended to i nd i crat.e that the analogous UNA sequence hybridizes to the same probe as the DNA sequence encoding the endoglucanase enzyme under certain specified conditions which are described in detail in the Materials and Methods section hereinafter. The oligonucleotide probe to be used is the DNA sequence corresponding to the endoglucanase encoding part of the DNA sequence shown in SEQ ID NO 1, 4, 6, 8, 10, 12, or 16 respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae, DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively.
—5^e homology referred to in iii) above is determined as the degree of identity between the two sequences indicat¬ing a derivation of the first sequence from the second.

:he homology may suitably be determined by means of com-5uter programs known in the art such as GAP provided in :he GCG program package (Needleman, S.B. and Wunsch, :.D., Journal of Molecular Biology, AB_: 443-453, 1970). Jsing GAP with the following settings for polypeptide sequence comparison: GAP creation penalty of 3.0 and GAP extension penalty of 0.1, the polypeptide encoded by an analogous DNA sequence exhibits a degree of identity pre¬ferably of at least 55%, more preferably at least 60%, nore preferably at least 65%, even more preferably at least 70%, more preferably at least 80%, especially at least 90%, with the enzyme encoded by a DNA construct :::omprising the DNA sequence shown in SEQ ID No.l, 4, 6, 3, 10, 12, 16 or 19, respectively, or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae, DSM 9770, DSM 10082, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively.
In connection with property iv) above it is intended to indicate an endoglucanase encoded by a DNA sequence iso¬lated txom strain Saccharomyces cerevisiae, DSM 9T70, DSM 100B2, DSM 10080, DSM 10081, Escherichia coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively, arid produced in a host organism transformed with said DNA sequence or the corresponding endoglucanase naturally produced by Myceliophthora thermophila, Acremonium sp., Thielavia terrestris, Macrophomina phaseolina, Crinipellis scabella, Volutella colletotrichoides, or Sordaria fimicola, respectively. The immunological reac¬tivity may be determined by the method described in the Materials and Methods section below.
In further aspects the invention relates to an expression
vector harbouring a DNA construct of the invention,—a
cell comprising the DNA construct or expression vector and a method of producing an enzyme exhibiting

vative amino acid substitutions that do not significantly affect the folding or activity of the protein, small deletions, typically of one to about 30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue, a small linker peptide of up to about 20-25 residues, or a small extension that facilitates purification, such as a poly-histidine tract, an antigenic epitope or a binding domain. See in general Ford et al. , Protein Expression and Purification 2: 95-107, 1991. Examples of conservative substitutions are within the group of basic amino acids (such as arginine, lysine, histidine), acidic amino acids (such as glutamic acid and aspartic acid), polar amino acids (such as glutamine and asparagine), hydrophobic amino acids (such as leucine, isoleucine, valine), aromatic amino acids (such as phenylalanine, tryptophan, tyrosine) and small amino acids (such as glycine, alanine, serine, threonine, methionine).
It will be apparent to persons skilled in the art that such substitutions can be made outside the regions criti¬cal to the function of tiie laolecule and still result in an active enzyme. Amino acids essential to the activity of the enzyme of the invention, and therefore preferably not subject to substitution, may be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham, 1989). In the latter technique mutations are introduced at every residue in the molecule, and the resultant mutant molecules are tested for cellulytic activity to identify amino acid residues that are critical to the activity of the molecule. Sites of ligand-receptor interaction can also be determined by analysis of crystal structure as determined by such techniques as nuclear magnetic resonancBr;—Crystallography or photoaffinity labelling. See, for example, de Vos et al., 1992; Smith et al., 1992. Wlodaver et al., 1992.
The homologue may be an allelic variant, i.e. an alterna¬tive form of a gene that arises through mutation, or an altered enzyme encoded by the mutated gene, but having substantially the same activity as the enzyme of the invention Hence mutations can be silent (no change in the encoded enzyme) or may encode enzymes having altered amino acid sequence.
The homologue of the present enzyme may also be a genus or species homologue, i.e. an enzyme with a similar activity derived from another species.
A homologue of the enzyme may be isolated by using the procedures described herein.
Molecular screening and cloning bv polymerase chain reaction fact)
Molecular screening for DNA sequences of the invention may be carried out by polymerase chain reaction (PCR) using genomic DNA or double—strained cDNA isolated from a sraitable source, such as any of the herein mentioned organisms, and synthetic oligonucleotide primers prepared on the basis of the DNA sequences or the amino acid sequences disclosed herein. For instance, suitable oligonucleotide primers may be the primers described in the Materials and Methods section.
In accordance with well-known procedures, the PCR fragment generated in the molecular screening may be isolated and suctioned into a suitable vector. The PCR fragment may be used for screening DNA libraries by e.g. colony or plaque hybridization.

Expression cloning in yeast
The DNA sequence of the invention encoding an enzyme exhibiting endoglucanase activity may be isolated by a general method involving
cloning, in suitable vectors, a DNA library from a
suitable source, such as any of the herein
mentioned organisms
transforming suitable yeast host cells with said
vectors,
culturing the host cells under suitable conditions
to express any enzyme of interest encoded by a
clone in the DNA library,
screening for positive clones by determining any
endoglucanase activity of the enzyme produced by
such clones, and
isolating the enzyme encoding DNA from such
clones.
The general method is further disclosed in WO 94/14953
the contents of which are hereby incorporated by
rejference. A acre detailed description of the screening
method is given in Example 1 belov.
The DNA sequence coding for the enzyme may for instance be isolated by screening a cDNA library of Macrophomina phaseolina, Crinipellis scabella, Sordaria flmicola or Volutella colletotricboides, and selecting for clones ex¬pressing the appropriate enzyme activity (i.e. endoglucanase activity) or from Escherichia coli DSM 10512 deposited under the Budapest Treaty on 2 February, 1996, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany), or- from Escherichia coli DSM
lOSl-l—dapcsited under the Budapest Treaty on 2 February,
1996, at DSK, or from Escherichia coli DSM 10576, deposited under the Budapest Treaty on 12 March, 1996, at

DSM; or from Escherichia -coli DSM 10571 deposited under the Budapest Treaty on 6 March, 19S6, at DSM; or by screening a cDNA library of Myceliphthora thermophila, CBS 117.55, Acremonium sp., CBS 473.94, or Thielavia terrestris, NRRL 8126, and selecting for clones expres¬sing the appropriate enzyme activity (i.e. endoglucanase activity) or from Saccharomyces cerevisiae DSM 9770 deposited under the Budapest Treaty on 24 February, 1995, at DSM (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 16, D-38124 Braunschweig, Germany), or from Saccharomyces cerevisiae DSM 10032 deposited under the Budapest Treaty on 30 June, 1995, at DSM, from Saccharomyces cerevisiae DSM lOOSO deposited under the Budapest Treaty on 30 June, 1995, or from Saccharomyces cerevisiae DSM 10081 deposited under the Budapest Treaty on 30 June, 19S5, at DSM. The appropriate DNA sequence may then be isolated from the clone by s~ar.dari proced"ai"es, e-g- as aescrj-sed m Example 1.
WTJcIeic acid construct
As used herein the term '"nucleic acid construct" is intended to indicate any nucleic acid molecule of cDNA, genomic DNA, syn-hetic DNA or RNA origin. The term "construct" is intended to indicate a nucleic acid segment which may be single- or double-stranded, and which may be based on a complete or partial naturally occurring nucleotide sequence encoding an enzyme of interest. The construct may optionally contain other nucleic acid segments.
The nucleic acid construct encoding the enzyme of the invention may suitably be_Qi_genomic orcDNA origin, for instance obtained by preparing a genomic or cDNA library and screening for DNA sequences coding for all or part of

the enzyme by hybridization using synthetic oligonucleotide probes in accordance with standard techniques (cf. Sambrook et al., 1989).
The nucleic acid construct encoding the enzyme may also be prepared synthetically by established standard methods, e.g. the phosphoamidite method described by Beaucage and Caruthers, (1981), or the method described by Matthes et al., (1984). According to the phospho¬amidite method, oligonucleotides are synthesized, e.g. in an automatic DNA synthesizer, purified, annealed, ligated and cloned in suitable vectors.
Furthermore, the nucleic acid construct may be of mixed synthetic and genomic, mixed synthetic and cDNA or mixed genomic and cDNA origin prepared by ligating fragments of s%Tithetic, genomic or cDNA origin (as appropriate), the fragmentLS corresponding to various parts cf rhe entire nucleic acid construct, in accordance with standard tech¬niques.
The nncleic acid construct nay also be prepared by polymerase chain reaction using specific priaers, for instance as described in US 4,683,202 or Saiki et al., (1988) .
The nucleic acid construct is preferably a DNA construct which term will be used exclusively in this specification and claims.
Recombinant vector
A recombinant vector comprising a DNA construct encoding the enzyme of the invention may be any -vector which s.^y conveniently be subjected to recombinant DNA procedures, and the choice of vector will often depend on the host

cell into which it is to be introduced. Thus, the vector may be an autonomously replicating vector, i.e. a vector which exists as an extrachromosomal entity, the replication of which is independent of chroinosomal replication, e.g. a plasmid. Alternatively, the vector may be one which, when introduced into a host cell, is integrated into the host cell genome and replicated together with the chromosome(s) into which it has been integrated.
The vector is preferably an expression vector in which the DNA seguence encoding the enzyme of the invention is operably linked to additional segments required for transcription of the DNA. In general, the expression vector is derived from plasmid or viral DNA, or may contain elements of both. The term, "operably linked" indicates that the segments are arranged so that they function in concert for their intended purposes, e.g. transcription initiates in a promoter and proceeds through the DNA sequence coding for the enzyme.
The prnmnter may be any DNA sequence which shows transcriptional activity in the host cell of choice and may be derived from genes encoding proteins either homologous or heterologous to the host cell.
Examples of suitable promoters for use in yeast host cells include promoters from yeast glycolytic genes (Hitzeman et al., J. Biol. Chem. 255 (1980), 12073 -12080; Alber and Kawasaki, J. Mol. APPI. Gen. 1 (1982), 419 - 434) or alcohol dehydrogenase genes (Young et al., in Genetic Engineering of Microorganisms for Chemicals (Hollaender et al, eds.), Plenum Press, New York, 1982), or the TPIl (US 4,599,311) or ADH2-4C (Russell et al.. Nature 3 0^ rT98 3). 652 - 654) promoters.
Examples of suitable promoters for use in filamentous

fungus host cells are, for instance, the ADH3 promoter (McKnight et al.. The EMBO J. i (1985), 2093 - 2099) or the tpJA promoter. Examples of other useful promoters are those derived from the gene encoding A. oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, A. niger neutral a-amylase, A. niger acid stable a-amylase, A. niger or A. avamori glucoamylase (gluA), Rhizomucor miehei lipase, A. oryzae alkaline protease, A. oryzae triose phosphate isomerase or A. nidulans acetamidase. Preferred are the TAKA-amylase and gluA promoters.
Examples of suitable promoters for use in bacterial host
cells include the promoter of the Bacillus
stearothermophilus maltogenic amylase gene, the Bacillus
licheniformis alpha-amylase. gene, the Bacillus . : . ,
ajuyloliquefaciens BAN amylase gene, the Bacillus subtilis alkaline protease gen, or the Bacillus pumilus xylcsidase gene, cr by the phage Lambda PR or P^ promoters or rhe E. coli lac, trp or tac promoters.
Th-e DNA sequence encoding th-e enzyme of the invention may also. i:f necessary, be operably connected to a suitable
terminator.
The recombinant vector of the invention may further comprise a DNA sequence enabling the vector to replicate in the host cell in question.
The vector may also comprise a selectable marker, e.g. a gene the product of which complements a defect in the host cell, such as the gene coding for dihydrofolate reductase (DHFR) or the Schizosaccharomyces pombe TPI gene (described by P.R. Russell, Gene 40, 19S5, pp. 125-130). For filamentous fungi, selectable markers include amdS7~PVrG. argB, niaP, sC.
To direct an enzyme of the present invention into the

secretory pathway of the host cells, a secretory signal sequence (also known as a leader sequence, prepro sequence or pre sequence) rr.ay be provided in the recombinant vector. The secretory signal sequence is joined to the DNA sequence encoding the enzyme in the correct reading frame. Secretory signal sequences are commonly positioned 5' to rhe DliA sequence encoding the enzyme. The secretory signal sequence may be that normally associated with the enzyme or may be from a gene encoding another secreted protein.
For secretion from yeast cells, the secretory signal sequence may encode any signal peptide which ensures efficient direction of the expressed enzyme into the secretory pathway of the cell. The signal peptide may be naturally occurring signal peptide, or a functional part thereof, or it may be a syntheric peptide. Suitable signal peptides have been fO'jnd to be che a-facror signal peptide (cf. US 4,370,COS), the signal peptide of mouse salivary amylase (of. O. Kagencuchle et al., Nature 289, 1981, pp. 643-646), a modified carbcxypeptidase signal peptide (cf. L.A. Vails er al.. Cell 48, 19S7, pp. 887-897), the yeast BARl signal peptide (cf. WO 87/02670), or the yeast aspartic protease 3 (YAP3) signal peptide (cf. M. Egel-Mitani et al., Yeast 6, 1990, pp. 127-137).
For efficient secretion in yeasr, a sequence encoding a leader peptide may also be inserted downstream of the signal sequence and upstream of the DNA sequence encoding the enzyme. The function of the leader peptide is to allow the expressed enzyme to be directed from the endoplasmic reticulum to the Golgi apparatus and further to a secretory vesicle for secrerion into the culture medium, (i.e. exportation of the enzyme across the cell wail or at least through the cellxrXar m.embrane into the periplasmic space of the yeast cell). The leader peptide may be the yeast a-factor leader (the use of which is

described in e.g. US 4,546,082, EP 16 201, EP 123 294, EP 123 544 and EP 163 529). Alternatively, the leader peptide may be a synthetic leader peptide, which is to say a leader peptide not found in nature. Synthetic leader peptides may, for instance, be constructed as described in WO S9/02463 or WO 92/11378.
For use in filamentous fungi, the signal peptide may conveniently be derived from a gene encoding an Aspergillus sp. amylase or glucoaraylase, a gene encoding a Rhizomucor miehei lipase or protease, a Humicola lanuginosa lipase. The signal peptide is preferably derived from, a gene encoding .4. oryzae TAKA amylase, A. niger neutral a-amylase, .-.. niger acid-stable amylase, or A. niger glucoam.ylase. ^ -
The procedures used to ligate the DN'A sequences coding for the present enzyme, the promoter and optionally the terminator and/or secretory signal sequence, respectively, and to insert them into suitable vectors containing the information necessary for replication, are well known to persons skilled in the art (cf., for instance, Sambrook et al., op.cit.).
Host cells
The DNA sequence encoding the present enzyme introduced into the host cell may be either homologous or heterologous to the host in question. If homologous to the host cell, i.e. produced by the host cell in nature, it will typically be operably connected to another promoter sequence or, if applicable, anoTiher secretory signal sequence and/or terminator sequence than in its natural environment. The term "homologous" is intended to include a cDNA sequence encoding an enzyme native to the host organism in question. The term "heterologous" is

intended to include a DNA sequence not expressed by the host cell in nature. Thus, the DNA sequence may he from another organism, or it may be a synthetic sequence.
The host cell into which the DNA construct or the recombinant vector of the invention is introduced may be any cell which is capable of producing the present enzyme and includes bacteria, yeast, fungi and higher eukaryotic cells.
Examples of bacterial host cells which, on cultivation, are capable of producing the enzyme of the invention are gram-positive bacteria such as strains of Bacillus, such as strains of B. subtilis, B. licheniformis, 5. ler.tus, B. brevis, B. stearothermophilus, B. alkalophilus, 3. amyloliquefacier.s, B. coagulans, B. circvlans, B. lautus, B. megatherium or 3. tburingier.sis, or strair.s cf Szrepzoi?.yces, such as S. livida::s or S. jnurinus, or gram-negative bacteria such as Echerichia coli. The transformation of the bacteria may be effected by protoplast transformation or by using comperent cells in a manner known per se (cf. Sambrook et al., suora).
When expressing the enzyme in bacteria' such as E. coli, the enzyme may be retained in the cytoplasm, typically as insoluble granules (known as inclusion bodies), or may be directed to the periplasmic space by a bacterial secretion sequence. In the for»-er case, the cells are lysed and the granules are recovered and denatured after which the enzyme is refolded by diluting the denaturing agent. In the latter case, the enzyme may be recovered from the periplasmic space by disrupting the cells, e.g. by sonication or osmotic shock, to release the contents of the periplasmic space and recovering the enzyme.
Examples of suitable yeasts cells include cells of Saccharomyces spp. or Schizosaccharomyces spp., in

pa:.-ticular strains of Saccharomyces cerevisiae or Saccharomyces kluyveri. Methods for transforming yeast cells with heterologous DNA and producing heterologous enzymes therefrom are described, e.g. in US 4,599,311, US 4,931,373, US 4,870,008, 5,037,743, and US 4,845,075, all of which are hereby incorporated by reference. Transformed cells are selected by a phenotype determined by a selectable marker, commonly drug resistance or the ability to grow in the absence of a particular nutrient, e.g. leucine. A preferred vector for use in yeast is the POTl vector disclosed in US 4,931,373. The DNA sequence encoding the enzyme of the invention may be preceded by a signal sequence and optionally a leader sequence , e.g. as described above. Further examples of suitable yeasr cells are strains of Kluyveromyces, such as .^'. lactis, Hansenula, e.g. H. polymorpha, or Pichia, e.g. P. pastoris (cf. Gleescn er al., J. Gen. Microbiol. 132, 1986, pp. 3455-3465; US 4,882,275).
Examples cf other fungal cells are cells of filamentcus fungi, e.g. Aspergillus spp., Neurospora spp., Fusarium spp. cr Trichoderma spp., in particular strains of A. oryzae, A. nldulans, A. niger, or Fusarium graminearum. The use of Aspergillus spp. for the expression of pro¬teins is described in, e.g., EP 272 277, EP 230 023. The transformation of F. oxysporuw may, for instance, be carried out as described by Malardier et al., 1989, Gene 73: 147-156.
When a filamentous fungus is used as the host cell, it may be transformed with the DNA construct of the invention, conveniently by integrating the DNA construct in the host chromosome to obtain a recombinant host cell. This integration is- generally considered to be an advantage as the DNA sequence is more likely to be stably maintained in the cell. Integration of the DNA constructs into the host chromosome may be performed

according to conventional methods, e.g. by homologous or heterologous recombination.
The transformed or transfected host cell described above is then cultured in a suitable nutrient medium under conditions permitting the expression of the present enzyme, after which the resulting enzyme is recovered from the culture.
The medium used to culture the cells may be any conventional medium suitable for growing the host cells, such as minimal or complex media containing appropriate supplements. Suitable media are available from commercial suppliers or may be prepared according to published recipes (e.g. in catalogues of the American Type Culture Collection) . The enz^ine produced by the cells may then be recovered from the culture medium by conventional procedures including separating the host cells from z'r.e. medium by csnrrifugarion or fiitra-icn. precipiraning zr.e proteinaceous components of the supernatant or filtrate by means of a salt, e.g. ammonium sulphate, purification by a variety of chromarographic procedures, e.g. ion exchange chromatography, gel filtration chromiatography, affinity chromatography, or the like, dependent on the type of enzyme in question.
In a still further aspect, the present invention relares to a method of producing an enzyme according to the invention, wherein a suitable host cell transformed with a DNA sequence encoding the enzyme is cultured under conditions permitting the production of the enzyme, and the resulting enzyme is recovered from the culture.
Enzyme Screening driven by taxonomy as well as ecology:
A powerful tool like the molecular screening disclosed herein, designed to detect and select said type of

interesting enzymes, can still not stand on its own. In order to maximize the chances of making interesting discoveries the molecular screening approach was in the present investigation combined with careful selection of which fungi to screen. The selection was done through a thorough insight in the identification of fungi, in taxonom.ical classification and in phylogenetic relationships.
A taxonomic hot spot for production of cellulytic enzymes can further only be fully explored if also the ecological approach is included. Thorough knowledge about the adaptation to various substrates (especially saprotrophic, necrotrophic or biotrophic degradation of •plant m-aterials) are prerequisites for designing an intelligent screening and for managing a successful selection of strains and ecological niches to be searc¬hed .
Both the taxonomy and the ecological approach disclosed herein aim at maximizing discovery" of said enzymes in the molecular screening program. However, still several hundreds (or if all preliminary work is included) several thousand fungi have been brought in culture in order to detect the 53 hits of said type of cellulytic enzy^m.e here reported.
The screening and cloning may be carried out using the following:
MATERIALS AND METHODS
List of organisms:
Saccharomyces cerevisiaB, -USiL. 917 0, DSM 10082, DSM 10080, DSM 10081, or Escherichia coli, DSM 10512, DSM 10511, DSM 10571, DSM 10576, respectively, containing the plasmid

comprising the full length DNA sequence, coding for the endoglucanase of the invention, in the shuttle, vector pYES 2.0.
Escherichia coli DSM 10583, 10584, 10585, 10586, 10587, and 10588.
Diplodia gossypir.a Cooke Deposit of Strain, Ace No: CBS 274.96 Classification: Ascomycota , Loculoascorr.ycetes , Dothideales, Cucurbitariaceae
Ulospora bilgramii {Hawksw. et al.) Hawksw. et al . Ace No of strain: NK3C 1444, Nippon University, (Prof. 'Tubaki collection) Classification: Ascorriycota , Loculoascomycetes, Dothideales, (family unclassified)
Microsphaeropsis sp.
Isolated from: Leaf of Camellia japonica (Theaceae,
Guttifsrales), grown in Kunming Botanical garden, Yunnan
Province, China
Classification: Ascomycota, Loculoascomycetes,
Dothideales, (family unclassified)
Macrophomina phaseolina (Tassi) Goidannich
Syn: Rhizoctonia baraticola
Deposit of Strain, Ace No.:CBS 281.96
Isolated from seed of Glycine max (Leguminosa), cv CMM
60, grown in Thailand, 1990
Classification: Ascomycota, Discomycetes, Rhytism.atales,
Rhytismataceae
Ascojbolus stictoideus Speg.
Isolated from goose dung, Svalbard, "TTorway
Classification: Ascomycota, Discomycetes, Pezizales,
Ascobolaceae

Saccobolus dilutellus (Fuck.) Sacc. Deposit of strain: Ace No CBS 275.96
Classification: Ascomycota, Discomycetes, Pezizales, Ascobolaceae
Penicillium verruculosum Peyronel
Ex on Ace No of species: ATCC 62396
Classification: Ascomycota, Plectomycetes, Eurotiales,
Trichocomaceae
Penicillium ckrysogenum Thorn
Ace No of Strain: ATCC 9480
Classification: Ascomycota, Plectomycetes, Eurotiales,
Trichocomaceae
Thermomyces verrucosus Pugh et ai Deposit of Strain, Ace No.: CES 235.96
Classification: Aseor.yecra, PlectcmyceTies, Eurcriales, (family unclassified; affiliation based on ISS RNA, sequencing and homologies)
Xylaria hypcxylcn L. ex Greville
Deposit of srrain, Ace No: CBS 2S4.96
Classification: Ascomycota, Pyrenomycetes, Xylariales,
Xylariaceae
Poronia punctata (Fr.ex L.} Fr.
Classification: Ascomycota, Pyrenomycetes, Xylariales,
Xylariaceae
Nodulisporum sp
Isolated from leaf of Camellia reticulata (Theaeeae,
Guttiferales), grown in Kunming Botanical Garden, Yunnan
Province, China
Classification: Ascomycota-,—Pyrenomycetes, Xylariales,
Xylariaceae

Cylindrocarpon sp
Isolated from marine sample, the Bahamas
Classification: Ascomycota, Pyrenomycetes, Hypocreales
(unclassified)
Acremonium so
Deposit of Strain, Ace. No.: CBS 478.94
Classification: Ascomycota, Pyrenomycetes, Hypocreales,
Hypocreaceae
Fusarium anguioides Sherbakoff
Ace No of strain: IFO 4467
Classification: Ascomycota, Pyrenomycetes, Hypocreales,
H>'pocr eaceae
Fusarium poae (Peck) Wr.
Ex on Ace No of species: ATCC 6OS S3
i_iassir ica~ ion: Ascomyccta , Pyrenomyce'ces , nypocreaJ.es,
Hypocreaceae
Fusariun solani (Mart.)Sacc.emnd,Snyd 5 Hans.
Ace No of strain: IMI 107.511
Classification: Ascomycota, Pyrenomycetes, Hypocreales,
Hypocreaceae
Fusariuui oxysporum ssp lycopersici (Sacc .) Sr.yd . & Har.s .
Ace No of strain: CBS 64 5.7 3
Classification: Ascomycota, Pyrenomyeeres, Hypocreales,
Hypocreaceae
Fusarium oxysporum ssp passiflora
hoc No of strain: CBS 7 4 4.79
::lassif ication: Ascomycota, Pyrenomycetes, Hypocreales,
-lypocreaceae
Gliocladium catenulatum Gillman & Abbott Ace. No. of strain: CBS 227.48

classification: Ascoinycota, Pyrenomycetes, Hypocreales, Hypocreaceae
Nectria pinea Dingley
Deposit of Strain, Ace. No. CBS 279.96
Classification: Ascomycota, Pyrenomycetes, Hypocreales,
Nectriaceae
Volutella colletotrichoides
Ace No of Strain: CBS 400.53
Classification: Ascoinycota, Pyrenomycetes, Hypocreales
(unclassified)
Sordaria macrospcra Auerswala
Ex on Ace No of species: ATCC 60255
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Sordariaceae
Sordaria fimicola (Roberge) Cesati ez De '^otaris Ex on Ace. No. for the species: ATCC 52644 Isolat.ed from dung by H.Dissi.ng, ISP, KU, Denmark Classification: Ascomycota, Py^enonycetes, Sordariales, Sordariaceae
Humicola grisea Traeen
ex on Ace No for the species: ATCC 2 2726
Source: Hatfield Polytechnic
Classification: Ascomycota, Pyrenomycetes, Sordariales,
(fam. unclassified)
Humicola nigrescens Omvlk
Ace No of strain: CBS S19.73
Classification: Ascomycota, Pyrenomycetes, Sordariales,
(fam. unclassified)
Scytalidium thermophilum (Cooney et Emerson) Austwick Ace No of strain: ATCC 28085

Classification: Ascomycota, PyrenomycGtes, Sordariales, (fam. unclassified)
Thielavia thermophila Fergus et Sinden I (syn Corynascus thermophilus)
Ace No of strain: CBS 174.70, IMI 145.136
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Chaetomiaceae
Isolated from Mushroom compost
Thielavia terrestris (Appinis) Malloch et Cain
Ace No of strain: NRRL3126
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Chaetomiaceae
Cladorrhinum foecundissimum Saccardo et Marchal
Ex on Ace No of species: ATCC 62 27 3
Ciassif icatio.n: A.sccr.ycora , ?'j"rencmyce"ces , Sordariales,
Lasiosphaeriaceae
Isolared from leaf of Selandin sp. (Ccmpositaceae,
A.sterales) , Dallas .Mountain, Jamaica
Syspastospora boninensis
Ace No of strain: NKBC 1515 (Nippon University, profe
Tubaki Collection)
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Cerastomataceae
Chaetomium cuniculorum Fuckel
Ace. No. of strain: CBS 799.83
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Chaetomiaceae
Chaetomium brasiliense Batista et Potual
Ace No of strain; CBS -L22^ 65
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Chaetomiaceae

Chaetojp.ium murorum Corda
Ace No of strain: CES 163.52
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Chaetomiaceae
Chaetowium virescens (von Arx) Udagawa
Ace.No. of strain: CBS 5 4 7.75
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Chaetomiaceae
Myceliophthora tbermophlla (Apinis) Oorschot
Deposit of Strain, Ace No:CBS 117.65
Classification: Ascomycota, Pyrenomycetes, Sordariales,
Chaetomiaceae
Nigrospora sp
Deposit of strain. Ace No: CBS 272.96
Isolated from leaf of Artocarp'uS aitiiis, Moraceae,
Urticales qrowv, in Christiana, Jamaica
Classification: Ascomycota, pyrenomycetes,
Trichcsphaeriales, (family unclassified)
Nigrospora sp
Isolated from leaf of Pinus yuannanensis, Botanical
Garden, Kuning, Yunnan.
Classification: A.scomycota, Pyrenomycetes,
Triehosphaeriales, Abietaceae, Pinales.
Diaporthe syngenesia
Deposit of strain, .Ace No: CBS 278.96
Classification: .Ascomycota, Pyrenomycetes, Diaporthales,
Valsaceae
Colletotrichum lagenarium (Passerini) Ellis at Halsted
syn Glomerella cinguiata var orbiculare Jenkins et
Winstead
Ex on ace No of species: ATCC 52609

Classification: Ascomycota, Pyrenomycetes, Phyllachorales
Exidia glandulosa Fr.
Deposit of Strain, Ace No: CBS 277.96 Classification: Basidiomycota, Hymenoiriycetes, Auriculariales, Exidiaceae
Crinipellis scabella (Alb.&Schw.:Fr.)Murr
Deposit of strain: Ace No CBS 280.96
Classification: Basidiomycota, Hymenomycetes, Agaricales,
Panaeolus retivugis (Fr.) Gill.
Ace.No. of strain: CBS 275.47
Classification: Basidiomycota, Kynenc-ycetes, Agaricales,
Coprinaceae
Fojnes fomentarius (L.) Fr.
Deposit of strain: Ace No. CBS 2 7 6.96
Classification: Basidiorr.ycora, Hymenomycetes,
Aphyllophorales,
Fomitaceae
Spongipellis sp.
Deposit of Strain: Ace No CBS 283.96
Classification: Basidiomycota, Hymenomycetes,
Aphyllophorales,
Bjerkanderaceae (identified and affiliated taxonomieally
by ISS sequence and homology)
Trametes sanguinea (Fr.) Lloyd
syn: Polyporus sanguineus; Pycnoporus sanguineus (L.:Fr.)
Murrill
Ace No of strain: AKU 5062 (Kyoto University Culture
Collection)
Classification: Basidiomycota, Aphy-Llophorales,
Polyporaceae

Schizophyllum commune Fr
Ace. No. of species: ATCC 38548
Classification: Basidiomycota, Aphyllophorales,
Schizophyllaceae
Rhizophlyctis rosea (de Bary & Wor) Fischer Deposit of Strain: Ace No.: CBS 282.96 Classification: Chytridiomycota, Chytridiomycetes, Spizellomycetales, Spizellomycetaceae
Rhizomucor pusillus (Lindt) Schipper
syn: Mucor pusillus
Ace No of strain: IFO 4 578
Ex en Ace No of species: ATCC 468S3
Classification: Zygomycota, Zygomycetes, Mucorales,
Mucoraceae
Phycomyces nitens (Kur.ze) van Tieghem & Le Monnier
Ace No of strain: IFO.4314
Ex en Kco No of species: ATCC 16327
Classification: Zygomycota, Zygomycetes, Mucorales,
Mucoraceae
Chaetostylum fresenii van Tieghem & Le Monnier
syn. Helicostylum fresenii
Ace No of strain NRRL 2 3 05
Classification: Zygomycota, Zygomyceres, Mucorales,
Thamnidiaceae
Unclassified:
Trichothecium roseum
Ace No of strain: IFO 5372
Coniothecium sp
Endophyte, isolated from leaf of unidentifed higher

plant, growing in Kunming, Yunnan, China
Unclassified and Un-identified:
Deposit of strain, Ace No.: CBS 271.96
Isolated from leaf of Artocarpus altilis (Moraceae,
Urticales), grown in Christiana, Jamaica
Deposit of strain. Ace No.: CBS 273.96
Isolated from leaf of Pimenta dioica (Myrtaceae,
Myrtales) grown in Dallas Mountain, Jamaica
Deposit of strain: CBS 270.96
Isolated from leaf of PseudocalyroiBa alliaceum - -. ..-
(Bignoniaceae, Solanales) growing in Dallas Mountain,
u s m a 1 c a
Other strains:
Escherichia coli MC1061 and DKIOB.
Yeast strain: The Saccharomyces csrsvisiae strain used was W3124 (MATa; ura 3-52; leu 2-3, 112; his 3-D200; pep 4-1137; prcl::HIS3; prbl:: LEU2; cir+).
Plasmids:
The Aspergillus expression vector pHD414 is a derivative of the plasmid p775 (described in EP 238 023). The construction of pHD414 is further described in WO 93/11249.
pYES 2.C (Invitrogen)
pA2C477, PA2C193, pA2C357, pA2C371, pA2C385, pA2C475, pA2C438, p/v2C502 (See example 1, 2, 2 and 4).

Isolation of the DNA sequence shown in SEQ ID No. l, 4, 6, 8, 10, 12, 16, or 19 respectively:
The full length DNA sequence, comprising the cDNA sequence shown in SEQ ID No. 1, 4, 6, 8, 10, 12, 16 or 19, respectively, coding for the endoglucanase of the invention, can be obtained from the deposited organism S. cerevisiae, DSM 9770, DSM 100S2, DSM 10080, DSM 10081, E. coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively, by extraction of plasmid DNA by methods known in the art (Sambrook et al. (1989) Molecular cloning: A laboratory manual. Cold Spring Harbor lab.. Cold Spring Harbor, NY).
PCR primers for molecular screening of cellulases of the present invention:
The four degenerate, deoxyinosine-conraining cligcnucieotide primers (sense; s and antisense; asl, as2 and as3) corresponding to four highly conserved amino acid regions fou.nd in the deduced amino acid sequences of Thielavia terrestris cellulase, Myceliophthora thermophllum cellulase, and two cellulases frcs Acremonlum sp. The residues are numbered according to the Myceliophthora thermophilum sequence. The deoxyinosines are depicted by an I in the primer sequences, and the restriction sites are underlined.
(klippe-klistre fra SKa

Molecular screening by polymerase chain reaction (PCR):
In vitro amplification of genomic DNA and double-stranded cDNA.
j Directional, double-stranded cDNA was synthesized from 5 /ig of poly(A)*RNA as described below. Genomic DNA was isolated according to Yelton et al.
Approximately 10 to 20 ng of double-stranded, cellulase-induced cDNA or 100 to 200 ng of genomic DNA from a selection of fungal strains was PCR amplified in PCR buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 1.5 mM MgClj, 0.01 % (w/v) gelatin) containing 200 ^M of each dNTF and 100 pmoi of each degenerate primer in three combinations:
1)
sense,
D — V-'v_'^ _ .-..^. vj c i lA'-—! /(-oiiA' / JTGLJGA /-[TG /[-i'lj /-[/"..-i /,; /(-C—-i
antisense 1,
5'- CTAGTCTAGATA\',;CAIGC\/J-;C.VV,;CACC -3'; or
2) sense,
5 ' - CCCCAAGCTTAClVrGITA'-7TTGGGA'7TTG^/TTG'7T^-A'^/n^/cC-3 ' antisense 2, CTAGTCTAGA.-J'.IAV.;/'^ICCIAV'7"ICCICCICCIGG -3 ■ ; or
3) sense,
5' - CCCCAAGCTTACIVCC-ITA^7TTGGGA'7TTG'7TTG'7T^"V\/,/'/,.C-3 ■
antisense 3,
5 ■ - CTAGTCTAGAIA;^.CCA\/nTCA'\/n-7'TAIC a DNA thermal cycler (Landgraf, Germany) and 2.5 units of Taq polymerase (Perkln-Elmer, Cetus, USA). Thirty cycles of PCR were performed using a cycle profile of denaturation at 94 °C for 1 min, annealing at 64 °C for 2

min, and extension at 72 °C for 3 min. Ten-/xl aliquots of the amplification products were analyzed by electrophoresis in 3 % agarose gels (NuSieve, FMC) with Haelll-digested ^XIV^ RF DNA as a size marker.
Direct sequencing of the PCR products. Eighty-/il aliquots of the PCR products were purified using the QIAquick PCR purification kit (Qiagen, USA) according to the manufacturer's instructions. The nucleotide sequences of
I the amplified PCR fragments were determined directly on the purified PCR products by the dideoxy
chain-termination method, using 50-150 ng template, the Taq deoxy-terminal cycle sequencing kit (Perkin-Elmer, US.A) , fluorescent labeled terminators and 5 pmol of the
i sense primer: 5'- CCCCAAGCTTACI"/(-GITA'\'TTGGGA'~/-T-
G'^/T-TG'/-AA'^/G'/(-C-3 ' . Analysis of the sequence data were perforned according to Devereux et al.
Cloning by polymerase chain reaction (PCR): Su±)cloning of PCR fragments.
Twentyfive-^i aliquots of the PCR products generared as described above were electrophoresed in 0.8 % low gelling temperature agarose (SeaPlaque GTG, FMC) gels, the relevant fragments were excised from the gels, and recovered by agarase treatment by adding 0.1 vol of 10 x agarase buffer (New England Biolabs) and 2 units per 100 jj.1 molten agarose to the sample, followed by incubation at 45 °C for 1.5 h. The sample was phenol and chloroform extracted, and precipitated by addition of 2 vols of 96 % EtOH and 0.1 of 3 M NaAc, pH 5.2. The PCR fragments were recovered by centrifugarion, washed in 70 % EtOH, dried and resuspended in 20 fxl of restriction enzyme buffer (10 mM Tris-HCl, 10 mM MgCl2, 50 mM NaCl, 1 mM DTT) . The fragments were digested witrh—^indlll and Xbal, phenol and chloroform extracted, recovered by precipitation with 2 vols of 96 % EtOH and 0.1 of 3 M

NaAc, pH 5.2, and subcloned into HindIII/Xi>aI-cleaved pYES 2.0 vector.
Screening of cDNA libraries and characterization of the positive clones. cDNA libraries in S. cerevisiae or E. coli, constructed as described below, were screened by colony hybridization (Sambrook, 1989) using the corresponding randoiTi-primed (Feinberg and Vogelstein) "p-labeled (>1 x lo"cpm/^q) PCR products as probes. The hybridizations were carried out in 2 x SSC (Sambrook, 1989), 5 X Denhardfs solution (Sambrook, 1989), 0.5 % (w/v) SDS, 100 fig/ml denatured salmon sperm DNA for 20 h at 65^C followed by washes in 5 x SSC at 25°C (2 x 15 min), 2 X SSC, 0.5 % SDS at 65°C (30 min), 0.2 x SSC, 0.5 % SDS at- 65°C (30 min) and finally in 5 x SSC ( 2 x 15 ' ' min) at 25=C. The positive cDNA clones were characterized by sequencing rhe ends of the cD.N'.A inserts with pYES 2.0 polylinker primers (Invitrogen, USA.), and by derermining the nucleotide seuence of the longest cDNA from both strands by the dideoxy chain termination method (Sanger et al.) using fluorescent labeled terminators. Qiagen purified plasmid D.^^'.-. (Qiagen, USA) was sequenced with the Taq deoxy terminal cycle sequencing kit (Perkin Elmer, USA) and either pYES 2.0 polylinker primers (Invitrogen, US.A) or synthetic oligonucleotide primers using an Applied Biosystems 373A automated sequencer according to the ma.nufacturers instructions. Analysis of the sequence data was performed according to Devereux et al.
Extraction of total RNA was performed with guanidinium thiocya.nate followed by ultracentrifugation through a 5.7 M CsCl cushion, and isolation of poly(A)"RNA was carried out by oligo(dT)-cellulose affinity chromatography using the procedures described in WO 94/14953.
cDNA synthesis: Double-stranded cDNA was synthesized from 5 /ig poly (A)* RNA by the RNase H method (Gubler and

Hoffman (1983) Gene 25:263-269, Sambrook et al. (1989) Molecular cloning: A laboratory manual, Cold Spring Harbor lab.. Cold Spring Harbor, NY) using the hair-pin modification developed by F. S. Hagen (pers. comm.). The poly(A)' RNA (5 Mg in 5 ^1 of DEPC-treated water) was heated at 70°C for 8 min. in a pre-siliconized, RNase-free Eppendorph tube, quenched on ice and combined in a final volume of 50 IJ.1 with reverse transcriptase buffer (50 mM Tris-Cl, pH 8.3, 75 mM KCl, 3 mM MgCl,, 10 mM DTT, Bethesda Research Laboratories) containing 1 mM of dATP, dGTP and dTTP and 0.5 mM 5-methyl-dCTP (Pharmacia), 40 units human placental ribonuclease inhibitor (RNasin, Promega) , 1.45 ug cf cligc (dT) ,,-Not I prim.er (Pharmacia) and 1000 units Superscript II RNase H reverse - trans-criptase (Bethesda Research Laboratories) --First- '• . , strand cDNA was synthesized by incubating the reacrion mixture at 4 5"C fcr 1 hour. After synthesis, rhe mirture was gelfij.'craTec ~hrougn a MicroSpin S-4 00 HR (Pharmacia) spin column according ro the manufacturer's instructions.
After rhe gelfiltration, the hybrids were diluted in 2 50 /il second strand buffer (20 mM Tris-Cl, pH 7.4, 90 miM KCl, 4.6 mM'MgCl,, 10 mM (NH4)2S04, 0.16 mM iSViAD^) containing 200 ^M of each dNTP, 60 units I. coli DNA polymerase I (Pharmacia), 5.25 units RNase K (Promega) and 15 units E. coli DNA ligase (Boehringer Mannheim). Second strand cDNA synTihesis was performied by incubating the reaction tube at 16°C for 2 hours and additional 15 min. at 25°C. The reaction was stopped by addition of EDTA to a final concentration of 2 0 inM followed by phenol and chloroform extractions.
Mung bean nuclease treatment: The double-stranded cDNA was precipitateti—at -20°C for 12 hours by addition of 2 vols 96% EtOH, 0.2 vol 10 M NH4AC, recovered by centrifugation, washed in 70% EtOH, dried and resuspended

in 30 Ml Mung bean nuclease buffer (30 mM NaAc, pH 4.6, 300 mM NaCl, 1 rc.M ZnSO,, 0.35 mM DTT, 2% glycerol) containing 25 units Mung bean nuclease (Pharmacia). The single-stranded hair-pin DNA was clipped by incubating the reaction at 30^C for 30 min., followed by addition of 70 /il 10 mM Tris-Cl, pH 7.5, 1 mM EDTA, phenol extraction and precipitation with 2 vols of 96% EtOH and 0.1 vol 3 M NaAc, pH 5.2 on ice for 30 min.
Blunt-ending with T4 DNA polymerase: The double-stranded cDNAs were recovered by centrifugation and blunt-ended in 30 fil T4 DNA polymerase buffer (20 mM Tr is-acetate, pH 7.9, 10 mM MgAc. 50 mM KAc, 1 mM DTT) containing 0.5 ir.M of each dNTP and 5 units T4 DNA polymerase (N'ew England Biolabs) by incubating, the reaction mixture at 16°C for 1 hour. The reaction was stopped by addition of EDTA to a final coficentrarion of 20 mM, followed by phenol and chlcrcfcr:?. e>rcrac-icns, and precipitarion for 12 hours at -20-C by adding 2 vols 96% EtOH and 0.1 vol 3 M NaAc pH
Adapter ligation/ Not I digestion and size selection:
After the fill-in reaction the cDNAs were recovered by centrifugation, washed in 70% EtOH and dried. The cDNA pellet was resuspended in 25 /il ligation buffer (30 mM Tris-Cl, pH 7.S, 10 -uM MgCl,, 10 mM DTT, 0. 5 mM ATP) containing 2.5 ^g non-palindromic BstXI adaptors (Invitrogen) and 30 units T4 ligase (Promega) and incubated at 16=C for 12 hours. The reaction was stopped by heating at 65°C for 20 min. and then cooling on ice for 5 min. The adapted cDNA was digested with Not I restriction enzvTue by addition of 2 0 ul water, 5 ul ICx Not I restriction enzyme buffer (New England Biolabs) and 50 units Not I (New England Biolabs), followed by incubation for 2.5 hours' at 37='C. The reaction was stopped by heating at 65°C for 10 min. The cDNAs were size-fractionated by gel electrophoresis on a 0.8%

SeaPlaque GTG low melting temperature agarose gel (FMC) in Ix TBE to separate unligated adaptors and small cDNAs. The cDNA was size-selected with a cut-off at 0.7 kb and rescued from the gel by use of /J-Agarase (New England Biolabs) according to the manufacturer's instructions and precipitated for 12 hours at -20°C by adding 2 vols 96% EtOH and 0.1 vol 3 M NaAc pH 5.2.
Construction of libraries: The directional, size-selected cDNA was recovered by centrifugation, washed in 70% EtOH, dried and resuspended in 30 ^1 10 mM Tris-Cl, pH 7.5, 1 mM EDTA. The cDNAs were desalted by gelfiltration through a MicroSpin S-300 HR (Pharmacia) spin coluinn according to the manufacturer's instructions. Three test ligations were carried out in 10 /il.ligation buffer (30 mM Tris-Cl, pH 7.8, 10 mM MgCl., 10 mM DTT, 0.5 nsM ATP) containing 5 til double-stranded cDNA (reacricn rubes -1 and #'2), 15 units T4 iigase (Fromega) and 50 ng (zube fl), 40 ng (tube #2) and 40 ng (tube #3, the vector background control) of BstXI-NctI cleaved pYES 2.0 vector. The ligation reactions were perfcruied by incubation at 16'C for 12 hours, heating at 70'C for 20 ir.in. and addition of 10 fil water to each tube. 1 ^1 of each ligation mixture was electroporated into 40'^1 electrocompetent E. coli DHIOB cells (Bethesda research Laboratories) as described (Sambrook et al. (1939) Molecular cloning: A laboratory manual, Cold Spring Harbor lab., Cold Spring Harbor, NY). Using the optimal conditions a library was established in E. coli consisting of pools. Each pool was made by spreading transformed E. coli on LB+ampicillin agar plates giving 15.000-30.000 colonies/plate after incubation at 37°C for 24 hours. 20 ml LB+ampicillin was added to the plate and the cells were suspended herein. The cell suspension was shaked in a 50 mi tube for 1 hour at 37°C. Plasmid DNA was isolated from the cells according to the manufacturer's instructions using QIAGEN plasmid kit and stored at -20°C.

1 ^1 aliquots of purified plasmid DNA (100 ng//il) from individual pools were transformed into S. cerevisiae W3124 by electroporation (Becker and Guarante (1991) Methods Enzymol. 194:182-137) and the transformants were plated on SC agar containing 2% glucose and incubated at 30°C.
Identification of positive colonies: After 3-5 days of growth, the agar plates were replica plated onto a set of SC + galactose-uracil agar plates containing 0.1% AZCL HE cellulose. These plates were incubated for 3-7 days at 30°C. Endoglucanase positive colonies were identified as colonies surrounded by a blue halo.
Cells from enz-v'-rrie-positive colonies were spread for single colony isolation on agar, and an enzy-e-producing single colony was selected for each of the endoglucanase-
Characterization of positive clones: The positive clones were obtained as single colonies, the cDNA inserts were aaplified direcrly from the yeast colony using biotinylated polylinker primers, purified by magnetic beads (Dynabead M-280, Dynal) system and characterized individually by sequencing the 5"-end of each cDNA clone using the chain-termination method (Sanger et al. (1977) Proc. Natl. Acad. Sci. U.S.A. 74:54 63-5467) and the Soquenase system (United States Biochemical).
The nucleotide sequence was determined of the longest cDNA from, both strands by the dideoxy chain termination -cethod (Sanger et al.) using fluorescent labeled terminators. Plasmid DNA was rescued by transformation into E. coll as described below. Qiagen purified plasmid DNA (Qiagen, USA) was sequenced with the Tag deoxy terminal cycle sequencing kit (Perkin Elmer, USA) and either pYES 2.0 polylinker primers (Invitrogen, USA) or

synthetic oligonucleotide primers using an Applied Biosystems 373A automated sequencer according to the manufacturers instructions. Analysis or the sequence data was performed according to Devereux et al .
'■)
Isolation of a cDNA gene for expression in Aspergillus: An endoglucanase-producing yeast colony was inoculated into 20 ml YPD broth in a 50 ml glass test tube. The tube was shaken for 2 days at 30°C. The cells were harvested
I by centrifugation for 10 min. at 3000 rpm.
DNA was isolated according to WO 94/14953 and dissolved in 50 /il water. The DNA was transformed into E. coli by standard procedures. Flasmid DNA was isolated from E. coli using standard procedures, and analyzed by restriction enzyme analysis. The cDNA insert was excised using appropriate restriction enzymes and ligated into an Aspergillus expression vector.
Transformation of Aspergillus oryzae or Aspergillus niger
Protoplasts may be prepared as described in WO 95/0204 2.. p. 16, line 21 - page 17, line 12, which is hereby incorporated by reference.
100 ^1 of protoplast suspension is mixed with 5-25 ug of the appropriate DNA in 10 ,ul of STC (1.2 M sorbitol, 10 my. Tris-HCl, pK = 7.5, 10 mM CaCl-,) . Protoplasts are mixed with p3SR2 (an A. nidulans amdS gene carrying plasmid). The mixture is left at room temperature for 25 minutes. 0.2 ml of 60% PEG 4000 (BDH 29576), 10 ir.M CaCl, and 10 muM Tris-HCl, pH 7.5 is added and carefully mixed (twice) and. finally 0.85 ml of the same solution is added and carefully mixed. The mixture is left at room temperature . f-cy^ 2 5 minutes, spun at 2 500 g for 15 minutes and the pellet is resuspended in 2 ml of 1.2 M sorbitol. After one more sedimentation the protoplasts are spread

on mininial plates (Cove, Biochem. Biophys. Acta 113 (1966) 51-56) containing 1.0 M sucrose, pH 7.0, 10 mM acetamide as nitrogen source and 20 mM CsCl to inhibit background growth. After incubation for 4-7 days at 37=C spores are picked and spread for single colonies. This procedure is repeated and spores of a single colony after the second reisolation is stored as a defined transformant.
Test of A. oryzae transformants
Each of the transformants were inoculated in 10 ml YPM and propagated. After 2-5 days of incubation at 37°C, 10 ml supernatant was removed. The endoglucanase activity was identified by AZCL HE cellulose as described above.
Hybridization conditions (to be used in evaluating
property ii) of the DN'A construct of the invention): Suirable ccnditicns for determining hybridizarion between a nucleotide probe and a homologous DNA or RNA sequence involves presoaki.ng of the filter containing the DN.-. fragments or RN.A. to hybridize in 5 x SSC (standard saline citrate) for 10 min, and prehybridization of the filter in a solution of 5 x SSC (Sambrook et al. 1939), 5 x Denhardt's solution (Sambrook et al. 1989), 0:5 % SDS and 100 ^g/ml of denatured sonicated salmon sperm DNA (Sambrook et al. 19S9), followed by hybridization in the same solution containing a random-primed (Feinberg, A. ?. and Vogelstein, B. (19o3) Anal. Biochem. 132:6-13), 'P-dCTP-labeled (specific activity > 1 x lO' cpm/^g ) probe for 12 hours at ca. 45»C. The filter is then washed two times for 30 minutes in 2 x SSC, 0.5 % SDS at preferably not higher than 50°C, more preferably not higher than 55°C, more preferably not higher than 60°C, more preferably not higher than 65°C, even more preferably not "higher than 70°C, especially not higher than 75s€-,— The nucleotide probe to be used in the hybridization is the DNA sequence corresponding to the endoglucanase

encoding part of the DNA sequence shown in SEQ ID No. l, 4, 6, 8, 10, 12, or 16, resepctively, and/or the DNA sequence obtainable from the plasmid in S. cerevisiae, DSM 9770, DSM 10082, DSM 10030, DSM 10081, E. coli, DSM 10512, DSM 10511, DSM 10571 or DSM 10576, respectively.
Immunological cross-reactivity: Antibodies tc be used in determining imrriunological cross-reactivity may be prepared by use of a purified cellulase. More specifically, antiserum against the cellulase of the invention may be raised by immunizing rabbits (or other rodents) according to the procedure described by N. Axelsen ejt a 1. in: A Manual of Quantitative Immunoelec¬trophoresis , Blackvell Scientific Publications, 1973, Chapter 2 3, or A. Johnstone and R. Thorpe, Immunochemistrv in Practice. Blackwell Scientific Publications, 1932 fr.cre specifically pp. 27-21). Purified im-unociooulins may be obtained from the anti-sera, for example by salt precipitation ( (NH/) , SOj) , followed by dialysis and ion exchange c.hromiatography, e.g. on DEAE-Sephadex. IiruEunochemicai characterization cf proteins may be done either by Gutcherlony double-diffusion analysis (O. Cuchterlony in: Handbook of Experimental I~munologv (D.M. Weir, Ed.), Blackwell Scientific Publications, 1967, pp. 655-706), by crossed immunoelectrophoresis (N. .Axelsen et al. , supra, Chapters 3 and 4), or by rocket immiunoelectrophoresis (N. Axelsen et al., Chapter 2).
Media
YPD: 10 g yeast extract, 20 g peptone, H^O to 900 m^l. Autoclaved, 100 ml 20% glucose (sterile filtered) added.
YPM: 10 g yeast extract, 20 g peptone, H^O to 900 ml. Autoclaved-,—;-ao ml 20% maltodextrin (sterile filtered) added.

10 X Basal salt: /5 g yeast nitrogen base, 113 g succinic acid, 63 g NaOH, H,0 ad 1000 ml, sterile filtered.
SC-\JPJ\: 100 ml 10 X Basal salt, 28 ml 20% casamino acids 'j without vitamins, 10 ml 1% tryptophan, H^O ad 900 ml,
autoclaved, 3.6 mi 5% threonine and 100 ml 20% glucose or 20% galactose added.
SC-URA agar: SC-URA, 20 g/1 agar added.
PD agar: 39g potato dextrose agar, DIFCO 0013; add deionized water up to 1000ml; autoclave (12l"C for 15-20 min) .
PC agar: Potatoes and carrots (grinded, 20 g of each) and water, added up to IGOGm.l, are boiled for 1 hr; agar (20g/l of Merck 16:4); autoclave (12l"C for 20 m.in)
PC liquid broth: as PC agar but without the Agar
?D liquid bro~h: 24g po~ax.o dextrose broth, Difco 0545, deionized water up to lOOOml; autoclave (121'~C for 15-20 min)
PC and PD liquid broth with cellulose: add 30 g Solcafloc
(Dicacel available from, Dicalite-Eurcpe-Nord, 9000 Gent, Belgium) per lOOOmI
PB-9 liquid broth: 12 g Rofec (Roquette 101-0441) and 24 g glucose are added to lOOOmI water; pH is adjusted to 5.5; 5ml mineral oil and 5 g CaCo, are added per lOOOr.l. autoclave (121'C for 40 min)
YPG liquid broth: 4g yeast extract (Difco 0127), Ig KK^PO, (Merck4873), 0.5g MgS04.7H20 Merck 5886, 15g Dextrose, Roquette 101-0441, 0.1ml Pluronic (101-3088); deionized water up to lOOOml; autoclave (20min at 121°C)

Dilute salt solution (DS): Make up two stock solutions:
P-stock: 13.61g KH.POj,- 13.21g (NHJ2P0j, 17.4 2g KH,POj;
deionized water up to 100ml
Ca/Mg stock: 7.35g CaCl,, 2H2O, 10.17g MgCl,, 6H_,0,
deionized water up to lOOml; pH adjusted to 7.0;
autoclaving (121'C; 2 0min)
Mix O..51T1I P-stock with 0.1ml Ca/Mg stock
add deionized water up to 1000ml
AZCL HE cellulose (Megazyme, Australia).
Uses
During washing and wearing, dyestuff from dyed fabrics cr garment will conventionally bleed from the fabric which then looks faded and worn. Removal of surface fibers from the fabric will partly restore the original colours and looks cf the fabric. By the term "colour clarification", as used herein, is meant the partly restoration cf the initial colours of fabric cr garment throughout m.ultiple washing cycles.
The term "de-pilling" denotes removing of pills fro- the fabric surface.
The term "soaking liquor" denotes an aqueous liquor in which laundry may be immersed prior to being subjected to a conventional washing proccoc. The soaking liquor may contain one or m.ore ingredients conventionally used in a washing or laundering process.
The term, "washing liquor" denotes an aqueous liquor in which laundry is subjected to a washi.ng process, i.e. usually a combined chemical and mechanical action either manually or in a washing machine. Conventionally, the

washing liquor is an aqueous solution of a powder or liquid detergent composition.
The term "rinsing liquor" denotes an aqueous liquor in which laundry is immersed and treated, conventionally immediately after being subjected to a washing process, in order to rinse the laundry, i.e. essentially remove the detergent solution from the laundry. The rinsing liquor may contain a fabric conditioning or softening composition.
The laundry subjected to the method of the present inven¬tion may be conventional washable laundry. Preferably, the major part of the laundry is sewn or unsewn fabrics, including knits, wover.s,_ denims, yarns, and toweling,' made from cotton, cotton blends or natural or manmade cellulosics (e.g. originating frr- xylan-containing cel¬lulose fibers s_ch as from wood pulp) or blends rhereoi. Examples of blends are blends of cotton or rayon/viscose with one or -ore companion material such as wool, syn¬thetic fibers (e.g. pclyamide fibers, acrylic fibers, polyester fibers, polyvinyl alcohol fibers, polyvinyl chloride fibers, pclyvinylidene chloride fibers, polyurethane fibers, polyurea fibers, aramid fibers), and cellulose-containing fibers (e.g. rayon/viscose, ramie, flax/linen, jute, cellulose acetate fibers, lyocell).
Detergent Compositions
According to one aspect of the present invention, the present enaoglucanases m.ay typically be componenrs of a detergent composition. As such, rhey may be included in the detergent comiposition in the form of a non-dusting granulate, a stabilized JJ-quid, or protected enzymes. Non-dusting granulates may be produced, e.g., as dis¬closed in US 4,106,991 and 4,661,452 (both to Novo

Industri A/S) and may optionally be coated by methods known in the art. Examples of waxy coating materials are poly(ethylene oxide) products (polyethyleneglycol, PEG) with mean molecular weights of 1000 to 20000; ethoxylated nonylphenols having from 16 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon aroms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglycerides of fatty acids. Examples of film-forming coating materials suitable for application by fluid bed techniques are given in patent GB 1483591. Liquid enzyme preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Other enzyme stabilizers are well known in the art. Protected enzymes may be prepared according to the method disclosed in EP
The detergent composition of the invention may be in any convenient form, e.g. as powder, granules, paste or liquid. .-. liquid detergent may be aqueous, typically con¬taining up to 70% water and 0-30% organic solvent, or nonaqueous.
The detergent composition comprises one or m.ore surf¬actants, each of which may be anionic, nonionic, cationic, or zwittericnic. The detergent will usually contain 0-50% of anionic surfactant such as linear alkylbenzenesulfonate (LAS), alpha-olefinsulfonate (AOS), alkyl sulfate (fatry alcohol sulfate) (AS), alcohol ethoxysulfate (.AEOS or AES) , secondary alkanesulfonates (SAS), alpha-sulfo fatty acid methyl esters, alkyl- or alkenylsuccinic acid, or soap. It may also contain 0-40% of nonionic surfaxrtant such as alcohol ethoxylate (AEO or AE), carboxylated alcohol ethoxylates, nonyiphenol ethoxylate, alkylpolyglycoside, alkyldimethylamine oxide.

ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, or polyhydroxy alkyl fatty acid amide (e.g. as described in WO 92/06154).
The detergent composition may additionally comprise one or more other enzymes such as amylase, lipase, cutinase, protease, peroxidase, and oxidase, e.g. laccase.
The detergent may contain 1-65% of a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, citrate, nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTMPA), alkyl- or =Ikenylsuccinic acid, soluble silicates or layered silicates (e.g. SKS-6 from.. Hoechst) . The detergent may also be unbuilt, i.e. essentially free of detergent JDUiider.
The detergent may comprise one or more polymers. Examiples are carboxymethylcellulose (CMC), poly(vinylpyrrolidone) (?'/?) , polyethyleneglycol (PEG; , pcly(vir.yl alcohol) (PVA), polycarboxylates such as pclyacrylates, maleic/acrylic acid copolymers and lauryl methacrylate/acrylic acid 'copolymers.
The detergent m.ay contain a bleaching system which may comprise a H;0; source such as perborate or percarbonare which -ay be combined with a peracid-forming bleach activator such as tetraacetylethylenediamine (TAED) or nonajnoyloxybenzenesulfonate (NOBS) . Alternatively, the bleaching system may comprise peroxyacids of, e.g., the amide, imide, or sulfone type.
The enzymes of the detergent composition of the invention may be stabilized using convfiai_i.onal stabilizing agents, e.g. a polyol such as propylene glycol or glycerol, a sugar or sugar alcohol, lactic acid, boric acid, or a

boric acid derivative such as, e.g., an aromatic borate ester, and the composition may be formulated as described in, e.g., WO 92/19709 and WO 92/19708.
The detergent may also contain other conventional deter¬gent ingredients such as, e.g., fabric conditioners in¬cluding clays, foam boosters, suds suppressors, anti-corrosion agents, soil-suspending agents, anti-soil-redeposit ion agents, dyes, bactericides, optical brighteners, or perfume.
The pH (measured in aqueous solution at use concentra¬tion) will usually be neutral or alkaline, e.g. in the ranee of 7—li.
Particular forms of detergent compositions within the scope of -he invention include:
1} .A deterge.nt composition formulated as a granulate having a bulk density of at least 600 g/l comprising

00 01 - 0.1"

2) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising
3) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising

detergent liquid comprising a liquid nonionic surfactant such as, e.g., linear alkoxylated primary alcohol, a builder system (e.g. phosphate), enzyme and alkali The detergent may also corr.prise anionic surfactant and/or a bleach system.
The endoglucanase may be incorporated in concentrations conventionally employed in detergents. It is at present contemplated that, in the laundry composition of the in¬vention, the cellulase may be added in an amount corre¬sponding to 0.0001-10 mg (calculated as pure enzyme protein) of cellulase per liter of wash liquor.
According to yet another aspect of the present invention, e.ndoglucanase may typically be a component of a conditioning or softener composition. Examples of softener co-positions are disclosed in e.g. EP 0
textile applications
In another embodiment, the present invention relates to use of the endoglucanase of the invention in the bio-polishing process. Bio-Polishing is a specific treatment of the yarn surface which improves fabric quality with respect to handle and appearance without loss of fabric wet ability. The most important effects of Bio-Polishing can be characterized by less fuzz and pilling, increased gloss/luster, improved fabric handle, increased durable softness and altered water absorbency. Bio-Polishing usually takes place in the wet processing of the manufacture of knitted and woven fabrics. Wet processing comprises such steps as e.g. desisting, scouring, bleaching, washing, dying/printing and finishing. During each of these steps, the fabric is more or less subjected to mechanical action. In general, after the textiles have

been knitted or woven, the fabric proceeds to a desizing stage, followed by a scouring stage, etc. Desizing is the act of removing size from textiles. Prior to weaving on mechanical looms, warp yarns are often coated with size starch or starch derivatives in order to increase their tensile strength. After weaving, the size coating must be removed before further processing the fabric in order to ensure a homogeneous and wash-proof result. It is known that in order to achieve the effects of Bio-Polishing, a combination of cellulytic and mechanical action is required. It is also known that "super-softness" is achievable when the treatment with a cellulase is combined with a conventional treatment with softening agents. It is contemplated that use of the endoglucanase of . the invention for bio-polishing of -cellulosic fabrics is advantageous, e.g. a more thorough polishing can be achieved. Bio-polishing may be obtained cy applying zhe -ernod described e.g. in WO S3/'2027£.
Stone-washing
It is known to provide a "stone-washed" look (localized abrasion of zhe colour) in dyed fabric, especially in denim fabric or jeans, either by washing the denim or jeans made from such fabric in the presence of pumice stones to provide the desired localized lightening of the colour of the fabric or by treating the fabric enzymatic ally, in particular with cellulytic enzymes. The treatment with an endoglucanase of the present invention may be carried out either alone such as disclosed in US 4,832,864, together with a smaller amount of pumice than required in the traditional process, or together with prelate such as disclosed in WO 95/09225.

Pulp and paper applications
In the papermaking pulp industry, the endoglucanase of the present invention may be applied advantageously e.g. as follows:
- For debarking: pretreatraent with the endoglucanase may degrade the cambium layer prior to debarking in mechanical drums resulting in advantageous energy savings.
- For defibration: treatment of a material containing cellulosic fibers with the endoglucanase prior to refin¬ing or beating may result in reduction of the energy consumption due to the hydrolysing effect of the cellulase on the interfere surfaces. Use of the endoglucanase may result in improved energy savings as compared to the use of known enzymes, since i~ is believed that the enzyme composition of the invention may possess a higher ability to penetrate fibre walls.
- For fibre modification, i.e. improvemenr of fibre prop¬erties where partial hydrolysis across the fibre wall is needed which requires deeper penetrating enzymes (e.g. in order to make coarse fibers more flexible). Deep treatment of fibers has so far not been possible for high yield pulps e.g. mechanical pulps or mixtures of recycled pulps. This has been ascribed to the nature of the fibre wall structure that prevents the passage of enzyme molecules due to physical restriction of the pore matrix of the fibre wall. It is contemplated that the present endoglucanase is capable of penetrating into the fibre wall.
- For drainage improvement. The — papermaking pulps may be improved by treatment of the pulp with hydrolysing enzymes, e.g. cellulose. Use of

the present endoglucanase may be more effective, e.g. result in a higher degree of loosening bundles of strongly hydrated micro-fibrils in the fines fraction (consisting of fibre debris) that limits the rate of drainage by blocking hollow spaces between fibers and in the wire mesh of the paper machine. The Canadian standard freeness (CSF) increases and the Schopper-Riegler drainage index decreases when pulp in subjected to cellulase treatment, see e.g. US patent 4,923,565; TAPPI T227, SCAN 019:65.ence.
- For inter fibre bonding. Hydrolytic enzymes are applied
in the manufacture of papermaking pulps for improving the
inter fibre bending. The enzymes rinse the fibre surfaces
for impurities e.g. cellulosic debris, thus enhancing the
area of exposed cellulose with attachment to the fibre
wall, thus improving the fibre-to-fibre hydrogen binding
capaci'cv. This -recess is also referred tr as
dehcrnification. Paper and board produced wirh a cellulase containing enzyme preparation r.ay have an improved strength or a reduced grammage,, a smoother surface and an improved printability.
- For enzymatic deinking. Partial hydrolysis of recycled
paper during or upon pulping by use of hydrolysing
enzymes such as cellulases are known to facilitate the
removal and agglomeration of ink particles. Use of the
present endoglucanse may give a more effective loosening
of ink from the surface structure due to a better pen¬
etration of the enzyme molecules into the fibrillar
matrix of the fibre wall, thus softening the surface
whereby ink particles are effectively loosened. The
agglomeration of loosened ink particles are also
improved, due to a more efficient hydrolysis of cellulo-
sic fragments found attached to ink particles or-i-ginating
from the fibres.

The treatment of lignocellulosic pulp may, e.g., be performed as described in WO 91/14819, WO 91/14822, WO 92/17573 and WO 92/18688.
Degradation of plant material
In yet another embodiment, the present invention relates to use of the endoglucanase and/or enzyme preparation according to the invention for degradation of plant material e.g. cell walls.
It is contemplated that the novel endoglucanase and/or enzyme preparation of the invention is useful in the pre¬paration of wine, fruit or vegetable juice in order to increase yield. Endoglucanases according to the invention may also be applied for enzymatic hydrolysis of various plant cell-wall derived materials cr waste materials, e.g. agricultural residues such as wheat-straw, corn cobs, whole corn plants, nut shells, grass, vegetable hulls, bean .hulls, spent grains, sugar beet pulp, and the like. The plant marerial may be degraded in order to improve different kinds of processing, facilitate purification or extraction of other components like purification of beta-glucan or beta-glucan oligomers from cereals, improve the feed value, decrease the water binding capacity, improve the degradability in waste v.'ater plants, improve the conversion of e.g. grass and corn to ensilage, etc.
The following examples illustrate the invention

EXAMPLE 1
Cellulytic enzymes from 4 fungi, belonging to 3 families under two orders within the Ascomycetes were detected by expression cloning; corresponding DNA sequences were determined; the enzymes heterologously expressed, and produced by liquid fermentation, characterized and demonstrated to give good performance in colour clarifi¬cation assays.
Isolate CBS 117.65, CBS 478.94, NRRL 8126, and ATCC 10523 were grown in shake flask cultures on cellulose enriched potato dextrose broth, incubated for 5 days at 2 6"c (shaking conditions, 150 rpm).
A. Cloning and expression of an endoglucanase from Myceliophthora tbermophila, Acremonlum sp.. and Thielavia terrssTiris and Volutella collstotrichcldes
mRNA was isolated from Myceliophthora thermophila, Acremcnium sp., Thielavia terrestris and"'/olu tell a colletotrichoides, respectively, crcwTi in a cellulose-containing fermentation medium with agitation to ensure sufficient aeration. Mycelia were harvested after 3-5 days' growth, immediately frozen in liquid nitrogen and stored at -80°C. Libraries from y.yceliophthora thermophila, Acreiuohium sp., Thielavia terrestris and Volutella colletotrichoides, respectively, each consisting of approx. 10* individual clones were constructed i.n E. coli as described with a vector background of 1%.
Plasmid DNA from some of the pools from each library was transformed into yeast, and 50-100 plates containing 250-400 yeast colonies were obtained—^om each pool.
Endoglucanase-positive colonies were identified and

isolated on SC-agar plates with the AZCL HE cellulose assay. cDNA inserts were amplified directly from the yeast colonies and characterized as described in the Materials and Methods section above.
The DNA sequence of the cDNA encoding the endoglucanase from Myceliophthora thermophila is shown in SEQ ID No. l and the corresponding amino acid sequence is also shown in SEQ ID No. 1. The cDNA is obtainable from the plasmid in DSM 9770.
The DNA sequence of the cDNA encoding the endoglucanase from Acremonium sp. is shown in SEQ ID No. 4 and the corresponding amino acid sequence is shown in SEQ ID No. 5. The cDN.A is obtainable from^ the pl3sm,id in DSM 10082.
The DNA sequence of the cDKA encoding the endoglucanase rrorT; __\nej.5V'i3 ^err^s^rxs is snovn m ^r-v XLJ HO. O anc the corresponding amino acid sequence is shown in SEQ ID No. 9. The cDNA is obtainable from the plasmid in DSM 10031.'
The DNA sequence of the cDNA encoding the endoglucanase from Volutella colletotrichoides is shown in SEQ ID No. 16 and the corresponding amino acid sequence is shown in SEQ ID No. 17. The cDNA is obtainable from the plasmid in DSM 10571.
Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanases in Aspergillus, the D.N'A was digested with appropriate restriction enzymes, size fractionated on gel, and a fragment corresponding to the endoglucanase gene from
Hyceliophthora thermophila, Acrewdnium sp., ThTWlavia
terrestris and Volutella colletotrichoides, respectively, was purified. The genes were subsequently ligated to

pHD414, digested with appropriate restriction enzymes, resulting in the plasmids pA2Cl93, pA2C357, pA2C385 and pA2C488, respectively.
After amplification of the DNA in E. coli the plasmids were transformed into Aspergillus oryzae as described above.
Test of A. oryzae transformants
Each of the transformants were tested for endoglucanase activity as described above. Some of the transformants had endoglucanase activity which was significantly larger than the Aspergillus oryzae background. This demonstrates efficient expression of the eridoglucanases in Aspergillus oryzae. The transformants with the highest endoglucanase activity were selected and inoculated in a 500 ml shake flask vith YPM r.edia. .After 3-5 days of fer-entation with sufficient agira~io- to ensure good aerarion, "he culture broth was centrifuged for 10 minutes at 20C0 g and the suoernatant recovered.
B. Determination of endoglucanase activity
The cellulytic activity of the endoglucanase may be determined relative to an analytical standard and expressed in the unit S-CEVU.
Cellulytic enzymes hydrolyse CMC, thereby decreasing the viscosity of the incubation mixture. The resulting reduction in viscosity may be determined by a vibration viscosimeter (e.g. MIVI 3000 from Sofraser, France).
Determination of the cellulytic activity, measured in terms of S-CEVU, may be determined according to the anal^Lsis method AF 301.1 which is available fro- the-Applicant upon request.

The S-CEV^U assay quantifies the amount of catalytic activity present in the sample by measuring the ability of the sample to reduce the viscosity of a solution of carboxy-methylcellulose (CMC). The assay is carried out at 40°C, pH 7.5 using a relative enzyme standard for reducing the viscosity of the CMC substrate.
Assay for determination of endoglucanase activity in terms of SAVI units using phosphoric-acid swollen cellu¬lose (PASC):
Definition:
1 SAVI-U is the amount of enzyme which form.s an amount of
reducing carbohydrates equivalent to 1 umol of glucose
per minute.
Assay condition:
Enzyme solution: 0,5 ml
4 g/1 PASC in 0,1 M Buffer: 2.0 ml
2 0 m,i.n, 40 ■ °C
Sensitivity:
Max 0.1 5AVIU/ml = approx. 1 S-CE\nj/mi (CMC viscosity)
Min 0.01 SAVIU/ml = approx. 0.1 S-CE'vTO/ml
Determination of for.~ation of reducing sugars:
The reducing groups assay was performed according to Lever, M. A new reaction for colormetric determination of carbohydrates. Anal. Biochem, 1972. Vol 47 (273-279). Reagent mixture was prepared by mixing 1,5 gram, p-hydroxybenzoic-acide hydracide (PHBAH) with 5 gram sodium tartrate in IGO ml 2 % sodium hydroxide.
Substrate:
PASC stock solution was prepared the following way using

ice cold acetone and phosphoric acid. 5 gram of cellulose (Avicel*) was moistered with water, and 150 ml ice cold 85% ortho-phosphoric acid was added. The mixture was placed in ice bath under slow stirring for 1 hr. Then 100 ml ice cold acetone was added with stirring. The slurry was transferred to a Buchner filter with pyrex sintered disc number 3 and then washed three times with 100 ml ice cold acetone, and sucked as dry as possible after each wash. Finally, the filter cake was washed twice with 500 ml water, sucked as dry as possible after each wash. The PASC was mixed with deionized water to a total volume of 300 ml, blended to homogeneity (using the Ultra Turrax Homogenizer) and stored in refrigerator (up to one month).
Substrate equilibration with buffer: 20 gram phosphoric acid swollen cellulose PASC stock solution was centrifu-ged for 20 r.in ar 5CC0 rpm. , rhe superna-an- was poured of; the sediment was resuspended in 30 ml of buffer and centrifuged for 20 -in. at 5000 rpm., the supernatant was poured of, and the sediment was resu^spended i.n buffer io a total of 60 g corresponding to a subsrrate concentra¬tion of 5 g cellulose/litre.
Buffer for pH S,5 determination: 0.1 M Barbital. Buffer for pH 10 determination: 0.1 M Glycine.
1. Dilution of enzyme samples
The enzyme solution is diluted in the same buffer as the substrate.
2. Enzyme reaction
The substrate in buffer solution is preheated for 5 min.
at 40*C (2 ml).
Then the enzyme solution (diluted to between 0.2 and 1 S-CEVU/ml) 0,5 ml is added and mixed for 5 sec. Enzymes

blanks are obtained by adding the stop reagent before enzyme solution. Incubate for 20 min. at 40 'c. The reaction is stopped by adding 0.5 ml 2% NaOH solution and mixing for 5 sec.
The samples are centrifuged for 20 min. at 5000 rpm. 1 ml supernatant is mixed with 0.5 ml PHBAH reagent and boiled for 10 min. The test tubes are cooled in a ice water bath.
3. Determination of reducing end groups:
The absorbancy at 410 nm is measured usi.ng a spectrophotom.eter. Blanks are prepared by adding sodium hydroxide before adding enzyme solution. k standard glucose curve was obtained by using glucose ccncentraticns of 5, 10, 15 and 25 mg/1 in the sane buffer and addin^ ?HH.-vH reacent before boilinc. The release of reducing glucose equivalent is calculated using this standard curve.
4. Calculation cf catalytic activity:
Measure absorbance at 410 nm
1) Standard curve
(Glucose) - (HpO) vs concentration of glucose
2) Enzyme sample
(Sample) - (Blank)
Calculate glucose concentration according to a standard

Activity (SAVIU/ml):
X (mg glucose/1) * Dilution
ISO.16 (MW of glucose) * 20 (min)
C. Purification and characterisation of the endoglucanase from M. thermophila
Aspergillus oryzae transformed with pA2C193 was grown on YPM medium for 4 days. The liquid was then centrifuged and sterile filtered.
-The.sample was.concentrated by ultrafilrration -on AMICON cells using a DOW membrane GR61P? with cut-off 20 kD. The Uf-concentrate was analyzed for S-CE'v'U/ml and SaviL'/mi
'—■ ^ ^ '^- V* X ;

UF-concentrate \ S-
i
; CE'.'TF/ml Saviu/ml
9.25 ml 1 570
1 41
Purification:
2 ml of the UF-concentrate was diluted 5 times to lower the ionic strength and filtered through 0.2 2 um disk filter. This sample was applied to a Mono Q* HR5/5 Phar¬macia column, equilibrated with 50 mM Tris/HCl buffer. pH 7.5, (buffer A) and a flow of 1 ml/min. After wash to baseline, with buffer h, the colum.n was eluted with a Tris/HCl buffer, pK 7.5, containing 1 M IJaCl (buffer B) , the elation gradient was 0-50% buffer B in 1 hour.
After 36 min. a peak complex showed up, 1 ml fractions were picked up and the first 10 fractions showed cellulase activity on CMC/Agarose/congo-red plates.

These fractions were pooled and concentrated, by ultrafiltration on AMICON cells using a DOW membrane GR61PP with cut-off 20 kD, to 3 ml.
This sample was applied to a HiLoad 26/60 Superdex 75"" prep grade Pharmacia column, equilibrated with 100 mM Na-Acetate buffer, pH 6.35, and a 1 ml/min flow.
After 82 min. a peak showed up, 1 ml fractions were picked up and.the first 10 fractions showed cellulase activity on CMC/Agarose/congo-red plates.
These fractions were pooled and the following results were obtained: A;so='^- 15, A2so/A26o=l-62 Mw(5DS:=2 2 kD pl=3.5-5
Purity on SDS-PAGE =100% S-CEVU/ml=28.5 S-CEVU/A:so=lS3 S-CEV~u/mg=4 3 6
Extinction ccefficient=54880 (calculated) •Mw(calculated)=22 kD
The Extinction coefficient is based on the content of tyrosine, tryptophane and cystein calculated fro- the sequence cf the enclcsad SEQ ID No. 1 (the amino acid sequence). SDS-Page was performed on NOVEX Pre-Cast Gels 4-20% Tris-Glycine Gel 1.0 mm x 10 Well
lEF was performed on Pharmacia FAGplate pH 3.5 - 9.5, the activity was visualized by CMC-Congored overlaying.
DPtPrmination of K^, & k^..,:
k„ and k„, was determined in the same manner as the
determination of SAVI Units at pH 8.5 with a substrate

concentration up to 8 g/1.
The following results were obtained:
k^.i 3 8 per sec.
>v„, 5 g/1,
phosporic acid swollen cellulose, pH S
Specific activity on CMC at pH 7.5: 435 S-CEVU per mg protein.
D. Determination of pH and temperature profile of the endoglucanase from M. thermophila
The pK profile was determined at the following conditions:
mixing G. IM Tri-sodium phosphate with O.IM cimc acid. Purified -endoglucanase was diluted to ensure rhe assay response to be within the linear range of the assay. The substrate was a 0.4% suspension of AZCL-HZ-cellulcse (MegaZyme) mixed 1:1 with the citrate/phosphate buffer to a final substrate concentration of 0.2% AZCL-HE-cellulose. 1 ml substrate in Eppendorf® 1.5-i polypropylene tubes were added 10 p.1 of enzyme solution and incubated for 15 minutes in Eppendorf^' temperature controlled Thermomixers before heat-inactivation of enzymes for 2U minutes at 95°C in a separate Thermomixer. The tubes were centrirugea and 200 /il of eacn supernatant was transferred to a well in a 96 well micrctiter plate and OD was measured at 620nm in an ELISA reader (Labsystems Multiskan® MCC/340).
For the pH optiraumi incubations took place an 30°C. For each pH value, three tubes were added enzyme and

incubated before heat-inactivation, whereas one tube (the blank) was added enzyme and heat-inactivated immediately. The mean value of the three incubated samples was calculated and the blank value was substracted.
The following pH profile was determined:

pH

Relative Activity

2.5

:10%

■'10%
3 . 5
4 . 5

5 100% 1
S
6.5 S6%
7 7S%
7.5
8 68%
S. 5 = ■ 3-_/ -t o
9 31%
10 lO-S
It is seen that the endoglucanase has more than 60% activity between pH 4.0 and 8.0 and optimal activity at pH 5.0-6.0. ~
Temperature profile:

The temperature optimum was determined in the same manner at pH 5.5. The temperatures ranged from 30^0 to 80°C. For each temperature three incubations were carried out and the mean calculated. Three blanks were produced by immediate heat-inactivation of enzyme and the mean was subtracted from the incubated sample values.
It is seen that the endoglucanase has optimal activity at 50-70°C.

Temp. {°C) 30 40 50 60 70 80
Relative
Activity 7 4^0 77% 99% 100% 93% 62%
The temperature stability was determined in the same manner at pH 5.5 and 3 0°C, and, further, the enzyme solu¬tions were preneate'.i _or 1 hour at une accuaj. cemperatur^ and cooled on ice. The residual activity is shown below in % of the activitv of a non-oreheated enzvme samnle:

40 50 60 70 80
Relative Activity 95% 84% 92% 86% 24%

E. Color clarification of Myceliophthora cellulase (SEQ ID No. 1) measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers

Apparatus
Liquid volume
Agitation
stirrer
Rinse time
Washing temp
Washing liqour
pH
Washing time
Repetitions
Enzymes
Dosage
Texrile
Drying Evaluation

Terg-o-tometer
100 rol
150 movements/min with vertical
5 min in tapwater 40°
0.05 M phosphate buffer 7.0
3 0 nin 2
Myceliophthora SEQ ID N'o. IB 500 and 2500 S-CEVU/1
2 swatches cf aged clack 100% ccrrcn 5x6 cm (0.9 gram) Tumble dry
The light re-issicn is -easured by a Datacolcr Elrepho Remission spectrophotometer. Rem.issicn is calculated as delta L (Hunter Lab-values) . When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker, and lower L values are obtained.

The sample is compared with a blind sample, i.e. washed without enzyme:
No cellulase 500 ECU/1 2500 ECU/i
0.00 -1.41 -1.3-3.—
Delta L-values compared to blind sample.

The data shows that Mvceliophthora cellulase without CBD gives good color clarification under the conditions tested.
F. Construction of the gene fusions between the endoglucanase from Myceliophthora thermophila and the 43k.D endoglucanase from Humicola insolens
The purpose of the two constructions was to make derivatives of the endoglucanase from M. thermophila with the linker and CBD from the 43kD endoglucanase from H. insolens (disclosed in WO 91/17243). The native endoglucanase from M. thermophila do not have a linker and/or a cellulose binding domain, CBD.
CMl: Construction 1 consist of the endoglucanase from .M. thermophila (225 amino acids) and the 72 C-ter~inai amiinc acids from the H. insolens 43kD endoglucanase.
CM2: Construction 2 consist of the endoglucanase from .M. thermophila (225 amino acids) and the S3 C-terminal amino acids from the H. insolens 43kD endoglucanase.
The 43kD endoglucanase cDNA from H. insolens was cloned into prtD414 in such a way that the endoglucanase gene was transcribed from the Taka-promoter. The resulting plasm.id v;as nam.ed pCaHj4 1S.
In a similar way the cDNA encoding the endoglucanase from M. thermophila was cloned into pHD414 and the resulting plasm.id was named pA2C193.

Primers:
primer 1: 5'-
CGGAGCTCACGTCCAAGAGCGGCTGCTCCCGTCCCTCCAGCAGCACCAGCTCTCCGG
-3'
primer 2: 5'
CCGGAGAGCTGGTGCTGCTGGAGGGACGGGAGCAGCCGCTCTTGGACGTGAGCTCCG
-3 '
primer 3: 5'-
CGGAGCTCACGTCCAAGAGCGGCTGCTCCCGTAACGACGACGGCAACTTCCCTGCCG
-3'
primer 4: 5'-
CGGCAGGGiiJvGTTGCCGTCGTCGTTACGGGAGCAGCCGCTCTTGC-ACGTGAGCTCCG
-3 '
Taka-pro. primer: 5' CAACATCACATCAAGCTCTCC -2'
AMG-term. primer: 5' CCCCATCCTTTAACTATAGCG -3'
The endoglucanase fusions were constructed by the PCR overlap-extension method as described by Higuchi et al. 1988.
Construction 1:
Reaction A: The Polymerase Chain Reaction (PCR) was used
to amplify the fragment of pCaKj418 between primer 1 and
;i>IG-term. primer (the linker and CBD from the 4 3kD
endoglucanase from H.insolens).
Reaction B: PCR amplification of the fragment between
Taka-pro. primer and primer 2 in pA2C193, the
endoglucanase gene from M.thermophila.
Reaction C: The two purified fragments were used in a
third PCR in the presence of the primers flanking the
total region, i.e. Taka-pro. primer and AMG-term. primer.
Cons^ifttction 2 :
The same procedure was used where primer 3 and primer 4

had replaced respectively primer 1 and primer 2.
The fragment amplified in reaction C was purified, digested with restriction enzymes Xba I and BsstE II. The purified digested fragment was ligated into pA2C193 digested with restriction enzymes Xba I and BsstE II.
Competent cells from E. coli strain DH5aF' (New England Biolabs.) were transformed with the ligated plasmid and colonies containing the gene fusion were isolated. The sequence of the cloned part was verified by DNA sequencing.
The sequence of the gene in the two constructs are shown in SEQ ID No. 2A and SEQ ID No. 3A.
Polymerase Chain Reactions were carried out under standard conditiicns, as reccmmended by Perkin-Elmer.
Reaction A and B started with 2 min. at 94°C followed by 20 cycles of {30 sec. at 94-C, 30 sec. at 50°C and 1 min. at 72°C) and end with 4 min. at 72 -C.
Reaction C started with (2 min. at 94=C, 1 min. at 52°C and 2 min. at 72°C), followed by 15 cycles of (30 sec. at 94°C, 30 sec. at 52°C and 90 sec. at 72°C) and end with 4 min. at 72°C.
The two constructs were transformed into Aspergillus ory-zae as described above.
G. Purification and characterisation of cloned cellulases with cellulose binding domains:
The cloned product is recovered after fermentation by separation of the extracellular fluid from the production

organism.
About one gram of cellulase is then highly purified by affinity chromatography using 150 gram of Avicel in a slurry with 20 mm Sodium- phosphate pH 7.5.
The Avicel is mixed with the crude fermentation broth which contain total about 1 gram of cellulase. .After mixing at 4 C for 20 rain the Avicel enzyme is packed into a column with a dimension of 50 times 200 mm about 400 ml total.
The column is washed with the 200 ml buffer, then washed with 0.5 M NaCl in the same buffer until no more protein elutes. Then washed with 500 ml 20 mir. Tris pH 8.5. Finally the pure full length enzyme is eluted with 1% triethylainine pH 11.8.
The eiured enzy~e solution is adjusted ro pH S and concentrated using a Amicon cell unit with a membrane DOW GR61?? (polypropylene with a cut off of 20 KD) to above 5
The purified cellulases were characterised as follow:
Mw pi Molar E.280 S-CEVU per
SDS-PAGE A.280
Myceliophthora

/4.950 13 5

Acremonium
(SEQ ID No.5) 40 kD 5 68.020 185
Thielavia
(SEC ID No.S) 3 5 kD 4.3 52.4 70 ~5

instructions.
Two of the cellulases were blocked, this is due to the N-terminal glutamine which form a pyroglutamate which can not be detected and which block for further sequencing. DSC Differential scanning calometry was done at neutral pH (7.0) using a MicroCalc Inc. MC calorimeter with a constant scan rate and raising the temperature from 20 to 90° at a rate of 90° per hour.
Raising antibody. The cellulases from Myceliophthora, Acremonium and Thielavia were used for raising antibody in rabits. 0.1 mg of the purified cellulase in 0.9 % NaCl solution mixed with Freunds adjuvant immediately prior to injection. The rabits were immunized 10 times with one week interval. The immunoglobulin G fraction (IgG) was purified by a-r.cnium sulfate precipitation (25% saturation), the precipitate was solubiiized in water and then dialyzed extensively against sodium acetate buffer (pH 5.0, 50 m-M) altering with deionized water. .A.fter filtration, the IgG fraction was stabilized with sodium" azide (0.01%) .
Using immunodiffusion in agar plates all three cellulases form a single immunoprecipitate with its homologous antiserum and no precipitate was seen between the 3 cloned cellulases and the sera raised against the other two cellulases.

H-I. Performance of endoglucanase of construction 1 (SEQ ID No. 2) measured in buffer as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers

Apparatus
Liquid volume
Agitation
Rinse time
Washing temp
Water Hardness
Washing liquor
PH
Washing rime
Repetitions
Textile
Drying

Terg-o-tometer
100 ml
150 movements/min (rpm)
5 min in tap water
40°C
1 mM CaCl,
0.05 M phosphate buffer 7 .0
30 min
2
2 swatches of aged black,
100% cotton 5x6 c-
Tumfcle dry

Evaluation:
The light "remission was measured by a Macberh Color Eye 7000 Remission spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn were removed by the cellulase, the surface appeared more bright, and lower L values were obtained.
Results:

2 50 1 1000
Inventive enzyme 0 -1.4 -1. 6
,
The data show that the enzyme of the invention gives very
good'color clarification under the conditions tested

a-II. Performance of cloned endoglucanase from Thielavia terrestris (SEQ ID No.9) in buffer measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers

Apparatus Liquid voluine Agitation
Rinse time
Washing temp
Washing liqour
pH
Washing time
Repetitions
Textile
Terg-o-tometer
100 ml
150 movements/min with vertical
stirrer
10 min in tapwater
40°
0.05 M phosphate buffer.
7.0
30 min
2
2 swatches of aged black cotton 5x6
cm (app. 150 g/m2)
:Lva j.uation
Tumble dry
~oior Elrepho
Remission- spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker and nicer, and lower L values are obtained.
Results

S-CEVU/1 0 50 2 00
Inventive enzyme 0 -0.56±0.10 -1.3210.06
The data show that the cellulase gives good color cLaxification under the conditions tested.

H-III. Performance of endoglucanase of Volutella colletrichoides (SEQ ID No. 17) measured in buffer as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers

Apparatus Liquid volume Agitation
Rinse time
Washing temp
Washing liqour
pH
Washing time
Repetitions
Dosage
Textile
Terg-o-tometer
100 ml
150 movements/min with vertical
stirrer
5 min in tapwater
40°
0.05 M phosphate buffer
7.0
30 min
2
2.5 S-CEVU/ml
2 swatches of aged black 100%
5x5 cm (0.5 gra.~)
Drying : Tumble dry
Evaluation:
The light remission is measured by a Datacoior Eirephc Remission spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker, and lower L values are obtained.
The sample is compared with a blind sample, i.e. washed without enzyme:
No cellulase With cellulase
O.CO -0.57
Delta L remission values compared to blind sample.
The data~"STTows that the Volutella colletrichoides cellulase gives good color clarification under the conditions tested.

H-IV. Performance of cloned cellulases from Thielavia terrestris and Acremonium sp. CBS 4 7 8.94 in high pH heavy duty detergent measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers

Apparatus Liquid volume Agitation
Rinse time Washing temp Washing liqour
Terg-o-toineter
150 ml
150 movements/min with vertical
stirrer
10 min in tapwater
35'C
l.D g/1 US type KDG
(zeolite/soda built, anionic/ncnionic
weight ratio > 2.5)
PH
Hardness
Washing time
Repetitions
Textile
10. 0
1.0 sM CaCl2
■-'.—' T iil^ -
jn -. >:6
2 swatches of aged black co:
Cm (app. 150 g/rri2)
2 swatches of heavy knitted co~zor.
5x6 cm (app. 600 g/in2)
Drying : Tumble dry
Evaluation :
The light remission is measured by a Datacolor Elrepho Remission spectrophotometer. Remission is caiculared as delta L (Hunter Lab-values). When the surface fibrils and fibcLti pruLruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker and nicer, and lower L values are obtained. Different dosages of cloned cellulases from Thielavia terrestris (SEQ ID No. 9) and Acremonium sp. CBS 478.94 (SEQ ID No.5), respectively, (denoted A and B, respectively) were tested.

Jesuits:

S-CEVU/1 0 500 2000
A 0 -2.09 ±0.22 -2 .36 ±0 . 19
B 0 -0. 60 ±0.36 -1.96 ±0. 23
The data show that both cellulases gives good color clarification under the conditions tested.
K-V. Performance of cellulases cloned from Tbielavia terrestris and Acremonium sp. CBS 47S.94, and construction 1 (SEQ ID No.2) measured as removal of surface fibrils and fibers protruding from the yarn of a
textile containing cellulosic fibers

.ppc
Liauid volume
Rinse time Washing temp Hardness
Washing liqour
Washing time
Repetitions
Textile

Terg-o-tometer 150 ml
150 mcvemencs/ir.in virh vertical stirrer
10 min in tapwater 35°C
1.0 mM CaCl2 0.3 4 mM MgC12
2.0 g/1 HDL {neutral, citrate built HDL, with nonionic/anionic weight ra¬tion > 0.5) 7 . 5
3 0 min 2 2 swatches of aged black cotton 5x6
cm (app. I5a g/m2} ,
2 swatches of heavy knitted cotton 4x7 cm (app. 600 g/m2)

Drying : Tumble dry-
Evaluation :
The light remission is measured by a Datacolor Elrepho Remission spectrophotometer. Remission is calculated as delta L (CIE Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker and nicer, and lower L values are obtained. Three different dosages Different dosages of cloned cellulases from Thielavia terrestris (SEQ ID No. 9) and Acremonium sp. CBS 478.94 (SEQ ID No.5) and the construction 1 (SEQ ID No.2), respectively, (denoted A and B and C, respectively) were tested. Results:

s-
CEVU/1 0 100 200 400
^
'^ -3 . 06 ±0. 24 -3. 15
±0. 27 -3.92 ±0.26
B c -1.75 ±0.27 -3 . OS ±0.32 -3.51 ±0.44
C 0 -1.84 ±0.39 -1.70 ±0.47 -2.30 ±0.61
The data show that all cellulases gives very good color clarification under the conditions tested.
I. Application of endoglucanases from Thielavia terre¬stris, Acremonium sp. and construction 1 (SEQ ID No. 2) in denim finishing

Experimental
Apparatus: Liquid volume: Fabric:

Washing machine Wascator FL 120
20 L
1.1 kg denim fabric, 14% oz 100
cotton
Desizing: 10 min, 55°C, pH 7
50 ml Aquazyme 120L 2.5 g/1 Phosphate buffer
Abrasion: 2 hours
pH and temperature varied according to the following table:
Enzyme Activity pH/temp Buffer system
SEQ ID
No. 2 1400 S-CEVU/g 6/55°C 2.5 g/1 phosphate
buffer No. 9 292 S-CEVU/g 5/65"C 1 g/1 citrate buffer No. 5 782 S-CEVU/g 7/45'C 2.5 g/1 phosphate
buffer
Inactivaricn: 15 -in, SO'C
1 g/ sodium carbonate Rinses: Three rinse cycles of 5 min in cold tap water
Evaluation:
Abrasion: The remission' from the fabric was determined at 420 nm using a Ticklish 2000 as a measure of the abrasion level.
The results from the treatment of the denim fabric with different endoglucanases of the invention is shown in the following table:

Enzyme Dosage Trial conditions Abrasion 42 0 mn
Blank 0 S-CEVU/g textile pH 6, 55'C 9 . 96
SEQ ID No. 2 10 S-CEVU/g textile pH 6, 55°C 14 . 37
Blank 0 S-CEVU/g textile pH 5, 65*C 9. 26
SEQ ID No. 9 10 S-CEVU/g textile pH 5, 65°C 16.86
Blank 0 S-CZ\Ti/g textile pH 7, 45*C 9 . 47
SEQ ID
No. 5
1 10 S-CEVU/g pH 7, 45*C 14 . 03
1 i
All rested cellulases show excellent performance in denim finishing, although each enz>Tne is unique in its own way. When applying the enzyme corresponding ro SEQ ID No. 2 for denim finishing it is possible to reach a high abrasion level with a minimum of strength loss. When treating denim with the enzyme corresponding to SEQ ilD No. 9, a very high wash down can be reached which leaves the fabric with an almost bleached appearance. Denim finishing with the enzyme corresponding to SEQ ID No. 5 gives a high abrasion level at a low temperature optimum which makes it possible to reduce the processing tempera¬ture and save energy.

Lyocell fibers which are sold under the trade name Te'

are spun from wood pulp cellulose in a more environmentally friendly waterbased solvent than is the case for normal viscose production). However, the fibers have a tendency to fibrillate when they are processed into textiles which is seen on the surface and denoted "fuzz". By using cellulases it is possible to permanently remove the exposed and fuzzy fibers and significantly improve the look of the finished fabric, the treatment generally known as Biopolishing. The endoglucanases of the present invention are especially suited for the removal of Lyocell surface fibers.
MATERIALS AND METHODS
The textile substrate was either 100 % woven or different kinds of jersey knitted dark blue Tencel. The dark -colour and jersey knit was preferred in order to enhance the visual effects which simplifed the evaluation. A woven 70/3C Tencel/Kayon blend was also used to a lesser extent.
The assays were either performed in 200 ml scale using a Launder-c-meter or in the 20 1 scale using a Wascator. The treatment time was 6Cmin at 55' C in Wascator and SO-SO min in LOM. The buffer was 2 g/i sodium acetate adjusted to pH 5 with acetic acid. The fabric to liquid ratio was 1:10 but in the Launder-o-meter 20 steel balls with a diameter of 14 mm (11 g each) was used to obtain sufficient mechanical abrasion. The biopolishing was immediately followed by inactivation using 2 g/lit sodium carbonate at 80° C for 15 min followed by rinsing in cold water.
The results were evaluated using a fuzz note scale from 1 - 5 were 1 is the fibrillated look of the starting material and 5 is a high quality look with no visible fibers on the surface. Since the performance of endocellulases is specific towards a'^snxface treatment the weightloss is below 2 % and is therefore not included in the evaluation. Two cellulases were evaluated: the

cellulases cloned from Acremonium sp. (SEQ ID No. 5) and from Thielavia terrestris (SEQ ID No. 9).
The two cellulases are able to defibrillate both Tencel and Tencel blended fabrics. By using an endoglucanase of the invention, only small fibrils are removed rather than whole fibers such as is the case when using acid cellulase mixtures from Trichoderma. The strength loss of the treated fabric is threrefore kept at a minimum when using endoglucanases of the present invention.
The following dosages gave a superior defibrillation, i.e. fuzz note 4 or above:
15 S-CEVU/g fabric of cellulase from Acrerao.iium sp (SEQ ID No. 5); and
10 S-CEVU/g fabric of cellulase from Thelavia terrestris (SEQ ID No.S).
EXAMPLE 2
A new cellulytic enzyme was by expression cloning as well as by PCR cloning detected to be produced by a plant pat¬hogen, isolated from soy bean seeds and identified as Ma-crophomina phaseolina.
Production of biomass for PCR and expression cloning procedures:
Isolate CBS 281.96 was grown in shake flask cultures on I cellulose enriched potato dextrose broth, incubated for 5 days at 260C (shaking conditions: 150 rpm).
A. Cloning and expression of an endoglucanase from Macrophomina phaseolina
mRNA was isolated from Macrophomina phaseolina, grown in a cellulose-containing fermentation medium with agitation

to ensure sufficient aeration. Mycelia were harvested after 3-5 days' growth, immediately frozen in liquid nitrogen and stored at -30°C. A library from Macrophomina phaseolina, consisting of approx. 10" individual clones was constructed in E. coli as described with a vector background of 1%.
Plasmid DNA from some of the pools was transformed into yeast, and 50-100 plates containing 250-400 yeast colonies were obtained from each pool.
Endoglucanase-positive colonies were identified and isolated on SC-agar plates with the AZCL HE cellulose assay. cDNA inserts were amplified directly from the yeast colonies and characterized as described in the Materials and Methods section above. The D^ The cDNA' is obtainable from rhe plasmid in DSM 10512.
Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanse in Aspergillus, the DNA was digested with appropriate restriction enzymes, size fractionated on gel, and a fragment corresponding to the endoglucanase gene was purified. The gene was subsequently ligated to pHD414, digested with appropriate restriction enzymes, resulting in the plasmid pA2C477.
After amplification of the DNA in E. coli the plas-.id was transformed into Aspergillus oryzaa as described above.
Screening of the cDNA library by hybridization and characterization of the positive clones. Approximately

6000 colony forming units (c.f.u.) from the Macrophomina phaseolina cDNA library in E. coli was screened by colony hybridization using a random-primed "P-labeled PCR product from M. phaseolina as probe. The PCR product was generated as described in the Materials and methods section. The positive cDNA clones were characterized by sequencing the ends of the cDNA inserts, and by determining the nucleotide seuence of the longest cDNA from both strands. The DNA sequence of the cDNA encoding the endoglucanase is shown in SEQ ID No. 10 and the corresponding amino acid sequence is shown in SEQ ID No. 11.
B. Construction of gene fusion between the endoglucanase from Macrophomina phaseolina and the 4 3 k.D endoglucanase
from Humicola insolens
One construction was prepared in order to make a derivative of the endoglucanase from M. phaseolina with the linker and C5D from the 4 3 kD endoglucanase from H. insolens (disclosed in WO 91/17243). The native endoglucanase from M. phaseolina does not have a linker and/or a cellulose binding domain, CBD.
The construction consists of the endoglucanase from, M. phaseolina (223 anino acids) and the 72 C-terminal amino acids from the H. insolens 4 3 kD endoglucanase.
The 43 kD endoglucanase cDNA from H. insolens is cloned into pHD414 in such a way that the endoglucanase gene is transcribed from the Taka-promoter. The resulting plasmid is named pCaHj4lS.
The'cDNA encodirrg"the endoglucanase from M. phaseolina is cloned into pYES2.0 as a BstX I/Not I fragment and the resulting plasmid is named pClC477.

Primers:
primer 1: 5'-
GGTCGCCCGGACTGGCTGTTCCCGTACCCCCTCCAGCAGCACCAGCTCTCCGG -3' primer 2: 5'
CCGGAGAGCTGGTGCTGCTGGAGGGGGTACGGGAACAGCCAGTCCGGGCGACC -3' pYES2.0 F.HT primer: 5' CGGACTACTAGCAGCTGTAATACG -3' AMG-term. primer: 5' CCCCATCCTTTAACTATAGCG -3'
The endoglucanase fusion is constructed by the PCR overlap-extension method as described by Higuchi et al. 1988.
Reaction A: The Pclymerase Chain Reaction (PCR) is used
to'amplify the fragment of pCaHj418 between printer 1 and
AMG-term. primer (the linker and CBD from the 4 3 kD
endoglucanase from H.insolens).
Reaction B: PC?, amplification of the fragmen" berwier.
pYES2.0 F.HT primer and primer 2 in pClC477, the
endoglucanase gene from M. p'naseolina.
Reaction C: The two purified fragments are used in a
third PCR in the presence of the primers flanking rhe
total region, i.e. pYES2.0 F.HT primer and AMG-term.
primer.
The fragment amplified in reaction C is purified, digested with restriction enzymes, e.g. Xba I and BamH I. The purified digested fragment is ligated into pHD4 14 digested with restriction enzymes, e.g. Xba I and BamH I.
Competent cells from E. coli strain DHSaF' (New England
Biolabs) are transformed with the ligated plasmid and
colonies containing the gene fusion are isolated. The
sequence of the cloned part was verified by DNA sequen¬
cing.
Polymerase Chain Reactions are carried out under standard

conditions, as recommended by Perkin-Elmer.
Reaction A and B start with 2 min. at 94°C followed by 20 cycles of (30 sec. at 94°C, 30 sec. at 52°C and 1 min. at 72°C) and ends with 4 min. at 72 =C.
Reaction C starts with (2 min. at 94°C, 1 min. at 52°C and 2 min. at 72°C), followed by 20 cycles of (30 sec. at 94°C, 30 sec. at 52°C and 90 sec. at 72°C) and ends with 4 min. at 72°C.
The construct may be transformed into Aspergillus oryzae as described above.
EXAMPLE 3
Cloning and expression of an endoglucanase froE Acremonlum sp. and Sordaria fimicola
Production of biomass for expression cloning procedures: Isolates CBS 473.94 and ATCC 52644, respec~iveiy, were grown in shake flask cultures on cellulose enriched pota¬to dextrose broth, incubated for 5 days at 260C (shaking conditions: 150 rpm).
mRNA was isolated from Acremonium sp., C5S 478.94, and Scrdaria flrrdcola, ATCC 52644, respectively, grown in a cellulose-containing fermentation medium with agitation to ensure sufficient aeration. Mycelia were harvested after 3-5 days' growth, immediately frozen in liquid nitrogen and stored at -80°C. Libraries from Acrer-:onlum sp., and Sordaria fimicola, respectively, each consisting of approx. lo' individual clones were constructed in E. jzpli as described with a vector background ofTT.
Plasmid DNA from some of the pools from each library was

transformed into yeast, and 50-100 plates containing 250-400 yeast colonies were obtained from each pool.
Endoglucanase-positive colonies were identified and isolated on SC-agar plates with the AZCL HE cellulose assay. cDNA inserts were amplified directly from the yeast colonies and characterized as described in the Materials and Methods section above.
The DNA sequence of the cDNA encoding the endoglucanase from Acremonium sp. is shown in SEQ ID No. 6 and the corresponding amino acid sequence is shown in SEQ ID No.7. The cDNA is obtainable from the plasmid in DSM
1003 0.
The partial DNA sequence of the cDNA encoding the endoglucanase from Sordaria fi~dccla is shown in SEQ ID No. 19 (Nucleotide sequence of rhe 5'-e.nd of the cDNA) and the corresponding amino acid sequence is shown in SEQ ID No. 20. The cDNA is obtainable from the plasmid in DSM
Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanase in Aspergrillus, the Dl^k was digested with appropriate restriction enzymes, size fractionated on gel, and a fragm.ent corresponding to the endoglucanase gene from Acremonium sp. and Sordaria fimicola, respectively, was purified. The genes were subsequently ligated to pHD414, digested with appropriate restriction enzymes, resulting in the plasmids pA2C371 and pA2C502, respectively.
After amplification of the DNA in E. coli the plasmids were trarrsformed into Aspergillus oryzae as described above.

EXAMPLE 4
A. Cloning by PCR an endoglucanase from Crinipellis sca-bella, CBS 280.96
Isolate CBS 280.96 was grown in static flask cultures, holding wheat bran medium (per flask: 300g wheat bran added 450 ml salt solution), incubated for 6 days at 26C. After incubation the wheat bran was extracted with destilled water (300ml per flask) and the extract tested for endoglucanase activity (0.1% AZCL-HE-Cellu]ose (megazyme) in 1% agarose (Litex agarose, Medinova). Activity was observed on the plates holding pH of 3.0, 7.0 and S.5.
mRN-ii was isolated from Crinipellis scabella grown as describe above. Mycelia were harvested after 3-5 days' growxh, luiTTiediatiely froze.n m liquid nitrogen and stored at -60'C. A library from Crinipellis scabella, consisting of apprcx. iO' individual clones was constructed in I. coli as described with a vector background of 1%.
Approximately 10 000 colony forming units (c.f.u.) from the Crinipellis scabella cDNA library in E. coli was screened by colony hybridization using a random-primed "P-labeled PCR product from C. scabella as probe. The PCR product was generated as described in the Materials and methods section. The positive cDNA clones were characterized by sequencing the ends of the cDNA inserts, and by determining the nucleotide seuence of the longest cDNA from both strands.
The DNA sequence of the cDNA encoding the endoglucanase is shown in SEQ ID No. 12 and the corresponding amino acid-ssqiience is shown in SEQ ID No. 13.
The cDNA is obtainable from the plasmid in DSM 10511.

Total DNA was isolated from a yeast colony and plasmid DNA was rescued by transformation of E. coli as described above. In order to express the endoglucanse in Aspergillus, the DNA was digested with appropriate restriction enzymes, size fractionated on gel, and a fragment corresponding to the endoglucanase gene was purified. The gene was subsequently ligated to pHD414, digested with appropriate restriction enzymes, resulting in the plasmid pA2C475.
After amplification of the DNA in E. coli the plasmid was transformed into Aspergillus oryzae as described above.
Construction of two gene fusions between the endoglucanase from Crinipellis scahella and the linker/CBD region of the 43 kDa endoglucanase from
cusj.ccxa x~i.sox^ns.
The native endoglucanase from Crinipellis scabella neit¬her has a linker nor a cellulose binding domain (CBD). In addition, the full-length cDNA contains an ATG start codon upstream from the endoglucanase encoding open reading frame (ORF), presumably resulting in scrambled translation initiation upon heterologous expression of the cDNA, such as in the yeast Saccharomyces cerevisiae and the filamentous fungus Aspergillus oryzae. Thus, two gene fusions between the endoglucanase from Crinipellis scabella and the linker/CBD region of the 43kD endoglucanase from Humicola insolens (disclosed in WO 91/17243) has been constructed using splicing by overlap extension (SOE) (Horton et al, 1S39).
Construction 1 consists of the cDNA encoding the 226-re-sidue fendoglucanase from C. scabella fused by PCR with the 3'-end cDNA of H. insolens coding for the linker and CBD region (72 amino acids) at the COOH-terminus of the

H. insolens 43kD endoglucanase. The second hybrid construct is identical to the abovementioned gene fusion, except that the first five residues from the putative signal peptide have been deleted by PCR resulting in a shorter signal, which starts with the second in-frame ATG start codon.
Plasmid constructs
The plasmid pClC475 contains the full-length cDNA from C. scabella, cloned into BstXI/Notl-cut yeast expression vector pYES 2.0, the plasmid pClC144 contains the full-length cDNA from H. insoiens, cloned into the BstXI site of pYES 2.0. ■■
Splicing by overlap extension
Two PCR fragments encoding the core region of the endog¬lucanase from C. scahella were generated in PCR buffer (10 mM Tris-HCl, pH 8.3, 50 inM"KCl, 1.5 r^M MgCl-,, 0.01 % gelatin; containing 200 .LLM each dNTP), usi.ng 50-100 ng of pClC475 as template, and 250 pmol of the reverse primer (5'-GACCGGAGAGCTGGTGCTGCTGGAGGGTTTACGAACACAGCCCGAGATATTAG TG- 3*) in two combinations with 300-350 pmol of each forward primer (forward no. 1 5'-
CCCCA„AGCTTGACTTGGAACCAJ^TGGTCCATCC-3 " , forward no. 2 5"-CCCCAAGCTTCCATCCAAACATGCTTAAAACGCTCG- 3"), a DNA thermal cycler (Landgraf, Germany) and 2.5 units of Tag polymerase (Perkin-Elmer, Cetus, USA). Thirty cycles of PCR were performed using a cycle profile of denaturation at 94 °C for 1 min, annealing at 55 °C for 2 m.in, and extension at 72 °C for 3 m.in. The PCR' fragffient coding for the linker and CBD of the endoglucanase of H. insolens was generated in PCR- buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 1.5 mM MgCl,, 0.01 % gelatin; containing 200 ^M each dNTP) using 100 ng of the pClC144 template, 250 pmol

forward primer (5'-CACTAATATCTCGGGC-
TGTGTTCGTAAACCCTCCAGCAGCACCA-GCTCTCCGGTC-3') , 250 pmol of the pYES 2.0 reverse primer
(5'-GGGCGTGAATGTAAGCGTGACATA-3■) a DNA thermal cycler (Landgraf, Germany) and 2.5 units of Tag polymerase (Perkin-Elmer, USA). Thirty cycles of PCR were performed as above. The PCR products were electrophoresed in 0.7 % low gelling temperature agarose gels (SeaPlaque, FMC), the fragments of interest were excised from the gel and recovered by treatment with agarase (New England Biolabs, USA) according to the manufacturer's instructions, followed by phenol extraction and ethanol precipitation at - 20 °C for 12 h by adding 2 vols of 96 % EtOH and 0. 1 vol of 3.M NaAc.
The recombinant hybrid genes between the endoglucanase from Crinipallis scabella and the linker/CBD region of the 4 3 kD endoglucanase from Humicola insolsr.s were generated by combining the overlapping PCR fragments from above (ca. 50 ng of each template) in two combinations in PCR buffer (10 mM Tris-HCl, pH S.3, 50 ir^M KCl, 1.5 mM MgCl-,, C.Ol % gelatin; containing 200 uM each dNT?) . The SOE reaction was carried out using the DNA thermal cycler (Landgraf, Germany) and 2.5 units of Taq polymerase (Perkin-Elmer, Cetus, USA). Two cycles of PCR were performed using a cycle profile of denaturation at S4 °C for 1 min, annealing at 55 °C for 2 min, and extension at 72 °C for 3 min, the reaction was stopped, 2 50 pmol of each end-primer (forward no. 1
5'-CCCCAAGCTTGACTTGGAACCAATGGTCCATCC-3', forward no. 2 5 ' -CCCCAAGCTTCCATCC.ILAJ\CATGCTTAAAACGCTCG-3 ' , reverse primer 5 '-GGGCGTGAATGT.AAGCGTGACATA-3 ' ) was added to rhe reaction mixture, and an additional 30 cycles of PCR were performed using a cycle profile of denaturation at 94 °C for 1 min, annealing at 55 'tnor 2 min, and extension at 72 °C for 3 min.

Construction of the expression cassettes for heterologous expression in Aspergillus oryzae
The PCR-generated, recombinant fragments were electrophoresed in a 0.7 % low gelling temperature agarose gel (SeaPlaque, FMC), the fragments of interest were excised from the gel and recovered by treatment with agarase (New England Biolabs, USA) according to the manufacturer's instructions, followed by phenol extraction and ethanol precipitation at - 20 °C for 12 h. The DNA fragments were digested to completion with Hindlll and Xbal, and ligated into Hi/idlll/A'bal-cleaved pHD414 vector followed by electroporation of the constructs into £. coli DHIOB cells according to the manufacturer's instructions (Life Technologies, -USA). -
The nucleotide sequence of the resulting gene fusions were cererTrined froir, born strands as described in rne Materials and methods section, SEQ ID no. 14A and 15A. The constructs may be transformed into A. oryzae as described.
EXAMPLE 5
PCR facilitated detection of said type of cellulytic en¬zyme from 46 filamentous and monocentric fungi, representing 32 genera, from 23 families, belonging to 15 orders of 7 classes, covering all in all all four groups of the true Fungi: Ascomycetous, Basidiomycetous, Chytri-diomycetous and Zygomycetous fungi
5.1 Materials
l.Diplodia gossypina Cooke
Deposit of Strain, Ace No: CBS 274.S6
2.Ulospora bilgramii (Hawksw. et al.) Hawksw. et al. Ace No of strain: NKBC 1444,

3. Microsphaeropsis sp
4. Ascobolus stictoideus Speg.
Ace No of strain: Q026 (Novo Nordisk collection) Isolated from goose dung, Svalbard, Norway
5. Saccobolus dilutellus (Fuck) Sacc. Deposit of strain: Ace No CBS 275.96
6. Penicillium verruculosum Peyronel Ex on Ace No of species: ATCC 62396
7. Penicillium chrysogenum Thorn Ace No of Strain: ATCC 9480
8. Thermomyces verrucosus Pugh et al Deposit of Strain, Ace No.: CES 2 3 5.56
9. Xylaria hypoxylon L. ex Greville Deposit of Strain, Ace No: CBS 2 3 4.96

10. Poronia punctata (Fr.ex L.) Fr. Ref:A.Munk: Danish Pyrenomyeetes, Dansk Botanisk Arkiv, Voll7,1 1957
11. Nodulisporum sp
Isolated from leaf of Camellia reticulata (Theaceae,
Guttiferales),
Kunming Botanical Garden, Yunnan Province, China
12. Cylindrocarpon sp
Isolated from marine sample, the Bahamas
13. Fusarium anguioides Sherbakoff
Ace No of strain: IFO 4467

14. Fusarium poae (Peck) Wr.
Ex on Ace No of species: ATCC 50883
15. Fusarium solani (Mart.)Sacc.emnd.Snyd & Hans. Ace No of strain: IMI 107.511
16. Fusarium oxysporum ssp lycopersici (Sacc.)Snyd. & Hans.
Ace No of strain: CBS 645.78
17. Fusarium oxysporum ssp passiflora
Ace No of strain: CBS 744.79
IS. Glioeladium catenulatum Gillman & Abbott Ace. No. of strain: ATCC 10523
19. Nectria pinea Dingley
Deposit of Surain, Ace. No. CBS 279.95
20. Sordaria macrospora .a.uersvald Ex on Ace No of species: .a.TCC 602 55
21. Humicola grisea Traeen
ex on Ace No for the species: ATCC 22726
22. Humicola nigrescens Omvik Ace No of strain: CBS 819.73
23. Scytalidium thermophilum (Cooney et Emerson) Austwick Ace No of strain: ATCC 28085
24. T.hielavia thermophila Fergus et Sinden (syn Corynascus thermophiius)
Ace No of strain: CBS 174.70, IMI 145.136
25. Cladorrhinum foecundissimum Saccardo et Marchal Ex on Ace No of species: ATCC 62373

26. Syspastospora boninensis
Ace No of strain: NKBC 1515 (Nippon University, profe Tubaki Collection)
27. Chaetoir.ium cuniculorum Fuckel Ace. No. of strain: CBS 799.83
28. Chaetomium brasiliense Batista et Potual Ace No of strain: CBS 122.65
29. Chaetomium murorum Corda Aec No of strain: CBS 163.52
30. Chaetomium virescens (von Arx) Udagawa Ace.No. of strain: C5S 547.75
31. Nigrcspora sp
Deposit of strain, Ace No: CBS 2"2.96
32. Nigrospora sp Isolated from:
33. Diaport-he syngenesia
Deposit of strain, Aec No: CBS 278.96
34. Colletotrichum lagenarium (Passerini) Ellis et
Kalsted
syn Glomerella cingulata var orbiculare Jenkins et
Winstead
Ex on a^^c No u£ species: ATCC 52609
35. Exidia glandulosa Fr.
Deposit of Strain, Aec No: CBS 277.96
36. Pomes fomentarius (L.) Fr.
Deposit of strain: Aec No. CBS 276.96

37. Spongipellis (?)
Deposit of Strain: Ace No CBS 233.96
38. Rhizophlyctis rosea (de Bary & Wor) Fischer Deposit of Strain: Ace No.: CBS 282.96
39. Rhizomucor pusillus (Lindt) Schipper syn: Mucor pusillus
Ace No of strain: IFO 4578
40. Phycomyces nitens (Kunze) van Tieghem & Le Monnier
Ace No of strain: IFO 4814
41 Chaetostylum fresenii van Tieghem & Le Monnier syn. Helicostylum fresenii Ace No of strain NRRL 2 3 05
D'.neciuiTi rcseum, Ace No or s-crain: xru o j, /:
43. Coniothecium sp
Endophyte, isolated from leaf of flevering plant, Kunming , Yunnan, China
44. Deposit of strain, Ace No.: CBS 271.96 Coelomycete, Isolated from leaf of Artocarpus altilis (Moraceae, Urticales), Christiana, Jamaica
45. Deposit of strain. Ace No.: CBS 273.96 Coelomycete, isolated from leaf of Pimenta dioica (Myrtaceae, Myrtales), Dallas Mountain, Jamaica
45. Deposit of strain: CBS 270.96
Coelomycete, isolated from leaf of Pseudocalymma
alliaceum (Bignoniaceae, Solanales) growing in Dallas
Mountain, Jamaica "

JO

5.2 Procedure
Maintenance of strains and production of biomass:
The strains were maintained on agar in petrie dishes (9cm) or on slants (see list of Media: PCA and PDA). 44 of the strains were grown in shake flasks under the following growth conditions: general fungal media as PC, PD and PB 9 or YPG (see list of media); incubation time from 3 to 9 days; temperature 26"C; rpm between 150 and 175 . Strain No 14 (F.poae) was grown on wheat bran for 15 days (26"C; static). Strain No 38 was grown in dilute salt solution (DS/2), added 1 cm- pieces of autoclaved filter paper.
Activity test:
Activity was tested on 0.1% AZCL-HE-Cellulose (Megazyme) plates (14 era Petrie dishes), made up in 1% agarose (HSE, Lirex Agarose, Medmova). .Ail tests were done in triplicate, viz. A2CL-HE-Cellulose dissolved in three buffers, adjusted ro pH 3, 7 or S.5 (using various proportions of t:he following two ingredients Citric acid sonohydrat, Merck art. No 100244 (21.0 g) dissolved in water, making a total of 1000 ml; 0.IM tri-Sodium dodeca-brohydrate, Merck art.no. 6578 (38 g), dissolved in wa¬ter, making a total of 1000 ml. The mixing is done imrnidiately before use.
Harvesting of Biomass:
The biomass was harvested by filtering (mesh adjusted to the growth of the fungus, the finest used for fungi which have highly sporulating mycelium as e.g. Fusarium spp.). The biomass on the filter was scraped into a sterile plastic bag and imrnidiately frozen (by submerging into liquid nitrogen).

5.3 Results
I. Using the PCR screening and amplification techniques described in Materials and Methods the following partial cDNA sequences were obtained:
Saccobolus dilutellus (Fuck) Sacc, CBS 275.96: SEQ ID
No. 21 (and the deduced amino acid sequence in SEQ ID No.
22) ;
Thermomyces verrucosus, CBS 285.96: SEQ ID No. 23 (and
the deduced amino acid sequence in SEQ ID No. 24);
Xylaria hypoxylon, CBS 284.96: SEQ ID No. 25 (and the
deduced amino acid sequence in SEQ ID No. 26);
Fusarium oxysporum ssp lycopersici, CBS 645.78: SEQ ID
No. 27 (and the deduced amino acid sequence in SEQ ID No.
28);
Nectria pinea, CBS 279.96: SEQ ID No. 29 (and the deduced
amino acid sequence in SEQ ID No. 30);
Humicola grisea, ATCC 2272 6: SEQ ID No. 31 (and rhe
deduced amino acid sequence in SEQ ID No. 32);
Humicola nigrescens, C3S 819.73: SEQ ID No. 33 (and the
deduced amino acid sequence in SEQ" ID No. 34);
Cladorrhinum foecundissimum, ATCC 62373: SEQ ID No. 35
(and the deduced amino acid sequence in SEQ ID No. 36);
Syspastospora boninensis, NKBC 1515: SEQ ID No. 37 (and
the deduced amino acid sequence in SEQ ID No. 38);
Nigrospora sp., CBS 272.96: SEQ ID No. 39 (and the
deduced amino acid sequence in SEQ ID No. 40);
Chaetostylum fresenii: SEQ ID No. 41 (and the deduced
amino acid sequence in SEQ ID No. 42);
Exidia glandulosa, CBS 277.96: SEQ ID No. 43 (and the
deduced amino acid sequence in SEQ ID No. 44);
Coniothecium sp.: SEQ ID No. 45 (and the deduced amino
acid sequence in SEQ ID No. 46);
Deposition No. CBS 271.96: SEQ ID No. 47 (and the deduced
amino acid sequence in S£i2—LD No. 48);
Deposition No. CBS 270.96: SEQ ID No. 49 (and the deduced
amino acid sequence in SEQ ID No. 50);

Diplodia gossypina, CBS 274.96: SEQ ID No. 51 (and the
deduced amino acid sequence in SEQ ID No. 52);
Ulospora bilgramii, NKBC 1444: SEQ ID No. 53 (and the
deduced amino acid sequence in SEQ ID No. 54);
Penicillium verruculosum, ATCC 62396: SEQ ID No. 55 (and
the deduced amino acid sequence in SEQ ID No. 56);
Poronia punctata: SEQ ID No. 57 (and the deduced amino
acid sequence in SEQ ID No. 58);
Fusarium anguioides, IFO 4467: SEQ ID No. 59 (and the
deduced amino acid sequence in SEQ ID No. 60);
Thielavia thermophila, CBS 174.70: SEQ ID No. 61 (and the
deduced amino acid sequence in SEQ ID No. 62);
Chaetomium cuniculorum, CBS 799.83: SEQ ID No. 63 (and
the deduced amino acid sequence in SEQ ID No. 64);
Chaetomium virescens: SEQ ID No. 65 (and the deduced
amino acid sequence in SEQ ID No. 66);
Colletctrichum lagenarium: SEQ ID No. 67 (and the deduced
amino acid sequence in SEQ ID No. 65);
Phycomyces nitens: SEQ ID No. 69 (and the deduced amino
acid sequence in SEQ ID No. 70); and
Trichothecium roseum: SEQ ID No. 71 (and the deduced
amino acid sequence in SEQ ID No. 72);
II. Using the PCR screening and amplification techniques
described in Materials and Methods partial cDNA encoding
partially for the enzyme of the invention was obtained
and the plasmid was deposited according to the Budapest
Treaty:
Escherichia coli, DSM 10583, deposition date 13 March,
1996;
cDNA from Trichothecium roseum;
Escherichia coli, DSM 10584, deposition date 13 March,
1996;
cDNA from Syspastospora boninensis;
Escherichia coli, DSM 10585, depbsitrTon date 13 March,
1996;
cDNA from Cheatomium murorum

Escherichia coli, DSM 10587, deposition date 13 March,
1996;
cDNA from Sordaria fimicola;
Escherichia coli, DSM 10588, deposition date 13 March,
1996;
cDNA from the unidentified strain CBS 273.96;
Escherichia coli, DSM 10586, deposition date 13 March,
1996;
cDNA from Spongipellis sp.
Color clarification of crude supernatants
During normal wash the fabric will often fade. However, the fabric appearance is improved and the original colours are much better preserved or maintained if the fabric is washed with a cellulase giving color clarificarion. Color clarification is measured as removal of surface fibrils and fibers protruding from the yarn of a textile containing cellulosic fibers.

Apparatus Liquid volume Agitation
Rinse time
Washing temp
Washing liqour
PH
Washing time
Repetitions
Enzymes
Dosage
Textile cotton

Terg-o-tometer
100 ml
150 movements/min with vertical
stirrer
5 min in tapwater
40°
0.05 M phosphate buffer
7.0
30 min
2
Crude supernatants from the
strains shown below.
Two dosages from : 200, 500,
1000 or 2500—S=CEVU/1
2 swatches of aged black 100%
5x6 cm (0.9 gram)

Drying : Tumble dry-
Evaluation:
The light remission is measured by a Datacolor Elrepho Remission spectrophotometer. Remission is calculated as delta L (Hunter Lab-values). When the surface fibrils and fibers protruding from the yarn are removed by the cellulase, the surface of the black fabric appears darker, and lower L values are obtained.
The samples are compared with a blind sample, i.e. washed without enzyme. Below is shown the delta L remission values compared to a blind sample.
SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT
(A) NAME: Novo Nordisk A/S
(B) STREET: Novo Alle
(C) CITY: DK-2880 Bagsvaerd
(E) COUNTRY: Denmark
(F) POSTAL CODE (ZIP): DK-2880
(G) TELEPHONE: +45 44 88 88
(H) TELEFAX: +45 44 49 32 56
(ii) TITLE OF INVENTION: NOVEL ENDOGLUCANASES (iii) NU'MBER OF SEQUENCES: 72
(iv) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk
(B) COMPUTER: IBM PC compatible
(C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: Patentin Release #1.0, Version
#1.30 (EPO)
(2) INFORMATION FOR SEQ ID NO: lA:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 891 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA

(vi) ORIGINAL SOURCE:
(A) ORGANISM: Saccharomyces cerevisiae
(B) STRAIN: DSM 9770
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: lA:
AAAGAAAGGC TCTCTGCTGT CGTCGCTCTC GTCGCTCTCG TCGGCATCCT CCATCCGTCC 60
GCCTTTGATA ACCCGCTCCC CGACTCAGTC AAGACGACGC ATACTTGGCA CCATGCATCT 12 0
CTCCGCCACC ACCGGGTTCC TCGCCCTCCC GGTCCTGGCC CTGGACCAGC TCTCGGGCAT 180
CGGCCAGACG ACCCGGTACT GGGACTGCTG CAAGCCGAGC TGCGCCTGGC CCGGCAAGGG 2 4 0
CCCCTCGTCT CCGGTGCAGG CCTGCGACAA GAACGACAAC CCGCTCAACG ACGGCGGCTC 3 00
CACCCGGTCC GGCTGCGACG CGGGCGGCAG CGCCTACATG TGCTCCTCCC AGAGCCCCTG 3 60
GGCCGTCAGC GACGAGCTGT CGTACGGCTG GGCGGCCGTC AAGCTCGCCG GCAGCTCCGA 4 2 0
GTCGCAGTGG TGCTGCGCCT GCTACGAGCT GACCTTCACC AGCGGGCCGG TCGCGGGCAA 480
GAAGATGATT GTGCAGGCGA CCAACACCGG TGGCGACCTG GGCGACAACC ACTTTGACCT 540
GGCCATCCCC GGTGGCGGTG TCGGTATTTT CAACGCCTGC ACCGACCAGT ACGGCGCTCC 600
CCCGAACGGC TGGGGCGACC GCTACGGCGG CATCCATTCC AAGGAAGAGT GCGAATCCTT 660

CCCGGAGGCC CTCAAGCCCG GCTGCAACTG GCGCTTCGAC TGGTTCCAAA ACGCCGACAA 72 0
CCCGTCGGTC ACCTTCCAGG AGGTGGCCTG CCCGTCGGAG CTCACGTCCA AGAGCGGCTG 780
CTCCCGTTAA GAGGGAAGAG AGGGGGCTGG 7AGGACCGAA AGATTCAACC TCTGCTCCTG 84 0
CTGGGGAAGC TCGGGCGCGA GTGTGAAACT GGTGTAAATA TTGTGGCACA CACAAGCTAC 900
TACAGTCCGT CTCGCCGTCC GGCTAACTAG CCTTGCTGCG GATCTGTCCA AAAAAAAAAA 960
(2) INFORMATION FOR SEQ ID NO: IB:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 225 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: IB:
Met His Leu Ser Ala Thr Thr Gly Phe Leu Ala Leu Pro Val Leu Ala
15 10 15
Leu Asp Gin Leu Ser Gly lie Gly Gin Thr Thr Arg Tyr Trp Asp Cys
20 25 30
Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Gly Pro Ser Ser Pro Val

35 40 45
Gin Ala Cys Asp Lys Asn Asp Asn Pro Leu Asn Asp Gly Gly Ser Thr
50 55 60
Arg Ser Gly Cys Asp Ala Gly Gly Ser Ala Tyr Met Cys Ser Ser Gin
65 70 75 80
Ser Pro Trp Ala Val Ser Asp Glu Leu Ser Tyr Gly Trp Ala Ala Val
85 90 95
Lys Leu Ala Gly Ser Ser Glu Ser Gin Trp Cys Cys Ala Cys Tyr Glu
100 105 110
Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met lie Val Gin
115 120 125
Ala Thr Asn Thr Gly Gly Asp Leu Gly Asp Asn His Phe Asp Leu Ala
130 135 140
lie Pro Gly Gly Gly Val Gly lie Phe Asn Ala Cys Thr Asp Gin Tyr
145 150 155 160
Gly Ala Pro Pro Asn Gly Trp Gly Asp Arg Tyr Gly Gly lie His Ser
165 170 175

Lys Glu Glu Cys Glu Ser Phe Pro Glu Ala Leu Lys Pro Gly Cys Asn
180 185 190
Trp Arg Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Ser Val Thr Phe
195 200 205
Gin Glu Val Ala Cys Pro Ser Glu Leu Thr Ser Lys Ser Gly Cys Ser
210 215 220
Arg 225
2) INFORMATION FOR SEQ ID NO: 2A:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 894 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: "Construction 1"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2A:
\TGCATCTCT CCGCCACCAC CGGGTTCCTC GCCCTCCCGG TCCTGGCCCT GGACCAGCTC 60
rCGGGCATCG GCCAGACGAC CCGGTACTGG GACTGCTGCA AGCCGAGCTG CGCCTGGCCC 120
GGCAAGGGCC CCTCGTCTCC GGTGCAGGCC TGCGACAAGA ACGACAACCC GCTCAACGAC

180
GGCGGCTCCA CCCGGTCCGG CTGCGACGCG GGCGGCAGCG CCTACATGTG CTCCTCCCAG 240
AGCCCCTGGG CCGTCAGCGA CGAGCTGTCG TACGGCTGGG CGGCCGTCAA GCTCGCCGGC 300
AGCTCCGAGT CGCAGTGGTG CTGCGCCTGC TACGAGCTGA CCTTCACCAG CGGGCCGGTC 360
GCGGGCAAGA AGATGATTGT GCAGGCGACC AACACCGGTG GCGACCTGGG CGACAACCAC 420
TTTGACCTGG CCATCCCCGG TGGCGGTGTC GGTATTTTCA ACGCCTGCAC CGACCAGTAC 480
GGCGCTCCCC CGAACGGCTG GGGCGACCGC TACGGCGGCA TCCATTCCAA GGAAGAGTGC 540
GAATCCTTCC CGGAGGCCCT CAAGCCCGGC TGCAACTGGC GCTTCGACTG GTTCCAAAAC 600
GCCGACAACC CGTCGGTCAC CTTCCAGGAG GTGGCCTGCC CGTCGGAGCT CACGTCCAAG 660
AGCGGCTGCT CCCGTCCCTC CAGCAGCACC AGCTCTCCGG TCAACCAGCC TACCAGCACC 720
AGCACCACGT CCACCTCCAC CACCTCGAGC CCGCCAGTCC AGCCTACGAC TCCCAGCGGC 780
TGCACTGCTG AGAGGTGGGC TCAGTGCGGC GGCAATGGCT GGAGCGGCTG CACCACCTGC 840
GTCGCTGGCA GCACTTGCAC GAAGATTAAT GACTGGTACC ATCAGTGCCT GTAG 894

INFORMATION FOR SEQ ID NO: 2B:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 297 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2B:
Met His Leu Ser Ala Thr Thr Gly Phe Leu Ala Leu Pro Val Leu Ala
15 10 15
Leu Asp Gin Leu Ser Gly lie Gly Gin Thr Thr Arg Tyr Trp Asp Cys
20 25 30
Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Gly Pro Ser Ser Pro Val
35 40 45
Gin Ala Cys Asp Lys Asn Asp Asn Pro Leu Asn Asp Gly Gly Ser Thr
50 55 60
Arg Ser Gly Cys Asp Ala Gly Gly Ser Ala Tyr Met Cys Ser Ser Gin
65 70 75 80
Ser Pro Trp Ala Val Ser Asp Glu Leu Ser Tyr Gly Trp Ala Ala Val
85 90 95

Lys Leu Ala Gly Ser Ser Glu Ser Gin Trp Cys Cys Ala Cys Tyr Glu
100 105 110
Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met lie Val Gin
115 120 125
Ala Thr Asn Thr Gly Gly Asp Leu Gly Asp Asn His Phe Asp Leu Ala
130 135 140
lie Pro Gly Gly Gly Val Gly lie Phe Asn Ala Cys Thr Asp Gin Tyr
145 150 155 160
Gly Ala Pro Pro Asn Gly Trp Gly Asp Arg Tyr Gly Gly He His Ser
165 170 175
Lys Glu Glu Cys Glu Ser Phe Pro Glu Ala Leu Lys Pro Gly Cys Asn
180 185 190
Trp Arg Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Ser Val Thr Phe
195 200 205
Gin Glu Val Ala Cys Pro Ser Glu Leu Thr Ser Lys Ser Gly Cys Ser
210 215 220
Arg Pro Ser Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr
225 230 235 240

Ser Thr Thr Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr
245 250 255
Thr Pro Ser Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn
260 265 270
Gly Trp Ser Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys
275 280 285
lie Asn Asp Trp Tyr His Gin Cys Leu
290 295
(2) INFORMATION FOR SEQ ID NO: 3A:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 927 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: "Construction 2"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3A:
ATGCATCTCT CCGCCACCAC CGGGTTCCTC GCCCTCCCGG TCCTGGCCCT GGACCAGCTC 60
TCGGGCATCG GCCAGACGAC CCGGTACTGG GACTGCTGCA AGCCGAGCTG CGCCTGGCCC 120
GGCAAGGGCC CCTCGTCTCC GGTGCAGGCC TGCGACAAGA ACGACAACCC GCTCAACGAC

180
GGCGGCTCCA CCCGGTCCGG CTGCGACGCG GGCGGCAGCG CCTACATGTG CTCCTCCCAG 240
hGCCCCTGGG CCGTCAGCGA CGAGCTGTCG TACGGCTGGG CGGCCGTCAA GCTCGCCGGC 300
AGCTCCGAGT CGCAGTGGTG CTGCGCCTGC TACGAGCTGA CCTTCACCAG CGGGCCGGTC 360
GCGGGCAAGA AGATGATTGT GCAGGCGACC AACACCGGTG GCGACCTGGG CGACAACCAC 420
TTTGACCTGG CCATCCCCGG TGGCGGTGTC GGTATTTTCA ACGCCTGCAC CGACCAGTAC 480
GGCGCTCCCC CGAACGGCTG GGGCGACCGC TACGGCGGCA TCCATTCCAA GGAAGAGTGC 540
GAATCCTTCC CGGAGGCCCT CAAGCCCGGC TGCAACTGGC GCTTCGACTG GTTCCAAAAC 600
GCCGACAACC CGTCGGTCAC CTTCCAGGAG GTGGCCTGCC CGTCGGAGCT CACGTCCAAG 660
AGCGGCTGCT CCCGTAACGA CGACGGCAAC TTCCCTGCCG TCCAGATCCC CTCCAGCAGC 720
ACCAGCTCTC CGGTCAACCA GCCTACCAGC ACCAGCACCA CGTCCACCTC CACCACCTCG 780
AGCCCGCCAG TCCAGCCTAC GACTCCCAGC GGCTGCACTG CTGAGAGGTG GGCTCAGTGC 840
GGCGGCAATG GCTGGAGCGG CTGCACCACC TGCGTCGCTG GCAGCACTTG CACGAAGATT 900
AATGACTGGT ACCATCAGTG CCTGTAG 927

) INFORMATION FOR SEQ ID NO: 3B:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3 08 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3B:
Mer His Leu Ser .Ala Thr,Thr,Gly P.he Leu Ala Leu Pro Val Leu Ala
15 10 15
Leu Asp Gin Leu Ser Gly lie Gly Gin Thr Thr Arg Tyr Trp Asp Cys
20 25 30
Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Gly Pro Ser Ser Pro Val
35 40 45
Gin Ala Cys Asp Lys Asn Asp Asn Pro Leu Asn Asp Gly Gly Ser Thr
50 55 50
Arg Ser Gly Cys Asp Ala Gly Gly Ser Ala Tyr Met Cys Ser Ser Gin
65 70 75 80
Ser Pro Trp Ala Val Ser Asp Glu Leu Ser Tyr Gly Trp Ala Ala Val

85

90

95

Lys Leu Ala Gly Ser Ser Glu Ser Gin Trp Cys Cys Ala Cys Tyr Glu

100

105

110

Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met lie Val Gin

115

120

12 5

Ala Thr Asn Thr Gly Gly Asp Leu Gly Asp Asn His Phe Asp Leu Ala

130

135

140

lie Pro Gly Gly Gly Val Gly lie Phe Asn Ala Cys Thr Asp Gin Tyr

145

150

155

160

Gly Ala Pro Pro Asn Gly Trp Gly Asp Arg Tyr Gly Gly lie His Ser

165

170

175

Lys Glu Glu Cys Glu Ser Phe Pro Glu Ala Leu Lys Pro Gly Cys Asn

180

185

190

Trp Arg Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Ser Val Thr Phe

195

200

205

Gin Glu Val Ala Cys Pro Ser Glu Leu Thr Ser Lys Ser Gly Cys Ser

210

215

220

Arg Asn Asp Asp Gly Asn Phe Pro Ala Val Gin lie Pro Ser Ser Ser

225 230 235 240
Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr Ser Thr Thr Ser Thr
245 250 255
Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser Gly Cys
260 265 270
Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser Gly Cys
275 2S0 285
Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys lie Asn Asp Trp Tyr
290 255 300
His Gin Cys Leu 305
(2) INFORMATION FOR SEQ ID NO: 4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 888 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Saccharoinyces cerevisiae, DSM 10082

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:
CCAGTGTGCT GGAAAGCCTT CGTGCTGTCC CCGACGTATC CCTGACCGCC ATGCGTTCCA 60
CCAGCATCTT GATCGGCCTT GTTGCCGGCG TCGCTGCTCA GAGCTCTGGC TCTGGCCATA 120
CAACCAGGTA CTGGGACTGC TGCAAGCCCT CATGCGCCTG GGATGAGAAG GCGGCTGTCA 180
GCCGGCCGGT CACAACATGC GACAGGAACA ACAGCCCCCT TTCGCCCGGC GCTGTGAGCG 240
GCTGCGACCC CAACGGCGTT GCATTCACCT GCAACGACAA CCAGCCTTGG GCCGTAAACA 300
ACAATGTCGC CTACGGTTTT GCGGCTACCG CCTTCCCTGG TGGCAATGAG GCGTCGTGGT 360
GCTGTGCCTG CTATGCTCTT CAATTCACAT CCGGCCCCGT TGCTGGCAAG ACGATGGTTG 420
TGCAATCCAC CAACACTGGC GGAGATCTCA GCGGCACTCA CTTCGATATC CAGATGCCCG 480
GTGGAGGTCT CGGCATCTTC GACGGCTGCA CCCCGCAGTT CGGCTTCACG TTCCCCGGCA 540
ACCGCTACGG CGGTACCACG AGCCGCAGCC AGTGCGCCGA GCTGCCCTCC GTCCTCCGTG 600
ACGGCTGCCA CTGGCGTTAC GACTGGTTCA ACGATGCCGA CAACCCCAAC GTCAACTGGC 660
GCCGCGTCCG ATGCCCGGCG GCCCTCACGA ACCGCTCCGG CTGCGTCCGC AACGACGACA 720
ACAGCTACCC CGTCTTCGAG CCCGGCACGG GCACCCCGCC GACCCCCACG ACCACGACTA 780
CCAGCTCCCC TCCTCAGCCC ACCAACGGCG GAGGCGGCGG CACTTCTCCT CACTGGGGCC 840

AGTGCGGCGG CCAGGGCTGG TCTGGCCCGA CGGCCTGTGC CGGTGGGTCG ACCTGCAACC 900
TGATCAACCC GTGGTACTCC CAGTGCATTC CCAACTAAGT GATCCGGGCA TTGCGGTCGA 960
AAGGGGACCG TTAGTCGACA AGGCCCAGCC AGACCTCAGG CAGGTGGCTG CCATGGCAGA
1020
TTGTATATAG TCTTCCGAGT ACATACTATT GAATGAAAAT AAGAGCGGCT CGGACCATGA
1080
GCAGATGCCA TTTGATAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
1140
AAAAAAAAAA AAAA
1154
(2) INFORMATION FOR SEQ ID NO: 5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 295 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS; single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:
Met Arg Ser Thr Ser lie Leu lie Gly Leu Val Ala Gly Val Ala Ala
15 10 15
Gin Ser Ser Gly Ser Gly His Thr Thr Arg Tyr Trp Asp Cys Cys Lys
20 25 30

Pro Ser Cys Ala Trp Asp Glu Lys Ala Ala Val Ser Arg Pro Val Thr

35

40

45

Thr Cys Asp Arg Asn Asn Ser Pro Leu Ser Pro Gly Ala Val Ser Gly

50

55

60

Cys Asp Pro Asn Gly Val Ala Phe Thr Cys Asn Asp Asn Gin Pro Trp

65

70

75

80

Ala Val Asn Asn Asn Val Ala Tyr Gly Phe Ala Ala Thr Ala Phe Pro

85

90

95

Gly Gly Asn Glu Ala Ser Trp Cys Cys Ala Cys Tyr Ala Leu Gin Phe

100

105

110

Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val Gin Ser Thr Asn

115

120

125

Thr Gly Gly Asp Leu Ser Gly Thr His Phe Asp lie Gin Met Pro Gly

130

135

140

Gly Gly Leu Gly lie Phe Asp Gly Cys Thr Pro Gin Phe Gly Phe Thr

145

150

155

160

Phe Pro Gly Asn Arg Tyr Gly Gly Thr Thr Ser Arg Ser Gin Cys Ala

165

170

175

Glu Leu Pro Ser Val Leu Arg Asp Gly Cys His Tjrp Arg Tyr Asp Trp
180 185 190
Phe Asn Asp Ala Asp Asn Pro Asn Val Asn Trp Arg Arg Val Arg Cys
195 200 205
Pro Ala Ala Leu Thr Asn Arg Ser Gly Cys Val Arg Asn Asp Asp Asn
210 215 220
Ser Tyr Pro Val Phe Glu Pro Gly Thr Gly Thr Pro Pro Thr Pro Thr
225 230 235 240
Thr Thr Thr Thr Ser Ser Pro Pro Gin Pro Thr Asn Gly Gly Gly Gly
245 250 255
Gly Thr Ser Pro His Trp Gly Gin Cys Gly Gly Gin Gly Trp Ser Gly
260 265 270
Pro Thr Ala Cys Ala Gly Gly Ser Thr Cys Asn Leu lie Asn Pro Trp
275 280 285
Tyr Ser Gin Cys lie Pro Asn
290 295
(2) INFORMATION FOR SEQ ID NO: 6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1423 base pairs

(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(Vi) ORIGINAL SOURCE:
(A) ORGANISM: Saccharomyces cerevisiae, DSM 10080
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:
AAAGTTCTGG CCGGAACAGA TCTCCGTTGT CGATCTTCGA TTTTCCAGAC TCAGTCTGTG 60
ACACTCCTTC AATCCACATT CCTTTACTTC TTCGTCACTC ATTCACATCA TGATTTCAGC 120
TTGGATTCTC CTGGGGCTGG TAGGCGCCGT GCCCTCCTCC GTCATGGCCG CCTCGGGCAA 180
AGGCCACACC ACCCGCTACT GGGATTGCTG CAAGACTTCT TGCGCATGGG AGGGCAAGGC 240
ATCCGTCTCC GAGCCTGTCC TGACCTGTAA CAAGCAGGAC AACCCCATCG TCGATGCCAA 300
CGCCAGAAGC GGCTGCGACG GCGGCGGGGC ATTTGCCTGT ACCAACAATT CCCCTTGGGC 360
CGTGAGCGAG GACCTGGCCT ACGGATTTGC TGCCACAGCC CTCAGCGGCG GCACTGAGGG 420
CAGCTGGTGC TGCGCGTGTT ACGCCATCAC ATTCACGAGT GGCCCTGTGG CTGGCAAGAA 480
GATGGTCGTC CAGTCCACGA ACACGGGAGG CGACCTGTCC AACAACCACT TTGACCTGAT 540
GATTCCCGGT GGAGGCCTCG GCATCTTTGA CGGTTGCTCG GCTCAGTTCG GACAACTTCT 600

TCCCGGCGAG CGTTACGGAG GTGTTTCGTC CCGCTCTCAA TGCGATGGCA TGCCCGAGCT 660
CTTGAAAGAC GGTTGCCAGT GGCGCTTCGA CTGGTTCAAG AACTCAGACA ACCCTGACAT 720
CGAGTTCGAG CAGGTCCAGT GTCCCAAAGA GCTCATTGCG GTCTCTGGGT GCGTCCGTGA 780
CGACGATAGC AGCTTTCCCG TCTTCCAAGG TTCGGGCTCA GGAGATGTCA ACCCACCTCC 840
CAAGCCGACT ACGACTACGA CCTCGTCAAA GCCGAAAACA ACCTCTGCAC CATCCACTCT 900
CTCGAACCCA TCCGCCCCTC AACAGCCAGG GAACACTGAT AGACCTGCCG AGACAACCAC 960
TACCAAGCTG CCTGCCCTGC CGGCCACGAC GAGCAGCCCT GCTGTCTCAG TTCCTTCGTC 1020
CAGCGCTCGC GTGCCTTTGT GGGGGCAATG CGACTCGGAA GCTTCATGGG ACGCACCTAA 1080
GAAGTGTGCA AAGGGCACCA AGTGTGTCTA CGTCAACGAC TGGTACTCTC AATGCCAGCC 1140
GAAGAACTCT TGTGCTTGAG AAGCAATGCT CACAGCATGT CCTCTTGTCA CCCCTTCTTT 1200
TCATTCCCAA ACATACTTAC TGTATTATTA TTTCCGATGC TTCATTTCTT GCTTGTTTCT 1260
GTCTTTCCTG CACGCAGCTT TCAACGATAC CCTTCATGCG ATTGCCCTAC GATCAGATGA 1320
TGGGCACGAC ATGGAGGATG GTTTGGGCAC TCACGCGTTC AGGACGGGAA AATTTATTAG 1380
GGCTGAGATC CGTGAATTGA CTTCATTTCG GCGGAATGTC TGC 1423

INFORMATION FOR SEQ ID NO: 7:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 349 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7:
Met lie Ser Ala Trp lie Leu Leu Gly Leu Val Gly Ala Val Pro Ser
15 10 15
Ser Val Met Ala Ala Ser Gly Lys Gly'His Thr Thr Arg Tyr Trp Asp
20 25 30
Cys Cys Lye Thr Ser Cys Ala Trp Glu Gly Lys Ala Ser Val Ser Glu
35 40 45
Pro Val Leu Thr Cys Asn Lys Gin Asp Asn Pro lie Val Asp Ala Asn
50 55 60
Ala Arg Ser Gly Cys Asp Gly Gly Gly Ala Phe Ala Cys Thr Asn Asn
65 70 75 80
Ser Pro Trp Ala Val Ser Glu Asp Leu Ala Tyr Gly Phe Ala Ala Thr
85 90 95

Ala Leu Ser Gly Gly Thr Glu Gly Ser Trp Cys Cys Ala Cys Tyr Ala
100 105 110
He Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met Val Val Gin
115 120 125
Ser Thr Asn Thr Gly Gly Asp Leu Ser Asn Asn His Phe Asp Leu Met
130 135 140
He Pro Gly Gly Gly Leu Gly He Phe Asp Gly Cys Ser Ala Gin Phe
145 150 155 160
Gly Gin Leu Leu Pro Gly Glu Arg Tyr Gly Gly Val Ser Ser Arg Ser
165 170 175
Gin Cys Asp Gly Met Pro Glu Leu Leu Lys Asp Gly Cys Gin Trp Arg
180 185 190
Phe Asp Trp Phe Lys Asn Ser Asp Asn Pro Asp He Glu Phe Glu Gin
195 200 205
Val Gin Cys Pro Lys Glu Leu He Ala Val Ser Gly Cys Val Arg Asp
210 215 220
Asp Asp Ser Ser Phe Pro Val Phe Gin Gly Ser Gly Ser Gly Asp Val
225 230 235 240

Asn Pro Pro Pro Lys Pro Thr Thr Thr Thr Thr Ser Ser Lys Pro Lys
245 250 255
Thr Thr Ser Ala Pro Ser Thr Leu Ser Asn Pro Ser Ala Pro Gin Gin
250 265 270
Pro Gly Asn Thr Asp Arg Pro Ala Glu Thr Thr Thr Thr Lys Leu Pro
275 280 285
Ala Leu Pro Ala Thr Thr Ser Ser Pro Ala Val Ser Val Pro Ser Ser
290 295 300
Ser Ala Arg Val Pro Leu Trp Gly Gin Cys Asp Ser Glu Ala Ser Trp
305 310 315 320
Asp Ala Pro Lys Lys Cys Ala Lys Gly Thr Lys Cys Val Tyr Val Asn
325 330 335
Asp Trp Tyr Ser Gin Cys Gin Pro Lys Asn Ser Cys Ala
340 345
(2) INFORMATION FOR SEQ ID NO: 8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1174 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Saccharomyces cerevisiae, DSM
; 10081
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8 I
GAGCAGCACC CCTCAAGCTG TACAGTTTCC ACCCCGCTCT CTTTTCTTCG GCCCCCAGGA 60
TGCGCTCTAC TCCCGTTCTT CGCACAACCC TGGCOGCTGC ACTTCCTCTG GTCGCCTCCG 120
CGGCCAGTGG CAGTGGCCAG TCCACXJAGAT ACTGGGACTG CTGC3AGCCG TCGTGCGCTT 180
GGCCCGGGAA GGCCGCCGTC AGCCAACCGG TCTACGCGTG CGATGCCAAC TTCCAGCGCC 240
TGTCCGACTT CAATGTCCAG TCGGGCTGCA ACGGCGGCTC GGCCTACTCC TGCGCCGACC 300
AGACTCCCTG GGCGGTGAAC GACAATCTCG CCTACGGCTT CGCCGCGACG AGCATCGCCG 360
GCGGGTCCGA ATCCTCGTGG TGCTGCGCCT GCTACGCGCT CACCTTCACT TCCGGTCCCG 420
TCGCCGGCAA GACAATGGTG GTGCAGTCAA CGAGCACTGG CGGCGACCTG GGAAGTAACC 480
AGTTCGATAT CGCCATGCCC GGCGGCGGCG TGGGCATCTT CAACGGCTGC AGCTCGCAGT 540
TCGGCGGCCT CCCCGGCGCT CAATACGGCG GCATTTCGTC GCGCGACCAG TGCGATTCCT 600
TCCCCGCGCC GCTCAAGCCC GGCTGCCAGT GGCGGTTTGA CTGGTTCCAG AACGCCGACA 660

ACCCGACGTT CACGTTCCAG CAGGTGCAGT GCCCCGCCGA GATCGTTGCC CGCTCCGGCT 720
GCAAGCGCAA CGACGACTCC AGCTTCCCCG TCTTCACCCC CCCAAGCGGT GGCAACGGTG 780
GCACCGGGAC GCCCACGTCG ACTGCGCCTG GGTCGGGCCA GACGTCTCCC GGCGGCGGCA 840
GTGGCTGCAC GTCTCAGAAG TGGGCTCAGT GCGGTGGCAT CGGCTTCAGC GGATGCACCA 900
CCTGTGTCTC TGGCACCACC TGCCAGAAGT TGAACGACTA CTACTCGCAG TGCCTCTAAA 960
CAGCTTTTCG CACGAGGTGG CGGGAC»GAG CAAGGAGACC GTCAACTTCG TCATGCATAT 1020
TTTTTGAGCG CTCAATACAT ACATAACCTT CGATTCTTGT ACATAGCACG CCGGTACACA 1080
TCTCACACCG ACTTTGGGGG CGGAATCAGG CCCGTTTTAA AAAAAAAAAA AAAAAAAAAA 1140
AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAA 1174
(2) INFORMATION FOR SEQ ID NO: 9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 299 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:
Met Arg Ser Thr Pro Val Leu Arg Thr Thr Leu Ala Ala Ala Leu Pro
15 10 15

Leu Val Ala Ser Ala Ala Ser Gly Ser Gly Gin Ser Thr Arg Tyr Trp
20 25 30
Asp Cys Cys Lys Pro Ser Cys Ala Trp Pro Gly Lys Ala Ala Val Ser
35 40 45
Gin Pro Val Tyr Ala Cys Asp Ala Asn Phe Gin Arg Leu Ser Asp Phe
50 55 60
Asn Val Gin Ser Gly Cys Asn Gly Gly Ser Ala Tyr Ser Cys Ala Asp
65 70 75 80
Gin Thr Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala
85 90 95
Thr Ser lie Ala Gly Gly Ser Glu Ser Ser Trp Cys Cys Ala Cys Tyr
100 105 110
Ala Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val
115 120 125
Gin Ser Thr Ser Thr Gly Gly Asp Leu Gly Ser Asn Gin Phe Asp lie
130 135 140
Ala Met Pro Gly Gly Gly Val Gly lie Phe Asn Gly Cys Ser Ser Gin

145

150

155

160

Phe Gly Gly Leu Pro Gly Ala Gin Tyr Gly Gly lie Ser Ser Arg Asp

165

170

175

Gin Cys Asp Ser Phe Pro Ala Pro Leu Lys Pro Gly Cys Gin Trp Arg

180

185

190

Phe Asp Trp Phe Gin Asn Ala Asp Asn Pro Thr Phe Thr Phe Gin Gin

195

200

205

Val Gin Cys Pro Ala Glu lie Val Ala Arg Ser Gly Cya Lys Arg Asn

210

215

220

Asp Asp Ser Ser Phe Pro Val Phe Thr Pro Pro Ser Gly Gly Asn Gly

225

230

235

240

Gly Thr Gly Thr Pro Thr Ser Thr Ala Pro Gly Ser Gly Gin Thr Ser

245

250

255

Pro Gly Gly Gly Ser Gly Cys Thr Ser Gin Lys Trp Ala Gin Cys Gly

260

265

270

Gly lie Gly Phe Ser Gly Cys Thr Thr Cys Val Ser Gly Thr Thr Cys

275

280

285

Gin Lys Leu Asn Asp Tyr Tyr Ser Gin Cys Leu
290 295

(2) INFORMATION FOR SEQ ID NO: 10:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 913 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Escherichia coli, DSM 10512
(Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10:
GCACTATTCT CAGCTCCATT CTCCCTTGAA GTAATTCACC ATGTTCTCTC CGCTCTGGGC 60
CCTGTCGGCT CTGCTCCTAT TTCCTGCCAC TGAAGCCACT AGCGGCGTGA CAACCAGGTA 120
CTGGGACTGC TGCAAGCCGT CTTGTGCTTG GACGGGCAAA GCATCCGTCT CCAAGCCCGT 180
CGGAACCTGC GACATCAACG ACAACGCCCA GACGCCGAGC GATCTGCTCA AGTCGTCCTG 240
TGATGGCGGC AGCGCCTACT ACTGCAGCAA CCAGGGCCCA TGGGCCGTGA ACGACAGCCT 300
TTCCTACGGC TTCGCTGCCG CCAAGCTGTC CGGAAAGCAG GAGACTGATT GGTGCTGTGG 360
CTGCTACAAG CTCACATTCA CCTCCACCGC CGTTTCCGGC AAGCAAATGA TCGTGCAAAT 420
CACGAACACG GGCGGCGhCC TCGGCAACAA CCACTTCGAC ATCGCCATGC CGGGCGGCGG 480
CGTCGGCATC TTCAACGGGT GCTCCAAGCA ATGGAACGGC ATCAATCTGG GCAACCAGTA 540

TGGCGGCTTC ACTGACCGCT CGCAATGTGC GACGCTCCCG TCCAAGTGGC AGGCCAGCTG 600
CAACTGGCGC TTCGACTGGT TCGAGAATGC CGACAACCCC ACCGTCGATT GGGAGCCTGT 660
CACTTGCCCA CAGGAATTGG TCGCCCGGAC TGGCTGTTCC CGTACCTAAG TGGGGGTGGA 720
ACCTCCATGT GAATTGGTGT ATATAGCTCC TGCCTGAGCA TCCACCAGTT CGCATGTGTT 780
6ATCAGGAGT TGTGTTGCCT TGCTAGGAAA GACTTTGTTG GAAACTTGCG TGTTTATTCC 840
AATTGAATAA CCCTGTATAG ACCGGTCACA TTTTTCTCTG AAAAAAAAAA AAAAAAAAAA 900
AAAAAAAAAA AAA
913
(2) INFORMATION FOR SEQ ID NO: 11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 222 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:
Met Phe Ser Pro Leu Trp Ala Leu Ser Ala Leu Leu Leu Phe Pro Ala
1 5 10 15
Thr Glu Ala Thr Ser Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys
20 25 30

Pro Ser Cys Ala Trp Thr Gly Lys Ala Ser Val Ser Lys Pro Val Gly
35 40 45
Thr Cys Asp lie Asn Asp Asn Ala Gin Thr Pro Ser Asp Leu Leu Lys
50 55 60
Ser Ser Cys Asp Gly Gly Ser Ala Tyr Tyr Cya Ser Asn Gin Gly Pro
65 70 75 80
Trp Ala Val Asn Asp Ser Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu
85 90 95
Ser Gly Lys Gin Glu Thr Asp Trp Cys Cys Gly Cys Tyr Lys Leu Thr
100 105 110
Phe Thr Ser Thr Ala Val Ser Gly Lys Gin Met He Val Gin He Thr
115 120 125
Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp He Ala Met Pro
130 135 140
Gly Gly Gly Val Gly He Phe Asn Gly Cys Ser Lys Gin Trp Asn Gly
145 150 155 160
He Asn Leu Gly Asn Gin Tyr Gly Gly Phe Thr Asp Arg Ser Gin Cys
165 170 175

Ala Thr Leu Pro Ser Lys Trp Gin Ala Ser Cys Asn Trp Arg Phe Asp
180 185 190
Trp Phe Glu Asn Ala Asp Asn Pro Thr Val Asp Trp Glu Pro Val Thr
195 200 205
Cys Pro Gin Glu Leu Val Ala Arg Thr Gly Cys Ser Arg Thr
210 215 220
2) INFORMATION FOR SEQ ID NO: 12:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 808 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Escherichia coli, DSM 10511
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12:
CCGCTGCTGG GTATATAATG CTCAGACTTG GAACCAATGG TCCATCCAAA CATGCTTAAA 60
ACGCTCGCTC CATTGATCAT CTTGGCCGCC TCGGTCACAG CGCAAACAGC AGGAGTTACG 120
ACCCGCTACT GGGACTGCTG CAAGCCAAGC TGTGGATGGA GTGGAAAGGC TTCTGTTTCT 180
GCTCCAGTCA GAACTTGCGA TCGTAATGGA AATACACTTG GCCCAGACGT GAAAAGCGGA 240

TGTGATAGCG GTGGAACGTC ATTCACTTGC GCGAACAATG GTCCATTTGC GATTGACAAT 300
AACACTGCAT ATGGTTTTGC TGCAGCCCAC TTAGCGGGCT CTAGCGAAGC A6CCTGGTGT 360
TGCCAGT6CT ACGAATTGAC GTTTACGAGT GGACCCGTAG TTGGGAAGAA ACTGACCGTT 420
CAAGTCACAA ACACGGGAGG TGACCTCGGA AATAATCACT TTGACCTGAT GATCCCCGGT 480
GGAGGTGTTG GCCTCTTCAC ACAAGGATGT CCTGCTCAGT TTGGGAGCTG GAACGGGGGT 540
GCTCAATACG GGGGTGTGTC CAGCCGTGAC CAATGCTCCC AACTTCCAGC AGCTGTGCAA 600
GCTGGATGTC AATTCCGTTT CGACTGGATG GGTGGCGCGG ATAACCCCAA CGTCACCTTC 660
CGACCTGTGA CCTGCCCAGC GCAGCTCACT AATATCTCGG GCTGTGTTCG TAAATGATTC 720
f ACGAATATGT AGTGTCGAAT ATGTACATGT GTATGTACTA TAGCTTCAAA GATGGAGGGT 780
CTGTTTAAAA AAAAAAAAAA AAAAAAAA 808 )
(2) INFORMATION FOR SEQ ID NO: 13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 226 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13:
Met Val His Pro Asn Met Leu Lys Thr Leu Ala Pro Leu lie lie Leu
15 10 15
Ala Ala Ser Val Thr Ala Gin Thr Ala Gly Val Thr Thr Arg Tyr Trp
20 25 30
Asp Cys Cys Lys Pro Ser Cys Gly Trp Ser Gly Lys Ala Ser Val Ser
35 40 45
Ala Pro Val Arg Thr Cys Asp Arg Asn Gly Asn Thr Leu Gly Pro Asp
50 55 60
Val Lys Ser Gly Cys Asp Ser Gly Gly Thr Ser Phe Thr Cys Ala Asn
65 70 75 80
Asn Gly Pro Phe Ala lie Asp Asn Asn Thr Ala Tyr Gly Phe Ala Ala
85 90 95
Ala His Leu Ala Gly Ser Ser Glu Ala Ala Trp Cys Cys Gin Cys Tyr
100 105 110
Glu Leu Thr Phe Thr Ser Gly Pro Val Val Gly Lys Lys Leu Thr Val
115 120 125
Gin Val Thr Asn Thr Gly Gly Asp Leu Gly Asn Aen His Phe Asp Leu

130 135 140
Met lie Pro Gly Gly Gly Val Gly Leu Phe Thr Gin Gly Cys Pro Ala
145 150 155 160
Gin Phe Gly Ser Trp Asn Gly Gly Ala Gin Tyr Gly Gly Val Ser Ser
165 170 175
Arg Asp Gin Cys Ser Gin Leu Pro Ala Ala Val Gin Ala Gly Cys Gin
180 185 190
Phe Arg Phe Asp Trp Met Gly Gly Ala Asp Asn Pro Asn Val Thr Phe
195 200 205
Arg Pro Val Thr Cys Pro Ala Gin Leu Thr Asn lie Ser Gly Cys Val
210 215 220
Arg Lys 225
(2) INFORMATION FOR SEQ ID NO: 14-A:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1048 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: CDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14-A:
GACTTGGAAC CAATGGTCCA TCCAAACATG CTTAAAACGC TCGCTCCATT GATCATCTTG 60
GCCGCCTCGG TCACAGCGCA AACAGCAGGA GTTACGACCC GCTACTGGGA CTGCTGCAAG 120
CCAAGCTGTG GATGGAGTGG AAAGGCTTCT GTTTCTGCTC CAGTCAGAAC TTGCGATCGT 180
AATGGAAATA CACTTGGCCC AGACGTGAAA AGCGGATGTG ATAGCGGTGG AACGTCATTC 240
ACTTGCGCGA ACAATGGTCC ATTTGCGATT GACAATAACA CTGCATATGG TTTTGCTGCA 300
GCCCACTTAG CGGGCTCTAG CGAAGCAGCC TGGTGTTGCC AGTGCTACGA ATTGACGTTT 360
ACGAGTGGAC CCGTAGTTGG GAAGAAACTG ACCGTTCAAG TCACAAACAC GGGAGGTGAC 420
CTCGGAAATA ATCACTTTGA CCTGATGATC CCCGGTGGAG GTGTTGGCCT CTTCACACAA 480
GGATGTCCTG CTCAGTTTGG GAGCTGGAAC GGGGGTGCTC AATACGGGGG TGTGTCCAGC 540
CGTGACCAAT GCTCCCAACT TCCAGCAGCT GTGCAAGCTG GATGTCAATT CCGTTTCGAC 600
TGGATGGGTG GCGCGGATAA CCCCAACGTC ACCTTCCGAC CTGTGACCTG CCCAGCGCAG 660
CTCACTAATA TCTCGGGCTG TGTTCGTAAA CCCTCCAGCA GCACCAGCTC TCCGGTCAAC 720
CAGCCTACCA GCACCAGCAC CACGTCCACC TCCACCACCT CGAGCCCGCC AGTCCAGCCT 780
ACGACTCCCA GCGGCTGCAC TGCTGAGAGG TGGGCTCAGT GCGGCGGCAA TGGCTGGAGC 840

G6CTGCACCA CCTGCGTCGC TGGCAGCACT TGCACGAAGA TTAATGACTG GTACCATCAG 900
TGCCTGTAGA CGCAGGGCAG CTTGAGGGCC TTACTGGTGG CGCAACGAAA TGACACTCCC 960
AATCACTGTA TTAGTTCTTG TACATAATTT CGTCATCCCT CCAGGGATTG TCACATAAAT 1020
GCAATGAGGA ACAATGAGTA CAGAATTC 1048
(2) INFORMATION FOR SEQ ID NO: 14-B:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 298 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY- linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14-B:
Met Val His Pro Asn Met Leu Lys Thr Leu Ala Pro Leu lie lie Leu
15 10 15
Ala Ala Ser Val Thr Ala Gin Thr Ala Gly Val Thr Thr Arg Tyr Trp
20 25 30
Asp Cys Cys Lys Pro Ser Cys Gly Trp Ser Gly Lys Ala Ser Val Ser
35 40 45
Ala Pro Val Arg Thr Cys Asp Arg Asn Gly Asn Thr Leu Gly Pro Asp
50 55 60

Val Lys Ser Gly Cys Asp Ser Gly Gly Thr Ser Phe Thr Cys Ala Asn
65 70 75 80
Asn Gly Pro Phe Ala lie Asp Asn Asn Thr Ala Tyr Gly Phe Ala Ala
85 90 95
Ala His Leu Ala Gly Ser Ser Glu Ala Ala Trp Cys Cys Gin Cys Tyr
100 105 110
Glu Leu Thr Phe Thr Ser Gly Pro Val Val Gly Lys Lys Leu Thr Val
115 120 125
Gin Val Thr Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp Leu
130 135 140
Met lie Pro Gly Gly Gly Val Gly Leu Phe Thr Gin Gly Cys Pro Ala
145 150 155 160
Gin Phe Gly Ser Trp Asn Gly Gly Ala Gin Tyr Gly Gly Val Ser Ser
165 170 175
Arg Asp Gin Cys Ser Gin Leu Pro Ala Ala Val Gin Ala Gly Cys Gin
180 185 190
Phe Arg Phe Asp Trp Met Gly Gly Ala Asp Asn Pro Asn Val Thr Phe

195 200 205
Arg Pro Val Thr Cys Pro Ala Gin Leu Thr Asn lie Ser Gly Cys Val
210 215 220
Arg Lys Pro Ser Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser
225 230 235 240
Thr Ser Thr Thr Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro
245 250 255
Thr Thr Pro Ser Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly
260 265 270
Asn Gly Trp Ser Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr
275 280 285
Lys lie Asn Asp Trp Tyr His Gin Cys Leu
290 295
(2) INFORMATION FOR SEQ ID NO: 15-A:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1031 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15-A:
CCATCCAAAC ATGCTTAAAA CGCTCGCTCC ATTGATCATC TTGGCCGCCT C6GTCACAGC 60
GCAAACAGCA GGAGTTACGA CCCGCTACTG GGACTGCTGC AAGCCAAGCT GTGGATGGAG 120
TGGAAAGGCT TCTGTTTCTG CTCCAGTCAG AACTTGCGAT CGTAATGGAA ATACACTTGG 180
CCCAGACGTG AAAAGCGGAT GTGATAGCGG TGGAACGTCA TTCACTTGCG CGAACAATGG 240
TCCATTTGCG ATTGACAATA ACACTGCATA TGGTTTTGCT GCAGCCCACT TAGCGGGCTC 300
TAGCGAAGCA GCCTGGTGTT GCCAGTGCTA CGAATTGACG TTTACGAGTG GACCCGTAGT 360
TGGGAAGAAA CTGACCGTTC AAGTCACAAA CACGGGAGGT GACCTCGGAA ATAATCACTT 420
TGACCTGATG ATCCCCGGTG GAGGTGTTGG CCTCTTCACA CAAGGATGTC CTGCTCAGTT 480
TGGGAGCTGG AACGGGGGTG CTCAATACGG GGGTGTGTCC AGCCGTGACC AATGCTCCCA 540
ACTTCCAGCA GCTGTGCAAG CTGGATGTCA ATTCCGTTTC GACTGGATGG GTGGCGCGGA 600
TAACCCCAAC GTCACCTTCC GACCTGTGAC CTGCCCAGCG CAGCTCACTA ATATCTCGGG 660
CTGTGTTCGT AAACCCTCCA GCAGCACCAG CTCTCCGGTC AACCAGCCTA CCAGCACCAG 720
CACCACGTCC ACCTCCACCA CCTCGAGCCC GCCAGTCCAG CCTACGACTC CCAGCGGCTG 780
CACTGCTGAG AGGTGGGCTC AGTGCGGCGG CAATGGCTGG AGCGGCTGCA CCACCTGCGT 840

CGCTGGCAGC ACTTGCACGA AGATTAATGA CTGGTACCAT CAGTGCCTGT AGACGCAGGG 900
CAGCTTGAGG GCCTTACTGG TGGCGCAACG AAATGACACT CCCAATCACT GTATTAGTTC 960
TTGTACATAA TTTCGTCATC CCTCCAGGGA TTGTCACATA AATGCAATGA GGAACAATGA 1020
GTACAGAATT C 1031
(2) INFORMATION FOR SEQ ID NO: 15-B:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 293 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15-B:
Met Leu Lya Thr Leu Ala Pro Leu lie lie Leu Ala Ala Ser Val Thr
15 10 15
Ala Gin Thr Ala Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys Pro
20 25 30
Ser Cys Gly Trp Ser Gly Lys Ala Ser Val Ser Ala Pro Val Arg Thr
35 40 45
Cys Asp Arg Asn Gly Asn Thr Leu Gly Pro Asp Val Lys Ser Gly Cys
50 55 60

Asp Ser Gly Gly Thr Ser Phe Thr Cys Ala Asn Asn Gly Pro Phe Ala
65 70 75 80
lie Asp Asn Asn Thr Ala Tyr Gly Phe Ala Ala Ala His Leu Ala Gly
85 90 95
Ser Ser Glu Ala Ala Trp Cys Cys Gin Cys Tyr Glu Leu Thr Phe Thr
100 105 110
Ser Gly Pro Val Val Gly Lys Lys Leu Thr Val Gin Val Thr Asn Thr
115 120 125
Gly Gly Asp Leu Gly Asn Asn His Phe Asp Leu Met lie Pro Gly Gly
130 135 140
Gly Val Gly Leu Phe Thr Gin Gly Cys Pro Ala Gin Phe Gly Ser Trp
145 150 155 160
Asn Gly Gly Ala Gin Tyr Gly Gly Val Ser Ser Arg Asp Gin Cys Ser
165 170 175
Gin Leu Pro Ala Ala Val Gin Ala Gly Cys Gin Phe Arg Phe Asp Trp
180 185 190
Met Gly Gly Ala Asp Asn Pro Asn Val Thr Phe Arg Pro Val Thr Cys

195 200 205
Pro Ala Gin Leu Thr Asn He Ser Gly Cys Val Arg Lys Pro Ser Ser
210 215 220
Ser Thr Ser Ser Pro Val Asn Gin Pro Thr. Ser Thr Ser Thr Thr Ser
225 230 235 240
Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser Gly
245 250 255
Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser Gly
260 265 270
Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys He Asn Asp Trp
275 280 285
Tyr His Gin Cys Leu 290
(2) INFORMATION FOR SEQ ID NO: 16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1132 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECtTLE TYPE: cDNA

(vi) ORIGINAL SOURCE:
(A) ORGANISM: Escherichia coli, DSM 10571
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16:
CAACAGTTCA AACACCTACA AGGTCCCGTG CCCTGTAGAC CATGCGTTCC TCTGCAGTCC 60
TCATCGGCCT CGTGGCCGGT GTGGCCGCCC AGTCCTCTGG CACCGGCCGC ACCACCAGAT 120
ACTGGGACTG CTGCAAGCCG TCCTGCGGGT GGGACGAAAA GGCCTCCGTC AGCCAGCCCG 180
TCAAGACGTG CGATAGGAAC AACAACCCTC TCGCGTCCAC GGCCAGGAGC GGCTGCGATT 240
CCAACGGCGT CGCCTACACG TGCAACGATA ACCAGCCGTG GGCTGTCAAC GATAACCTGG 300
CCTATGGTTT TGCTGCCACG GCTTTCAGTG GTGGATCGGA GGCCAGCTGG TGCTGTGCCT 360
GCTATGCCCT TCAGTTCACC TCCGGCCCTG TTGCGGGAAA GACCATGGTC GTCCAGTCGA 420
CAAACACCGG CGGCGACCTC AGCGGCAACC ACTTTGACAT CCTCATGCCC GGCGGCGGCC 480
TGGGCATCTT CGACGGCTGC ACCCCGCAAT GGGGCGTCAG CTTCCCCGGA AACCGCTACG 540
GCGGCACCAC CAGCCGCAGC CAGTGCTCCC AAATCCCCTC GGCCCTGCAG CCCGGCTGCA 600
ACTGGCGGTA CGACTGGTTC AACGACGCCG ACAACCCCGA CGTCTCGTGG CGCCGCGTCC 660
AGTGCCCCGC CGCACTCACC GACCGCACCG GCTGCCGCCG CTCCGATGAC GGGAACTATC 720
CCGTCTTCCA GCCCGGTCCG CCCCCGGCCA CGACGATCAG GACATCGACT ACCATCACAG 780

CCTCATCGTC GTCTTCGTCT TCGTCGTCGT CGACTACGGC TGGTAGCCCG CCTGTGCCGA 840
CTGGTGGTGG TAGTGGGCCA ACGTCGCCTG TCTGGGGACA GTGCGGCGGT CAGGGATGGA 900
GTGGTCCTAC GCGTTGTGTT GCTGGGTCGA CATGCAGTGT GGTCAACCCG TGGTACTCGC 960
AGTGTTTTCC TTAAGGAGCC TCTGGCTGAG CAGATCCTTT CGAAGAGGAG GGTCTCTCTG 1020
CTCTTTCAGT CTGTTCAGGG AACGGCCGTC TCGGCTACAT TGTACATATC CCACCTCGTA 1080
TATAGCTAGC TCATCTACAC TTGTGATCTC CAAAAAAAAA AAAAAAAAAA AA
1132
(2) INFORMATION FOR SEQ ID NO: 17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 310 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17:
Met Arg Ser Ser Ala Val Leu lie Gly Leu Val Ala Gly Val Ala Ala
15 10 15
Gin Ser Ser Gly Thr Gly Arg Thr Thr Arg Tyr Trp Asp Cys Cys Lys
20 25 30
Pro Ser Cys Gly Trp Asp Glu Lys Ala Ser Val Ser Gin Pro Val Lys

35

40

45

Thr Cys Asp Arg Asn Asn Asn Pro Leu Ala Ser Thr Ala Arg Ser Gly

50

55

60

Cys Asp Ser Asn Gly Val Ala Tyr Thr Cys Asn Asp Asn Gin Pro Trp

65

70

75

80

Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Ala Phe Ser

85

90

95

Gly Gly Ser Glu Ala Ser Trp Cys Cys Ala Cys Tyr Ala Leu Gin Phe

100

105

110

Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val Gin Ser Thr Asn

115

120

125

Thr Gly Gly Asp Leu Ser Gly Asn His Phe Asp He Leu Met Pro Gly

130

135

140

Gly Gly Leu Gly He Phe Asp Gly Cys Thr Pro Gin Trp Gly Val Ser

145

150

155

160

Phe Pro Gly Asn Arg Tyr Gly Gly Thr Thr Ser Arg Ser Gin Cys Ser

165

170

175

Gin He Pro Ser Ala Leu Gin Pro Gly Cys Asn Trp Arg Tyr Asp Trp

180

185

190

Phe Asn Asp Ala Asp Asn Pro Asp Val Ser Trp Arg Arg Val Gin Cyj

195

200

205

Pro Ala Ala Leu Thr Asp Arg Thr Gly Cys Arg Arg Ser Asp Asp Gly

210

215

220

Asn Tyr Pro Val Phe Gin Pro Gly Pro Pro Pro Ala Thr Thr lie Arg

225

230

235

240

Thr Ser Thr Thr lie Thr Ala Ser Ser Ser Ser Ser Ser Ser Ser Ser

245

250

255

Ser Thr Thr Ala Gly Ser Pro Pro Val Pro Thr Gly Gly Gly Ser Gly

260

265

270

Pro Thr Ser Pro Val Trp Gly Gin Cys Gly Gly Gin Gly Trp Ser Gly

275

280

285

Pro Thr Arg Cys Val Ala Gly Ser Thr Cys Ser Val Val Asn Pro Trp

290

295

300

Tyr Ser Gin Cys Phe Pro
305 310

(2) INFORMATION FOR SEQ ID NO: 18-A:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 885 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: CDNA
(vi) ORIGINAL SOURCE:
"Construction from Macrophomina"
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..885
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18-A:
ATG TTC TCT CCG CTC TGG GCC CTG TCG GCT CTG CTC CTA TTT CCT GCC
48
Met Phe Ser Pro Leu Trp Ala Leu Ser Ala Leu Leu Leu Phe Pro Ala
145 150 155
ACT GAA GCC ACT AGC GGC GTG ACA ACC AGG TAC TGG GAC TGC TGC AAG
96
Thr Glu Ala Thr Ser Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys
160 165 170
CCG TCT TGT GCT TGG ACG GGC AAA GCA TCC GTC TCC AAG CCC GTC GGA 144
Pro Ser Cys Ala Trp Thr Gly Lys Ala Ser Val Ser Lys Pro Val Gly
175 180 185
ACC TGC GAC ATC AAC GAC AAC GCC CAG ACG CCG AGC GAT CTG CTC AAG
192
Thr Cyg Asp He Asn Asp Asn Ala Gin Thr Pro Ser Asp Leu Leu Lys
190 195 200 205
TCG TCC TGT GAT GGC GGC AGC GCC TAC TAC TGC AGC AAC CAG GGC CCA

240
Ser Ser Cys Asp Gly Gly Ser Ala Tyr Tyr Cys Ser Asn Gin Gly Pro
210 215 220
TGG GCC GTG AAC GAC AGC CTT TCC TAC GGC TTC GCT GCC GCC AAG CTG 288
Trp Ala Val Asn Asp Ser Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu
225 230 235
TCC GGA AAG CAG GAG ACT GAT TGG TGC TGT GGC TGC TAC AAG CTC ACA 336
Ser Gly Lys Gin Glu Thr Asp Trp Cys Cys Gly Cys Tyr Lys Leu Thr
240 245 250
TTC ACC TCC ACC GCC GTT TCC GGC AAG CAA ATG ATC GTG CAA ATC ACG 384
Phe Thr Ser Thr Ala Val Ser Gly Lys Gin Met lie Val Gin lie Thr
255 260 265
i AAC ACG GGC GGC GAC CTC GGC AAC AAC CAC TTC GAC ATC GCC ATG CCG 432
Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp He Ala Met Pro
270 275 280 285
GGC GGC GGC GTC GGC ATC TTC AAC GGG TGC TCC AAG CAA TGG AAC GGC 480
Gly Gly Gly Val Gly He Phe Asn Gly Cys Ser Lys Gin Trp Asn Gly
290 295 300
I ATC AAT CTG GGC AAC CAG TAT GGC GGC TTC ACT GAC CGC TCG CAA TGT 528
He Asn Leu Gly Asn Gin Tyr Gly Gly Phe Thr Asp Arg Ser Gin Cys
305 310 315
GCG ACG CTC CCG TCC AAG TGG CAG GCC AGC TGC AAC TGG CGC TTC GAC
576
Ala Thr Leu Pro Ser Lys Trp Gin Ala Ser Cys Asn Trp Arg Phe Asp
320 325 330
TGG TTC GAG AAT GCC GAC AAC CCC ACC GTC GAT TGG GAG COT GTC ACT
624
Trp Phe Glu Asn Ala Asp Asn Pro Thr Val Asp Trp Glu Pro Val Thr
335 340 345

TGC CCA CAG GAA TTG GTC GCC CGG ACT GGC TGT TCC CGT ACC CCC TCC
672
Cys Pro Gin Glu Leu Val Ala Arg Thr Gly Cys Ser Arg Thr Pro Ser
350 355 360 365
AGC AGC ACC AGC TCT CCG GTC AAC CAG CCT ACC AGC ACC AGC ACC ACG 720
Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr Ser Thr Thr
370 375 380
TCC ACC TCC ACC ACC TCG AGC CCG CCA GTC CAG CCT ACG ACT CCC AGC
768
Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser
385 390 395
GGC TGC ACT GCT GAG AGG TGG GCT CAG TGC GGC GGC AAT GGC TGG AGC
816
Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser
400 405 410
GGC TGC ACC ACC TGC GTC GCT GGC AGC ACT TGC ACG AAG ATT AAT GAC 864
Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys lie Asn Asp
415 420 425
TGG TAC CAT CAG TGC CTG TAG
885
Trp Tyr His Gin Cys Leu *
430 435
(2) INFORMATION FOR SEQ ID NO: 18-B:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 295 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18-B:
Met Phe Ser Pro Leu Trp Ala Leu Ser Ala Leu Leu Leu Phe Pro Ala

15 10 15
Thr Glu Ala Thr Ser Gly Val Thr Thr Arg Tyr Trp Asp Cys Cys Lys
20 25 30
Pro Ser Cys Ala Trp Thr Gly Lys Ala Ser Val Ser Lys Pro Val Gly
35 40 45
Thr Cys Asp lie Asn Asp Asn Ala Gin Thr Pro Ser Asp Leu Leu Lys
50 55 60
Ser Ser Cys Asp Gly Gly Ser Ala Tyr Tyr Cys Ser Asn Gin Gly Pro
65 70 75 80
Trp Ala Val Asn Asp Ser Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu
85 90 95
Ser Gly Lys Gin Glu Thr Asp. .Trp Cys Cys Gly Cys Tyr Lys Leu Thr
100 105 110
Phe Thr Ser Thr Ala Val Ser Gly Lys Gin Met lie Val Gin lie Thr
115 120 125
Asn Thr Gly Gly Asp Leu Gly Asn Asn His Phe Asp lie Ala Met Pro
130 135 140
Gly Gly Gly Val Gly lie Phe Asn Gly Cys Ser Lys Gin Trp Asn Gly
145 150 155 160
lie Asn Leu Gly Asn Gin Tyr Gly Gly Phe Thr Asp Arg Ser Gin Cys
165 170 175
Ala Thr Leu Pro Ser Lys Trp Gin Ala Ser Cys Asn Trp Arg Phe Asp
180 185 190
Trp Phe Glu Asn Ala Asp Asn Pro Thr Val Asp Trp Glu Pro Val Thr
195 200 205
Cys Pro Gin Glu Leu Val Ala Arg Thr Gly Cys Ser Arg Thr Pro Ser
210 215 220
Ser Ser Thr Ser Ser Pro Val Asn Gin Pro Thr Ser Thr Ser Thr Thr
225 230 235 240

Ser Thr Ser Thr Thr Ser Ser Pro Pro Val Gin Pro Thr Thr Pro Ser
245 250 255
Gly Cys Thr Ala Glu Arg Trp Ala Gin Cys Gly Gly Asn Gly Trp Ser
260 265 270
Gly Cys Thr Thr Cys Val Ala Gly Ser Thr Cys Thr Lys He Asn Asp
275 280 285
Trp Tyr His Gin Cys Leu *
290 295
(2) INFORMATION FOR SEQ ID NO: 19:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 425 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Escherichia coli, DSM 10576
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:l..4 25
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19:
CAAGATACAA TATGCGTTCC TCCACTATTT TGCAAACCGG CCTGGTGGCC GTTCTCCCCT 60
TCGCCGTCCA GGCCGCCTCA GGATCCGGCA AGTCCACCAG ATATTGGGAC TGCTGCAAAC 120
CATCTTGTGC CTGGTCCGGC AAGGCTTCTG TCAACCGCCC TGTTCTCGCC TGCAACGCAA 180

ACAACAACCC GCTGAACGAC GCCAACGTCA AGTCAGGATG TGATGGCGGT TCTGCATACA 240
CCTGTGCCAA CAACTCTCCC TGGGCAGTGA ATGACAATCT GGCCTACGGC TTCGCGGCCA 300
CAAAACTCAG CGGGGGGACC GAGTCATCTT GGTGCTGCGC CTGTTATGCC CTCACATTCA 360
CATCGGGTCC TGTTTCTGGC AAAACCTTGG TTGTCCAGTC TACCAGTACC GGTGGTGATC 420
TTGGC 425
(2) INFORMATION FOR SEQ ID NO: 20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 141 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20:
Met Arg Ser Ser Thr He Leu Gin Thr Gly Leu Val Ala Val Leu Pro
15 10 15
Phe Ala Val Gin Ala Ala Ser Gly Ser Gly Lys Ser Thr Arg Tyr Trp
20 25 30
Asp Cys Cys Lys Pro Ser Cys Ala Trp Ser Gly Lys Ala Ser Val Asn
35 40 45
Arg Pro Val Leu Ala Cys Asn Ala Asn Asn Asn Pro Leu Asn Asp Ala

Asn Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys Ala Asn
65 70 75 80
Asn Ser Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala
85 90 95
Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser Trp Cys Cys Ala Cys Tyr
100 105 110
Ala Leu Thr Phe Thr Ser Gly Pro Val Ser Gly Lys Thr Leu Val Val
115 120 125
Gin Ser Thr Ser Thr Gly Gly Asp Leu Gly
130 135
2) INFORMATION FOR SEQ ID NO: 21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 108 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Saccobolus dilutellus
(B) STRAIN: CBS 275.96

(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:!..108
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21:
TCG GOT TGC GAT AAC GGT GGT GGC ACT GCA TAC ATG TGT GCC AGC CAG
Ser Ala Cys Asp Asn Gly Gly Gly Thr Ala Tyr Met Cys Ala Ser Gin
is 10 15
GAG CCG TGG GCA GTG AGC TCC AAC GTC GCG TAC GGC TTT GCT GCA GTT
Glu Pro Trp Ala Val Ser Ser Asn Val Ala Tyr Gly Phe Ala Ala Val
20 25 30
AGA ATC AGC GGA
Arg lie Ser Gly 35
(2) INFORMATION FOR SEQ ID NO: 22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 36 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22:
Ser Ala Cys Asp Asn Gly Gly Gly Thr Ala Tyr Met Cys Ala Ser Gin
15 10 15
Glu Pro Trp Ala Val Ser Ser Asn Val Ala Tyr Gly Phe Ala Ala Val
20 25 30
Arg He Ser Gly 35

(2) INFORMATION FOR SEQ ID NO: 23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 99 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Thermomyces verrucosus
(B) STRAIN: CBS 285.96
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:!..99
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 23:
GCC TGC AAC GCA AAC TTC CAG CGC ATC AGT GAC CCC AAC GCC AAG TCG Ala Cys Asn Ala Asn Phe Gin Arg lie Ser Asp Pro Asn Ala Lys Ser
GGC TGC GAT GGT GGC TCG GCC TTC TCT TGC GCC AAA CAA ACC CCT TGG Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Lys Gin Thr Pro Trp
GCC
Ala
(2) INFORMATION FOR SEQ ID NO: 24:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 33 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24:
Ala Cys Asn Ala Asn Phe Gin Arg lie Ser Asp Pro Asn Ala Lys Ser
1 5 10 15
Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Lys Gin Thr Pro Trp
20 25 30
Ala
(2) INFORMATION FOR SEQ ID NO: 25:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 225 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Xylaria hypoxylon
(B) STRAIN: CBS 284.96
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..225
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25:
GAC CAG CCG CTC GGC GGA CAA CGG ACG CGA CCA AGG AGC GCG TGC GAC
48
Asp Gin Pro Leu Gly Gly Gin Arg Thr Arg Pro Arg Ser Ala Cys Asp
35 40 45
AAT GGC GGC TCT GCA TAG ATG TGC AGC AAC CAG AGC CCG TGG GCC GTC

96
Asn Gly Gly Ser Ala Tyr Met Cys Ser Asn Gin Ser Pro Trp Ala Val
50 55 60 65
GAC GAT TCT CTC AGT TAG GGA TGG GCT GCC GTT AGG ATC TAT GGA CAT
144
Asp Asp Ser Leu Ser Tyr Gly Trp Ala Ala Val Arg lie Tyr Gly His
70 75 80
ACC GAA ACT ACT TGG TGC TGC GCT TGC TAC GAG TTG ACT TTT ACC AGC
192
Thr Glu Thr Thr Trp Cys Cys Ala Cys Tyr Glu Leu Thr Phe Thr Ser
85 90 95
GGT CCG GTT AGC GGC AAG AAG ATG ATT GTT CAG
225
Gly Pro Val Ser Gly Lys Lys Met lie Val Gin
100 105
(2) INFORMATION FOR SEQ ID NO: 26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 75 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26:
Asp Gin Pro Leu Gly Gly Gin Arg Thr Arg Pro Arg Ser Ala Cys Asp
15 10 15
Asn Gly Gly Ser Ala Tyr Met Cys Ser Asn Gin Ser Pro Trp Ala Val
20 25 30
Asp Asp Ser Leu Ser Tyr Gly Trp Ala Ala Val Arg lie Tyr Gly His
35 40 45
Thr Glu Thr Thr Trp Cys Cys Ala Cys Tyr Glu Leu Thr Phe Thr Ser
50 55 60

Gly Pro Val Ser Gly Lys Lys Met lie Val Gin
65 70 75
(2) INFORMATION FOR SEQ ID NO: 27:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 177 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Fusarium oxysporum ssp lycopersici
(B) STRAIN: CBS 645.78
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:!..177
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27:
AGA AAC GAC AAC CCC ATC TCC AAC ACC AAC GCT GTC AAC GGT TGT GAG
48
Arg Asn Asp Asn Pro lie Ser Asn Thr Asn Ala Val Asn Gly Cys Glu
30 35 40 45
GGT GGT GGT TCT GCT TAT GCT TGC ACC AAC TAC TCT CCC TGG GCT GTC
96
Gly Gly Gly Ser Ala Tyr Ala Cys Thr Asn Tyr Ser Pro Trp Ala Val
50 55 60
AAC GAT GAG CTT GCC TAC GGT TTC GCT GCT ACC AAG ATC TCC GGT GGC
144
Asn Asp Glu Leu Ala Tyr Gly Phe Ala Ala Thr Lys lie Ser Gly Gly

65 70 75
TCC GAG GCC AGC TGG TGC TGT GCC TGC TAT CTA 177
Ser Glu Ala Ser Trp Cys Cys Ala Cys Tyr Leu
80 85
(2) INFORMATION FOR SEQ ID NO: 28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 59 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28:
Arg Asn Asp Asn Pro lie Ser Asn Thr Asn Ala Val Asn Gly Cys Glu
15 10
15
Gly Gly Gly Ser Ala Tyr Ala Cys Thr Asn Tyr Ser Pro Trp Ala Val
20 25 30
Asn Asp Glu Leu Ala Tyr Gly Phe Ala Ala Thr Lys lie Ser Gly Gly
35 40 45
Ser Glu Ala Ser Trp Cys Cys Ala Cys Tyr Leu
50 55
(2) INFORMATION FOR SEQ ID NO: 29:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 63 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: LINEAR
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Nectria pinea
(B) STRAIN: CBS 279.96
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..63
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 29:
AGC GGC TGT GAC GGT GGT TCT GCC TAC GCC TGT GCA AAC AAC TCC CCT 4 8
Ser Gly Cys Asp Gly Gly Ser Ala Tyr Ala Cys Ala Asn Asn
Ser Pro
60 65 70
75
TGG GCT GTC AAC GAT
63 Trp Ala Val Asn Asp
80
(2) INFORMATION FOR SEQ ID NO: 30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid

(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 30:
Ser Gly Cys Asp Gly Gly Ser Ala Tyr Ala Cys Ala Asn Asn Ser Pro
15 10
15
Trp Ala Val Asn Asp 20
(2) INFORMATION FOR SEQ ID NO: 31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 177 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Humicola grisea Traeen
(B) STRAIN: ATCC 22726
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..177
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 31:
AAC CAG CCT GTC TTC ACT TGC GAC GCC AAA TTC CAG CGC ATC
ACC GAC 48
Asn Gin Pro Val Phe Thr Cys Asp Ala Lys Phe Gin Arg lie

Thr Asp
25 30 35
CCC AAT ACC AAG TCG GGC TGC GAT GGC GGC TCG GCC TTT TCG TGT GCT 96
Pro Asn Thr Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala
40 45 50
GAC CAA ACC CCC TGG GCT CTG AAC GAC GAT TTC GCC TAT GGC TTC GCT 144
Asp Gin Thr Pro Trp Ala Leu Asn Asp Asp Phe Ala Tyr Gly Phe Ala
55 60 65
GCC ACG GCT ATT TCG GGT GGA TCG GAA GCC TCG
177
Ala Thr Ala lie Ser Gly Gly Ser Glu Ala Ser
70 75 80
(2) INFORMATION FOR SEQ ID NO: 32:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 59 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 32:
Asn Gin Pro Val Phe Thr Cys Asp Ala Lys Phe Gin Arg lie Thr Asp
15 10

15
Pro Asn Thr Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala
20 25 30
Asp Gin Thr Pro Trp Ala Leu Asn Asp Asp Phe Ala Tyr Gly Phe Ala
35 40 45
Ala Thr Ala lie Ser Gly Gly Ser Glu Ala Ser
50 55
(2) INFORMATION FOR SEQ ID NO: 33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 153 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Humicola nigrescens Oinvik
(B) STRAIN: CBS 819.73
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:!..15 3
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 33: GTC TAC GCC TGC AAC GCA AAC TTC CAG CGC ATC ACC GAC GCC

AAC GCC 48
Val Tyr Ala Cys Asn Ala Asn Phe Gin Arg lie Thr Asp Ala
Asn Ala
60 65 70
75
AAG TCC GGC TGC GAT GGC GGC TCC GCC TTC TCG TGC GCC AAC CAG ACC 96
Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Asn Gin Thr
80 85
90
CCG TGG GCC GTG AGC GAC GAC TTT GCC TAC GGT TTC GCG GCT ACG GCG 144
Pro Trp Ala Val Ser Asp Asp Phe Ala Tyr Gly Phe Ala Ala Thr Ala
95 100 105
CTC GCC GGC
153 Leu Ala Gly 110
(2) INFORMATION FOR SEQ ID NO: 34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 51 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein .
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 34:
Val Tyr Ala Cys Asn Ala Asn Phe Gin Arg lie Thr Asp Ala

Asn Ala
15 10
15
Lys Ser Gly Cys Asp Gly Gly Ser Ala Phe Ser Cys Ala Asn Gin Thr
20 25 30
Pro Trp Ala Val Ser Asp Asp Phe Ala Tyr Gly Phe Ala Ala Thr Ala
35 40 45
Leu Ala Gly 50
(2) INFORMATION FOR SEQ ID NO: 35:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 181 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Cladorrhinum foecundissimum
(B) STRAIN: ATCC 62373
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..181
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 35;

GTC AAC CGC CCT GTC CTC GCC TGC GAC GCA AAC AAC AAC CCT CTG ACC 48
Val Asn Arg Pro Val Leu Ala Cys Asp Ala Asn Asn Asn Pro Leu Thr
15 10
15
GAC GCC GGC GTC AAG TCC GGA TGT GAT GGC GGT TCT GCA TAC ACC TGT 96
Asp Ala Gly Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys
20 25 30
GCC AAC AAC TCC CCA TGG GCA GTG AAC GAC CAG CTC GCC TAC GGC TTT 144
Ala Asn Asn Ser Pro Trp Ala Val Asn Asp Gin Leu Ala Tyr Gly Phe
35 40 45
GCC GCC ACC AAA CTG AGC GGC GGA ACT GAG TCG TCA
180
Ala Ala Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser
50 55 60
(2) INFORMATION FOR SEQ ID NO: 36:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 60 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 36;

Val Asn Arg Pro Val Leu Ala Cys Asp Ala Asn Asn Asn Pro Leu Thr
15 10
15
Asp Ala Gly Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys
20 25 30
Ala Asn Asn Ser Pro Trp Ala Val Asn Asp Gin Leu Ala Tyr Gly Phe
35 40 45
Ala Ala Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser
50 55 60
(2) INFORMATION FOR SEQ ID NO: 37:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 64 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Syspastospora boninensis
(B) STRAIN: NKBC 1515
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:!..64

(xi) SEQUENCE DESCRIPTION; SEQ ID NO: 37:
GGC TGC GAC GGC GGC AGC GCC TTC ACC TGC TCC AAC AAC TCT CCA TGG 48
Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser Pro Trp
GCT GTG AAC GAA GAT
63 Ala Val Asn Glu Asp
(2) INFORMATION FOR SEQ ID NO: 38:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 38:
Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser Pro Trp
Ala Val Asn Glu Asp
(2) INFORMATION FOR SEQ ID NO: 39:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 153 base pairs

(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Nigrospora sp
(B) STRAIN: CBS 272.96
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1.-153
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 39:
ACA AGA AAC GAC GGG CCC CTG TCC AGC CCC GAT GCC GCC TCC GGC TGT 48
Thr Arg Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser Gly Cys
25 30 35
GAT GGC GGC GAA GCC TTT GCC TGT TCT AAT ACC TCG CCT TGG GCC GTC 95
Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp Ala Val
40 45 50
AGC GAC GAG CTC GCG TAG GGA TAG GTC GCC ACG TCC ATC TCC GGC GGC 144
Ser Asp Gin Leu Ala Tyr Gly Tyr Val Ala Thr Ser lie Ser Gly Gly
55 60 65

ACC GAG TCA
153 Thr Glu Ser 70
(2) INFORMATION FOR SEQ ID NO: 40:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 51 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 40:
Thr Arg Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser Gly Cys
15 10
15
Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp Ala Val
20 25 30
Ser Asp Gin Leu Ala Tyr Gly Tyr Val Ala Thr Ser lie Ser Gly Gly
35 40 45
Thr Glu Ser 50
(2) INFORMATION FOR SEQ ID NO: 41:
(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 159 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(Vi) ORIGINAL SOURCE:
(A) ORGANISM: Chaetostylum fresenii
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..159
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 41:
GTC CGA ACG TGT AGT GCC AAC GAC TCG CCC TTG TCC GAC CCA AAT GCC 48
Val Arg Thr Cys Ser Ala Asn Asp Ser Pro Leu Ser Asp Pro Asn Ala
55 60 65
CCA AGT GGG TGT GAC GGT GGT AGC GCC TTC ACT TGT TCC AAC AAC TCC 9 6
Pro Ser Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser
70 75 80
CCG TGG GCA GTC GAT GAC CAG ACA GCT TAT GGC TTT GCG GCA ACA GCC 14 4
Pro Trp Ala Val Asp Asp Gin Thr Ala Tyr Gly Phe Ala Ala Thr Ala
85 90 95

ATC AGT GGC CAG TCC
159 lie Ser Gly Gin Ser 100
(2) INFORMATION FOR SEQ ID NO: 42:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 53 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 42:
Val Arg Thr Cys Ser Ala Asn Asp Ser Pro Leu Ser Asp Pro Asn Ala
15 10
15
Pro Ser Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys Ser Asn Asn Ser
20 25 30
Pro Trp Ala Val Asp Asp Gin Thr Ala Tyr Gly Phe Ala Ala Thr Ala
35 40 45
lie Ser Gly Gin Ser 50
(2) INFORMATION FOR SEQ ID NO: 43;
(i) SEQUENCE CHARACTERISTICS;

(A) LENGTH: 153 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Exidia glandulosa
(B) STRAIN: CBS 277.96
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..153
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 43:
TGT GAG AAG AAC GAC AAC CCC TTA GCT GAC TTC AGC ACG AAA TCC GGG 48
Cys Glu Lys Asn Asp Asn Pro Leu Ala Asp Phe Ser Thr Lys Ser Gly
55 60 65
TGT GAA AGC GGA GGT TCG GCT TAT ACG TGT AAC AAC CAA TCA CCA TGG 96
Cys Glu Ser Gly Gly Ser Ala Tyr Thr Cys Asn Asn Gin Ser
Pro Trp
70 75 80
85
GCC GTC AAT GAC TTG GTG TCG TAT GGC TTC GCC GCC ACA GCG ATC AAT 144
Ala Val Asn Asp Leu Val Ser Tyr Gly Phe Ala Ala Thr Ala lie Asn
90 95

100
GGT GGC AAT
153 Gly Gly Asn
(2) INFORMATION FOR SEQ ID NO: 44:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 51 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 44:
Cys Glu Lys Asn Asp Asn Pro Leu Ala Asp Phe Ser Thr Lys Ser Gly
15 10
15
Cys Glu Ser Gly Gly Ser Ala Tyr Thr Cys Asn Asn Gin Ser Pro Trp
20 25 30
Ala Val Asn Asp Leu Val Ser Tyr Gly Phe Ala Ala Thr Ala lie Asn
35 40 45
Gly Gly Asn 50
(2) INFORMATION FOR SEQ ID NO: 45:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 171 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Coniothecium sp
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:l..171
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 45:
AGG GGC CCC GTC GGA ACC TGC AAG AGG AAC GAG AAC CCC CTC TCC GAC 48
Ser Arg Pro Val Gly Thr Cys Lys Arg Asn Asp Asn Pro Leu Ser Asp
55 60 65
CCC GAT GCC AAG TCC GGC TGC GAC GGC GGC GGC GCC TTC ATG TGC TCC 96
Pro Asp Ala Lys Ser Gly Cys Asp Gly Gly Gly Ala Phe Met Cys Ser
70 75 80
ACC CAG CAG CCG TGG GCC GTC AAC GAC AAT CTG GCA TAT GGC TTC GCC 144
Thr Gin Gin Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala
85 90 95

GCC ACG GCC ATC AGC GGC GGC AAC GAG
171
5 Ala Thr Ala lie Ser Gly Gly Asn Glu
100 105
(2) INFORMATION FOR SEQ ID NO; 46: )
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 57 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 46:
Ser Arg Pro Val Gly Thr Cys Lys Arg Asn Asp Asn Pro Leu Ser Asp
15 10
15
Pro Asp Ala Lys Ser Gly Cys Asp Gly Gly Gly Ala Phe Met Cys Ser
20 25 30
Thr Gin Gin Pro Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala
35 40 45
Ala Thr Ala lie Ser Gly Gly Asn Glu
50 55
(2) INFORMATION FOR SEQ ID NO: 47:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 159 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: CBS 271.96
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:!..159
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 47:
ACT TGC AAC AAG AAC GAC GGG CCC CTG TCG AGC CCC GAT GCC I GCC TCC 4 8
Thr Cys Asn Lys Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser
60 65 70
GGC TGT GAT GGC GGC GAA GCC TTT GCC TGT TCT AAT AGC TCG CCT TGG 96
Gly Cys Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp
75 80 85
GCC GTC AGC GAC GAG CTC GCG TAG GGA TAG CTC GCC ACG TCC ATC TCC 144
Ala Val Ser Asp Gin Leu Ala Tyr Gly Tyr Leu Ala Thr Ser
lie Ser
90 95 100

105
GGC GGC ACC GAG TCG
159 Gly Gly Thr Glu Ser
110
(2) INFORMATION FOR SEQ ID NO: 48:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 53 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 48:
Thr Cys Asn Lys Asn Asp Gly Pro Leu Ser Ser Pro Asp Ala Ala Ser
15 10
15
Gly Cys Asp Gly Gly Glu Ala Phe Ala Cys Ser Asn Thr Ser Pro Trp
20 25 30
Ala Val Ser Asp Gin Leu Ala Tyr Gly Tyr Leu Ala Thr Ser He Ser
35 40 45
Gly Gly Thr Glu Ser 50
(2) INFORMATION FOR SEQ ID NO: 49

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 84 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: CBS 270.96
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..84
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 49:
CCA GTT TTC TCC TGT GAC AAG TAC GAC AAC CCT CTA CCT GAC GCC AAT 4 8
Pro Val Phe Ser Cys Asp Lys Tyr Asp Asn Pro Leu Pro Asp Ala Asn
55 60 65
GCT GTG TCC GGG TGT GAC CCC GGA GGT ACT GCC TTC
84
Ala Val Ser Gly Cys Asp Pro Gly Gly Thr Ala Phe
70 75 80
(2) INFORMATION FOR SEQ ID NO: 50:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 28 amino acids
(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 50:
Pro Val Phe Ser Cys Asp Lys Tyr Asp Asn Pro Leu Pro Asp 5 Ala Asn
15 10
15
Ala Val Ser Gly Cys Asp Pro Gly Gly Thr Ala Phe
20 25
(2) INFORMATION FOR SEQ ID NO: 51:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 147 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Diplodia gossypina, CBS 274.96
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 51:
ACCTGCGACG CCTGCGACAG CCCCCTCAGC GACTACGACG CCAAGTCCGG CTGCGACGGC 60
GGTAGCGCAT ACACCTGCAC CTACTCTACC CCCTGGGCCG TCGACGACAA CCTCTCCTAC 120
GGTTTCGCCG CCGCCAAGCT GAGCGGA 147

(2) INFORMATION FOR SEQ ID NO: 52:
(i) SEQUENCE CPiARACTERISTICS:
(A) LENGTH: 49 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 52:
Thr Cys Asp Ala Cys Asp Ser Pro Leu Ser Asp Tyr Asp Ala Lys Ser
15 10
15
Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys Thr Tyr Ser Thr Pro Trp
20 25
30
Ala Val Asp Asp Asn Leu Ser Tyr Gly Phe Ala Ala Ala Lys Leu Ser
35 40 45
Gly
(2) INFORMATION FOR SEQ ID NO: 53:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 13 5 base pairs
(B) TYPE: nucleic acid

(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Ulospora bilgramii, NKBC 1444
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 53:
CCACTAGCAG ATTTCACCGG TGGAACCGGC TGTAATGGCG GTTCGACATT CTCATGCTCA 60
AACCAACAAC CATGGGCGGT CAACGACACA TTCTCGTACG GCTTTGCGGG CATCTTTATC 120
ACAGGCCATG TCGAG 135
(2) INFORMATION FOR SEQ ID NO: 54:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 4 5 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 54:
Pro Leu Ala Asp Phe Thr Gly Gly Thr Gly Cys Asn Gly Gly Ser Thr
15 10

15
Phe Ser Cys Ser Asn Gin Gin Pro Trp Ala Val Asn Asp Thr Phe Ser
20 25
30
Tyr Gly Phe Ala Gly He Phe He Thr Gly His Val Glu
35 40 45
(2) INFORMATION FOR SEQ ID NO: 55:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 114 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Penicillium verruculosum, ATCC 62396
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 55:
GCCAAATCTG GATGTGATGC TGGTGGAGGT CAAGCCTACA TGTGCTCCAA CCAACAACCT 60
TGGGTAGTCA ACGACAACCT CGCCTACGGT TTCGCCGCAG TCAACATTGC
CGGC 114
(2) INFORMATION FOR SEQ ID NO: 56:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 38 amino acids

(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 56:
Ala Lys Ser Gly Cys Asp Ala Gly Gly Gly Gin Ala Tyr Met Cys Ser
15 10
15
Asn Gin Gin Pro Trp Val Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala
20 25
30
Ala Val Asn lie Ala Gly 35
(2) INFORMATION FOR SEQ ID NO: 57:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 113 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Poronia punctata

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 57:
TTCGACGTCC GGGTGCGACA ATGGCGGCAG CGCCTTCATG TGCTCTAACC AAAGCCCCTG 60
GGCCGTCAAC GACGATCTGG CCTACGGCTG GGCCGCCGTC TCAATCGCGG
GCC 113
(2) INFORMATION FOR SEQ ID NO: 58:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 37 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 58:
Ser Thr Ser Gly Cys Asp Asn Gly Gly Ser Ala Phe Met Cys Ser Asn
15 10
15
Gin Ser Pro Trp Ala Val Asn Asp Asp Leu Ala Tyr Gly Trp Ala Ala
20 25
30
Val Ser lie Ala Gly 35
(2) INFORMATION FOR SEQ ID NO: 59:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 177 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Fusariura anguioides, IFO 4467
(XI) SEQUENCE DESCRIPTION: SEQ ID NO: 59:
[■CAACACCGG TGCAGACGTG CGACCGCAAC GACAACCCGC TCTACGACGG :GGGTCGACG 6O
'.GGTCCGGCT GCGACGCCGG CGGCGGCGCC TACATGTGCT CGTCGCACAG ICCGTGGGCC 120
TCAGCGACA GCCTCTCGTA CGGCTGGGCG GCCGTCCGCA TCGCCGGCCA
TCCGAG 177
2) INFORMATION FOR SEQ ID NO: 60:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 59 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 60:

Ser Thr Pro Val Gin Thr Cys Asp Arg Asn Asp Asn Pro Leu Tyr Asp'
15 10
15
Gly Gly Ser Thr Arg Ser Gly Cys Asp Ala Gly Gly Gly Ala Tyr Met
20 25
30
Cys Ser Ser His Ser Pro Trp Ala Val Ser Asp Ser Leu Ser Tyr Gly
35 40 45
Trp Ala Ala Val Arg lie Ala Gly Gin Ser Glu
50 55
(2) INFORMATION FOR SEQ ID NO: 61:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 150 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Thielavia thermophila, CBS 174.70
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 61: AACGACAACC CCATCTCCAA CACCAACGCT GTCAACGGTT GTGAGGGTGG

TGGTTCTGCT 60
TACGCTTGCT CCAACTACTC TCCCTGGGCT GTCAACGATG ACCTTGCCTA CGGTTTCGCT 12 0
GTTACCAAGA TCTCCGGTGG CTCCGAGGCC 150
(2) INFORMATION FOR SEQ ID NO: 62:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 50 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 62:
Asn Asp Asn Pro lie Ser Asn Thr Asn Ala Val Asn Gly Cys Glu Gly
15 10
15
Gly Gly Ser Ala Tyr Ala Cys Ser Asn Tyr Ser Pro Trp Ala Val Asn
20 25
30
Asp Asp Leu Ala Tyr Gly Phe Ala Val Thr Lys lie Ser Gly Gly Ser
35 40 45

Glu Ala 50
(2) INFORMATION FOR SEQ ID NO: 63:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 180 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Chaetomium cuniculoruin, CBS 799-83
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 63:
GTCAATCAGC CCATCCGAAC GTGTAGTGCC AACGACTCGC CCTTGTCCGA CCCAAATACC 60
CCAAGTGGCT GTGACGGTGG TAGCGCCTTC ACTTGTTCCA ACAACTCCCC GTGGGCAGTC 120
GATGACCAGA CAGCTTATGG CTTTGCGGCA ACAGCCATCA GTGGCCAGTC CGAGAGCAGC 180
(2) INFORMATION FOR SEQ ID NO: 64:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 60 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 64:
Val Asn Gin Pro lie Arg Thr Cys Ser Ala Asn Asp Ser Pro Leu Ser
15 10
15
Asp Pro Asn Thr Pro Ser Gly Cys Asp Gly Gly Ser Ala Phe Thr Cys
20 25
30
Ser Asn Asn Ser Pro Trp Ala Val Asp Asp Gin Thr Ala Tyr Gly Phe
35 40 45
Ala Ala Thr Ala lie Ser Gly Gin Ser Glu Ser Ser
50 55 60
(2) INFORMATION FOR SEQ ID NO: 65:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 159 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: CDNA
(iv) ORIGINAL SOURCE: Chaetomium virescens, CBS 547.75

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 65:
ACCTGCGACA AGAAGGACAA CCCCATCTCT GATGCCAACG CCAAGAGCGG CTGTGATGGC 60 5
GGTTCTGCTT TCGCCTGCAC CAACTACTCT CCCTTCGCCG TCAACGACAA CCTCGCCTAC 120
GGTTTCGCTG CCACCAAGCT TGCTGGAGGC TCCGAGGCT
I 159
(2) INFORMATION FOR SEQ ID NO: 66:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 53 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 66:
Thr Cys Asp Lys Lys Asp Asn Pro lie Ser Asp Ala Asn Ala Lys Ser
15 10
15
Gly Cys Asp Gly Gly Ser Ala Phe Ala Cys Thr Asn Tyr Ser Pro Phe
20 25
30
Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Lys Leu Ala

35 40 45
Gly Gly Ser Glu Ala 50
(2) INFORMATION FOR SEQ ID NO: 67:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 81 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Colletotrichum lagenarium
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 67:
ACCTGCTACG CCAATGACCA GCGCATCGCC GACCGCAGCA CCAAGTCCGG CTGCGACGGC 60
GGCTCGGCCT ACTCCTGTTC T 81
(2) INFORMATION FOR SEQ ID NO: 68:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 68:
Thr Cys Tyr Ala Asn Asp Gin Arg lie Ala Asp Arg Ser Thr Lys Ser
15 10
15
Gly Cys Asp Gly Gly Ser Ala Tyr Ser Cys Ser
20 25
(2) INFORMATION FOR SEQ ID NO: 69:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 160 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(B) STRAIN: Phycomyces nitens
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 69:
ACCTGTGACA AGAAGGACAA CCCCATCTCA AACTTGAACG CTGTCAACGG TTGTGAGGGT 60
GGTGGTTCTG CCTTCGCCTG CACCAACTAC TCTCCTTGGG CGGTCAATGA CAACCTTGCC 12 0
TACGGCTTCG CTGCAACCAA GCTTGCCGGT GGCTCCGAGG 160

(2) INFORMATION FOR SEQ ID NO: 70:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 53 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 70:
Thr Cys Asp Lys Lys Asp Asn Pro lie Ser Asn Leu Asn Ala Val Asn
15 10
15
Gly Cys Glu Gly Gly Gly Ser Ala Phe Ala Cys Thr Asn Tyr Ser Pro
20 25
30
Trp Ala Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Lys Leu
35 40 45
Ala Gly Gly Ser Glu 50
(2) INFORMATION FOR SEQ ID NO: 71:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 165 base pairs
(B) TYPE: nucleic acid

5372

(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(iv) ORIGINAL SOURCE: Trichothecium roseum, IFO
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 71:

CCAGTAGGCA CCTGCGACGC CGGCAACAGC CCCCTCGGCG ACCCCCTGGC CAAGTCTGGC 60
TGCGAGGGCG GCCCGTCGTA CACGTGCGCC AACTACCAGC CGTGGGCGGT CAACGACCAG 120
CTGGCCTACG GCTTCGCGGC CACGGCCATC AACGGCGGCA CCGAG 165
(2) INFORMATION FOR SEQ ID NO: 72:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 55 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 72:
Pro Val Gly Thr Cys Asp Ala Gly Asn Ser Pro Leu Gly Asp Pro Leu
1 5 10
15

Ala Lys Ser Gly Cys Glu Gly Gly Pro Ser Tyr Thr Cys Ala Asn Tyr
20 25
30
Gin Pro Trp Ala Val Asn Asp Gin Leu Ala Tyr Gly Phe Ala Ala Thr
35 40 45
Ala lie Asn Gly Gly Thr Glu
50 55

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GB-A-1368599
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WO 91/17243
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WO 95/26398
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Dalbcsge, H., and Heldt-Hansen, H. (1994) Mol. Gen. Genet. 243, 253-260
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WE CLAIM;
1. An enzyme preparation exhibiting endoglucanase activity, wherein the enzyme is
a) encoded by a DNA construct comprising the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, or
b) encoded by a DNA construct comprising an analogue of the DNA sequence shown in SEQ ID No. 8 or a DNA construct comprising the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSM 10081, which DNA sequence has at least 75% identity with the DNA sequence shown in SEQ ID No. 8 or the DNA sequence obtainable from the plasmid in Saccharomyces cerevisiae DSN 10081, or
c) a polypeptide which has at least 70% identity with the amino acid sequence shown
_ in SEQ ID No. 9.
2. The enzyme preparation according to claim 1, wherein it said enzyme.
3. The enzyme preparation according to claim 1 or 2, wherein it additionally comprises one or more enzymes selected from the group consisting of galactanases, xylanases, arabinanases, pectin acetyl esterases, polygalacturonases, rhamnogalacturonases, pectin lyases, pectate lyases, endogucanases, pectin methylesterases, proteases, lipases, amylases, cutinases, peroxidases, laccases, cellobiohydrolases and transglutaminases.
4. A detergent composition comprising the enzyme preparation according to any of
the claims 1 to 3, and a compound selected from the group consisting of a surfactant, a
builder compound, and a fabric softening agent.
5. The composition according to claim 4, wherein the surfactant is a nonionic,
anionic, cationic, zwitterionic, ampholytic or amphoteric surfactant.
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6. The composition according to claim 4, wherein the fabric softening agent is a cationic or nonionic softening agent, preferably a quaternary ammonium compound, and which optionally further comprises one or more compounds selected from a surfactant, an electrolyte, a buffer, an antioxidant and a liquid carrier.
7. A method of providing colour clarification of laundry, wherein said method comprises
treating the laundry with a soaking, washing or rinsing liquor comprising an enzyme preparation according to any of claims 1 to 3 or
with a detergent composition according to any one of the claims 4 to 6.
8. The method according to claim 7, wherein the laundry is treated in a washing machine.
9. The method according to claim 7 or 8, wherein the endoglucanase is present in the soaking, washing, or rinsing liquor in an effective amount of between 1 and 1000 S— CEVU, preferably between 5 and 200 S-CEVU, per liter of liquor during machine cycle use conditions.

10. The method according to any of the claims 7 to 9, wherein the pH of the soaking, washing, or rinsing liquor is between 4 and 11, preferably between 6 and 10.5.
11. The method according to any of the claims 7 to 10, wherein the temperature is between 15°C and 60°C.
12. A method for biopolishing of a cellulosic fabric which method comprises treating the yam surface of said fabric with the enzyme preparation according to any of claims lto3.
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13. The method according to claim 12, wherein it takes place in the wet processing of the manufacture of knitted and woven fabrics.
14. A method for "stone-washing" which method comprises treating a dyed cellulosic fabric with the enzyme preparation according to any of claims 1 to 3.
15. An enzyme preparation exhibiting endoglucanase activity substantially as herein described and exemplified.

16. A detergent composition comprising the enzyme preparation substantially as herein described and exemplified.
17. A method of providing colour clarification substantially as herein described and exemplified.

18. A method for biopolishing of a cellulosic fabric substantially as herein described and exemplified.
19. A method for "stone-washing" substantially as herein described and exemplified.

Documents:

0919-che-2003 abstract duplicate.pdf

0919-che-2003 description (complete) duplicate-1.pdf

0919-che-2003 description (complete) duplicate-2.pdf

0919-che-2003 description (complete) duplicate-3.pdf

0919-che-2003 description (complete) duplicate-4.pdf

0919-che-2003 description (complete) duplicate.pdf

0919-che-2003 description (complete)-1.pdf

0919-che-2003 description (complete)-2.pdf

0919-che-2003 description (complete)-3.pdf

0919-che-2003 description (complete)-4.pdf

0919-che-2003 form-26.pdf

919-che-2003-abstract.pdf

919-che-2003-assignement.pdf

919-che-2003-claims.pdf

919-che-2003-correspondnece-others.pdf

919-che-2003-correspondnece-po.pdf

919-che-2003-description(complete).pdf

919-che-2003-drawings.pdf

919-che-2003-form 1.pdf

919-che-2003-form 26.pdf

919-che-2003-form 3.pdf

919-che-2003-otherdocument.pdf

919-che-2003-pct.pdf

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Patent Number

219745

Indian Patent Application Number

919/CHE/2003

PG Journal Number

27/2008

Publication Date

04-Jul-2008

Grant Date

13-May-2008

Date of Filing

10-Nov-2003

Name of Patentee

NOVOZYMES A/S

Applicant Address

Inventors:

#	Inventor's Name	Inventor's Address
1	LASSEN, SOREN FLENSTED
2	SCHULEIN MARTIN
3	NIELSEN, RUBY ILIM
4	TAKAGI SHINOBU
5	IHARA MICHIKO
6	ANDERSEN, LENE NONBOE
7	LANGE, LENE
8	KAUPPINEN, MARKUS SAKARI

PCT International Classification Number

C12N15/56

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA