Title of Invention

"A PROCESS FOR BIOFINISHING OF DENIMS IN TEXTILE INDUSTRY"

Abstract The present invention describes a process for biofinishing of denims in textile industry. It comprises of soaking of the denim fabric in a mixture of water and cellulase enzyme. The cellulase enzyme was found to be effective in biofinishing of denim fabric with respect to loss of hairiness with minimum weight loss, wash down effect, back staining, impartation of softness and colour contrast. The cellulase enzyme used is obtained as a extracellular culture filtrate of Thermomonospora sp. Produced. The cellulase enzyme possesses a complement of characteristics that can replaces pumice stones used to achieve the faded effect of denim fabric with minimum wear of the fabric and blockage of the machinery. The novelty of this process exists in the effectiveness of the said cellulase in biofinishing of denim under non - buffering conditions using tap water.
Full Text Field of Invention
The present invention relates to the process for biofinishing of denims in textile industry. More particularly it relates to eco-friendly application of the cellulase produced by a novel alkalothermophilic actinomycete in the biofinishing of denim processes of textile industry. More particularly, it relates to the preparation of the said alkali stable cellulase using the alkalothermophilic Thermomonospora sp. deposited at MTCC and has been allotted accession number MTCC 5117. Still more particularly, it relates to reducing hairiness with marginal total weight loss and impartation of softness of cotton fabrics especially denims during the biofinishing process. Background of the Invention.
Cellulases are enzymes involved in the degradation of cellulose to soluble sugars. They are made up of three major components namely endo ß-1,4 glucanases, exo p-1,4 glucanases and ß- glucosidases. These three enzyme act synergistically in the hydrolysis of crystalline cellulose. Endo P-1,4 glucanases hydrolyse cellulose chains in a random fashion; thereby converting long chains to oligosaccharides while exo p-1,4 glucanases remove cellobiose or glucose from the non-reducing end of the polymer. P-glucosidases hydrolyse cellobiose to glucose units (Mandel M. Annual Report of Fermentation Proceedings (1982) Vol. 5, 35-78). Wide varieties of microorganisms belonging to bacteria and fungi are reported to produce cellulases (Singh A. and Hayashi K. (1995) In Advances in Applied Microbiology Vol. 40, pp.
1-44, Academic press Inc.). Cellulases have been investigated mainly due to their potiential application for the bioconversion of agricultural biomass resources in to useful products. Cellulases and hemicellulases are used in clearing fruit juices and beer, extraction of juice and oil from fruit and seed pulp (Deshpande V. V. and M. B. Rao (1998) In: Microbes: For health, wealth and sustainable environment. Ajit Verma (editor) Malhotra publishing house, New Delhi, pp. 293-306). Cellulases are also useful in paper industry (Kim T. J., Ow S. S. K. and T. J. Eom (1991) Proceedings of the 1991 TAPPI pulping conference, pp. 1023-1027) and in animal feed as additive for monogastric animals (Cowan W. D. In: Godfrey T., West S. (editors) Industrial Enzymology. 2nd ed. London, MacMillan Press, 1996 pp. 360-371). Recently, endoglucanases have been successfully introduced in to laundry reagents along with alkaline proteases (Kang M. K. and Rhee Y. H. (1995) Biotechnology letters. Vol. 17, 507). Cellulases play a crucial in developing ecofriendly technologies due to environmental pressures imposed on the textile industry and the need to minimize water contamination (Horikoshi K., Nakao M., Kurono Y. and Sashihara N. 1984. Canadian Journal of Microbiology. Vol. 30, 774). The traditional use of pumice stone in a water-loaded tumbling machine produced severe wear and loss of tensile strength when used to achieve high degree of indigo fading. Treatment of denim garments with cellulase enzymes has become a standard finishing procedure for this type of clothes (Tyndall R. M. (1992) Textile Chemistry and Color. Vol. 24, 23-26). As a result of such treatment, the fabrics acquire fashionable looks preferred by most customers. Cellulase could replace the use of pumice stones in the manufacture of stone washed denims. It could improve fabric appearance by reducing fuzz or piling
or enhance softness, luster and color brightening of cotton fabrics. A common problem associated with treatment of such cotton containing fabrics with the cellulase solution is that the treated fabrics exhibit significant strength loss as compared to the untreated fabric. Strength loss arises because the cellulase hydrolyzes P-1,4 glucan linkages of cellulose, which in turn can result in a breakdown of a portion of the polymer of the cotton polymer. As more and more cotton polymer are disrupted the tensile strength of the fabric is reduced. Accordingly it would be particularly desirable to modify such cellulase treatment methods so as to provide reduced strength loss while still achieving the desired enhancements in the treated cotton-containing fabric arising from treatment with cellulase as compared to the fabric prior to treatment. Clarkson et. al has described a process of treating cotton containing fabric with cellulase free from exocellobiohydrolase component. (Clarkson K. A., Larenas E. and G. L. Weiss (1993) US Patent 5,246,853). Clonning and expression of full-length tuncated cellulase gene in to E. coli from actinomycete and its use in textile industry for biofinishing has also been demonstrated (Jones B. E., Van Der K., Wilhelmus A. H., Van S. P., Weyler W. (2001) US Patent 6,287, 839). It has been reported that the use of a fungal cellulase at pH 6.8 comprising of a combination of exo-cellobiohydrolase I and endoglucanase wherein the exo-cellobiohydrolase I component is relatively enriched showed enhancement in the feel, appearance and softening during biofinishing of denims (Clarkson K. A., Collier K., Lad P. J. and Weiss G. L. (1997) US Patent 5,668,009).
Homology search and literature survey has shown that the actinomycete used in this invention has 97% homology to Thermomonospora mesouviformis and no citations have been observed on production of cellulase by this organism. Also, the cultural conditions for the growth of alkalothermophilic Thermomonospora sp. is pH 9 and 50 C different from the reported Thermomonospora sp. The highlights of the present invention are: i. that, the extracellular culture filtrate shows endoglucanase and xylanase activity, ii. that, the culture filtrate is used for biofinishing of denim at pH 8 resulting in the
reduction of hairiness causing fuzz with marginal weight loss, iii. that, the culture filterate is used to replace the pumice stone used to give the faded look to the denim fabric thus minimizing the wear of the fabric and blockage of the machinery, iv. that, the culture filtrate applied for biofmishing of denim is effective under non-buffering conditions using tap water of hardness 50 parts per million (ppm).
In spite of the knowledge in art related to cellulase compositions, have desirable properties, the quest for new cultures secreting cellulases having varying spectrum of characteristics useful in the biofmishing of denim continued. Applicants have invented a process for using cellulases from novel alkalothermophilic Thermomonospora sp., which possesses such a complement of characteristics useful in biofinishing of denims. Detailed description of the invention:
Accordingly the present invention provides a process for biofinishing of denims in textile industry, which comprises, soaking the denim in a mixture of water and cellulase enzyme obtained from novel alkalothermophilic Thermomonospora sp for a

period of 60-70 minutes, washing the treated denim successively with water for atleast three times and air drying to get biofinished denims.
In one of the embodiments of the present invention the soaking of denim
fabric is done at 40 to 55°C.
In yet another embodiment the water used is of hardness in the range of 30 to 50 ppm.
In yet another embodiment, the concentration of enzyme in water solution may be 0.5 to 3.0 percent.
In yet another embodiment the enzyme is used under non-buffering conditions.
In still another embodiment a material to liquor ratio of 1:50 is maintained.
In yet another embodiment the cellulase enzyme is obtained as per process claimed and described in our co-pending patent application no.NCL-15/ 2004.
In still another feature, the cellulase is evaluated for reducing hairiness of denim fabric. The enzyme is found to be effective in reducing hairiness of denim fabric in the enzyme concentration range of 0.5 to 3.0% with respect to the weight of fabric; in a pH range of 7-9. Soaking the fabric in the enzyme reduced the hairiness of the fabric by removing the protruding fibers from the surface of the fabric giving it a clean and clearer look from its original fuzzy look.
In another feature, the cellulase is found to be less effective in reducing the total weight of the fabric with an enzyme concentrations ranging from 0.5% to 1.5% with respect to the weight of the fabric, after incubation for 1-2 hours at 50°C in a pH varying from 7-9. Weight loss measurement studies of enzyme indicated lower
weight loss of the fabric with considerable loss of hairiness of the fabric indicating that the enzyme causes less harm to the fabric.
In yet another feature, the cellulase is found to be effective in imparting softness to the fabric when incubated in an enzyme concentration varying from 0.5 to 3.0% with respect to the weight of fabric at 50°C for 1-2 hours. Visual assessment using 3 experts indicated the impartation of better softness of the fabric as compared to the control with out enzyme treatment.
In another feature, the cellulase is found to show better wash down effect as compared to the control, which is not treated with the enzyme. Treatment of the denim fabric with cellulase enzyme of concentration varying from 0.5 to 3% at 50°C incubated for a period of 1 to 2 hours at a pH range varying from 7 to 9 indicated a better wash down effect when assessed by 3 experts separately.
In yet another feature, the cellulase treated fabric showed less back staining as compared to the control that is not treated with cellulase. The denim fabric treated with cellulase enzyme in a concentration varying from 0.5% to 3% at 50°C incubated for a period of 1 to 2 hours at a pH range varying from 7 to 9 indicated less back staining of the denim fabric during washing.
The process of the present invention is described herein below with examples, which are illustrative only and should not be constructed to limit the scope of the present invention in any manner whatsoever.
Example 1
This example illustrates the preparation of cellulase for application in textile industry. Spores of alkalothermophilic Thermomonospora sp. from 3 day old Luria
Bertani agar slant with 1%CMC at pH 9.0 is inoculated and incubated for 48 hours at 50°C in to the inoculum media consisting of (g/L) KH2PO4 - 2, (NH4)2S04-7, Urea-1.5, MgS04- 0.3, Yeast extract- 10, Peptone- 1.25, in trace salt solution containing (µg/ml) FeSO4-50, MnS04- 15.6, ZnS04- 14, C0CI2- 20 with 2% wheat bran and 1ml Tween 80. After 48 hours 10% of the inoculum is transferred in to fermentation media containing 4% cellulose paper powder instead of wheat bran in the above inoculum media. This is incubated at 50 C for 120 hours on a shaker adjusted to 200 rpm. The culture supernatant is harvested by centrifugation at 5000 to 7000rpm for 10 minutes. The activity in the cell free broth, after 120 hours is 7.0 lU/ml. The culture filtrate also showed xylanase activity of 120 lU/ml.
Example 2
This example illustrated the application of alkali stable cellulase produced by Thermomonospora sp. having MTCC No. MTCC 5117 for biofinishing of denim. Initially the efficiency of the application of cellulase in textile industry is determined in terms of viscometric units. 1.5% of the enzyme when incubated with 2% CMC paste at 50°C for 10 minutes showed 250 cps.
Example 3
This example illustrates the application of enzyme for biofinishing of denim fabric. 200g of denim fabric is treated with 0.5 to 1.5% of enzyme with respect to weight of fabric. A material to liquor ratio of 1:50 is maintained at 55°C at pH 8. The hardness of water is maintained at 50ppm. Incubation is continued for 1 hour, thereafter the cloth is rinsed thoroughly thrice with normal tap water, and the fabric is air-dried.
Results: Measurement of hairiness and total weight loss exhibited desirable observations as compared to the control. About 50 % loss of hairiness is visible on treatment of the fabric with 0.5% of enzyme with respect to the weight of the fabric for one hour with marginal loss of weight of 0.65% (Table 1 and 2). Increasing concentration of the enzyme showed increased loss of hairiness with marginal loss of weight of the fabric.
Table 1
(Table Removed)
Example 4
This example illustrates the application of cellulase for biofinishing of denim fabric. 200g of denim fabric are treated with 0.5 to 1.5% with respect to weight of fabric. A material to liquor ratio of 1:50 is maintained at 55 C in normal tap water whose hardness of water is maintained at 50ppm. Incubation is continued for 1 hour and thereafter the cloth is rinsed thoroughly thrice with normal tap water and the fabric is air-dried.
Results: During the incubation period the efficiency of the enzyme is checked with respect to the following parameters - Impartation of softness, wash down effect and back staining. Visual monitoring of the treated fabrics is done by three experts. The panel rated the treated fabric as 1 for Best, 2 for second best, and like that (Table 3). NCL developed enzyme showed good efficiency with respect to the above parameters under this non-buffering condition, which would be most preferred in an industry.
Table 3

(Table Removed)
Where a= Impartation of softness, b= wash down effect, c= back staining










We claim:
1. A process for biofinishing of denims in textile industry, which comprises, soaking the denim in a mixture of water and cellulase enzyme obtained from novel alkalothermophilic Thermomonospora sp for a period of 60-70 minutes, washing the treated denim successively with water for atleast three times and air drying to get biofinished denims.
2. A process as claimed in claim 1, wherein the soaking of denim fabric is done at 40 to 55°C.
3. A process as claimed in claim 1 and 2, wherein the water used is of hardness in the range of 30 to 50ppm.
4. A process as claimed in claims 1 to 3, wherein the enzyme concentration in water solution is 0.5 to 3.0 percent.
5. A process as claimed in claims 1 to 4, wherein a material to liquor ratio of 1:50 is maintained.
6. A process for biofinishing of denims in textile industry substantially as herein described with reference to the examples and drawings accompanying the specifications.

Documents:

1299-del-2004-Abstract-(06-12-2010).pdf

1299-del-2004-abstract.pdf

1299-del-2004-Claims-(06-12-2010).pdf

1299-del-2004-claims.pdf

1299-del-2004-Correspondence-Others-(06-12-2010).pdf

1299-del-2004-correspondence-others.pdf

1299-del-2004-Description (Complete)-(06-12-2010).pdf

1299-del-2004-description (complete).pdf

1299-del-2004-Form-1-(06-12-2010).pdf

1299-del-2004-form-1.pdf

1299-del-2004-form-18.pdf

1299-del-2004-Form-2-(06-12-2010).pdf

1299-del-2004-form-2.pdf

1299-del-2004-Form-3-(06-12-2010).pdf

1299-del-2004-form-3.pdf

1299-del-2004-form-5.pdf


Patent Number 245202
Indian Patent Application Number 1299/DEL/2004
PG Journal Number 02/2011
Publication Date 14-Jan-2011
Grant Date 07-Jan-2011
Date of Filing 15-Jul-2004
Name of Patentee COUNCIL OF SCIENTIFIC & INDUSTRIAL RESEARCH
Applicant Address RAFI MARG, NEW DELHI-110001, INDIA
Inventors:
# Inventor's Name Inventor's Address
1 ANISH RAMAKRISHNAN AHMEDABAD TEXTILE INDUSTRIES RESEARCH ASSOCIATION, AHMEDABAD, 380 018, GUJARAT,INDIA
2 ABSAR AHMAD AHMEDABAD TEXTILE INDUSTRIES RESEARCH ASSOCIATION, AHMEDABAD, 380 018, GUJARAT,INDIA
3 RAO MALA BALACHANDRA AHMEDABAD TEXTILE INDUSTRIES RESEARCH ASSOCIATION, AHMEDABAD, 380 018, GUJARAT,INDIA
PCT International Classification Number C12N;D06M16/00; D06P5/02
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA