Title of Invention


Abstract An enzyme modified oat cereal suspension comprising maltose and maltodextrin units, intact B-glucans, and proteins, said enzyme modified oat cereal suspension prepared by the method which comprises the steps of: (ij providing an oat cereal substrate suspension, (ii) providing an enzyme composition comprising B-amylase and a-amylase, and (iii) treating said oat cereal substrate suspension (i) with said enzyme composition (ii) wherein said β-amylase and a-amylase enzymes are introduced simultaneously for accelerated enzymatic hydrolysis, reduction of viscosity, and regulation of sugar content formation of said modified oat cereal suspension, said method allowing use of lesser amounts of said enzymes than otherwise needed when said enzymes are used separately.
Full Text FORM 2
[39 OF 1970]
[See Section 10; rule 13]

CEREAL BASE CEBA AB, of Von Troils vag 1, S-205 03 Malmo, Sweden,
The following specification particularly describes the nature of the invention and the manner in which it is to be performed:-

25-4-2006 1

Cross Reference To Refated Applications This application claims the benefit of.U.S. Provisional Application No. 60/106,224-filed.October 3O, 1998 andUIS'Provisionai Appiicarion No.60/1.04i706 3edOcrober l9, 1998.
Fieid of the Invention. The present invention relates to. enzyme suspensions, comprising enzyme preparations, for
errzymaric. hydrolysis of cereal starch: In another, aspect it relates, to methods of producing
homogeneous.andstable cereal suspensions prepared by using the: enzyme, preparations..
Background of the Invention
The- benencial heaith. effects of dietary fibers are well-known. In this..cantext there.has been
a growing-interest in food products ; made from grains, such as oats: and bariey.
In. many respect oats: are; dirrerent than, othergrains. They have; higher protein; and fat
coment .than.comparable: cereals. as. well as a high β-glucan content.
In recent years,, there has been a. growing-interest, in.food products., made from oats..
The; main reason for this. is, that oar fibers have been found to have a. wholesome effect by.
lowering the serum.choiesterol level of hypercholesteremic individuals.. Another reasoan is that
oats contain protein, of high; food value as. well as; a.considerable. proportion of mono and
poiyunsarnrated-fats. In 'addition,. oats contain many essential amino acids and minerals.
A. great, advantage- of oats, is that, the- whole grain, caa be- used for making various
products; oncethe hull has.been removed. In oats;. the- most nutritious: substances, are
distributted fairly- evenly in the whole grain. In other grains the nutritious substances: are
frequentlyconcentrated in: spenffc parts:c£the.grain.

The nutritional aspects of oat, components have prompted.the introduction of oats or parts thereof into several different food products. For instance, U.S. Patent No. 4,996,063 (G.F. Ihgletr.) discloses the, preparation of water-soluble dietary fibre compositions by treating 'ground oat products with a-amylases. The a-amylase serves to thin the oat starch, and any a-amylase. may thus, be used. The produced pulverulent dietary fibre compositions are used, as additives in food products, such as fat substitutes. However, these products not only lack desirable aromatics of natural, oats, but are also deprived of agreeable natural oat flavorings.
U.S. Patent.No. 5,686,123 (to L. Lihdahl.et.al) discloses ahomogeneous and stable cereaLsuspensionhaving the.taste and aroma of natural oats. The disclosed cereal suspension is prepared by treating a, suspension of oatmeal, with β-amylase, which has no giucanase and proteinase activity, in .a.first enzymetreatment step, which.specifically generates ; maltose- and maltodextrin units. Then the. suspension is treated with a-amylase, which also: has. no giucanase and proteinase, activity, in a. second enzyme, treatment step which specifically generates maltose units. This, oat suspension is a, milky product which can be used, as an. alternative to milk, especially for lactose-intolerant people.. It may also be used.as the basis of oran additive in the manufacture, of ice-cream, gruel, yogurt;, milkshakes, health drinks and snacks. However, this process is time consuming'because of the sequential, treatments with, different hydrolases,, thereby increasing the costof production. Moreover, sequential enzyme treatments eliminate any possible positive synergic, effects that may occur when combining of eazymes.. Furthermore, the. overall viscosity and/or sugar: content: of the cereal suspension, cannot, be efficiently controlled or manipulated


In view of these shortcomings, there is a need for enzyme, preparations thar hydrolyze cereai. starch in a. more cost-efficient and timely manner while producing a cereai suspension product' that retains, the flavoring and aromatic, qualities; of natural cereai and in which the viscosity, sugar content, and overall texture can be regulated or modified fora preferred end product.
Summary of the Invention
For purposes, of this, invention, the. terms, and. expressions below appearing: in the specification; and claims are intended.to have: the:.following. meanings:
"Preprocessed cereal suspension" as used herein..means, a.product that has been previously processed by themethod. disclosed in U.S. Batent.No.. 5,686,123.
"Cereal, substrate' as. used herein means, a. suspension selected from the. group consisting.of cereal meal. suspension,, preprocessed cereal suspension. andmixtures.there of
"Oatmeal,suspension"'as usedherein.means,asuspension;comprismg oat flour;and/or rolled oats-
Accordingly, it is a principal, object of the present invention to provide, an enzyme modified suspension comprising a. cereai. substrate suspension, and an enzyme preparation for enzymatic hydrolysis, of constituents kthe cereal. substrate,suspension whereby thespecific properties of the-cereal, substrate, suspension, such as viscosity and/or sugar ratio content modified and/or manipulated, by combinations of various hydrolases and isomerases-
It. is another object of the- present invention to. provide: an efficient, selective and economical method for producing' a cereal, suspension, having the aroma, and/or flavor of

narurai cereals and having the unexpected improvement of regulating the viscosity and/or sugar content, of the cerealsuspension. through treatment with a. spectrum of new enzyme preparations.
It is.an. additional object o£ the. present invention to provide homogeneous and stable cereal suspension having, the. aroma, and/or flavor of natural cereals and. containing intact β -giucans that are. components, of acereal substrate, suspension.
A still furtherobjectof the.presentinvention is;to provide, a. method, for preparing the. homogeneous, and stable.- cereal.suspension havingj the.aroma;.and: taste.of .narurai cereals and containing, intact β-glucans.
All of the above objects may be: accomplished by. an. enzyme modified, suspension comprismga-cered. substrate. suspension,andan.enzyme:preparation for anzmatic hydrolisis
of'constituents in the. cereal substrate suspension,,. The:enzyme .preparatiori comprises, at least one. hydrolase, with, the ability to. frydroiyze α-glycosidic: bonds of the constituents of a. cereal substrate suspension, and optionally,. combining the. at least one hydrolase, with an. isomerase. The constituents: may include,, but are- not limited, to,, amylose,.amylopedin,.,maltodextrins, maltose maitotriose-and a.mixture thereof. More, specifically, the. above objects may be accomplished by an enzyme modified:suspension.comprising.a.cereal substrate, suspension and an enzyme preparation wherein, the; enzyme., preparation comprises at least one hydrolase having the. ability to hydrolyze. α-glycosidia bonds of constituents: of a. cereal, substrate suspension- It: is preferred that, the enzyme preparation has no giucanase and/or proteinase, activity. The preferred, hydrolaser may be selected, from, the group consisting of β-amyIase; 5

a-amyiase, amyiogiuco sidase and puiiuianase, with the proviso that when the enzyme preparation comprises β-amylase or a-amylase. there is a mixture of at least one other of the named, α-glycosidic hydrolases., ft is further preferred, mat whea the, enzyme preparation, comprises both (3-amyiase and a-amylase they are combined simultaneously in the cereal substrate suspension. In addition, to the above-identified, hydrolases, the enzyme preparations of the presentinvention may further comprise an isomerase,. such as glucose isomerase.
The previously discussed, enzyme, preparations may be. utilized, to produce a. homogeneous.and stable, improved, cereal, suspension having the: aroma.and taste, of natural cereals and.containing bract p-glucans, the improved cereal suspension prepared by the method, comprisingthe steps:
a) providing- axereaL substrate suspension;
b) treatmg the cereal,substrate;suspensioriof step (a) with an enzyme-preparatfon.as. previously discussed, namely an. enzyme preparation, for enzymatic-hydrolysis of a cereai starch, in the suspensioa comprising, at. least: one hydrolase enzyme having the ability to hydrolyze a-giycosidic bonds. Preferably, a a-glycosidic hydrolase is selected .from the, group consisting of P-amylase, a-amyiase, amyioglucosidase; puilulanase. and. mixtures thereof; with theproviso that when the. enzyme preparation comprises -amylase or α-amylase there is a. mixture of at least, one other of the named, a-glycosidic hydrolases.
It is preferred, that when, the-enzyme preparation comprises both J3-amylase and α-amylase, the enzymes are introduced simultaneously to the. cereai substrate suspension.

The method of making the homogeneous, and stable improved cereal suspension may also include performing at least one. finishing process step on the enzyme, treated suspension of. step (b) for making the cereaL suspension..
The, finishing process step which, may improve die shelf-life of the cereal suspension or end. product can. include: removing coarse, particles by centrifuging or decanting; homogenizing: the. enzyme treated suspension; and/or subjecting the.product to Ultra Hgh. Temperature:(UHT) treatments disclosed.in FdodEngineering-and.Dairy Technology, KG. Kessier, Verlay A.. Kessler, 1981, Chapter 6, pp:. 13.9-207, the-contents of which..are incorporated, by reference herein. After UHT me product-may. be- aseptically packed. Additionai processes for.improved shelf life, may includepasteurizing:. and refrigeration until used;, orthe endproductmay be evaporated and subsequently spray dried to yield a stable powder Preferably,.the enzyme treated suspension is homogenized.:and.subjectedt oUHT and. aseptically packed.
Enzyme activity may be terminated or removed fromme.-enzyme treated, suspension before processing;forimproved shelf life... As, an-alternative,. the.enzyme activity, may be terrninated during-some, of the. processes that improve the.- self-life of the product,, such as the. UHT process.
Optionally, theenzymetreatedsuspension of step (b) may be-treated sequentially with a. second enzyme-preparation comprising at least, one hydrolase: enzyme, having the. ability to hydrolyzect-giycosidic bonds: seiected from the group consisting of β-amylase, α-amylase, amylogiucosidase,-. pullulanase. and a mixture.- thereof: Further in. this respect;, the- above-

improved cereal substrate suspensions may be treated by combining an isomerase, such- as glucose isomerase with the hydrolases of the enzyme preparations.
The. homogeneous and stable improved cereal suspension having the. aroma and taste, of natural cereals and containing intact (3-glucans may also be prepared by the method comprising:the steps:
a) preparing a. cereal meal suspension;
b) treating. the cereal meal suspension of step (a) with. (3-amylase and followed sequentially with the. introduction of a-amylase;
c) treating the enzyme, treated suspension-ofstep (b) with-an. .enzyme preparation..as discussedpreviousiy herein, above comprising at.leastone. hydrolase, enzyme, having the ability to. hydroiyze a-glycosidic. bonds.. Preferably, an. a-giycosidic: hydrolase is selected-from-, the group consisting; of β-amylaser.α-aniylase,, amyIoglucosidase,„pullulanase. and.a..mixture thereof withthe proviso that when the enzyme preparation comprises Pramylase; or a-amylase there, is. a mixture, of at least one otherof the named argh/cosidic hydrolases...
It is preferred that when-theenzyme-. preparation comprises .both β-amylase. anda-amylase,. the enzymes are introduced simultaneously to the cereal, substrate suspension.
The enzyme- treated suspension of step (b) and (c) may be- further treated by discontinuing enzymatic, activity and/or further performing, a-fmishing process step. Detailed Description of the Preferred Embodiments
Accordingto the; invention, there is provided an enzyme, preparation, for enzymatic hydrolysis, of constituents: it. a. cereal substrate:, suspension comprising: at least one

hydrolase enzyme having the ability to hydrolyze a-giycosidic bonds. The hydrolases may be seieaed from. the. group consisting of P-amylase, aramylase, amyloglucosidase puiluianase andmixtures thereof- Preferably, when the enzyme preparation, comprises P-amylase-ora-amylase mere is a mixture of at least one other of me named a-glycosidic hydrolases, and more preferably, when there are two or more enzymes, combined, in the enzyme preparation, the enzymes are introduced simultaneously to the cereal, substrate suspension. In addition to the above-identified hydrolases, the enzyme, preparation may further comprise an isomerase, such as glucose isomerase.
ln preferred embodiments of the invention,, the. enzyme, preparations may comprise puiluianase solely; amyloglucosidase solely; or several different combinations of hydrolases including: amixture combining p-amylase with;puilulanase;. amixturexombining P-amyiase, puiluianase. and. amyloglucosidase; a mixture: combining: β-amylase-and aramylase;. and a mixture combining: a-amylase, P-amylase and .amyloglucosidase. Any of the above, enzyme preparations, comprising a. hydrolase, alone or in combination with another may further . comprise anisomerase, such, as .glucose, isomerase..
The enzyme- preparations, of the present invention, convert cereal starch, which comprises both amylase and amyiopectin,. to lai emolectilar wdght maltodaxtrins.andlow--molecular-weight compounds of various degrees of modification such: as maltotriose,. maltose and glucose. For instance, p-amylase hydrolyzes a-l-4-glycosidic bonds, sequentially from the non-reducing; terminal endofarnylose and amylopectm with adeaved productof maltose; a-amylase hydrolyzes intemala-l-4 glycosidicr bondson. both: amylose and amylopectin. with.

a cleaved maltodextrin; and. amyloglucosidase liydrolyzes a-L-4 and 1-6 glycosidic bonds on the non-reducing end of the starch, releasing glucose molecules. Therefore, a combination of hydrolases having, the. ability to hydrolyze a-glycosidic bonds will provide various ratios of maitodextrin/sugar in an enzyme treated cereaL suspension.
The choice of enzymes and the reaction time determine the degree, of degradation and product, spectra. Different species of di-and. mono-saccharides are produced, by using different enzyme preparations, which, include the combination of at least, one a-glycosidic. hydrolase and/or- an isomerase. The end products could include. the.di-saccharide,. maltose, and the. mono-saccharides fructose, and. glucose. For example, the debranching-.enzyme pulluianase together with β-amylase will accumulate, large amounts, of maltose. Amyloglucosidase and glucose isomerase may result in. the production of fructose- and glucose. When combining- β-amyiase and; a-amylase,. the. aramyiase promotes, the action of the [3-amyiase thereby obtaining maltose and..mdtodextrin.units.whiIe using a smailer amount of enzymes then if enzymes, are used separately. A. cereal starch may be converted entirely to low-molecuiar-weight compounds, such as glucoseby combining for example,.. (3-amyiase, aramylase and amyloglucosidase. Glucose isomerase, solely, will result in the.production of fructose when added.to acereaLsubstrate.thatalready contains glucose. In the. alternative, the enzyme preparation wherein. (3-amylase, a-amylase and amyloglucosidase are combined with glucose, isomerase may also producea.productwith.high.Ievels of fructose..
The enzyme preparations, that being a single enzyme or a mixture of enzymes may treat a.. cereaL substrate suspension, of the present invention, by introducing: free enzymes

directly into a cereal substrate suspension, or in the alternative introducing the cereal, substrate suspension to a vessel.containing immobilized enzymes.
Free, enzymes as used herein, means enzymes, which' are free to move in. the suspension and are not restricted by containment or affixed to a substrate. Usually, free enzymes or cells. are not re-used because they are too small to filter and recovery may become cost, prohibitive. Hence, elimination of the- biocatalytic activity of free, enzymes in the present invention is usually accomplished by denaturing" of the. enzyme;
Immobilized, enzymes:as used herein.means. free, enzymes that are-physically confined by different, methods including, but not. limited to,, semi-permeable; membranes, hollow bore fibers, or. ultrafiltration, membranes. Immobilized, enzymes, whether soluble-or insoluble, allow for the simultaneous immobilization of many enzymes,, selectively controilmg..substrates and products' through membrane, selectivity. As usedin. the present invention-immobilized enzymes provide for ease of loading and. treating the cereal meal suspension in a. continuous mode reactor.
The benefits of immobilized enzymes, include complete recovery of the. enzymes from the. reaction mixture whether used in a batch or continuous mode operation. Thus, the enzymes can. be used, repeatedly without any contamination of the final product and without the. need to heat the productso as to denature the enzyme. Also, larger concentrations of immobilized enzyme, can.be. utilized because the immobilized enzymes can.be recovered and re-used, resulting in a.shortening. of reaction time, and/orthe size of the vessel needed to carry out the reaction; Another advantage-is the virtual absenceof enzymeintheftnal productso

that.the enzyme: only has to be approved as a .food processing.aid and not as a.food.additive even when heating and subsequent enzyme inactivation is not. included, in the. process.
It is further contemplated, by the inventors that the enzymes used to prepare the enzyme-modified suspensions and/or the homogeneous and stable improved, cereal suspension may include, enzymes derived from whole ceils, organelles, or even microorganisms used, as biological catalysts in a.fermentation process.
In the present invention, the. overall, conditions including, temperature, pHand the. addition of other, substrates, such as enzyme cofactors or buffering, agents: will! determine me enzyme, activity, and therefore, the yield, and quality of the- end.product It; is known, that enzymes may be extracted, from different sources but can. catalyze, the. same reaction;. For instance aramylase from.the fungal organism. Aspergillus, oryzae has a: optimum pBC of 47 and an. optimum: temperature- of 50 0c, while a-amylase from the. bacterium: Bacillus lichenformis has a pH. optimum of 7 .5 and, an. optimum-, temperature of: 90°C. Thus, understood,, the. optimum, conditions, including the. amount: of enzyme, the-temperature of the slurry, the agitation, time, and the pH value, are optimized, to obtain: a final product of suitable viscosity. Techniques used for determining optimal parameters are well known.and. widely used. in. the. air
The. enzyme, preparations of the present: invention may be used to provide: a. homogeneous and stable, improved, cereal suspension having: the. aroma, and taste of natural, cereals and.containing.intact β-glucans. The-enzyme preparations are used to treata cereal substrate, which: may include a. cereal, meai suspension, a.preprocessed cereal suspension, or


a. mixture thereof.
In one embodiment, the cereal, substrate suspension is a-cereai. meal.suspension. The
cereal meal suspension is prepared by dry- or wetrgrinding rolled cereals, or otherwise,
hear- and water-treated cereals to-meal, and suspending the cereal meal in water to.form a.
cereal meal.suspension. Optionally, the suspension may be centrifuged ordecanted in order
to remove coarse fibre, particles before being treated with the: enzyme preparation.
Conveniently, the cereal meal suspension is prepared, on. the. basis of commercially produced,, pregelatinized rolled, oats' retaining the originaL taste, and.aroma.of the: oats. The rolled oats are ground to oatmeal by total, dry or wet grinding. In dry grinding, the oatmeal is suspended.in. water, preferably at a.temperatureof 50°-65°C. Also in wet grinding, water is: used preferably at a temperature, of 50°-65°C'.. Especially good results are; obtained if the: water has. been, deionized.
Fora, majority of starch contamed. within the.cereai meal. the. heating of the. suspension to a. temperature between 50° to 65°C gelatinizes the. cereal, starch; for easier hydrolyzation However, some oats contain esistant starches.that are not getatinized at these.temperatures, and therefore, are not easily hydrolyzed by the enzyme preparations ofthe. present invention. In this instance,, it. has been, found to be beneficial, to initially hydrolyze the. non-resistant starch in afirst enzyme treating'step with the enzyme preparations of the: present invention and then, subjecting; the- suspension to higher temperatures,, preferably above 100°C to gelatinize, the,1 resistant starch. The suspension is subsequently cooled to a workable temperature and standard conditions. The suspension is the retreated with, the enzyme
preparations of the present invention. Accordingly, the non-resistanr starch in the cereal meal, suspension may be.hydrolyzed.with the enzyme preparations of the present invention and then the suspension is heat, treated, to a. higher temperature to solubilize. the, resistant starch. The. suspension is cooled to a. temperature, that, is suitable for enzymatic activity and then, retreated with. an. enzyme- preparation of the present invention. This method, will allowed, for more compieterhydroiyzation. of substantially all the cereal, starch including the resistant starch in. the cereaL meal suspension.
Suitably, the slurry or suspension has a weight ratio of meal to water in the range of about 1:6 to about 1:9, which corresponds to.adrysolids. conten to fabout 10 to about 15 % w/v. The suspension is agitated until the meal has been dispersed. The. slurry should have. a.. pH of at.Ieast 5 to about 8. This pH range has been found to be effective when .adding the enzyme preparations of the present invention.. Withuithis. pKrangethe enzyme preparations, have acceptable, catalytic, activity and. can. avoidthe. use of additives to alter the pH.
In, order to remove the. coarse particles, the. suspension can then be centrifuged or decanted at 350-450 G. for about 10-15' minutes.
in a.fiuther ambodiment of the present.invention, a. preprocessed cereal suspension, may be used The. preprocessed. cereal suspension is defined as that which: is first prepared, according to the. methods disclosed in US Patent No. 5,686,123. The cereal, meal suspension is treated wkh.β-amylase in a first enzyme treatment step which specifically generates maltose units and. has no giucanase and proteinase activity, to a. viscosity of 3-0.1. Pas at the shear rate, of 10-100 s'1. Then the suspension.is treated with, a-amylase- in a.second enzyme-treatment

step which specifically generates maltose units and has no glucanase and proteinase activity, to a.viscosiiy of The cereal substrate: suspension is treated, with the enzyme, preparations under carefully regulated operating temperatures. The temperature, is. chosen to favor the enzymatic performance allowing both, fast hydroiytic rates and good enzyme, stability. A. temperature-from about 40°C to a temperature, below that which would denature, the enzyme or combination of: enzymes is generally employed, preferably from about 50° to about: 90°C depending on the enzyme. At lower temperatures the enzyme activity may be low and at higher temperatures me enzyme stability may be; low. Accordingly, the temperature, of the, cataiytic reacrion is chosen, to. optimize production of the end products while maintatning the stability of the. enzyme, preparation.. The present.invention -isvalso applicable to thermostable starch degrading enzymes, in which case the operation conditions, can. be adapted, to the. characteristics, of such enzymes..
A. hydrolase, and/or combination, of several hydrolases are introduced, to a, cereal substrate.suspension:hi.a-sufrIcient amountto hydrolyze a-glycosidic bonds of constituents in the cereal substrate suspension to provide an. end product with the desired viscosity. The combination of enzymes and amount, of each specific; enzyme results in suspensions, that contain differenr sugars,. but also m different amounts.. High content of low molecular sugars, such as. maltose, and glucose will yield an end product suspension, with. less, viscosity, In

contrast, a. higher content of maitodextrin, which is considered a higher molecular weight molecule yields a product with, greater viscosity that may be used, in soups or yogurts because of thicker consistency. Accordingly, varying the kind and/or the amount of enzymes in the-mixturewillyield, specially designed-products. Using a. specific combination of enzymes, helps to standardize the process so that the end product is related to the enzyme combination whereby reaction time and other process parameters do not greatly affect the. end product. In carrying outthe presentinvention,..it is generally advantageous to employ between about 1 to about 100 mL of enzyme preparation perlrilogram of oats or other grain material which make, up the. cereal, substrate suspension. The suspension, may be treated, with the, enzyme preparation to produce, a, final.product, which has. the- viscosity of. about.water or about. 10 mPas.to about several hundreds of mPas at a..sheer rate of about 500 to about 1.000s"1.
Generally, megrainthatis incorporated, into, the cereal meal, suspension: may be-any starting grain material including; but not limited, to oats, barley, rice, wheat; maizer rye, sorghum, triticaie and pearl millet Preferably, the grain is. oats. As stated above., oats have properties.that make them, especially desirable from a. consumer point of view because.of the large, amounts of high, molecular weight (3-gIucans which are natural, hydrocolloids. Ik suspensions produced, by enzymatic, hydrolysis widi the. enzyme preparations ofthe present invention, the (3-glucans found in. oats function as indigenous stabilizers. Therefore, the cereal suspensions of the present invention may be used, in food for thickening, gelling or emulsion, stabilizing, effects.


The sweetness of an. enzyme modified suspension, or a. homogeneous and stable improved cereai suspension can be regulated and/or manipulated using appropriate enzyme, preparations.. In fact;, the: enzyme preparations of the present invention may be introduced in several steps to tailor-make: me end product: For instance, an enzyme preparation comprising α- and β- amylase may produce a, high, level of maltose. With a second treatment of an enzyme preparation comprising, amylogiucosidase and/or glucose isomerase. the maltose can be converted, to glucose and. fructose. The. production, of glucose, and,, particularly, of fructose will, resultin. a sweeter: suspensionman one. comaining:mainly maltose.. A. suspension comprising fructose has the significant advantage that it. can be consumed by diabetics without adverse affects.,
The specifi'c..type of sugar affects, not only the properties of the suspension but also the organoleptic.properties of products producedby using.the: suspensions. By altering; the sugar profile; it is possible to tailor-make.: suspensions, that yield, end products- having functional properties, such: as viscosity; nutritional properties and sugar ratio content to meet the; needs of the final user.
Enzymatic activity may be. discontinuedor terminated in. the enzyme modified: cereai suspension, or the enzyme treated. cereaL suspension, by any method, well known, in. the art; including denaturation, centrifuge, chromatography for free enzymes and/or removal, of the suspension from contact: with immobilized enzymes. Preferably, The. enzyme reaction is terminated by heating, the cereai suspension to at least 80°C, and preferably, between about 80° to 90°C

As an alternative the. enzyme activity may be discontinued terminated, during the final process steps that improve the shelf-life of me. product. Representative final process steps may include: removing, coarse particles by centri&ging or decanting; homogenizing the enzyme treated, suspension at atemperature of about 42 to about 45°C, atapressure of about 200 to about 250 bar; or subjecting the.producr to Ultra High Temperature (UHT) treatments disclosed, in. Food. Engineering and. Dairy Technology, H.G. Kessler, VerlayA- Kessier, 1981, Chapter 6, pp. 139-207J the contents of which are incorporated by reference herein. After UHT the. product may be.aseptically packaged.. Additional-processes: for. improved sheif life may include pasteurizing and refrigeration until .used; or the. end product may be evaporated.and subsequently spray dried to yiekLa. stable powder.
Cereal suspensions, according to the invention, can be used in the.same. fields as the products disclosed, in. US. Patent No. 5,686,123, thatis.as amilksubstitute;. as. the. basis of or an additive in the manufacture of ica-cream,. gruel, yogurt;, milkshakes, and snacks.
The invention will now be described, in more detail, by. the following, non-limiting examples.
Pregeiatmized rolled oats: were wet-milled, at: atemperature of about 52° to about. 63°C. The concentration of the., suspension was about 10 to about 15% w/v. An, enzyme preparation according:to the. invention which comprised barley -amylase (Genencor Intl., Rochester, NY, USA;orRhodiaLtd,. Cheshire, UK) and pullulanase, adebranching enzyme, e.g. Promozyme (Novo Nordisk,. Bagsvaerd,. Denmark), was added, to the cereal meal

suspension at a. concentration- of approximately 2 ml per kg of oats and at a. temperature of about 58° to about 61°C. The concentration of the enzymes in the enzyme preparation was about 500 to about 1000 DP0 and. about 150 to about: 300 PU (pullulanase units) per ml respectively. The enzyme preparauon was. allowed, to act for-2 hours, or untii.the. viscosity ofthe suspension, dropped-to about20 to about 40 mPas at ashear rate of around 700 s'1. The product contained, large, amounts of maltose. The majority of starch (approximately 60% of the oats) was. converted, to maltose.
The suspension: was.then aheaied.to.about 85° to,.about90°C to.inactivate theenzymes. Theproductwas decanted:to remove the excess of.non-soluble fibre, and homogenized. Optionally the product: could be UHT treated: andaseptically packed, pasteurized, and. kept refrigerated, until., used, or it is evaporated, and. subsequently spray dried, to yield, a. stable-powder
EXAMPLE 2 Pregeiatinized. roiled, oats, were: milled, as. in Example. 1. An. enzyme preparation accordingto, the. invention- which comprised barley P-amylase.(GenencorIhtl,. Rochester;. NY, USA; orPvhodiaLtd, Cheshire,,UK), pullulanase,.a.debranching enzyme, e.g. Promozyme. (Novo Nordisk,. Bagsvaerd, Denmark) and amyloglucosidase, e.g:, AMG (Novo Nordisk,. Bagsvaerd,. Denmark) orQptidex (Genencor M., Rochester, NY, USA) was added to the-oat. meal suspension at a. concentration of about 3 to. about 4- mi per kg oats. and. at a. temperature of about 5 8° to about61°C. The concentration of these, enzymes in. the enzyme-preparation was about 400 to about. 700 DP0, about 100 to about 200 PU (pullulanase units)


and about 90 to about 110 AGU per ml, respectively. The enzyme preparation was. allowed to actfor about 1. to about 2. hours, or until me viscosity of the suspension dropped to about 20 to about 40 mPas at a shear rate of 700 s"\ The. product contained large amounts of glucose. Finally, the suspension was heated and treated as in Example 1.
EXAMPLE 3 Pregelatinized. rolled oats were milled, as. in. Example. 1. An. enzyme preparation according, to the. invention, which, comprised a., mixture of {3-amylase (Genencor Intl., Rochester, NY, USA;, or Rhodia. Ltd, Cheshire, UK) and. endo-acting a-amyiase, e.g. Fungamyl (Novo Nordisk, Bagsvaerdr Denmark) or Mycoiase (Genencor Intl., Rochester, NY, USA), was added to the.cereal.meal suspension at. a. concentration, of approximately 2 mi per kg of oats. and. at atemperature of about 54° to about57°C. The-concentration ofthese enzymes, in. the. enzyme, preparation was aboutl400 to about 1600. DP0 and about 30 to about. 70 AU (amylase units) per ml,, respectively. The enzyme preparation was. allowed to actfor about L hour, or until the viscosity of the suspension, dropped, to about 20 to about.40 mPas at. a. shear rate.of about 700 s -l. Most of the oat. starch (60-70%) was convertedto maltose and the. rest was present as maltodextrins (step 1) Then (in step 2), another exo-acting: enzyme was added, e.g;, amyloglucosidase AMG (Novo Nordisk, Bagsvaerd,. Denmark) or Optidex (Genencor IntL, Rochester, NY, USA), at a. dosage of approximately 600 AGU (amyloglucosidase. units) per kg. of oats. The. reaction, was terminated, when the. desired amount of glucose had been: produced. For example,. 30 minutes after the addition, of amyloglucosidase (giucoamylase), the suspension contained equal, amounts jof maltose-and

glucose while the: maltose, content was 50% of the suspension instep. 1. The maltose content was high in step 1, and. amyloglucosidase rapidly hydroiyzed diis substrate. As the maltose content decreased,, maitodextrin. became the preferred substrate and aiso became increasingly hydroiyzed. At full, conversion all the. starch was converted to glucose. Finally, the suspension was heated and treatedas inExample' 1.
EXAMPLE 4 Pregelatinized. rolled, oats were milled as: in. Example 1. An enzyme preparation, accordingto the invention. wMch.comprised.amixture.of barley (3-amyiase. (Genencor Intl., Rochester, NY, USA; or Rhodia. Ltd,. Cheshire, UK),, a-amylase, e.g; Fungamyl (Novo Nordisk,.Bagsvaerd, Denmark) or Mycolase (Genencorlhti., Rochester, NY, USA) and amyloglucosidase,. e.g:, AMG (Novo Nordisk, Bagsvaerd, Denmark) or Optidex (Genencor IhtL, Rochester, NY, USA) was. added to the suspension at a dosage.of.about 1 to about 4 ml.perkg.oats and at a. temperature- of about 54° to about57°C. The.concentration of these, enzymes in the: enzyme, preparation was about 700 to about 900 DP0, about. 1 to, about. 3 5 AU (aramylase.units) and about200 to about 350 AGU per mi, respectively. The enzyme preparationwas allowed to actfbrabout l-2.hours, or until the viscosity of'the suspension dropped.to about20 to about 40 mPas at a. shear rate, of about 700 s"1. Finally, thesuspension was heated.andtreatedas in Example. 1.

A suspension of oats was prepared as in U.S. Patent No. 5,686,123 is treated, with an enzyme preparation; according to the invention and. which comprised, barley {3 -amylase (GenencorlmL, Rochester; NT, USA; or Rhodia Ltd, Cheshire, UK) and pullulanase, a debranching enzyme, e.g. Promozyme (Novo Nordisk, Bagsvaerd,. Denmark), at a. concentratioa of approximately 2, ml per kg of oats. Alternatively, the suspension was. treated with. a. debranching;. enzyme;, such, as pullulanase,. e.g: Promozyme (Novo Nordisk, Bagsvaerd, Denmark), at. a concentration., of approximately 800 PU per kg of oats. Otherwise the. conditions, were, the same as in Example 1. The product, was high in. maltose and. contained essentially no maltodextrins.
A suspension, ofoats. was. prepared as in. U.S. Patent No.5',686,123. and was. treated, with the.same, enzyme preparation as in.Exampie.2, orwith amylogiucosidase as kExample 3 (as in. step 2). The product: contained a, decreasing; amount of maltodextrins. and an increasing amount of glucose as thehydrolytic. reaction, proceeded.
To any of the-products ofExamples 2,3', 4-and 6, i.e.r whichcontained glucose,, there, was added an enzyme preparation according tcr the invention which, comprised, amylogiucosidase of. about 50 to about. 60 AGU' per ml. e.g:, AMG (Novo Nordisk, Bagsvaerd,. Denmark) or Optidex; (Genencor Ind, Rochester, NY, USA) and glucose isomerase. (aboutSOOO G3XI per ml), ata. concentration; of about. 18 to about 70 ml per kg;

of oatsr or only giucose isomerase, e.g. Spezyme GI (Genencor Intl., Rochester, NY, USA) or Sweetzyme.(Novo Nordisk, Bagsvaerd, Denmark), at a concentration of about 50,000-to about 200,000 GH (glucose isomerase units). Within, two (2) hours, 25% of me glucose was convened to fructose.
EXAMPLES Pregelarinized. rolled, oats were milled, as. in Example. 1. An enzyme, preparation according to me invention which comprised, a. mixture- of J3-amylase (Genencor Intl., Rochester, NY,.USA;.or RhodiaLtd, Cheshire, UK), and endo-acting.a-amylase, e.g. Fungamyl (Novo Nordisk,. Bagsvaerd, Denmark) or Mycolase (Genencor Intl., Rochester, NY, USA) was added, to the cereaL meal, suspension.ata. concentration of approximately 2„ml per kg oats. and. at a. temperature of about 54-° to .about57°C The. concentration ofthese enzymes in the enzyme, preparation: was: about 1400. to. about 1600 DP°.and dabout L to. about 8 AU (a-amylas& units) perml,.respectiveiy. Theenzyme preparation was allowed to actfor approximaieiy 1 hours, or until, the, viscosity of the. suspension dropped to about45' to about 65 mPas at a shear rate of about 700 s'1. One third of the. starch, was convertedto maltose, whiie.the rest was present; as maitodextrin.. The productwas quite thickbecause of the high content of maitodextrin.. Finally the suspension, was. treated, as. in Example.-. 1.

We Claim:
1. An enzyme modified oat cereal suspension comprising maltose and maltodextrin units, intact B-glucans, and proteins, said enzyme modified oat cereal suspension prepared by the method which comprises the steps of:
(ij providing an oat cereal substrate suspension, (ii) providing an enzyme composition comprising B-amylase and a-amylase, and
(iii) treating said oat cereal substrate suspension (i) with said enzyme composition (ii)
wherein said β-amylase and a-amylase enzymes are introduced simultaneously for accelerated enzymatic hydrolysis, reduction of viscosity, and regulation of sugar content formation of said modified oat cereal suspension, said method allowing use of lesser amounts of said enzymes than otherwise needed when said enzymes are used separately.
2. The enzyme modified cereal suspension as claimed in Claim 1, wherein the enzyme preparation is free of glucanase or proteinase activity.
3. The enzyme modified suspension as claimed in Claim 1, wherein the enzyme modified oat cereal suspension has a viscosity from 45mPas to 65mPas at a sheer rate of 700s-1

4. The enzyme modified oat cereal suspension as claimed in Claim 1, wherein at least one finishing step is performed on the enzyme modified oat cereal suspension.
5 The enzyme modified oat cereal suspension as claimed in Claim 4, wherein the enzyme treated suspension of step (iii) has its enzymatic activity terminated or enzymes removed before performing the at least one finishing step.
6. The enzyme modified oat cereal suspension as claimed in Claim 4, wherein the at least one finishing step is selected from the group consisting of homogenization, ultra high temperature treatment, pasteurizing, refrigeration, evaporation and spray drying.
7. The enzyme modified oat cereal suspension as claimed in Claim 1, wherein the suspension of step (i) is further treated by removing coarse fiber particles therefrom.
8 The enzyme modified oat cereal suspension as claimed in Claim 1, comprising the step of heat treating the suspension of step (iii)to solubilize the resistant starch, cooling the suspension to a temperature suitable for enzymatic activity and treating with an enzyme preparation to further hydrolyze resistant starch.

9. The enzyme modified oat cereal suspension as claimed in Claim 1,
wherein the suspension comprises a mixture of low molecular weight
sugars and high molecular weight maltodextrins.
10. The enzyme modified oat cereal suspension as claimed in Claim 1,
wherein the enzyme composition of step (ii) comprises an isomerase.
11. The enzyme modified oat cereal suspension as claimed in Claim 1,
wherein the cereal substrate suspension is converted principally to
low molecular weight compounds.
12 The enzyme modified oat cereal suspension, wherein the final product
has a viscosity in the range of 10 mPas to several hundred mPas at a
sheer rate of 500 to 1,000s1
13 An enzyme modified oat cereal suspension substantially as herein
described with reference to the foregoing examples.
Dated this 1st Day of February, 2005.



99-mumnp-2005-cancelled pages(22-3-2006).pdf





99-mumnp-2005-form 1(1-2-2005).pdf

99-mumnp-2005-form 1(25-4-2006).pdf

99-mumnp-2005-form 18(4-8-2005).pdf

99-mumnp-2005-form 2(granted)-(25-4-2006).doc

99-mumnp-2005-form 2(granted)-(25-4-2006).pdf

99-mumnp-2005-form 3(31-1-2005).pdf

99-mumnp-2005-form 3(5-4-2006).pdf

99-mumnp-2005-form 4(25-4-2006).pdf

99-mumnp-2005-form 5(31-1-2005).pdf


99-mumnp-2005-petition under rule 137(7-04-2006).pdf

99-mumnp-2005-power of authority(22-3-2006).pdf

Patent Number 206656
Indian Patent Application Number 99/MUMNP/2005
PG Journal Number 42/2008
Publication Date 17-Oct-2008
Grant Date 04-May-2007
Date of Filing 01-Feb-2005
Name of Patentee CEREAL BASE CEBA AB
Applicant Address SCHEELEVAEGEN 18, 22363 LUND,
# Inventor's Name Inventor's Address
PCT International Classification Number A23L 1/00
PCT International Application Number PCT/IB99/01860
PCT International Filing date 1999-10-08
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 60/104,706 1998-10-19 U.S.A.
2 60/106,224 1998-10-30 U.S.A.
3 09/302,127 1999-04-29 U.S.A.