A process for preparing an OAT-derived base for OAT-based dairy alternative food products

Abstract

The present invention relates to a method for making an oat-derived base, comprising treating a mixture comprising plant-derived substrate comprising whole oat flour and vegetable protein isolate and:or vegetable protein concentrate with an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme. The present invention further relates to use of the oat-derived base to produce an oat-based acidified dairy alternative food product.

Description

[0001] A PROCESS FOR PREPARING AN OAT-DERIVED BASE FOR OAT-BASED DAIRY ALTERNATIVE FOOD PRODUCTS

[0002] Reference to sequence listing

[0003] This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference.

[0004] FIELD OF THE INVENTION

[0005] The present invention relates to the use of a beta glucanase and optionally additional enzymes for obtaining an oat-derived base with improved organoleptic and techno-functional properties.

[0006] BACKGROUND OF THE INVENTION

[0007] Vegetarian diets in general, and vegetarian sources of protein in particular, have increased in popularity as consumer interest in healthier and more eco-friendly eating habits has grown. Plant-based acidified dairy alternatives, such as plant-based dairy-alternative yogurts, offer an interesting alternative to traditional animal-derived yogurts, and additionally are viewed as healthier choices because of their reduced level of cholesterol and saturated fat. Finally, plant-based dairy-alternative yogurts are free of lactose.

[0008] Oat-based dairy-alternative yogurt has a high consumer demand. However, production of oat-based dairy-alternative yogurt has a number of challenges. Fermented oat milk alone cannot form a gel network analogous to that found in traditional yogurt. To overcome this, additional ingredients such as texturizers or thickeners are frequently added, thereby increasing the ingredient list of the product. However, a long ingredient list is frequently viewed by consumers as undesirable.

[0009] There is a need for effective, efficient solutions which can improve the texture of oat-based dairy-alternative yogurt without the need of additional texturizers.

[0010] SUMMARY OF THE INVENTION

[0011] The present invention is based on the surprising finding that treating a plant-derived substrate comprising whole oat flour and vegetable protein isolate and / or vegetable protein concentrate with an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme produces an oat-derived base that has superior organoleptic and technofunctional properties. The superior organoleptic qualities are evidenced by the viscosity and gelstrength of the resulting plant-based acidified dairy alternative food product. Oat-based dairy alternative food products that are produced from the oat-derived base of the invention have a texture that is surprisingly thicker and smoother, so that additional texturizers and / or thickeners are not required. In a first aspect, the present invention relates to a method for making an oat-derived base, comprising: a) obtaining a mixture comprising a plant-derived substrate comprising whole oat flour and vegetable protein isolate and / or vegetable protein concentrate; and b) treating the mixture of step a) with an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme; wherein the enzymatic activity on the mixture of step a) produces the plant-base.

[0012] The invention provides a method of preparing an oat-based product with improved organoleptic and techno-functional properties.

[0013] DETAILED DESCRIPTION OF THE INVENTION

[0014] In accordance with this detailed description, the following definitions apply. Note that the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0015] Unless defined otherwise or clearly indicated by context, all percentages are percentage by weight (percent w / w or “% (w / w)”).

[0016] Unless defined otherwise or clearly indicated by context, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0017] As used herein, the verb “comprise” as is used in this description and in the claims and its conjugations are used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, reference to an element by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements are present, unless the context clearly requires that there is one and only one of the elements. The indefinite article “a” or “an” thus usually means “at least one”.

[0018] As used herein, an "effective amount" is defined as an amount effective to achieve a desired biological result, such as reducing, preventing, or treating a disease or condition and / or inducing a particular beneficial effect. In some embodiments, the effective amount is the amount provided in a single serving probiotic beverage where consumption of multiple servings is required over time for the beneficial effect to be observed.

[0019] The present disclosure provides methods for making an oat-derived base by treating a mixture comprising plant-derived substrate comprising whole oat flour and vegetable protein isolate and / or vegetable protein concentrate with an enzyme composition comprising a beta- glucanase and optionally at least one additional enzyme. Many current methods for the production of a oat-based acidified dairy alternative food product rely on an oat-derived base which comprises oat-based milk (also referred to as “oat drink”), which is then processed into an acidified dairy alternative food product. The present invention is based on the unexpected finding that a superior oat-derived base could be produced from a mixture comprising whole oat flour. Beginning with a mixture comprising whole oat flour reduces the overall processing required to produce the oat-derived base, and therefore also reduces the processing required to produce the oat-based acidified dairy alternative food product. This reduction in processing results in both time and resource (including energy resource) savings, and increases the efficiency of production. Also, the oat-derived base of the invention provides an unexpectedly thicker and smooth texture when used to produce an oat-based acidified dairy alternative food product, so that additional texturizers and / or thickeners are not needed, producing a shorter, “cleaner” ingredient list for the food product.

[0020] The oat-derived base of the invention is produced from a mixture comprising a plant- derived substrate comprising whole oat flour and vegetable protein isolate and / or vegetable protein concentrate. Whole oat flour, also referred to as “oat flour”, is milled from whole grain oat groats and includes the germ, bran, and endosperm. In some embodiments, the oat flour is heat-treated.

[0021] Vegetable protein isolates and vegetable protein concentrates are produced by extracting some of the oil or starch present in the vegetable protein source. This process raises the relative amount of protein in the isolate or concentrate. For both, the amount of protein present may be measured by dry weight (w / w). Vegetable protein concentrates typically contain around 65% protein. Vegetable protein isolates typically contain about 90% protein. Vegetable protein isolate and / or vegetable protein concentrate may be added to a food product or to the preparation of a food product to increase its nutritional value and / or to give it specific functional properties. Functional properties include changing water retention, oil retention, foaming capacity, gelling capacity, and / or emulsion capacity.

[0022] In some embodiments, the vegetable protein isolate is derived from a legume, a nut, or any combination thereof. In further embodiments, the vegetable protein isolate is derived from peas, yellow peas, lentils, chickpeas, soybeans, beans, peanuts, fava beans, lima beans, mung beans, lupins, almonds, cashews, macadamia nuts, pistachios, pecans, walnuts, pili nuts, hazel nuts, sesame seeds, or any combination thereof. In some embodiments, the vegetable protein isolate a legume protein isolate and is derived from peas, lentils, chickpeas, soybeans, beans, peanuts, fava beans, lima beans, mung beans, lupins, or any combination thereof. In further embodiments, the legume protein isolate is derived from peas, soybeans, fava beans, or any combination thereof.

[0023] In some embodiments, the plant-derived substrate of the mixture of the invention comprises a ratio of whole oat flour and vegetable protein isolate. In some embodiments, the plant- derived substrate comprises a ratio of whole oat flour to vegetable protein isolate from 20:80 to 80:20 by dry weight. In further embodiments, the plant-derived substrate comprises a 20:80, 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, or 80:20 ratio of oat flour to vegetable protein isolate by dry weight. In further embodiments, the plant-derived substrate comprises a 70:30 to 30:70 ratio of oat flour to vegetable protein isolate by dry weight. In some embodiments, the plant-derived substrate comprises 55:45, 50:50, 45:55, 40:60, 35:65, 30:70, or 25:75 ratio of oat flour to vegetable protein isolate by dry weight. In some embodiments, the plant-based substrate of the invention may have been standardized and:or homogenized. In some embodiments, the plant-based substrate may have been pasteurized or otherwise heat-treated.

[0024] The level of protein provided by the whole oat flour and the vegetable protein isolate and:or vegetable protein concentrate dictates the amount of protein in the resulting oat-derived base, which then also plays a role in the level of protein in an oat-based dairy-alternative food product which may be prepared from the oat-derived base. In some embodiments, the oat-derived base comprises 2-15% protein content. In some embodiments, the oat-derived base comprises 2- 10%, 3-9%, 2-5%, 3-5%, 5-8%, 6-9%, 8-10%, 10-12%, or 12-15% protein content. In further embodiments, the oat-derived base comprises about 6-8% protein content, such as for example 5.5-8.5% protein content. In other embodiments, the oat-derived base comprises about 2-4% protein content, such as for example 1.5-4.5% protein content.

[0025] In other embodiments, the oat-based dairy-alternative food product prepared from the oat- derived base of the invention comprises 2-10% protein content, without additional protein added during the production of the oat-based dairy-alterative food product. In some embodiments, the oat-based dairy-alternative food product comprises 2-4%, 4-6%, 5-7%, 6-8%, 7-9%, 8-10%, or 10-12% protein content. In further embodiments, the oat-based dairy-alternative food product comprises about 6-10% protein content, such as for example 5.5-10.5% protein content. In other embodiments, the oat based dairy-alternative food product comprises about 2-4% protein content, such as for example 1.5-4.5% protein content.

[0026] The mixture of the invention comprises a plant-derived substrate comprising whole oat flour and vegetable protein isolate and:or vegetable protein concentration. In some embodiments, the mixture comprises 10-25% (w:w) solids. In further embodiments, the mixture comprises 6-20%, 8-20%, 12-18%, 13-17%, or 15-17% solids. In some embodiments, the mixture may comprise additional ingredients. The mixture may further comprise water, salt, sugar, fat, oil, flavor, flavor modifiers, or any combination thereof.

[0027] In some embodiments, the mixture may comprise oil, such as a plant oil, such as for example rapeseed oil, flaxseed oil, safflower oil, flaxseed oil, soybean oil, olive oil, sunflower oil, palm oil, coconut oil, canola oil, linseed oil, corn oil, peanut oil, vegetable oil, or any combination thereof. In some embodiments, the mixture may comprise fat, such as a plant-based fat, such as for example coconut fat (for example coconut oil), nut fat (for example almond oil, cashew nut oil, chestnut oil, hazelnut oil, macadamia oil, pecan oil, pistachio oil, walnut oil, or a combination of any thereof), or nut butter (for example almond butter, peanut butter, pistachio butter, walnut butter, sunflower butter, hazelnut butter, tahini, cashew butter, coconut butter, or a combination of any thereof). The mixture of the invention may comprise 0-10% (w:w) fat. In some embodiments, the mixture of the invention may comprise 0-2.5%, 2.5-5%, 5-7.5%, or 7.5-10% fat.

[0028] In some embodiments, the mixture may comprise salt. The salt may be sodium chloride, dicalcium carbonate, dicalcium phosphate, tricalcium phosphate, calcium carbonate and any combination thereof.

[0029] In some embodiments, the mixture may comprise flavor or flavor modifiers. Flavor, also referred to as flavorings, include for example vanilla, coconut, chocolate, cinnamon, almond, and citrus (such as lemon, lime, or orange) flavorings. In some embodiments, the flavorings are artificial. In other embodiments, the flavorings are plant-derived. A flavor modifier, also referred to as a taste modifier, alters sweetness quality (increases or reduces), saltiness (magnifies or provides umami flavors), enhances specific flavors (such as fruit or spice), or masks undesirable flavor notes.

[0030] In some embodiments, the mixture of the invention may have been standardized and:or homogenized before and:or after enzymatic treatment. In some embodiments, the mixture may have been pasteurized or otherwise heat-treated before and: or after enzymatic treatment.

[0031] Enzymes

[0032] In the methods of the invention, the mixture comprising the whole oat flour and vegetable protein isolate and:or vegetable protein concentrate is treated with a beta-glucanase. The term “beta-glucanase” encompasses polypeptides having exo- and:or -endo beta-1 , 3-glucanase activity, beta-1 , 6-glucanase activity, and:or beta-1 ,4 glucanase activity, and may possess other enzymatic activities as well. The term beta-glucanase further includes whatever auxiliary compounds may be necessary for the enzyme's catalytic activity, such as, e.g., an appropriate acceptor or cofactor, which may or may not be naturally present in the reaction system. An enzyme having exo- and:or -endo beta-1 , 3-glucanase activity, beta-1 , 6-glucanase activity, and:or beta-1 ,4 glucanase activity may be a member of a glycoside hydrolase family selected from GH5 (such as for example GH5_5 and GH5_15), GH7, GH16, or GH64.

[0033] In some embodiments, the beta-glucanase is derived from a fungus. In other embodiments, the beta-glucanase is derived from a bacterium. The endopeptidase may be extracellular. It may have a signal sequence at its N-terminus, which is cleaved off during secretion.

[0034] The beta-glucanase and optionally any additional enzyme may be derived from any of the sources mentioned herein. The term “derived” means in this context that the enzyme may have been isolated from an organism where it is present natively, i.e. the amino acid sequence of the endopeptidase is identical to a native polypeptide. The term “derived” also means that the enzyme may have been produced recombinantly in a host organism, the recombinantly produced enzyme having either an amino acid sequence which is identical to a native enzyme or having a modified amino acid sequence, e.g. having one or more amino acids which are deleted, inserted and:or substituted, i.e. a recombinantly produced enzyme which is a mutant of a native amino acid sequence. Within the meaning of a native enzyme are included natural variants. Furthermore, the term “derived” includes enzymes produced synthetically by, e.g., peptide synthesis. The term “derived” also encompasses enzymes which have been modified e.g. by glycosylation, phosphorylation etc., whether in vivo or in vitro. With respect to recombinantly produced enzymes the term “derived from” refers to the identity of the enzyme and not the identity of the host organism in which it is produced recombinantly.

[0035] The beta-glucanase and optionally any additional enzyme may be obtained from a microorganism by use of any suitable technique. For instance, an enzyme preparation may be obtained by fermentation of a suitable microorganism and subsequent isolation of a beta glucanase preparation from the resulting fermented broth or microorganism by methods known in the art. The beta-glucanase may also be obtained by use of recombinant DNA techniques. Such method normally comprises cultivation of a host cell transformed with a recombinant DNA vector comprising a DNA sequence encoding the beta-glucanase and the DNA sequence being operationally linked with an appropriate expression signal such that it is capable of expressing the enzyme in a culture medium under conditions permitting the expression of the enzyme and recovering the enzyme from the culture. The DNA sequence may also be incorporated into the genome of the host cell. The DNA sequence may be of genomic, cDNA or synthetic origin or any combinations of these, and may be isolated or synthesized in accordance with methods known in the art.

[0036] In some embodiments, the enzyme having beta-glucanase activity may be in a cellulolytic enzyme preparation. In further embodiments, the enzyme preparation may be obtained from Trichoderma reesei. Examples of enzyme preparations comprising a beta-glucanase or beta- glucanase activity include Celluclast®, Viscozyme® L, Ultraflo® Prime, or Ultraflo® Key each available from Novozymes A:S; Cellulase 13L (Biocatalysts Limit, Wales, UK), Depol® 793L (Biocatalysts Limit, Wales, UK), Depol® 385L (Biocatalysts Limit, Wales, UK), Validase® TRL (dsm- firmenich), Validase® BG (dsm-firmenich), and Crystalzyme® PML-MX (Valley Research).

[0037] SEQ ID NOs: 1-4 are beta-glucanases and are each members of EC 3.2.1.4. SEQ ID NOs: 1, 3, and 4 are GH5 family members. SEQ ID NO: 2 is a GH7 family member. In some embodiments, the beta-glucanase of the invention has an amino acid sequence which is at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NOs: 1 , 2, 3, or 4. In some embodiments, the beta-glucanase of the present invention has the amino acid sequence of SEQ I D NOs: 1 , 2, 3, or 4. The term “identity” is the relatedness between two amino acid sequences or between two nucleotide sequences. For purposes of the present invention, the degree of identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends in Genetics 16: 276-277), preferably version 3.0.0 or later. The optional parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle labelled “longest identity” (obtained using the -nobrief option) is used as the percent identity and is calculated as follows: (Identical Residues x 100):(Length of Alignment - Total Number of Gaps in Alignment)

[0038] The amino acid changes may be of a minor nature, that is conservative amino acid substitutions or insertions that do not significantly affect the folding and:or activity of the protein; small deletions, typically of 1-30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue; a small linker peptide of up to 20-25 residues; or a small extension that facilitates purification by changing net charge or another function, such as a polyhistidine tract, an antigenic epitope, or a binding domain.

[0039] Examples of conservative substitutions are within the groups of basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine and valine), aromatic amino acids (phenylalanine, tryptophan and tyrosine), and small amino acids (glycine, alanine, serine, threonine and methionine). Amino acid substitutions that do not generally alter specific activity are known in the art and are described, for example, by H. Neurath and R.L. Hill, 1979, In, The Proteins, Academic Press, New York. Common substitutions are Ala:Ser, Vai: I le, Asp:Glu, ThrSer, Ala:Gly, Ala:Thr, SerAsn, Ala:Val, SerGly, TyrPhe, Ala:Pro, Lys:Arg, Asp:Asn, Leu:lle, Leu:Val, Ala:Glu, and Asp:Gly.

[0040] In some embodiments, the beta-glucanase has at least 70% sequence identity, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even 100% sequence identity to SEQ ID NO: 1.

[0041] In another embodiment, the beta-glucanase has at least 70% sequence identity, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even 100% sequence identity to SEQ ID NO: 2.

[0042] In another embodiment, the beta-glucanase has at least 70% sequence identity, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even 100% sequence identity to SEQ ID NO: 3. In another embodiment, the beta-glucanase has at least 70% sequence identity, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even 100% sequence identity to SEQ ID NO: 4.

[0043] In some embodiments of the invention, the mixture is treated with an enzyme composition comprising a beta-glucanase and at least one additional enzyme. The additional enzyme may be a pectolytic enzyme, pectin methyl esterase, endo-polygalacturonase, pectin lyase, cellulase, hemicellulase, carbohydrase, beta-glucosidase, arabanase, .arabinofuranosidase, lipase, cyclodextrin glucanotransferase, endopeptidase, glucanase, pentosanase, protease, ligninmodifying enzyme, laccase, arabinoxylanase, ferulic acid esterase, xylanase, protein deamidase, transglutaminase, or any combination thereof.

[0044] In some embodiments, the additional enzyme is a pectolytic enzyme, pectin methyl esterase, endo-polygalacturonase, and:or pectin lyase.

[0045] In some embodiments, the additional enzyme is a hemicellulolytic enzyme, or hemicellulase. The term "hemicellulolytic enzyme" or "hemicellulase" means one or more (e.g., several) enzymes that hydrolyze a hemicellulosic material. See, for example, Shallom and Shoham, 2003, Microbial hemicellulases, Current Opinion In Microbiology 6(3): 219-228. Hemicellulases are key components in the degradation of plant biomass. Examples of hemicellulases include, but are not limited to, an acetylmannan esterase, an acetylxylan esterase, an arabinanase, an arabinofuranosidase, a coumaric acid esterase, a feruloyl esterase, a galactosidase, a glucuronidase, a glucuronoyl esterase, a mannanase, a mannosidase, a xylanase, and a xylosidase. The substrates of these enzymes, the hemicelluloses, are a heterogeneous group of branched and linear polysaccharides that are bound via hydrogen bonds to the cellulose microfibrils in the plant cell wall, crosslinking them into a robust network. Hemicelluloses are also covalently attached to lignin, forming together with cellulose a highly complex structure. The variable structure and organization of hemicelluloses require the concerted action of many enzymes for its complete degradation. The catalytic modules of hemicellulases are either glycoside hydrolases (GHs) that hydrolyze glycosidic bonds, or carbohydrate esterases (CEs), which hydrolyze ester linkages of acetate or ferulic acid side groups. These catalytic modules, based on homology of their primary sequence, can be assigned into GH and CE families marked by numbers. Some families, with overall similar fold, can be further grouped into clans, marked alphabetically (e.g., GH-A). An informative and updated classification of these and other carbohydrate active enzymes is available on the Carbohydrate-Active Enzymes (CAZy) database. Hemicellulolytic enzyme activities can be measured according to Ghose and Bisaria, 1987, Pure & Appl. Chern. 59: 1739-1752, at a suitable temperature, e.g., 50°C, 55°C, or 60°C.

[0046] In one embodiment, the hemicellulase is provided as a commercial hemicellulolytic enzyme preparation. Examples of commercial hemicellulolytic enzyme preparations suitable for use in the present invention include, for example, SHEARZYME™ (Novozymes A:S), CELLIC® HTec (Novozymes A:S), CELLIC® HTec2 (Novozymes A:S), VISCOZYME® L(Novozymes A:S), ULTRAFLO® (Novozymes A:S), PULPZYME(R) HC (Novozymes A:S), ACCELLERASE® XY (Genencor), ACCELLERASE® XC (Genencor), ECOPULP® TX-200A (AB Enzymes), DEPOL™ 333P (Biocatalysts Limit, Wales, UK), DEPOL® 740L. (Biocatalysts Limit, Wales, UK), and DEPOL® 762P (Biocatalysts Limit, Wales, UK).

[0047] In some embodiments, the additional enzyme is a protease. The protease may be from Bacillus, e.g., B. amyloliquefaciens. A suitable protease may be Neutrase®, Formea®, or Vertera ® Smooth, each available from Novozymes A:S.

[0048] In some embodiments, the additional enzyme is a cyclodextrin glucanotransferase (CGTase). A suitable CGTase may be the cyclodextrin glucanotransferase “Amano” (manufactured by Amano Enzyme Inc.) or Toruzyme® (Novozymes A:S).

[0049] Enzyme Treatment

[0050] The enzymes used in the methods of the invention may be added to the mixture comprising the plant-derived substrate in any suitable form, such as in the form of a liquid, in particular a stabilized liquid, or it may be added as a substantially dry powder or granulate. Granulates may be produced, e.g., as disclosed in US Patent No. 4,106,991 and US Patent No. 4,661 ,452. Liquid enzyme preparations may, for instance, be stabilized by adding a sugar or sugar alcohol or lactic acid according to established procedures. Other enzyme stabilizers are well-known in the art.

[0051] A beta-glucanase and optionally additional enzymes to be used in the process of the invention may be added at a concentration of 0.1-1000 mg enzyme protein per kg whole oat flour, 0.5-500 mg enzyme protein per kg whole oat flour, or 1-100 mg enzyme protein per kg whole oat flour. The dosage will depend on parameters such as the temperature, the incubation time and the dairy alternative recipe. The skilled person will know how to determine the optimal enzyme dosage.

[0052] In some embodiments, the beta-glucanase and optionally an additional enzyme(s) is provided at 0.05, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or 1.0% of plant-derived substrate (w:w). In further embodiments, the plant derived substrate may be or may comprise oat flour, whole oat flour, vegetable protein, vegetable protein isolate, or vegetable protein concentrate. In some embodiments, the beta-glucanase is provided at 0.01 %-5% of plant-derived substrate. In some embodiments, the beta-glucanase is provided at 0.01-0.05%, 0.05-0.1%, 0.1-0.3%, 0.3-0.7%, 0.7-1.2%, 1.2-2.0%, or 2.0-3.0% of plant substrate.

[0053] The enzymes may be added to the mixture comprising the plant-derived substrate in any suitable manner, such as individual components (separate or sequential addition of the enzymes) or addition of the enzymes together in one step or one composition, or any combination thereof. The enzymes are added to the mixture and the mixture is held at 20-65°C to allow for the enzymes to act on the plant-derived substrate.

[0054] In some embodiments, the mixture is held at a temperature between 20-40°C, 30-45°C, 35-50°C, 40-55°C, 50°C-60°C, or 50°C-65C. In further embodiments, the mixture is held at a temperature between 40-45°C, 45-50°C, 47-53°C, 50-55°C, 55-60°C, or 60-65°C. In some embodiments, the mixture is held at a temperature of about 20 °C, about 25°C, about 30°C, about 35°C, about 40°C, about 45°C, about 50°C, about 55°C, about 60°C, or about 65°C.

[0055] In some embodiments, the mixture with the added enzymes is held at 20-65°C for at least 10 minutes to allow for enzymatic treatment of the plant-derived substrate. In some embodiments, the mixture is held for at least 30, 40, 45, 50, 55, 60, 65, 70, or 75 minutes. In further embodiments, the mixture is held for about 10, about 15, about 20, about 25, about 30, about 60, about 120, about 180, or about 240 minutes to allow for enzymatic treatment of the plant-derived substrate. In some embodiments, the mixture is held for at least about 10, 30, 60, 90, or 120 minutes. In further embodiments, the mixture is held for 60, 90, or 120 minutes. In still further embodiments, the mixture is held at about 50°C for about 60, 90, or 120 minutes.

[0056] The process used, including temperature ranges, pH and the length of enzymatic treatment, will vary depending on the plant-derived substrate and the enzymes added to the slurry. The skilled person will know how to determine the best process parameters based, e.g., on the plant-derived substrate and enzymes used. Additionally, the enzyme dosage will depend on parameters such as the temperature, the incubation time and the dairy alternative recipe. The skilled person will know how to determine the optimal enzyme dosage.

[0057] After treatment with the enzymes, the enzymes may be inactivated. The enzymes may be inactivated at any step after enzymatic treatment. In some embodiments, the enzymes are inactivated by a heat treatment. In some embodiments, the heat treatment is 75-95°C for 5-30 minutes. In further embodiments, the heat treatment is 85-95°C for 10 minutes. In some em- bodiments, the heat treatment is 95°C for 5, 10, 15, 20, 25, or 30 minutes.

[0058] In some embodiments, the enzymes are inactivated by an Ultra High Temperature (UHT) treatment. The UHT treatment may be direct or indirect. In some embodiments, the UHT treatment is 135-154°C for 1-10 seconds. in further embodiments, the UHT treatment is 140-150°C for 3, 4, 5, 6, 7, 8, 9, or 10 seconds. In further embodiments, the UHT treatment is 140-145°C for 3, 4, 5, 6, 7, 8, 9, or 10 seconds. In some embodiments, the UHT treatment is 143°C for 4,

[0059] 5, 6, 7, or 8 seconds.

[0060] After enzyme inactivation, the treated mixture may be cooled. In some embodiments, the mixture is cooled to a temperature between 20-40°C.

[0061] In some embodiments, the oat-derived base of the invention is further processed to produce an oat-based dairy-alternative food product. The oat-based acidified dairy alternative food product of the invention is an oat-based food product which is produced by acidification and which is an oat-based alternative to an acidified dairy product produced by acidification of a milk substrate based on milk obtained from a mammal. In some embodiments, the oat-based acidified dairy alternative food product is a yogurt alternative, a set-type yogurt alternative, a stirred yogurt alternative, a strained yogurt alternative, a sour cream alternative, a greek-style yogurt alternative, a skyr alternative or a cream cheese alternative.

[0062] The oat-derived base of the invention has a reduced viscosity compared to an oat-derived base prepared by the same process but without addition of a beta-glucanase. The reduction in viscosity is related to the reduction of high molecular weight beta-glucan in the plant-derived or oat-derived substrate. Reduced viscosity may be critical for processing the oat-derived base into oat-based dairy-alternative food products. In some embodiments, protein may be added to the oat-derived base of the invention to produce a food product that has a higher amount of protein. It is known in the art that the addition of protein to a base for making a dairy-alterative food product can increase the viscosity and gelling capacity of the base. Therefore, a base with lower viscosity enables the addition of protein without negatively impacting the texture of the final food product.

[0063] The oat-derived base of the invention may be incubated with an acidifer to produce an oat-based acidified dairy alternative food product. Acidification may be carried out chemically or via fermentation. Chemical acidifiers include glucono delta-lactone (GDL) and food grade acids, such as for example vinegar, citric acid, tartaric acid, malic acid, fumaric acid, and lactic acid. Alternatively, acidification may also be carried out by incubation of the oat-derived base of the invention with a lactic acid bacterium. Lactic acid bacteria include bacteria of the genus Streptococcus, Lactococcus, Lactobacillus, Leuconostoc, Pseudoleuconostoc, Pediococcus, Propionibacterium, Enterococcus, Brevibacterium, Bifidobacterium or any combination thereof.

[0064] A stirred yogurt alternative may be produced by carrying out fermentation in fermentation tanks where the formed acid gel is disrupted e.g. by agitation after fermentation when the desired pH has been obtained. The stirred product may be partially cooled to 20-30°C and flavoring ingredients may be added. The stirred product is pumped to filling line and filled in retail containers. The stirred yogurt alternative may then be cooled and then stored.

[0065] A set yogurt alternative may be fermented in a retail container and not agitated after fermentation. After fermentation, a set yogurt alternative may be cooled and then stored. The cooling may be carried out in blast chiller tunnel or in a refrigerated storage room.

[0066] A strained yogurt alternative, such as a Greek yogurt alternative or a labneh alternative, is a yogurt alternative that has been strained to remove part of its aqueous phase, thus resulting in a thicker consistency than an unstrained yogurt alternative, while preserving yogurt's distinctive sour taste. The term "after fermentation" as used herein means when fermentation is ended and the desired pH obtained. In some embodiments, the pH after fermentation may be between 3.5 and 5.5. In further embodiments, the pH after fermentation may be between 4 and 5. A pasteurization step may be performed after fermentation. Pasteurization may be performed, e.g., at 80- 95°C for 1-30 minutes, such as at 80-85°C for 30 minutes or at 90-95°C for 2-15 minutes.

[0067] The methods of the invention produce an oat-derived base which may be further processed into an oat-based acidified dairy alternative food product. Unexpectedly, the oat-derived base of the invention produces an oat-based acidified dairy alternative food product with desirable levels of viscosity, cohesiveness, and stability, so that texturizers, thickeners, and:or stabilizers do not need to be added. Texturizers, thickeners, and stabilizers include carrageenan, pectin, sodium alginate, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, agar, gelatin, seaweed powder, xanthan gum, starch (such as, for example tapioca starch, corn starch and potato starch), modified starch, and gelling agents.

[0068] In a preferred embodiment, the oat-based acidified dairy alternative food product has a smoother texture or a less brittle texture compared to an oat-based acidified dairy alternative food product prepared by the same process but without addition of an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme. The texture may be visually evaluated after overnight storage at 4°C by placing a sample of the oat-based acidified dairy alternative food product on a black plastic spoon.

[0069] In a preferred embodiment, the oat-based acidified dairy alternative food product has a less grainy texture compared to a oat-based acidified dairy alternative food product prepared by the same process but without addition of an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme. The texture may be visually evaluated after overnight storage at 4°C by placing a sample of the plant-based acidified dairy alternative on a black plastic spoon.

[0070] In a preferred embodiment, the oat-based acidified dairy alternative food product has a thicker texture compared to a oat-based acidified dairy alternative food product prepared by the same process but without addition of an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme. The texture may be visually evaluated after overnight storage at 4°C by placing a sample of the plant-based acidified dairy alternative on a black plastic spoon. In particular, the thickness of the texture may be evaluated by how well the product sticks to the spoon when it is scooped with the spoon and the spoon is subsequently flipped.

[0071] It is known in the art that many plant-based acidified dairy alternative food products, such as some kinds of plant-based dairy alternative yogurt, require an additional processing step of going through a smoothing pump after fermentation. This smoothing pump improves the texture of the yogurt to make it smoother. The methods of the invention produce an oat-derived base which can be processed into an oat-based dairy alternative yogurt with a superior texture, such that a smoothing step is not needed.

[0072] PREFERRED EMBODIMENTS

[0073] 1 . A method for making an oat-derived base, comprising: a) obtaining a mixture comprising plant-derived substrate comprising whole oat flour and vegetable protein isolate and:or vegetable protein concentrate; and b) treating the mixture of step a) with an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme; wherein the enzymatic activity on the mixture of step a) produces the oat-derived base.

[0074] 2. The method of embodiment 1 , wherein the additional enzyme is a pectolytic enzyme, pectin methyl esterase, endo-polygalacturonase, pectin lyase, cellulase, hemicellulase, carbohydrase, arabanase, arabinofuranosidase, lipase, cyclodextrin glucanotransferase, endopeptidase, glu- canase, protease, lignin-modifying enzyme, laccase, arabinoxylanase, ferulic acid esterase, xy- lanase, protein deamidase, transglutaminase, or any combination thereof.

[0075] 3. A method for making an oat-derived base, comprising: a) obtaining a mixture comprising a plant-derived substrate comprising whole oat flour and vegetable protein isolate and:or vegetable protein concentrate; and b) treating the mixture of step a) with a beta-glucanase; wherein the beta -glucanase activity on the mixture of step a) produces the oat-derived base.

[0076] 4. The method of any one of the preceding embodiments, wherein the vegetable protein isolate or vegetable protein concentrate is derived from a legume or a nut.

[0077] 5. The method of any one of the preceding embodiments, wherein the vegetable protein isolate or vegetable protein concentrate is derived from peas, yellow peas, lentils, chickpeas, soybeans, beans, peanuts, fava beans, lima beans, mung beans, lupins, almonds, cashews, macadamia nuts, pistachios, pecans, walnuts, pili nuts, hazel nuts, sesame seeds, or any combination thereof.

[0078] 6. The method of any one of the preceding embodiments, wherein the vegetable protein isolate or vegetable protein concentrate is derived from pea, soybean, fava bean, or any combination thereof. 7. The method of any one of the preceding embodiments, wherein the plant-derived substrate comprises a ratio of whole oat flour to vegetable protein isolate from 20:80 to 80:20 by dry weight.

[0079] 8. The method of any one of the preceding embodiments, wherein the plant-derived substrate comprises a ratio of whole oat flour to vegetable protein isolate from 30:70 to 30:70 by dry weight.

[0080] 9. The method of any one of the preceding embodiments, wherein the plant-derived substrate comprises a 30:70 or a 50:50 ratio of whole oat flour to vegetable protein isolate by dry weight.

[0081] 10. The method of any one of the preceding embodiments, wherein the vegetable protein isolate is derived from pea.

[0082] 11. A method for making an oat-derived base, comprising: a) obtaining a mixture comprising plant-derived substrate comprising whole oat flour and pea protein isolate, wherein the ratio of whole oat flour to pea protein isolate is from 30:70 to 70:30 by dry weight; and b) treating the mixture of step a) with a beta-glucanase; wherein the beta-glucanase activity on the mixture of step a) produces the oat-derived base.

[0083] 12. The method of any one of the preceding embodiments, wherein the treatment with en- zyme(s) is a temperature between 20-65°C for at least 15 minutes.

[0084] 13. The method of any one of the preceding embodiments, where the treatment with enzyme(s) is a temperature between 40-45°C, 45-50°C, 47-53°C, 50-55°C, 55-60°C, or 60-65°C for at least 30, 45, 60, 75, 90, 105, or at least 120 minutes.

[0085] 14. The method of any one of the preceding embodiments, wherein the treatment with en- zyme(s) is about 50°C for at least 30, 45, 60, 75, 90, 105, or at least 120 minutes.

[0086] 15. The method of any one of the preceding embodiments, wherein the beta-glucanase comprises an amino acid sequence which is at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or is 100% identical to the amino acid sequence of SEQ I D NOs: 1 , 2, 3, or 4. 16. The method of any one of the preceding embodiments, wherein the mixture further comprises water, salt, sugar, fat, oil, flavor, flavor modifiers, or any combination thereof.

[0087] 17. The method of any one of the preceding embodiments, wherein the mixture further comprises rapeseed oil, flaxseed oil, safflower oil, flaxseed oil, soybean oil, olive oil, sunflower oil, palm oil, coconut oil, canola oil, linseed oil, corn oil, peanut oil, vegetable oil, or any combination thereof.

[0088] 18. The method of any one of the preceding embodiments, wherein the mixture further comprises dextrose, sucrose, fructose, honey, an artificial sweetener, a natural sweetener, or any combination thereof.

[0089] 19. The method of any one of the preceding embodiments, wherein the mixture further comprises natural flavoring, artificial flavoring, flavor modifiers, or any combination thereof.

[0090] 20. A method of producing an oat-based acidified dairy alternative food product, comprising the method of any one of the preceding embodiments and further the step of: c) acidification of the oat-derived base by incubation with a lactic acid bacterium or with a chemical acidifier, such as glucono delta-lactone (GDL), lactic acid, vinegar, citric acid, tartaric acid, malic acid, fumaric acid, or any combination thereof; and d) processing to produce an oat-based acidified dairy alternative food product.

[0091] 21. The method of embodiment 20, wherein the oat-based acidified dairy alternative food product is an oat-based fermented dairy alternative food product, and wherein the oat-based substrate is incubated with a lactic acid bacterium, preferably of the genus Streptococcus, Lactococcus, Lactobacillus, Leuconostoc, Pseudoleuconostoc, Pediococcus, Propionibacterium, Enterococcus, Brevibacterium, Bifidobacterium or any combination thereof.

[0092] 22. The method of embodiment 20 or 21 , wherein the oat-based acidified dairy-alternative food product is a yogurt alternative, a set-type yogurt alternative, a stirred yogurt alternative, a strained yogurt alternative, a sour cream alternative, a cream cheese alternative, a greek-style yogurt alternative or a skyr alternative.

[0093] 23. The method of embodiments 20-22, wherein no additional texturizers and: or thickeners are added to produce the oat-based acidified dairy-alternative food product.

[0094] 24. The method of any one of the preceding embodiments, wherein the viscosity and:or gel strength of the oat-derived base is improved compared to an oat-derived base prepared by the same process but without addition of an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme.

[0095] 25. An oat-derived base produced by the method of any one of embodiments 1-19 or 24.

[0096] 26. An oat-based acidified dairy-alternative food product produced by the method of any one of embodiments 1-24.

[0097] 27. The oat-based acidified dairy-alternative food product of embodiment 26, wherein no additional texturizers and:or thickeners are added to produce the oat-based acidified dairy-alternative food product.

[0098] 28. A composition for the production of an oat-derived base for an oat-based acidified dairyalternative, comprising whole oat flour, vegetable protein isolate and:or vegetable protein concentrate, and an enzyme composition comprising a beta-glucanase and optionally at least one additional enzyme.

[0099] 29. The composition of embodiment 28, wherein the enzyme composition comprises a beta- glucanase and no additional enzymes.

[0100] 30. The composition of embodiment 28 or 29, wherein the plant-derived substrate comprises a ratio of whole oat flour to vegetable protein isolate from 20:80 to 80:20 by dry weight.

[0101] 31. The composition of any one of embodiments 28-30, wherein the vegetable protein isolate is derived from peas, yellow peas, lentils, chickpeas, soybeans, beans, peanuts, fava beans, lima beans, mung beans, lupins, almonds, cashews, macadamia nuts, pistachios, pecans, walnuts, pili nuts, hazel nuts, sesame seeds, or any combination thereof.

[0102] 32. The composition of any one of embodiments 28-31 , wherein the composition further comprises water, salt, sugar, fat, oil, flavor, flavor modifiers, or any combination thereof.

[0103] 33. The composition of any one of embodiments 28-32, wherein the composition further comprises natural flavoring, artificial flavoring, flavor modifiers, or any combination thereof.

[0104] 34. The composition of any one of embodiments 28-33, wherein the additional enzyme is a pectolytic enzyme, pectin methyl esterase, endo-polygalacturonase, pectin lyase, cellulase, hemicellulase, carbohydrase, arabanase, arabinofuranosidase, lipase, cyclodextrin glucanotransferase, endopeptidase, glucanase, protease, lignin-modifying enzyme, laccase, arabinoxylanase, ferulic acid esterase, xylanase, protein deamidase, transglutaminase, or any combination thereof.

[0105] 35. The composition of any one of embodiments 28-34, wherein the enzyme composition comprises a beta-glucanase and no additional enzymes.

[0106] 36. The composition of any one of embodiments 28-35, wherein the beta-glucanase comprises an amino acid sequence which is at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or is 100% identical to the amino acid sequence of SEQ ID NOs: 1 , 2, 3, or 4.

[0107] 37. Use of a beta-glucanase and optionally at least one additional enzyme in the production of an oat-derived base.

[0108] 38. Use of a beta-glucanase and optionally at least one additional enzyme in the production of an oat-derived base to produce an oat-based acidified dairy alternative food product to improve viscosity, gel strength, texture and:or stability of the oat-based acidified dairy alternative food product.

[0109] EXAMPLES

[0110] Assays for determining enzymatic activity:

[0111] Beta-glucanase activity (FBG): One fungal beta glucanase unit (FBG) is the amount of enzyme, which, according to the standard conditions outlined below, releases reducible oligosaccharides or reduces carbohydrate with a reduction capacity equivalent to 1 mol glucose per minute. Fungal beta glucanase reacts with beta-glucan to glucose or reducing carbohydrate that is determined as reducing sugar according to the Somogyi Nelson method. The standard reaction conditions were: 0.5% barley beta-glucan substrate, 30°C, pH 5.0, and reaction time of 30 min.

[0112] Endopeptidase activity (KPRU): Trypsin-like and lysine-specific endopeptidases hydrolyse the chromophoric substrates Ac-Arg-p-nitro-anilide (Ac-Arg-pNA) and: or Ac-Lys-p-nitro-anilide (Ac- Arg-pNA). The liberated pNA produces an absorption increase at 405 nm, which is proportional to enzyme activity. One KPRU is equivalent to the amount of enzyme that produces 1 micromole p-nitroaniline per minute, when Ac-Arg-pNA or Ac-Lys-pNA is incubated with the enzyme at pH 8.0 at 37°C. The activity may be determined relative to a standard of declared strength. Example 1 : Vegetable protein isolate and whole oat flour ratios to produce an oat-based acidified dairy alternative food product

[0113] Mixtures comprising different ratios of pea protein isolate (PPI; PLIRIS P870 pea protein isolate, Puris Proteins, Minneapolis, Minnesota) and whole oat flour (colloidal whole oat flour; Grain Millers, Inc, Eden Prairie, Minnesota) were prepared with a final solid content of 16% by dry weight. Ratios of pea protein isolate to whole oat flour were 10:90; 30:70; 50:50; 70:30; and 90:10. Each mixture was treated with an enzyme composition comprising the beta-glucanase of SEQ ID NO: 1 (108.7 FBG:g) at 0.2% by weight of whole oat flour and then incubated at 50°C for 60 minutes. Following incubation, the enzyme composition was inactivated by a heat treatment of 90°C for 10 minutes and the mixture was cooled to room temperature.

[0114] To process the base, a trypsin-like endo-protease (13.00 KPRU:g; Novozymes A:S, Denmark) was added at 0.4% by weight of whole oat flour-PPI total protein content. Glucono delta lactone (Roquette America Inc., Geneva, Illinois) was also added by weight of whole oat flour-PPI total protein content. The target for acidification was a pH of 4.0-4.6. Table 1 depicts the components of the samples.

[0115] Table 1 : Sample composition

[0116] The samples were refrigerated overnight with no stirring. Following acidification, the samples were analyzed for texture, stirred with a spoon, and then analyzed for viscosity. Texture, also referred to as gel strength, was measured by a TA-XT2 Texture Analyzer (Stable Micro Systems, Surrey, UK). The gel strength was measured by the force required to attain a certain deformation and is considered as a measure of hardness of the yogurt. A 50 kg load cell was used for the gel strength analysis. Results are shown in Table 2. Samples comprising 100% PPI and samples of market available dairy-based skyr yogurt and stirred yogurt were included for comparison.

[0117] Table 2: Texture Analyzer results As shown in Table 2, the 30:70 (PPkwhole oat flour) sample has a gel strength comparable to the marketed dairy stirred yogurt as determined by the Texture Analyzer. The 70:30 sample has a gel strength comparable to the market dairy skyr yogurt.

[0118] Viscosity was measured by a DV2T viscometer (AMETEK Brookfield Engineering, Middle- boro Massachusetts, USA) in centipoise (cP). Samples comprising 100% PPI and 20% whole oat flour, with no PPI, were included for comparison.

[0119] Table 3: Viscosity

[0120] The textures of samples depended on the composition of the plant substrate in the initial mixture, with the viscosity of the sample higher when the mixture comprised more PPI. The 10:90 sample (PPI: whole oat flour) had decreased set properties, with the sample being fluid with no set structure. The 30:70 sample (PPkwhole oat flour) yielded a soft set more similar to the set of stirred yogurt, and had a homogenous texture. The 50:50 sample has a slightly lower set compared to the 70:30 sample, yet had a thick and homogenous texture. The 70:30 sample (PPkwhole oat flour) resulted in a product with homogenous texture and Greek yogurt like set without brittleness, and upon scooping remained intact on the spoon. Additionally, the 70:30 sample stayed on the spoon when scooped in spoon and flipped upside down. The 90:10 sample (PPkwhole oat flour) had a viscosity similar to Greek-style yogurt, but was brittle and less smooth.

[0121] Example 2: Whole oat flour as substrate for Beta-glucanase to produce an oat-based acidified dairy alternative food product

[0122] A mixture comprising 70:30 PPkwhole oat flour was prepared according to Table 4.

[0123] Table 4: Ingredients for Mixture The mixture was divided into five samples into which different amounts of enzyme were added: no enzyme (control); 0.2% (based on the amount of oat flour) of an enzyme composition comprising the beta-glucanase of SEQ ID NO: 1 (108.7 FBG:g); 0.02% (based on the amount of oat flour) of the beta-glucanase of SEQ ID NO: 2 (1815.64 FBG EX:g); 0.1% (based on the amount of oat flour) of the beta-glucanase of SEQ ID NO: 2; and 0.2% (based on the amount of oat flour) of the beta-glucanase of SEQ ID NO: 2. Samples were incubated at 50°C for 60 minutes, then heat-treated at 90°C for 10 minutes to inactivate the enzymes. The samples were then cooled to 30°C. Viscosity was measured by a DV2T viscometer (AMETEK Brookfield Engineering, Mid- dleboro Massachusetts, USA) in centipoise (cP). Table 5 shows the viscosity measurements of the samples. Viscosity of the oat-derived base is shown in Table 5

[0124] Table 5: Viscosity of oat-derived base

[0125] Samples where a beta-glucanase was added to the mixture have much lower viscosity compared to samples to which no beta-glucanase was added. An oat-derived base with an initial process viscosity as shown in Table 5 can be further processed to produce an oat-based acidified dairy alternative food product with desirable organoleptic and techno-functional properties, including a smooth texture without sliminess.

[0126] Example 3: Degradation of beta-glucan in an oat-derived base improves set of oat-based acidified dairy alternative food product

[0127] Beta glucanase activity as driver for the smooth textured acidified yogurt alternative was evaluated indirectly by measuring the difference in high molecular weight beta glucan content in formulations with and without enzymatic treatment. High molecular weight beta glucan was estimated in samples using an enzymatic spectrophotometry method (AOAC 995.16) with a limit of quantification of 0.05% (w:w). All enzyme treatments were on an oat-derived base comprising a blend of pea protein isolate and whole oat flour in a ratio of 70:30 (PPkwhole oat flour) at a 16% total solids in water. Sample 1 was the control, with no enzyme treatment. Sample 2 was treated with an enzyme composition comprising the beta-glucanase of SEQ ID NO: 1 , where the beta-glucanase was provided at 0.2% of whole oat flour. Sample 3 was treated with an enzyme composition comprising the beta-glucanase of SEQ ID NO: 4 and a xylanase, where the beta-glucanase was provided at 0.1 % of whole oat flour content. Sample 4 was treated with the purified beta-glucanase of SEQ ID NO: 4, where the beta-glucanase was provided at 0.1 % of whole oat flour content. The enzyme treatment was incubated with the oat-derived base at 50°C for 60 minutes, followed by an enzyme deactivation step at 90°C for 10 minutes.

[0128] Table 6: Beta-glucan in an oat-derived base

[0129] The high molecular weight beta glucan in Sample 1 is responsible for the high viscosity in oat based fermented food formulations and doesn’t allow enough of the oat flour to be in the base to give the needed set property without denseness and or associated sliminess. Samples 2, 3, and 4 yielded a product with less than 0.05% high molecular weight beta glucan. Sample 4 indicates that it is the activity of the beta-glucanase that is responsible for the decrease in beta-glucan in the oat-derived base.

[0130] An acidified dairy alternative product was prepared using the oat-derived base from samples 1-4. Oat-derived base from samples 2, 3, and 4 produced a smooth, light yogurt like product, which also allows for additional ingredients like protein without negatively affecting the texture. Oat-derived base from Sample 1 produced a product with relatively high viscosity and high gel strength and was not desirable.

Claims

CLAIMS1 . A method for making an oat-derived base, comprising: a) obtaining a mixture comprising a plant-derived substrate comprising whole oat flour and vegetable protein isolate and:or vegetable protein concentrate; and b) treating the mixture of step a) with a beta-glucanase; wherein the beta -glucanase activity on the mixture of step a) produces the oat-derived base.

2. The method of claim 1 , wherein the vegetable protein isolate and:or vegetable protein concentrate is derived from pea, soybean, fava bean, or any combination thereof.

3. The method of any one of the preceding claims, wherein the plant-derived substrate comprises a ratio of whole oat flour to vegetable protein isolate of from 30:70 to 70:30 by dry weight.

4. The method of any one of the preceding claims, wherein the mixture is treated with at least one additional enzyme, wherein the additional enzyme is a pectolytic enzyme, pectin methyl esterase, endo-polygalacturonase, pectin lyase, cellulase, hemicellulase, carbohydrase, arabinase, arabinofuranosidase, lipase, cyclodextrin glucanotransferase, endopeptidase, glucanase, protease, lignin-modifying enzyme, laccase, arabinoxylanase, ferulic acid esterase, xylanase, protein deamidase, transglutaminase, or any combination thereof.

5. The method of any one of the preceding claims, wherein the treatment with enzymes is at a temperature between 40-45°C, 45-50°C, 47-53°C, 50-55°C, 55-60°C, or 60-65°C for at least 30, 45, 60, 75, 90, 105, or at least 120 minutes.

6. The method of any one of the preceding claims, wherein the beta-glucanase comprises an amino acid sequence which is at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical, or has 100% sequence identity to the amino acid sequence of SEQ I D NOs: 1 , 2, 3, or 4.

7. A method of producing an oat-based acidified dairy alternative food product, comprising the method of any one of the preceding claims and further the step of:c) acidification of the oat-derived base by incubation with a lactic acid bacterium or with a chemical acidifier, such as glucono delta-lactone (GDL), lactic acid, vinegar, citric acid, tartaric acid, malic acid, fumaric acid, or any combination thereof; and d) processing to produce an oat-based acidified dairy alternative food product.

8. The method of claim 7, wherein the oat-based acidified dairy alternative food product is an oat-based fermented dairy alternative food product, and wherein the oat-based substrate is incubated with a lactic acid bacterium, preferably of the genus Streptococcus, Lactococcus, Lactobacillus, Leuconostoc, Pseudoleuconostoc, Pediococcus, Propionibacterium, Enterococcus, Brevi- bacterium, Bifidobacterium or any combination thereof.

9. The method of claim 7 or 8, wherein the oat-based acidified dairy-alternative food product is a yogurt alternative, a set-type yogurt alternative, a stirred yogurt alternative, a strained yogurt alternative, a sour cream alternative, a cream cheese alternative, a greek-style yogurt alternative or a skyr alternative.

10. The method of any one of the preceding claims, wherein the viscosity and:or gel strength of the oat-derived base is improved compared to an oat-derived base prepared by the same process but without the addition of a beta-glucanase.

11. An oat-derived base produced by the method of any one of claims 1-7 or 10.

12. An oat-based acidified dairy-alternative food product produced by the method of any one of claims 7-10.

13. A composition for the production of an oat-derived base for an oat-based acidified dairyalternative, comprising whole oat flour, vegetable protein isolate and:or vegetable protein concentrate, and an enzyme composition comprising a beta-glucanase.

14. Use of a beta-glucanase in the production of an oat-derived base.

15. Use of a beta-glucanase in the production of an oat-derived base to produce an oat-based acidified dairy alternative food product to improve viscosity, gel strength, texture and:or stability of the oat-based acidified dairy alternative food product.

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