Method for enhancing flavor and / or texture of plant-based products

Abstract

The present invention relates to methods, uses and processes for enhancing the flavor and / or texture of a plant-based product, such as a food or beverage. The present invention relates to the use of one or more alpha galactosidase enzymes for enhancing the flavor and / or texture of plant-based products. The one or more alpha galactosidase enzymes may be added to the plant-based product or in a process of making the plant-based product.

Description

[0001] References to sequence listings

[0002] This application contains a sequence listing in computer readable form. This computer readable form is incorporated herein by reference. Technical Field

[0003] The present invention relates to a method for improving the flavor and / or texture of a plant-based product. In particular, the present invention relates to a method for improving the flavor and / or texture of a plant-based product, such as food or beverage, using α-galactosidase. Background Art

[0004] Recently, consumers' interest in healthy foods has been increasing, making natural foods and functional foods popular. Among them, plant-based products made from legume pods contain high-quality protein and plant linoleic acid, and also contain a large number of nutrients such as vitamin E and lecithin.

[0005] For example, the amount of soy milk consumed in the form of functional food is increasing year by year. A lot of research has been done on plant-based products such as soy milk and methods for producing soy milk. However, soy milk is still not widely accepted as a beverage due to the soybean smell, texture, mouthfeel, raw taste, astringency, etc. that are unique to soybeans. Food additives (such as whey protein, palatinose or phosphate sugars) can be added to enhance the flavor of soy milk.

[0006] Inactivating lipoxygenase by heating the aqueous medium in which the seeds are crushed and / or changing its pH is another approach to improving the flavor of soy milk, however these treatments often render the soy protein insoluble, reducing soy milk yield and giving it a floury mouthfeel.

[0007] The existing methods for preparing soymilk without beany taste still need to be improved. Therefore, there is a need to prepare legume-based products with improved properties, such as sweetness, reduced astringency, richness, fattiness, sweet aftertaste, mouthfeel and texture. Summary of the invention

[0008] The present invention relates to the use of alpha-galactosidase to enhance the flavor and / or texture of plant-based products, such as food or beverages.

[0009] In one aspect, the present invention relates to the use of one or more alpha galactosidases for enhancing the flavor and / or texture of a plant-based product, wherein the plant-based product is a food and / or a beverage.

[0010] In another aspect, the present invention relates to a method for improving the flavor and / or texture of a product, which is carried out by adding one or more α-galactosidases to a plant-based material and / or the plant-based product and incubating the plant-based material and / or the plant-based product, wherein the plant-based product is a food and / or a beverage.

[0011] In yet another aspect, the present invention relates to a process for preparing a plant-based product with enhanced flavor and / or texture, wherein the plant-based product is a food and / or a beverage. The process comprises soaking one or more plant materials and grinding the one or more plant materials into a slurry. One or more alpha galactosidases are added to the slurry. Thereafter, the fibers are separated from the slurry to obtain a plant material milk base. The one or more alpha galactosidases are added to the plant material milk base to obtain the plant-based product.

[0012] Sequence Listing Overview

[0013] SEQ ID NO: 1 is the mature amino acid sequence of alpha-galactosidase derived from Aspergillus niger.

[0014] definition

[0015] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning consistent with their meaning in the context of the specification and the relevant art, and should not be interpreted in an idealized or overly formal sense unless explicitly defined herein. For the sake of brevity and / or clarity, well-known functions or structures may not be described in detail.

[0016] As used herein, the term α-galactosidase means a glycoside hydrolase that catalyzes the hydrolysis reaction of terminal non-reducing α-D-galactose residues in α-D-galactosides (including galacto-oligosaccharides, galactomannans and galactolipids). For the purposes of the present invention, α-galactosidase is an enzyme that breaks down carbohydrates in plant materials (such as grains, legumes and cruciferous vegetables) into simpler sugars while preparing plant-based products.

[0017] As used herein, the term bean texture means having a grainy feel with a nutty, rich aftertaste. For the purposes of the present invention, bean texture is the grainy, nutty, rich aftertaste of a plant-based product such as soy milk.

[0018] As used herein, the term legume texture means having a grainy feel with a nutty, rich aftertaste. For the purposes of the present invention, legume texture is the grainy, nutty, rich aftertaste of a plant-based product such as soy milk.

[0019] As used herein, the term astringent refers to the quality of tasting or smelling dry, sour, and slightly bitter.

[0020] As used herein, the term richness means having a large proportion of rich or fatty ingredients, such as butter or cream. For the purposes of the present invention, richness means a plant-based product that is rich and creamy, such as rich and creamy soy milk.

[0021] As used herein, the term sweet aftertaste refers to the taste that remains after consuming a sweetened product.

[0022] As used herein, the term mouthfeel refers to the distinctive taste of a food or drink perceived in the mouth.

[0023] As used herein, the term "raffinose oligosaccharides" (RFO) are α-1,6-galactosyl extended structures of sucrose, such as raffinose, stachyose, and verbascose.

[0024] As used herein, the term plant-based material means materials and products derived in part or in whole from plants. For the purposes of the present invention, plant-based materials mean materials and products derived in part or in whole from leguminous plant pods (such as soybeans, peas, chickpeas, lentils, beans and / or any leguminous plant pods containing raffinose oligosaccharides). For example, the plant-based material can be a leguminous plant pod base or a leguminous plant pod slurry, or a leguminous plant pod paste.

[0025] As used herein, the term plant material means parts of a plant, such as stems, roots, seeds and leaves. For the purposes of the present invention, plant material means plant seeds, preferably plant seeds of the Leguminosae family.

[0026] For purposes of the present invention, use Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J.Mol.Biol. [Journal of Molecular Biology] 48:443-453) to determine the output of sequence identity between two amino acid sequences as " longest identity ", this algorithm is as EMBOSS software package (EMBOSS:TheEuropean Molecular Biology Open Software Suite [European Molecular Biology Open Software Suite], Rice et al., 2000, Trends Genet. [Genetics trend] 16:276-277) (preferred 6.6.0 version or updated version) Ni Deer program is implemented.The parameter used is gap opening penalty 10, gap extension penalty 0.5 and EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.In order to make Ni Deer program report the longest identity, must specify non-simplified option in command line.The output calculation of " longest identity " of Ni Deer mark is as follows:

[0027] (number of identical residues x 100) / (length of alignment - total number of gaps in the alignment)

[0028] For purposes of the present invention, use Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, the same) to determine the output of sequence identity between two polynucleotide sequences as " longest identity ", this algorithm is implemented by the Needleman program of EMBOSS software package (EMBOSS:The European Molecular Biology Open Software Suite [European Molecular Biology Open Software Suite], Rice et al., 2000, the same) (preferred 6.6.0 version or updated version).The parameter used is gap opening penalty 10, gap extension penalty 0.5 and EDNAFULL (EMBOSS version of NCBINUC4.4) substitution matrix.In order to make Needleman program report the longest identity, non-simplified options must be specified in the command line.The output calculation of " longest identity " of Needleman mark is as follows:

[0029] (number of identical deoxyribonucleotides x 100) / (length of alignment – ​​total number of gaps in the alignment)

[0030] Although certain aspects of the present disclosure will be described hereinafter with reference to embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the claims. DETAILED DESCRIPTION

[0031] The present invention relates to methods, uses and products for improving the flavor and / or texture of plant-based products.

[0032] In order to provide plant-based products (such as soy milk) with enhanced flavor and / or texture, the inventors have found that when α-galactosidase is added to soy milk or during the production of soy milk, the texture and / or taste (such as sweetness, body, fat perception, sweet aftertaste and / or mouthfeel) is improved, and / or the bitterness, astringency and / or bean texture perception is reduced.

[0033] Plant-based products :

[0034] In an embodiment of the present invention, the plant-based product is prepared from one or more plant-based materials and / or plant materials. In an embodiment, the plant material is one or more legume pods. In another embodiment, the plant-based material is a legume pod slurry and / or a legume pod milk base. In an embodiment of the present invention, the plant-based product is prepared from one or more legume pods. In an embodiment, the one or more legume pods are selected from soybeans, peas, chickpeas, lentils and broad beans. In a further embodiment, the one or more legume pods are legume pods containing raffinose oligosaccharides (RFO). In a preferred embodiment, the one or more legume pods are soybeans. RFO is raffinose, stachyose and verbascose.

[0035] In an embodiment of the present invention, the plant-based product is a soy drink. In a preferred embodiment, the soy drink is soy milk.

[0036] α-galactosidase :

[0037] In an embodiment of the invention, the one or more alpha galactosidases are derived from Aspergillus niger. In another embodiment, the one or more alpha galactosidases hydrolyze raffinose oligosaccharides (RFO) to produce sucrose and galactose. In one embodiment, RFO is one or more selected from the group consisting of: raffinose, stachyose and verbascose.

[0038] In an embodiment of the invention, the one or more alpha galactosidases are polypeptides selected from the group consisting of:

[0039] (a) a polypeptide having at least 80% sequence identity to SEQ ID NO: 1,

[0040] (b) a polypeptide derived from SEQ ID NO: 1 having 1 to 30 alterations, e.g. substitutions, deletions and / or insertions, for example 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 alterations, in particular substitutions, at one or more positions,

[0041] (c) a polypeptide derived from the polypeptide of (a), wherein the N- and / or C-terminus has been extended by the addition of one or more amino acids, and

[0042] (d) A fragment of the polypeptide of (a).

[0043] In another embodiment, the polypeptide has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:1.

[0044] In a preferred embodiment of the present invention, the one or more alpha galactosidases are polypeptides comprising, consisting essentially of, or consisting of SEQ ID NO:1.

[0045] In embodiments, the polypeptide may have an N-terminal and / or C-terminal extension of one or more amino acids (eg, 1-5 amino acids).

[0046] In another aspect, the polypeptide is derived from SEQ ID NO: 1 by substitution, deletion or addition of one or several amino acids. In another aspect, the polypeptide is derived from the mature polypeptide of SEQ ID NO: 1 by substitution, deletion or addition of one or several amino acids. In some embodiments, the polypeptide is a variant of SEQ ID NO: 1 comprising substitutions, deletions and / or insertions at one or more positions. In one aspect, the number of amino acid substitutions, deletions and / or insertions introduced into the polypeptide of SEQ ID NO: 1 is up to 15, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15. The amino acid changes may be of a minor nature, i.e., conservative amino acid substitutions or insertions that do not significantly affect the folding and / or activity of the protein; typically small deletions of 1-30 amino acids; small stretches of amino or carboxyl terminal extensions, such as amino-terminal methionine residues; small linker peptides of up to 20-25 residues; or small extensions that facilitate purification by changing the net charge or another function (such as a polyhistidine stretch, antigenic epitope or binding module).

[0047] In one embodiment, the α-galactosidase has the following characteristics: the α-galactosidase is active in the pH range of 3-8, wherein the optimum value is around pH 3.5, the applied temperature range of the α-galactosidase is 4°C to 60°C, and the α-galactosidase is inactivated within 10 minutes at 90°C.

[0048] use

[0049] In one aspect, the present invention relates to the use of one or more alpha galactosidases for enhancing the flavor and / or texture of a plant-based product, wherein the plant-based product is a food and / or a beverage. In an embodiment, the plant-based material is prepared from one or more legume pods. The one or more legume pods are selected from soybeans, peas, chickpeas, lentils, beans, and any legume pods containing raffinose oligosaccharides. In a preferred embodiment, the one or more legume pods are soybeans.

[0050] In an embodiment of the present invention, the enhanced flavor and / or texture is one or more selected from the group consisting of improved sweetness, reduced bitterness, reduced astringency, improved body, improved lipid perception, improved sweet aftertaste, improved mouthfeel, and reduced legume / bean texture perception. In another embodiment, the enhanced texture is reduced legume / bean texture perception.

[0051] method:

[0052] In an embodiment of the present invention, a method for improving the flavor and / or texture of a plant-based product comprises adding one or more α-galactosidases to a plant-based material and / or the plant-based product; and incubating the plant-based product, wherein the plant-based product is a food and / or a beverage.

[0053] In an embodiment, a plant-based material is used to prepare a plant-based product. In a preferred embodiment, the plant-based product is prepared from one or more legume pods. The plant-based material can be a legume pod slurry and / or a legume pod milk base. In a preferred embodiment, the plant-based material is a soy slurry and / or a soy milk base. In an embodiment, the step of adding one or more alpha galactosidases to the plant-based product comprises adding 0.01% w / w to 1% w / w of one or more alpha galactosidases to the plant-based product. In another embodiment, the step of adding one or more alpha galactosidases to the plant-based product comprises adding one or more alpha galactosidases between 67 milligrams of enzyme protein / kilogram of legume pods (mgEP / kg) and 13.4 grams of enzyme protein / kilogram of legume pods (gEP / kg) to the plant-based product.

[0054] In further embodiments, the plant-based product is incubated at a temperature range between 4 degrees Celsius (° C.) and 60° C. In another further embodiment, the plant-based product is incubated for a period of 15 minutes to 24 hours.

[0055] In yet another embodiment, the method reduces raffinose oligosaccharides in the plant-based product and increases sucrose and galactose in the plant-based product.In yet another embodiment, the raffinose oligosaccharides are one or more selected from the group consisting of raffinose, stachyose and verbascose.

[0056] Technology :

[0057] In yet another aspect, the present invention relates to a process for preparing a plant-based product with enhanced flavor and / or texture, wherein the plant-based product is a food and / or beverage. The process comprises soaking one or more plant materials and grinding the one or more plant materials into a slurry. One or more alpha galactosidases are added to the slurry. Thereafter, fibers are separated from the slurry to obtain a plant material base. The one or more alpha galactosidases are added to the plant material base to obtain the plant-based product.

[0058] In an embodiment, the plant material is one or more legumes. In another embodiment, the plant material base is a legume milk base.

[0059] In embodiments, after adding the one or more alpha galactosidases to the slurry, the temperature is maintained in the range of 4 degrees Celsius (° C.) to 55° C. for 15 minutes to 20 hours.

[0060] In an embodiment, after adding the one or more alpha galactosidases to the legume milk base, the temperature is maintained in the range of 4 degrees Celsius (°C) to 55°C for 15 minutes to 20 hours.

[0061] In embodiments, the method further comprises exposing the legume milk base to a heat treatment to inactivate the one or more non-inactivated alpha galactosidases.

[0062] In a preferred embodiment, the process for preparing a soy-based product with an enhanced flavor and / or texture includes producing a soy base and producing soy milk. The soy base production process is diversified by applying or omitting various processing steps. In order to produce the soy base, whole or dehulled soybeans are used. Alkali blanching is also an option, which promotes the inactivation of endogenous proteases and lipoxygenases. The process includes soaking the soybeans and grinding the soybeans into a soy slurry. In addition, soaking the soybeans before grinding improves the grinding efficiency. The soybeans undergoing the grinding process produce a colloidal solution of soybeans in water, also known as soy slurry. The protein concentration of the soy slurry is determined by the ratio of beans to water at the inlet of the grinder. One or more alpha galactosidases are added to the soy slurry. Thereafter, the fiber is separated from the soy slurry to obtain a soy milk base. The one or more alpha galactosidases are added to the soy milk base to obtain a soy-based product.

[0063] In the soy slurry, one or more alpha galactosidases are added. The temperature to be applied is between 4°C and 55°C for 15 minutes to 20 hours.

[0064] In the next step, fibrous particles called okara are separated by using a decanter or centrifuge. The efficiency of the decanter determines the yield and quality of the soy milk. To optimize the yield, the water content in the okara should be low. The amount of sediment (dry matter) in the slurry should be low to avoid a powdery feel caused by insoluble particles in soy-based products.

[0065] To remove the undesirable volatile off-flavors that cause beany taste, the soy base is exposed to a heat treatment, such as direct steam injection (which heats to 140°C) followed by flash cooling. The heat treatment inactivates any endogenous or added enzymes (such as alpha galactosidase that has not been inactivated).

[0066] The soy base is standardized to the desired protein content and composition by adding water and other ingredients and further processed into soy-based products such as soy milk, soy drinks, soy yogurt and soy ice cream.

[0067] Different formulations can be added to the standardized soy base. The final soy milk appearance varies depending on the region or type of soybean. Soy milk typically contains about 3% protein, 2% oil and 2% carbohydrates. Soy milk can also be a beverage with reduced protein content but higher sugar levels. Soy milk beverages can be mineral-fortified or flavored.

[0068] In another preferred embodiment, the one or more alpha galactosidases are added to the soy base.The incubation temperature is in the range of 4°C to 55°C, and the incubation time is in the range of 15 minutes to 20 hours.

[0069] The formulated soy base is then homogenized and the soymilk is exposed to an ultra-heat treatment (UHT) process and aseptic filling, thereby obtaining soymilk with a long shelf life at room temperature. The UHT process applied can be direct or indirect. In the direct UFT process, steam is briefly injected into the product to be treated, followed by rapid flash cooling. In the indirect UHT process, the product to be treated is not in direct contact with the heat source, but is heated using a heat exchanger. The UHT process inactivates any enzymes added to the soymilk, such as alpha-galactosidase.

[0070] The following numbered paragraphs describe particular embodiments of the present disclosure:

[0071] 1. Use of one or more alpha galactosidases for improving the flavor and / or texture of a plant-based product, wherein the plant-based product is a food and / or a drink.

[0072] 2. The use according to paragraph 1, wherein the one or more alpha-galactosidases are derived from Aspergillus niger.

[0073] 3. The use according to paragraph 1 or 2, wherein the one or more alpha galactosidases hydrolyze raffinose oligosaccharides (RFO) to produce sucrose and galactose.

[0074] 4. The use according to paragraph 3, wherein the RFO is one or more selected from the group consisting of raffinose, stachyose and verbascose.

[0075] 5. The use according to any one of the preceding paragraphs, wherein the one or more alpha galactosidases are polypeptides selected from the group consisting of:

[0076] (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% sequence identity to SEQ ID NO: 1;

[0077] (b) a polypeptide derived from SEQ ID NO: 1 having 1-30 alterations, e.g. substitutions, deletions and / or insertions, e.g. 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 alterations, in particular substitutions, at one or more positions;

[0078] (c) a polypeptide derived from the polypeptide of (a), wherein the N- and / or C-terminus has been extended by the addition of one or more amino acids; and

[0079] (d) A fragment of the polypeptide of (a).

[0080] 6. The use according to any one of the preceding paragraphs, wherein the one or more alpha galactosidases is a polypeptide having the sequence of SEQ ID NO: 1.

[0081] 7. The use according to any of the preceding paragraphs, wherein the enhanced flavor and / or texture is one or more selected from the group consisting of improved sweetness, reduced bitterness, reduced astringency, improved body, improved lipid perception, improved sweet aftertaste, improved mouthfeel, and reduced legume / bean texture perception.

[0082] 8. The use according to any of the preceding paragraphs, wherein the enhanced texture is a reduced legume / bean texture.

[0083] 9. The use according to any one of the preceding paragraphs, wherein the plant-based product is prepared from one or more leguminous plant pods.

[0084] 10. The use according to any one of the preceding paragraphs, wherein the one or more legumes are selected from soybeans, peas, chickpeas, lentils, beans and any legumes containing raffinose oligosaccharides.

[0085] 11. The use according to any one of the preceding paragraphs, wherein the one or more leguminous plant pods are soybeans.

[0086] 12. The use according to any of the preceding paragraphs, wherein the plant-based product is a soy drink.

[0087] 13. The use according to paragraph 12, wherein the soy beverage is soy milk.

[0088] 14. A method for improving the flavor and / or texture of a plant-based product, the method comprising:

[0089] a. adding one or more α-galactosidases to the plant-based material and / or the plant-based product; and

[0090] b. incubating the plant-based material and / or the plant-based product,

[0091] The plant-based product is food and / or beverage.

[0092] 15. The method according to paragraph 14, wherein the one or more alpha-galactosidases are derived from Aspergillus niger.

[0093] 16. The method according to paragraph 14 or 15, wherein the one or more alpha-galactosidases hydrolyze raffinose family oligosaccharides (RFO) to produce sucrose and galactose.

[0094] 17. The method according to any one of paragraphs 14-16, wherein the one or more alpha galactosidases are polypeptides selected from the group consisting of:

[0095] (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% sequence identity to SEQ ID NO: 1;

[0096] (b) a polypeptide derived from SEQ ID NO: 1 having 1-30 alterations, e.g. substitutions, deletions and / or insertions, e.g. 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 alterations, in particular substitutions, at one or more positions;

[0097] (c) a polypeptide derived from the polypeptide of (a), wherein the N- and / or C-terminus has been extended by the addition of one or more amino acids; and

[0098] (d) A fragment of the polypeptide of (a).

[0099] 18. The method according to paragraphs 14-17, wherein the one or more alpha-galactosidases are polypeptides having the sequence of SEQ ID NO: 1.

[0100] 19. The method according to paragraphs 14-18, wherein the plant-based material is used to prepare the plant-based product.

[0101] 20. The method of any of paragraphs 14-19, wherein the enhanced flavor and / or texture is selected from the group consisting of improved sweetness, reduced bitterness, reduced astringency, improved body, improved lipid perception, improved sweet aftertaste, improved mouthfeel, and reduced legume / bean texture perception.

[0102] 21. The method of paragraph 20, wherein the enhanced texture is a legume / bean texture.

[0103] 22. The method of any of paragraphs 14-21, wherein the plant-based product is prepared from one or more leguminous plant pods.

[0104] 23. The method according to paragraphs 14-22, wherein the plant-based material is a legume berry pulp and / or a legume berry milk base.

[0105] 24. The method according to paragraph 22, wherein the one or more legumes are selected from soybeans, peas, chickpeas, lentils, beans and any legumes containing raffinose oligosaccharides.

[0106] 25. The method of paragraph 22, wherein the one or more legume pods are soybeans.

[0107] 26. The method according to paragraphs 14-25, wherein the plant-based material is a soy slurry and / or soy milk base.

[0108] 27. The method according to any of paragraphs 14-26, wherein the plant-based product is a soy drink.

[0109] 28. The method according to paragraph 27, wherein the soy beverage is soy milk.

[0110] 29. The method according to any of paragraphs 14-28, wherein the method comprises adding 0.01% w / w to 1% w / w one or more alpha galactosidases to the plant-based product.

[0111] 30. The method according to any of paragraphs 14-29, wherein the method comprises adding between 67 mg EP / kg and 13.4 g EP / kg of one or more alpha galactosidases to the plant-based product.

[0112] 31. The method of any of paragraphs 14-30, wherein the plant-based product is incubated at a temperature range between 4 degrees Celsius (°C) and 60°C.

[0113] 32. The method of any of paragraphs 14-31, wherein the plant-based product is incubated for a period of 15 minutes to 24 hours.

[0114] 33. The method according to any of paragraphs 14-32, wherein the method reduces raffinose oligosaccharides in the plant-based product and increases sucrose and galactose in the plant-based product.

[0115] 34. A process for preparing a plant-based product having enhanced flavor and / or texture, the process comprising:

[0116] a. soaking one or more plant materials;

[0117] b. grinding the one or more plant materials into a slurry;

[0118] c. adding one or more α-galactosidases to the slurry;

[0119] d. separating fibers from the slurry to obtain a plant material base;

[0120] e. adding one or more alpha galactosidases to the plant material base, wherein the plant based product is a food and / or a beverage.

[0121] 35. A process for preparing the plant-based product with enhanced flavor and / or texture according to paragraph 34, wherein after adding the one or more alpha-galactosidases to the slurry, the temperature is maintained in the range of 4 degrees Celsius (° C.) to 55° C. for 15 minutes to 20 hours.

[0122] 36. The process for preparing the plant-based product with enhanced flavor and / or texture according to paragraph 34 or 35, further comprising exposing the legume milk base to a heat treatment to inactivate the one or more non-inactivated alpha-galactosidases.

[0123] 37. A process for preparing the plant-based product with enhanced flavor and / or texture according to any one of paragraphs 34-36, wherein after adding the one or more alpha-galactosidases to the legume milk base, the temperature is maintained in the range of 4 degrees Celsius (°C) to 55°C for 15 minutes to 20 hours.

[0124] 38. The process according to any of paragraphs 34-37, wherein the one or more alpha-galactosidases are derived from Aspergillus niger.

[0125] 39. The process of any of paragraphs 34-38, wherein the one or more alpha-galactosidases hydrolyze raffinose oligosaccharides (RFO) to produce sucrose and galactose.

[0126] 40. The process of paragraphs 34-39, wherein the RFO is selected from the group consisting of raffinose, stachyose and verbascose.

[0127] 41. The process according to any of paragraphs 34-40, wherein the one or more alpha galactosidases are polypeptides selected from the group consisting of:

[0128] (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% sequence identity to SEQ ID NO: 1;

[0129] (b) a polypeptide derived from SEQ ID NO: 1 having 1-30 alterations, e.g. substitutions, deletions and / or insertions, e.g. 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 alterations, in particular substitutions, at one or more positions;

[0130] (c) a polypeptide derived from the polypeptide of (a), wherein the N- and / or C-terminus has been extended by the addition of one or more amino acids; and

[0131] (d) A fragment of the polypeptide of (a).

[0132] 42. The process according to paragraphs 34-41, wherein the one or more alpha-galactosidases is a polypeptide having the sequence of SEQ ID NO: 1.

[0133] 43. The process of any of paragraphs 34-42, wherein the enhanced flavor and / or texture is selected from the group consisting of improved sweetness, reduced bitterness, reduced astringency, improved body, improved lipid perception, improved sweet aftertaste, improved mouthfeel, and reduced legume / bean texture perception.

[0134] 44. The process of paragraph 43 wherein the enhanced texture is a legume / bean texture.

[0135] 45. The process according to any of paragraphs 34-44, wherein the plant material is one or more leguminous plant pods.

[0136] 46. ​​The process according to any of paragraphs 34-45, wherein the one or more legume pods are selected from soybeans, peas, chickpeas, lentils, beans and any legume pods containing raffinose oligosaccharides.

[0137] 47. The process of paragraph 45, wherein the one or more legume pods are soybeans.

[0138] 48. The process of paragraphs 34-47, wherein the plant material base is a leguminous plant pod milk base.

[0139] 49. The process of paragraph 48, wherein the leguminous plant legume milk base is a soy milk base.

[0140] 50. The process of any of paragraphs 34-49, wherein the plant-based product is a soy beverage.

[0141] 51. The process according to paragraph 50, wherein the soy beverage is soy milk.

[0142] Examples

[0143] The following examples are not intended to be an exhaustive list of all the different ways in which the present disclosure may be implemented or an exhaustive list of all the features that may be added to the present disclosure. It will be appreciated by those skilled in the art that various changes and additions may be made to the various embodiments without departing from the present disclosure. Therefore, the following description is intended to illustrate some specific embodiments of the present invention, rather than to exhaustively specify all permutations, combinations, and variations thereof.

[0144] Unless otherwise indicated, the percentages listed in the following examples are weight percentages based on the total weight of the composition.

[0145] Materials and methods

[0146] Experiments were performed in Denmark with commercially available soy milk from Nature Green and an alpha-galactosidase with SEQ ID NO: 1.

[0147] Example 1 - Sensory Analysis of α-Galactosidase Treated and Untreated Soymilk by 22 Untrained Evaluators

[0148] A. Sample production

[0149] In the test, commercial soy milk from Nature Green was used as substrate for the α-galactosidase incubation. The incubation was carried out in a Thermomix Vorwerk. The soy milk was briefly preheated to 40°C; α-galactosidase was added at a dose of 0.013 mg EP / 1 mL soy milk. In addition, a blank sample without α-galactosidase was also carried out in parallel. The incubation was carried out at 40°C for 1 hour. After the incubation, the Vorwerk temperature was increased to 90°C to inactivate the enzyme and the holding time was 10 minutes. The soy milk was cooled in ice cubes and transferred to pasteurized glass bottles and stored at 4°C until sensory analysis.

[0150] B. Sugar Analysis

[0151] The treated soymilk samples were analyzed on an HPLC (high pressure liquid chromatography) equipped with refractive index detection. The column used was an Aminex HPX-87P column (from Bio-Rad) placed in a column oven set at 80°C. Separation was performed at an isocratic level using milliQ water. A reflectivity (RI) detector (ThermoScientific TM RefractoMax 521 refractive index detector). The flow rate was 0.6 milliliters per minute (mL / min). Chemically pure standards of raffinose, stachyose, galactose and sucrose were used for calibration. Before HPLC analysis, 5 microliters (μL) of 50% acetic acid were added to the soy milk samples; the samples were centrifuged at 14000 rpm for 5 minutes and 10 minutes. Afterwards, sample cleanup was continued by adding 5 μL Carrez 1 and 5 μL Carrez 2 and centrifuging the samples under the above conditions. Finally, the samples were filtered through a 0.22 μm filter.

[0152] Table 1: Results of HPLC analysis

[0153]

[0154]

[0155] As shown in Table 1, all raffinose and stachyose were hydrolyzed into sucrose and galactose by α-galactosidase. Sucrose was measured to increase from 6.5 g / L to 9.6 g / L. Galactose increased from 0.25 g / L to 2.9 g / L.

[0156] C. Sensory analysis

[0157] Paired comparisons for sweetness were performed by 22 untrained assessors. Assessors were asked to identify the sweetest sample. In addition, they were asked to provide details of other differences between samples in the comments column. Sensory analysis was paper-based. Room temperature samples were provided in clear plastic cups with 3-digit labels. Each assessor received 30 ml (about 1.01 oz) of sample.

[0158] result

[0159] 20 out of 22 people thought that 0.02% α-galactosidase treated soy milk (0.013 mg EP / 1 mL soy milk) was the sweetest. α-galactosidase had a significant effect on the sensory perception of sweetness (binomial test). The assessors also supplemented the sensory test with written comments. α-galactosidase treated soy milk was found to be fattier, richer and more textured.

[0160] Example 2 - Two rounds of sensory analysis of α-galactosidase treated and untreated soy milk by 6 untrained assessors

[0161] A. Sample production

[0162] The same production method as described in Example 1

[0163] B. Sugar Analysis

[0164] Same sugar analysis method as described in Example 1

[0165] Table 2: Results of HPLC analysis

[0166]

[0167] As shown in Table 2, all raffinose and stachyose were hydrolyzed by α-galactosidase to sucrose and galactose. Sucrose was measured to increase from 4.8 g / L to 6.7 g / L. Galactose increased from 0.2 g / L to 3 g / L.

[0168] C. Sensory analysis

[0169] In this sensory analysis, there were 6 untrained assessors in the sensory panel. The sensory test was conducted in 2 rounds:

[0170] Round 1: A training session was conducted in which the assessors drank samples (blank and 0.013 mg EP / 1 mL soy milk) and identified the attributes to be scored: sweetness, bean taste and texture perception. During the training, a 9-point preference scale was used. The sample volume was 50 ml.

[0171] Round 2: A blind test was conducted with combined ratings. Evaluators used a paper-based 9-point preference scale. Samples were provided in 30 ml (approximately 1.01 oz) volumes. Samples were provided at room temperature in clear plastic cups with 3-digit labels.

[0172] As shown in Table 3, the assessors clearly noticed the sweetness produced by α-galactosidase, and the rating of the sweetness was significant (p<0.05).

[0173] Table 3:

[0174] Sample_ID Sweetness Rating blank 3 α-galactosidase_0.02pct 4.7

[0175] When looking at the texture and bean attribute scores - 4 assessors were able to differentiate between the blank and α-galactosidase treated soymilk samples. There was a trend for assessors to favor the α-galactosidase treated soymilk samples.

[0176] Example 3 - Sensory analysis of α-galactosidase treated and untreated soy milk by 23 untrained assessors

[0177] A. Sample production

[0178] The same production method as described in Example 1

[0179] B. Sugar Analysis

[0180] Same sugar analysis method as described in Example 1

[0181] Table 4: Results of HPLC analysis

[0182]

[0183] As shown in Table 4, all raffinose and stachyose were hydrolyzed by α-galactosidase to sucrose and galactose. Sucrose was measured to increase from 6.9 g / L to 9.2 g / L. Galactose increased from 0.08 g / L to 2.5 g / L.

[0184] C. Sensory analysis

[0185] Purpose:

[0186] The purpose of the sensory evaluation was to investigate whether there was a difference in sweetness between the two soymilk samples (with and without α-galactosidase). In addition, other differences between the samples were evaluated.

[0187] method:

[0188] Paired comparisons for sweetness were performed by 23 untrained assessors. In order to better focus on low sweetness levels, nose clips were provided, which reduced the impact of odor and allowed to focus on taste and mouthfeel. Assessors were asked to identify the sweetness samples but were also allowed to give equal ratings to both samples. In addition, assessors were asked to provide detailed information on other differences between samples in the comments column. Data were collected using the sensory software EyeQuestion.

[0189] The samples were provided at approximately 10°C in clear plastic cups with 3-digit labels. Each assessor received 30 ml (approximately 1.01 oz) of the sample.

[0190] result:

[0191] Comments by the evaluators who identified the sample containing α-galactosidase as the sweetest suggested that there may also be differences in mouthfeel, as the sample containing α-galactosidase was described as slightly creamier / mellower / rounder.

[0192] The results of the pairwise comparisons are shown in Table 5. Most of the evaluators selected the sample containing α-galactosidase as the sweetest sample. Based on the p-value of 0.077, it was concluded that there was a significant difference in sweetness between the soymilk containing α-galactosidase and the soymilk not containing α-galactosidase.

[0193] Table 5:

[0194] The sweetest sample Number of assessors No difference 7 Contains alpha-galactosidase 12 Does not contain alpha-galactosidase 4

Claims

1. Use of one or more alpha galactosidases for improving the flavor and / or texture of a plant-based product, wherein the plant-based product is a food and / or a drink.

2. Use according to claim 1, wherein the one or more alpha galactosidases are derived from Aspergillus niger.

3. The use according to claim 1 or 2, wherein the one or more alpha galactosidases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% sequence identity to SEQ ID NO: 1; (b) a polypeptide derived from SEQ ID NO: 1 having 1-30 alterations, e.g. substitutions, deletions and / or insertions, e.g. 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 alterations, in particular substitutions, at one or more positions; (c) a polypeptide derived from the polypeptide of (a), wherein the N- and / or C-terminus has been extended by the addition of one or more amino acids; and (d) A fragment of the polypeptide of (a).

4. Use according to any one of the preceding claims, wherein the one or more alpha galactosidases is a polypeptide having the sequence of SEQ ID NO:

1.

5. Use according to any one of the preceding claims, wherein the enhanced flavour and / or texture is one or more selected from the group consisting of improved sweetness, reduced bitterness, reduced astringency, improved body, improved lipid perception, improved sweet aftertaste, improved mouthfeel, and reduced perception of legume pods / beans texture.

6. Use according to any one of the preceding claims, wherein the plant-based product is prepared from one or more leguminous plant pods.

7. Use according to any one of the preceding claims, wherein the one or more leguminous plant pods are soybeans.

8. Use according to any one of the preceding claims, wherein the plant-based product is a soy drink, preferably soy milk.

9. A method for improving the flavor and / or texture of a plant-based product, the method include: a. adding one or more α-galactosidases to the plant-based material and / or the plant-based product; as well as b. incubating said plant-based material and / or said plant-based product, The plant-based product is a food and / or a beverage.

10. The method of claim 9, wherein the one or more alpha-galactosidases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, 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 100% sequence identity to SEQ ID NO: 1; (b) a polypeptide derived from SEQ ID NO: 1 having 1-30 alterations, e.g. substitutions, deletions and / or insertions, e.g. 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 alterations, in particular substitutions, at one or more positions; (c) a polypeptide derived from the polypeptide of (a), wherein the N- and / or C-terminus has been extended by the addition of one or more amino acids; and (d) A fragment of the polypeptide of (a).

11. The method according to claim 9 or 10, wherein the one or more alpha galactosidases are polypeptides having the sequence of SEQ ID NO:

1.

12. The method according to any one of claims 9 to 11, wherein the method comprises adding 0.01% w / w to 1% w / w of one or more alpha galactosidases to the plant-based product.

13. The method according to any one of claims 9 to 12, wherein the method comprises adding between 67 mg EP / kg and 13.4 g EP / kg of one or more alpha galactosidases to the plant-based product.

14. A process for preparing a plant-based product having improved flavor and / or texture, the process include: a. soaking one or more plant materials; b. grinding the one or more plant materials into a slurry; c. adding one or more α-galactosidases to the slurry; d. separating fibers from the slurry to obtain a plant material base; e. adding one or more alpha galactosidases to the plant material base, wherein the plant-based product is a food and / or a beverage.

15. The process according to claim 14, wherein the plant material base is a leguminous plant legume milk base, preferably a soy milk base.

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