Method of manufacturing processed plant-based milk

Abstract

The object of the present invention is to provide a processing technology that improves the dispersion stability and / or solubility of plant-based milks such as oat milk, walnut milk, and peanut milk. [Solution] By obtaining processed plant milk using a method that includes a step of treating plant-based milk such as oat milk, walnut milk, and peanut milk with a protein deamidation enzyme, the dispersion stability and / or solubility of the obtained processed plant milk can be improved.

Description

Technical Field

[0001] The present invention relates to a method for producing processed plant milk. More specifically, the present invention relates to a processing technology for enhancing the dispersion stability and / or solubility of plant milk such as peanut milk, oat milk, and almond milk.

Background Art

[0002] Beverages rich in nutrients such as protein have been widely favored by people because they can be easily consumed. On the other hand, in recent years, due to the increase in vegetarians, allergy problems, and religious reasons, soy milk made from soybeans rich in plant protein has become widely popular as an alternative to animal milk represented by cow's milk.

[0003] Regarding soy protein, various modification treatments have been studied for the purpose of improving its existing properties and providing foods with new taste characteristics.

[0004] For example, Patent Document 1 (JP2000-50887A) describes that the yield of soy protein from soy flour can be improved by treating soy flour with a protein deamidating enzyme. Also, Patent Document 2 (JP2008-283900A) describes that a polyglycerol fatty acid ester having a fatty acid with 12 to 22 carbon atoms as the main constituent fatty acid is effective as a dispersion stabilizer for soy milk. Furthermore, Patent Document 3 (JP2015-159765A) describes that by performing a deamidation treatment on soy milk with a cation exchange resin and / or a phytic acid removal treatment with an anion exchange resin, precipitation is less likely to occur with a coagulant.

[0005] On the other hand, from the perspective of meeting further diversification of preferences, there is a need for more options in addition to soy milk in food and beverages containing plant protein.

[0006] Therefore, as plant-based milks containing plant proteins other than soy protein, milks made from grains such as peas, rice, and oats, and milks made from nuts such as peanuts and walnuts are being developed.

[0007] Among these, oat milk has characteristics that differentiate it from other grain milks, as it is rich in lipids, β-glucans, and minerals in addition to protein, and its high nutritional value has attracted attention. For example, Patent Document 4 (US6,451,361B1) describes how treating an oat suspension with α-amylase and β-amylase solved the high viscosity problem and obtained an oat dispersion that maintained protein and β-glucans. Also, Patent Document 5 (CN101991163A) describes how treating oat with α-amylase, β-amylase, and transglucosidase produces maltooligosaccharides, thereby improving the prebiotic effect of oat beverages.

[0008] Walnut milk contains far more lipids than protein. Patent document 6 (WO2019 / 104971A1) describes how a low-fat, high-protein walnut milk with good texture and stability was obtained by separating and removing some of the oils and fats during the manufacturing process. Patent document 7 (CN109122876A) also describes how a highly stable walnut milk was obtained by using protease, lipase, and cellulase. [Overview of the project] [Problems that the invention aims to solve]

[0009] Oat milk, walnut milk, and peanut milk (a type of nut milk similar to walnut milk) are all highly nutritious and have high value as health foods. However, their dispersion stability and / or solubility have not been adequately studied.

[0010] The present invention aims to provide a processing technology that improves the dispersion stability and / or solubility of plant-based milks such as oat milk, walnut milk, and peanut milk. [Means for solving the problem]

[0011] Oat milk is characterized by its high viscosity and high nutritional value, and walnut milk and peanut milk are characterized by their high fat and high nutritional value. Therefore, it has been recognized that the characteristic composition of these plant-based milks, other than protein, has a very significant impact on the milk properties. However, as a result of diligent research by the present inventors, it has been found that the dispersion stability and / or solubility of these plant-based milks can be improved by treating them with protein deamidation enzymes. That is, the present invention provides the invention in the following embodiments.

[0012] Item 1. A method for producing processed plant-based milk, comprising the step of treating plant-based milk with a protein deamidation enzyme. Item 2. The method for producing processed plant milk according to Item 1, wherein the plant milk is selected from the group consisting of oat milk, walnut milk, and peanut milk. Item 3. A method for producing processed plant milk according to Item 1 or 2, wherein the protein deamidation enzyme is used at a rate of 0.5 U or more per gram of plant protein. Item 4. A method for producing processed plant milk according to any one of Items 1 to 3, wherein the plant milk is oat milk, and the protein deamidation enzyme is used at a rate of 1.5 U or more per gram of oat protein. Item 5. A method for producing processed plant milk according to any one of Items 1 to 3, wherein the plant milk is the oat milk, and α-amylase, hemicellulase, and / or maltotriosyltransferase are used in combination with the protein deamidase. Item 6. A method for producing processed plant milk according to any one of Items 1 to 3, wherein the plant-based milk is the walnut milk, the processed plant-based milk is walnut milk with improved dispersion stability than the walnut milk, and the protein deamidation enzyme is used at a rate of 3 U or more per gram of walnut protein. Item 7. A method for producing processed plant milk according to any one of Items 1 to 3, wherein the plant-based milk is the walnut milk, the processed plant-based milk is walnut milk with improved solubility than the walnut milk, and the protein deamidation enzyme is used at a rate of 1 U or more per 1 g of walnut protein. Item 8. A method for producing processed plant milk according to any one of items 1 to 3, wherein the plant milk is the walnut milk, and glucoamylase, α-amylase, β-amylase, transglucosidase, and / or glutaminase are used in combination with the protein deamidation enzyme. Item 9. A method for producing processed plant milk according to any one of Items 1 to 3, wherein the plant-based milk is the peanut milk, the processed plant-based milk is peanut milk with improved dispersion stability than the peanut milk, and the protein deamidation enzyme is used at a rate of 2 U or more per gram of peanut protein. Item 10. A method for producing processed plant milk according to any one of Items 1 to 3, wherein the plant-based milk is the peanut milk, the processed plant-based milk is peanut milk with improved solubility than the peanut milk, and the protein deamidation enzyme is used at a rate of 1 U or more per gram of peanut protein. Item 11. A method for producing processed plant milk according to any one of Items 1 to 3, wherein the plant-based milk is the peanut milk, the processed plant-based milk is peanut milk with improved texture than the peanut milk, and the protein deamidation enzyme is used at a rate of 2 U or more per gram of peanut protein. Item 12. A method for producing processed plant milk according to any one of items 1 to 3, wherein the plant milk is the peanut milk, and α-amylase, β-amylase, glucoamylase, and / or hemicellulase are used in combination with the protein deamidation enzyme. Item 13. Use of protein deamidase enzymes for the production of dispersion stability enhancers for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk. Item 14. Use of protein deamidase for the production of solubility enhancers for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk. Item 15. Use of protein deamidation enzymes for the manufacture of texture enhancers for peanut milk. Item 16. A dispersion stability enhancer for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk, comprising a protein deamidase. Item 17. A solubility enhancer for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk, comprising a protein deamidase. Item 18. A texture enhancer for peanut milk containing a protein deamide enzyme. [Effects of the Invention]

[0013] The present invention provides a processing technology for improving the dispersion stability and / or solubility of plant-based milks such as oat milk, walnut milk, and peanut milk. [Modes for carrying out the invention]

[0014] 1. Method for producing processed vegetable milk The present invention relates to a method for producing processed plant-based milk, characterized by comprising a step of treating plant-based milk with a protein deamidation enzyme. The method for producing processed plant-based milk according to the present invention will be described in detail below.

[0015] 1-1. Plant-based milk The plant-based milk used in this invention refers to a liquid in which crushed edible parts of a plant are dispersed in water. The crushing of the edible parts of the plant can be carried out by any method, such as pressing and / or grinding, and these crushing methods can preferably be carried out in water. In addition, in the plant-based milk, the crushed edible parts are dispersed, and components derived from the edible parts that have been exposed in water by extraction or the like may be partially or completely dissolved, dispersed, and / or emulsified. Furthermore, in the plant-based milk, insoluble matter derived from the peels, etc., of the edible parts may be removed as appropriate by any means such as centrifugal filtration, filtration, filter bags, sieves, etc.

[0016] The edible part of the plant used as the raw material of the plant milk used in the present invention is not particularly limited. In the present invention, preferably, the plant milk is selected from the group consisting of oat milk, almond milk and peanut milk.

[0017] The oat milk used in the present invention is not particularly limited, and general oat milk can be used. Examples of oat milk include liquid substances obtained by filtering heat-treated oat slurry (for example, porridge of oat powder, crushed product of oatmeal porridge, etc.). In the heat-treated oat slurry, the amount of water relative to 1 part by weight of oats is, for example, 2 to 10 parts by weight, preferably 3 to 8 parts by weight, more preferably 4 to 6 parts by weight, and even more preferably 4.5 to 5.5 parts by weight. The temperature of the heat treatment is, for example, 83 to 100 ° C, preferably 85 to 96 ° C, more preferably 88 to 93 ° C. The number of meshes of the sieve used for filtering the heat-treated oat slurry may be such that coarse insoluble fibers of oats can be removed, and examples thereof include 50 to 70 meshes, preferably 55 to 65 meshes.

[0018] The almond milk used in the present invention is not particularly limited, and general almond milk can be used. Examples of almond milk include heat-treated almond slurry (for example, heat-treated product of almond slurry without skin or its water dilution product). The temperature of the heat treatment is, for example, 83 to 100 ° C, preferably 85 to 96 ° C, more preferably 88 to 93 ° C. The amount of water relative to 1 part by weight of almonds in the almond milk is, for example, 1 to 10 parts by weight, preferably 2 to 8 parts by weight, more preferably 3 to 5 parts by weight, and even more preferably 3.5 to 4.5 parts by weight.

[0019] The peanut milk used in the present invention is not particularly limited, and general peanut milk can be used. Examples of peanut milk include heat-treated peanut slurries (e.g., boiled products of slurries of peeled roasted peanuts or water dilutions thereof). Examples of the temperature of the heat treatment include, for example, 90°C to the boiling temperature, preferably 95°C to the boiling temperature, more preferably the boiling temperature. The content of solids (peanut components) in the peanut milk is, for example, 2 to 15 v / w%, preferably 4 to 12 v / w%, more preferably 6 to 10 v / w%, still more preferably 7 to 9 v / w%, and even more preferably 7.5 to 8.5 v / w%. The content of peanut protein in the peanut milk is, for example, 0.8 to 4 w / v%, preferably 1.2 to 3 w / v%, more preferably 1.5 to 2.5 w / v%, still more preferably 1.8 to 2.2 w / v%. Also, the pH (25°C) of the peanut milk is, for example, 5.5 to 6.5, preferably 5.8 to 6.2.

[0020] These plant-based milks may be used alone or in combination of multiple types.

[0021] 1-2. Protein deamidation enzymes The protein deamide enzyme used in the present invention is not particularly limited in type or origin, as long as it is an enzyme that degrades the amide group-containing side chain of a protein without cleaving peptide bonds or crosslinking proteins. Examples of protein deamide enzymes include those from the genera Chryseobacterium, Flavobacterium, Empedobacter, Sphingobacterium, Aureobacterium, or Myroides, as disclosed in JP2000-50887A, JP2001-218590A, and WO2006 / 075772A1, as well as commercially available protein glutaminases from the genus Chryseobacterium. These protein deamide enzymes may be used individually or in combination of multiple types.

[0022] Among these protein deamide enzymes, from the viewpoint of further improving the dispersion stability and / or solubility of the above-mentioned plant-based milk, or further from the viewpoint of further improving the texture of peanut milk, protein deamide enzymes derived from the genus Chryseobacterium are preferred, more preferably protein glutaminases derived from the genus Chryseobacterium, and even more preferably protein glutaminases derived from the species Chryseobacterium proteolyticum.

[0023] Protein deamide enzymes can be prepared from the culture medium of the microorganism from which the above-mentioned protein deamide enzymes originate. Specific preparation methods include recovering the protein deamide enzyme from the culture medium or cells of the above-mentioned microorganisms. For example, when using a protein deamide enzyme-secreting microorganism, the cells can be recovered from the culture medium by filtration, centrifugation, etc., as needed, and then the enzyme can be separated and / or purified. When using a protein deamide enzyme-non-secreting microorganism, the cells can be recovered from the culture medium by filtration, centrifugation, etc., as needed, and then the cells can be crushed by pressurization, sonication, etc., to expose the enzyme, and then the enzyme can be separated and / or purified. The enzyme separation and / or purification method can be any known protein separation and / or purification method without particular limitation, such as centrifugation, UF concentration, salting out, and various chromatography methods using ion exchange resins. The separated and / or purified enzyme can be powdered by drying methods such as freeze-drying or vacuum drying, and can also be powdered using appropriate excipients and / or drying aids in the drying method.

[0024] Commercially available protein deamidation enzymes can also be used, and a preferred example of such an enzyme is "Amano" 500 protein glutaminase manufactured by Amano Enzyme Co., Ltd.

[0025] The titer of the protein deamidase is not particularly limited, but examples include 10 to 50,000 U, preferably 100 to 10,000 U, more preferably 200 to 800 U / g, even more preferably 300 to 700 U / g, even more preferably 400 to 600 U / g, and even more preferably 450 to 550 U / g.

[0026] The amount of protein deamidation enzyme used is not particularly limited, but examples of the amount used per gram of plant protein include 0.01 U or more, preferably 0.1 U or more, more preferably 0.5 U or more, and even more preferably 0.8 U or more.

[0027] When the plant-based milk is oat milk, from the viewpoint of further improving the dispersion stability and / or solubility of the oat milk, the amount of protein deamidase used per gram of oat protein is preferably 1.5 U or more, more preferably 2 U or more, even more preferably 2.5 U or more, even more preferably 3 U or more, even more preferably 4 U or more, and particularly preferably 4.5 U or more. There is no particular upper limit to the range of the amount of protein deamidase used per gram of oat protein, but examples include 25 U or less, 22 U or less, 17 U or less, 14 U or less, 10 U or less, 8 U or less, or 6 U or less.

[0028] When the plant-based milk is walnut milk, from the viewpoint of further improving the dispersion stability of the walnut milk, the amount of protein deamidation enzyme used per gram of walnut protein is preferably 3U or more, more preferably 4U or more, even more preferably 5U or more, even more preferably 7U or more, even more preferably 9U or more, and particularly preferably 10U or more.

[0029] When the plant-based milk is walnut milk, from the viewpoint of further improving the solubility of the walnut milk, the amount of protein deamidation enzyme used per gram of walnut protein is preferably 1 U or more, more preferably 2 U or more, even more preferably 4 U or more, even more preferably 6 U or more, even more preferably 8 U or more, and particularly preferably 10 U or more.

[0030] When the plant-based milk is walnut milk, there is no particular upper limit to the amount of protein deamide enzyme used per gram of walnut protein, but examples include 25U or less, 22U or less, 17U or less, 14U or less, or 12U or less.

[0031] When the plant-based milk is peanut milk, from the viewpoint of further improving the dispersion stability of the peanut milk, the amount of protein deamidation enzyme used per gram of peanut protein is preferably 2 U or more, more preferably 2.5 U or more.

[0032] When the plant-based milk is peanut milk, from the viewpoint of further improving the solubility of the peanut milk, the amount of protein deamidation enzyme used per gram of peanut protein is preferably 1 U or more, more preferably 1.5 U or more, even more preferably 2 U or more, and even more preferably 2.5 U or more.

[0033] When the plant-based milk is peanut milk, from the viewpoint of further enhancing the texture of the peanut milk, the amount of protein deamidation enzyme used per gram of peanut protein is preferably 2U or more, more preferably 2.5U or more.

[0034] When the plant-based milk is peanut milk, there is no particular upper limit to the amount of protein deamidation enzyme used per gram of peanut protein, but examples include 25U or less, 20U or less, 15U or less, 10U or less, 5U or less, or 3U or less.

[0035] Regarding the activity of protein deamidase enzymes, one unit (1U) is defined as the amount of enzyme that releases 1 μmol of ammonia per minute using benzyloxycarbonyl-L-glutaminylglycine (Z-Gln-Gly) as a substrate.

[0036] 1-3. Other enzymes In the production method of the present invention, when the plant-based milk is oat milk, α-amylase, hemicellulase, and / or maltotriosyltransferase can be used in combination with a protein deamidase to further enhance the dispersion stability and / or solubility of the oat milk. Among these combined enzymes, it is preferable to use at least α-amylase to further enhance the solubility of the oat milk.

[0037] In the production method of the present invention, when the plant-based milk is walnut milk, glucoamylase, α-amylase, β-amylase, transglucosidase, and / or glutaminase can be used in combination with the protein deamidation enzyme to further enhance the dispersion stability and / or solubility of the walnut milk.

[0038] In the manufacturing method of the present invention, the plant-based milk is peanut milk, and from the viewpoint of further improving the dispersion stability and / or solubility of the peanut milk, α-amylase, β-amylase, glucoamylase, and / or hemicellulase can be used in combination with the protein deamidation enzyme.

[0039] The origin of the α-amylase is not particularly limited, but examples include α-amylases from the Aspergillus genus (e.g., Aspergillus oryzae, Aspergillus niga, etc.) and the Bacillus genus (e.g., Bacillus amyloriquefaciens, Bacillus subtilis, Bacillus licheniformis, etc.), preferably α-amylases from the Bacillus genus, and more preferably α-amylases from the Bacillus amyloriquefaciens species.

[0040] Regarding the amount of α-amylase used, for example, 10 to 10,000 U, preferably 50 to 5,000 U, and more preferably 100 to 1,000 U per 100g of plant-based milk solids (referring to components derived from the edible parts of the plant used as raw material; the same applies hereinafter). Furthermore, regarding the amount of α-amylase used when processing oat milk, for example, 50 to 5,000 U, preferably 100 to 1,000 U, more preferably 200 to 500 U, and even more preferably 400 to 500 U per 100g of oat milk solids. Regarding the amount of α-amylase used when processing walnut milk, for example, 50 to 5,000 U, preferably 100 to 1,000 U, and more preferably 200 to 500 U per 100g of walnut milk solids. Regarding the amount of α-amylase used in processing peanut milk, for example, 50 to 10,000 U, preferably 100 to 8,000 U, and more preferably 200 to 7,000 U per 100 g of peanut milk solids.

[0041] Regarding α-amylase activity, one unit (1U) is defined as the amount of enzyme that, using soluble starch as a substrate, produces an increase in reducing power equivalent to 10 mg of glucose in 30 minutes.

[0042] The origin of the hemicellulase is not particularly limited, but examples include hemicellulases from the genera Trichoderma (e.g., Trichoderma longiblatiatum), Aspergillus, Aspergillus (e.g., Aspergillus oryzae, Aspergillus niga, etc.), and Bacillus (e.g., Bacillus amyloriquefaciens, Bacillus subtilis, Bacillus licheniformis, etc.). Preferably, hemicellulases from the genus Aspergillus are preferred, and more preferably, hemicellulases from Aspergillus niga are preferred.

[0043] Regarding the amount of hemicellulase to be used, for example, 100 to 100,000 U, preferably 1,000 to 50,000 U, and more preferably 5,000 to 20,000 U per 100g of plant-based milk solids. Furthermore, regarding the amount of hemicellulase to be used when processing oat milk, for example, 100 to 100,000 U, preferably 1,000 to 50,000 U, and more preferably 4,000 to 20,000 U per 100g of oat milk solids. Regarding the amount of hemicellulase to be used when processing peanut milk, for example, 100 to 100,000 U, preferably 1,000 to 50,000 U, and more preferably 8,000 to 20,000 U per 100g of peanut milk solids.

[0044] Regarding hemicellulase activity, one unit (1U) is defined as the amount of enzyme that uses xylose as a substrate and produces a reducing sugar equivalent to 1 mg of xylose per minute.

[0045] The origin of the maltotriosyltransferase is not particularly limited, but preferably it is a maltotriosyltransferase derived from the genus Aeribacillus.

[0046] Regarding the amount of maltotriosyltransferase used, for example, 10 to 5000 U, preferably 50 to 1000 U, is used per 100 g of plant-based milk solids. Furthermore, when used in the processing of oat milk, the amount of maltotriosyltransferase used is for example, 10 to 5000 U, preferably 50 to 1000 U, and more preferably 100 to 500 U, per 100 g of oat milk solids.

[0047] For maltotriosyltransferase activity, one unit (1U) is defined as the amount of enzyme that produces 1 μmol of glucose per minute using maltotetraose as a substrate.

[0048] The origin of the β-amylase is not particularly limited, but examples include β-amylase derived from plants (wheat, soybeans) and β-amylase derived from the genus Bacillus. Preferably, β-amylase derived from the genus Bacillus is preferred, and more preferably, β-amylase derived from the species Bacillus flexus is preferred.

[0049] Regarding the amount of β-amylase used, for example, 1 to 1000 U, preferably 5 to 500 U, and more preferably 10 to 150 U per 100 g of plant-based milk solids. Furthermore, regarding the amount of β-amylase used when processing walnut milk, for example, 5 to 500 U, 10 to 150 U, 10 to 60 U, preferably 20 to 50 U, and more preferably 30 to 40 U per 100 g of walnut milk solids. Regarding the amount of β-amylase used when processing peanut milk, for example, 5 to 500 U, 10 to 300 U, 30 to 150 U, preferably 40 to 120 U, more preferably 50 to 90 U, and even more preferably 60 to 70 U per 100 g of peanut milk solids.

[0050] Regarding β-amylase activity, one unit (1U) is defined as the amount of enzyme that produces an increase in reducing power equivalent to 1 mg of glucose per minute, using potato starch as a substrate.

[0051] The origin of the transglucosidase is not particularly limited, but preferably it is a transglucosidase derived from the genus Aspergillus, and more preferably it is a transglucosidase derived from the species Aspergillus niga.

[0052] Regarding the amount of transglucosidase used, for example, 100 to 300,000 U, preferably 1,000 to 100,000 U, and more preferably 5,000 to 50,000 U per 100g of plant-based milk solids. Furthermore, regarding the amount of transglucosidase used when processing walnut milk, for example, 1,000 to 100,000 U, preferably 5,000 to 50,000 U, and more preferably 10,000 to 30,000 U per 100g of walnut milk solids.

[0053] Regarding the activity of transglucosidase, one unit (1U) is defined as the amount of enzyme that produces 1 μg of glucose per minute using α-methyl-D-glucoside as a substrate.

[0054] The origin of the glutaminase is not particularly limited, but examples include glutaminases from the genera Cryptococcus, Bacillus, and Aspergillus. Preferably, glutaminases from the genus Bacillus are used, and more preferably, glutaminases from the species Bacillus amyloriquefaciens are used.

[0055] Regarding the amount of glutaminase to use, for example, 0.1 to 1000 GTU, preferably 0.2 to 100 GTU, and more preferably 0.5 to 20 GTU per 100 g of plant-based milk solids. Furthermore, regarding the amount of glutaminase to use when processing walnut milk, for example, 0.2 to 100 GTU, preferably 0.5 to 20 GTU, more preferably 1 to 10 GTU, even more preferably 3 to 8 GTU, and even more preferably 4 to 6 GTU per 100 g of walnut milk solids.

[0056] Regarding glutaminase activity, one unit (1 GTU) is defined as the amount of enzyme that produces 1 μmol of L-glutamic acid per minute using L-glutamine as a substrate.

[0057] The origin of the glucoamylase is not particularly limited, but examples include glucoamylases derived from the Aspergillus or Rhizopus genera. Preferably, glucoamylases derived from Aspergillus niga or Rhizopus esp are used, and glucoamylases derived from Rhizopus esp are particularly preferred.

[0058] Regarding the amount of glucoamylase to be used, for example, 1 to 10,000 U, preferably 10 to 5,000 U, more preferably 20 to 2,000 U, and even more preferably 30 to 800 U per 100 g of plant-based milk solids. Furthermore, regarding the amount of glucoamylase to be used when processing walnut milk, for example, 10 to 2,000 U, preferably 20 to 1,000 U, more preferably 30 to 200 U, even more preferably 50 to 150 U, even more preferably 60 to 120 U, and even more preferably 70 to 90 U per 100 g of walnut milk solids. Regarding the amount of glucoamylase to be used when processing peanut milk, for example, 20 to 2,000 U, preferably 100 to 800 U, more preferably 120 to 600 U, even more preferably 140 to 400 U, and even more preferably 150 to 350 U per 100 g of peanut milk solids.

[0059] Regarding glucoamylase activity, one unit (1U) is defined as the amount of enzyme that produces an increase in reducing power equivalent to 1 mg of glucose per minute, using potato starch as a substrate.

[0060] 1-4. Reaction conditions, etc. In the process of treating plant-based milk with a protein deamidation enzyme, a plant-based milk composition containing plant-based milk and a protein deamidation enzyme, or plant-based milk, a protein deamidation enzyme, and other enzymes, is prepared by adding the protein deamidation enzyme to the plant-based milk, and optionally adding the protein deamidation enzyme and other enzymes to the plant-based milk. The enzymatic treatment reaction can then be carried out by maintaining the plant-based milk composition under heating conditions.

[0061] The heating temperature (enzyme treatment reaction temperature) of the plant-based milk composition is not particularly limited and can be appropriately determined by those skilled in the art depending on the optimal temperature of the enzyme used and / or the thermal properties of the plant-based milk, but examples include 40 to 70°C.

[0062] For example, if the plant-based milk is oat milk, the heating temperature of the oat milk composition is preferably 50-70°C, more preferably 55-65°C, and even more preferably 58-62°C. If the plant-based milk is walnut milk, the heating temperature of the walnut milk composition is preferably 40-60°C, more preferably 45-55°C, and even more preferably 48-52°C. If the plant-based milk is peanut milk, the heating temperature of the peanut milk composition is preferably 40-60°C, more preferably 45-55°C, and even more preferably 48-52°C.

[0063] The enzyme treatment reaction time for the plant-based milk composition is not particularly limited and can be appropriately determined according to the production scale of the composition, etc., but for example, 0.5 hours or more, preferably 1 hour or more. There is no particular upper limit to the range of the enzyme treatment reaction time, but for example, 24 hours or less, 12 hours or less, 8 hours or less, or 6 hours or less.

[0064] The enzyme treatment reaction can be terminated by enzyme inactivation treatment using high heat. Examples of enzyme inactivation treatment temperatures include 85°C or higher, preferably 90°C or higher, and examples of enzyme inactivation treatment times include 5 to 25 minutes, preferably 10 to 20 minutes.

[0065] The plant-based milk composition after enzyme treatment can be further processed, such as by filtration, as needed, to obtain processed plant-based milk. Compared to the plant-based milk before enzyme treatment, processed plant-based milk can be obtained with improved dispersion stability and / or solubility.

[0066] 2. Uses and applications of protein deamidase enzymes As described above, protein deenzyme amides can improve the dispersion stability and / or solubility stability of plant-based milks. Accordingly, the present invention also provides the use of protein deenzyme enzymes for the production of dispersion stability improvers for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk; and the use of protein deenzyme enzymes for the production of solubility improvers for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk, and also provides a dispersion stability improver for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk, comprising a protein deenzyme enzyme; and a solubility improver for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk, comprising a protein deenzyme enzyme.

[0067] Furthermore, protein deenzyme amides can improve the texture of peanut milk. Therefore, the present invention also provides the use of protein deenzyme enzymes for the production of peanut milk texture enhancers, and also provides peanut milk texture enhancers containing protein deenzyme enzymes.

[0068] The types and amounts of ingredients used in the above-mentioned usage, dispersion stability enhancers, solubility enhancers, and texture enhancers are as shown in section 1, "Method for Manufacturing Processed Vegetable Milk." [Examples]

[0069] The present invention will be described in detail below with reference to examples, but the present invention is not limited to the following examples.

[0070] Enzymes used Details of the enzymes used in the following test examples are as follows:

[0071] [Table 1]

[0072] The activity of protein deamidation enzymes (protein glutaminases) was measured using the following method. (1) 1 ml of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly was mixed with 0.1 ml of aqueous solution containing protein deamidase, incubated at 37°C for 10 minutes, and then 1 ml of 0.4 M TCA solution was added to stop the reaction. As a blank, 1 ml of 0.4 M TCA solution was mixed with 1 ml of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly, followed by 0.1 ml of aqueous solution containing protein deamidase, and incubated at 37°C for 10 minutes. (2) The amount of ammonia produced in the reaction solution was measured using the ammonia tester Wako (Wako Pure Chemical Industries) on the solution obtained in (1). The ammonia concentration in the reaction solution was determined from a calibration curve showing the relationship between ammonia concentration and absorbance (630 nm) prepared using ammonia standard solution (ammonium chloride). (3) The activity of the protein deamide enzyme was calculated using the following formula, with the amount of enzyme that produces 1 μmol of ammonia per minute defined as 1 unit (1 U). In the formula, the volume of the reaction solution is 2.1, the volume of the enzyme solution is 0.1, and Df is the dilution ratio of the enzyme solution. Also, 17.03 is the molecular weight of ammonia.

[0073] [ka]

[0074] Evaluation of dispersion stability and solubility The dispersion stability and solubility of the processed plant-based milk prepared in the following test examples were evaluated using Brix, A280, A660, CSR, and instability indices measured by LUMiSizer651. Brix represents the total amount of soluble components, including solubilized proteins and other soluble components, and is used as an evaluation index for "solubility." A280 represents the amount of soluble protein and is used as an evaluation index for "solubility." A660 represents the amount of insoluble components, more specifically, the amount of insoluble components that did not precipitate in the supernatant after centrifugation of plant-based milk, and is used as an evaluation index for "dispersion stability" as it is considered to have a positive correlation with "dispersion stability." CSR represents the amount of insoluble components that precipitated by centrifugation and is used as an evaluation index for "dispersion stability" as it is considered to have a negative correlation with "dispersion stability." The instability index measured by LUMiSizer651 is an index based on the behavior of particles during centrifugation and is used as an evaluation index for "dispersion stability." Details of the measurement conditions for each evaluation item are described in each test example.

[0075] Test Example 1 (1) Preparation of oat milk 300g of oats were mixed with 1200ml of 80°C hot water and processed in a colloid mill for 30 minutes to obtain an oat slurry. Warm water was added to the oat slurry to make up 1800g (total amount of water to 1 part by weight of oats is 5 parts by weight), and it was heated at 90°C for 15 minutes. After that, coarse fibers were removed by passing it through a 60-mesh sieve, and it was cooled to 60°C to prepare oat milk. The prepared oat milk was divided into smaller portions while stirring.

[0076] (2) Enzyme treatment The enzymes shown in Tables 2 and 3 were added in the indicated amounts and reacted at 60°C for 3 hours. After enzyme inactivation treatment at 90°C for 15 minutes, the mixture was stirred and filtered through a sieve (100 mesh) to obtain processed oat milk.

[0077] (3) Evaluation The processed oat milk obtained was subjected to measurements of pH (25°C), presence or absence of thermal coagulation, Brix, A280, A660, CSR, and instability index, and further sensory evaluation regarding sweetness was conducted. The results are shown in Tables 2 and 3.

[0078] <Presence or absence of thermal coagulation> Processed oat milk was heat-treated at 95°C for 15 minutes, then allowed to cool to room temperature. The presence or absence of coagulation in the processed oat milk was visually checked.

[0079] <brix> The Brix (%) of processed oat milk was measured.

[0080] <a280> Processed oat milk was centrifuged at 4000 rpm for 10 minutes, the supernatant was diluted 100-fold, and the absorbance at 280 nm was measured.

[0081] <a660> Processed oat milk was centrifuged at 4000 rpm for 10 minutes, the supernatant was diluted 10-fold, and the absorbance at 660 nm was measured.

[0082] <csr> 1.6 ml of processed oat milk was centrifuged at 4000 rpm for 10 minutes, and the weight of the precipitate was measured.

[0083] <Instability index> The instability index was measured using a LUMiSizer 651 under the following conditions: 4000 rpm (RCA 2100g), 25°C, 865 nm, 300 profiles, interval 10 s, and light factor 1. A smaller instability index indicates better dispersion stability.

[0084] <Sensory evaluation regarding sweetness> The sweetness of processed oat milk was evaluated based on the following evaluation criteria. -: Lower sweetness than the processed oat milk in Comparative Examples 1-2 (no sweetness detected). +: Similar sweetness to the processed oat milk in Comparative Examples 1-2 (slightly sweet). ++: Sweeter than the processed oat milk in Comparative Example 1-2 (sweeter)

[0085] [Table 2]

[0086] [Table 3]

[0087] Test Example 2 (1) Preparation of walnut milk 350g of walnuts were soaked in a 1 w / w% sodium hydroxide solution and heated (above 70°C) for 10 minutes while stirring. The sodium hydroxide solution was discarded, the walnuts were peeled, washed with water, and drained. 1000ml of hot water (80°C) was added to 300g of the peeled walnuts, and the mixture was treated in a colloid mill for 30 minutes to obtain a walnut slurry. The walnut slurry was heated at 90°C for 15 minutes, then cooled to 50°C, and diluted with warm water to a total volume of 1500g to prepare walnut milk. The prepared walnut milk was divided into smaller portions while stirring.

[0088] (2) Enzyme treatment The enzymes shown in Tables 4 and 5 were added in the indicated amounts and reacted at 50°C for 3 hours. After enzyme deactivation at 90°C for 15 minutes, the mixture was stirred and filtered through a sieve (100 mesh) to obtain processed walnut milk.

[0089] (3) Evaluation The pH (25°C), Brix, A280, and instability index of the obtained processed walnut milk were measured. The results are shown in Tables 4 and 5.

[0090] <brix> Processed walnut milk was centrifuged at 14,000 rpm for 10 minutes, and the Brix (%) of the supernatant was measured.

[0091] <a280> Processed walnut milk was centrifuged at 14,000 rpm for 10 minutes, the supernatant was diluted 50-fold, and the absorbance at 280 nm was measured.

[0092] <Instability index> The instability index was measured using a LUMiSizer 651 under the following conditions: 4000 rpm (RCA 2100g), 25°C, 865 nm, 300 profiles, interval 10 s, and light factor 1.

[0093] [Table 4]

[0094] [Table 5]

[0095] Test Example 3 (1) Preparation of peanut milk 350g of shelled peanuts were placed in 1400mL of brine (0.8w / v%) and boiled for 5 minutes. After cooling and draining, foreign matter (such as skins) was removed. The shelled peanuts were roasted in an electric oven at 150°C for 40 minutes, then cooled and foreign matter was removed. 300g of the obtained peanuts were weighed, 1800mL of warm water (50°C) was added, and the mixture was treated in a colloidal mill for 40 minutes to obtain a peanut slurry. 1700mL of water was added to the peanut slurry, boiled for 5 minutes, cooled to room temperature, adjusted to pH 6.0 (0.5M citric acid), and then diluted to 3750mL with warm water. This prepared peanut milk (pH 6.0) with a protein content of 2w / v% and a solids content of 8w / v%. The prepared peanut milk was divided into 500ml portions while stirring.

[0096] (2) Enzyme treatment The enzymes shown in Table 6 were added in the indicated amounts and reacted at 50°C for 1 hour, 3 hours, or 6 hours. After enzyme inactivation treatment at 90°C for 15 minutes, the mixture was stirred and filtered through a sieve (100 mesh) to obtain processed peanut milk.

[0097] (3) Evaluation The pH (25°C), soluble protein concentration (mg / mL), protein solubilization rate (%), Brix (%), A280, instability index, and viscosity (cp) of the obtained processed peanut milk were measured. The results are shown in Table 6.

[0098] <Soluble protein concentration> Processed peanut milk was centrifuged (16000 × g, 10 minutes), and the soluble protein concentration was measured using the Lowry method with the supernatant filtrate (filtered through a 0.45 μm filter).

[0099] <Protein solubilization rate> The percentage of soluble protein (by weight) was calculated, assuming that the total protein content of processed peanut milk was 100% by weight.

[0100] <brix> The Brix (%) of processed peanut milk was measured.

[0101] <a280> Processed peanut milk was centrifuged at 16,000 g for 10 minutes, the supernatant was diluted 100-fold, and the absorbance at 280 nm was measured.

[0102] <Instability index> Using a LUMiSizer 651, the instability index was obtained as a measurement taken at 96 seconds (equivalent to 56 hours elapsed: 96 seconds × 2100(g) / 3600s) at 4000 rpm, (RCA 2100g), 25℃, 470nm, 180 profiles, interval 10s, and light factor 1.

[0103] <Viscosity> The viscosity of processed peanut milk was measured using an automatic viscometer, RVA techmaster, at 25°C and 800 rpm.

[0104] [Table 6]

[0105] Test Example 4 The peanut milk prepared in the above-mentioned Test Example 3, Example 3-2, was evaluated for any improvement in texture (smoothness) compared to Comparative Example 3-1. As a result, an improvement in the texture of the peanut milk was observed. < / brix> < / brix> < / csr> < / brix>

Claims

1. A method for producing processed plant-based milk, comprising the step of treating plant-based milk with a protein deamidation enzyme.

2. The method for producing processed plant-based milk according to claim 1, wherein the plant-based milk is selected from the group consisting of oat milk, walnut milk, and peanut milk.

3. A method for producing processed plant milk according to claim 1 or 2, wherein the protein deamidation enzyme is used at a concentration of 0.5 U or more per gram of plant protein.

4. The method for producing processed plant milk according to claim 1, wherein the plant milk is the oat milk, and the protein deamidation enzyme is used at a rate of 1.5 U or more per gram of oat protein.

5. A method for producing processed plant milk according to claim 1 or 2, wherein the plant milk is the oat milk, and α-amylase, hemicellulase, and / or maltotriosyltransferase are used in combination with the protein deamidase.

6. The method for producing processed plant milk according to claim 1, wherein the plant-based milk is the walnut milk, the processed plant-based milk is walnut milk with improved dispersion stability than the walnut milk, and the protein deamidation enzyme is used in amounts of 3 U or more per gram of walnut protein.

7. The method for producing processed plant milk according to claim 1, wherein the plant-based milk is the walnut milk, the processed plant-based milk is walnut milk with improved solubility than the walnut milk, and the protein deamidation enzyme is used at a rate of 1 U or more per gram of walnut protein.

8. A method for producing processed plant milk according to claim 1, wherein the plant milk is the walnut milk, and glucoamylase, α-amylase, β-amylase, transglucosidase, and / or glutaminase are used in combination with the protein deamidation enzyme.

9. The method for producing processed plant milk according to claim 1, wherein the plant-based milk is the peanut milk, the processed plant-based milk is peanut milk with improved dispersion stability than the peanut milk, and the protein deamidation enzyme is used at a rate of 2 U or more per gram of peanut protein.

10. The method for producing processed plant milk according to claim 1, wherein the plant-based milk is the peanut milk, the processed plant-based milk is peanut milk with improved solubility than the peanut milk, and the protein deamidation enzyme is used at a rate of 1 U or more per gram of peanut protein.

11. The method for producing processed plant milk according to claim 1, wherein the plant-based milk is the peanut milk, the processed plant-based milk is peanut milk with improved texture than the peanut milk, and the protein deamidation enzyme is used at a rate of 2 U or more per gram of peanut protein.

12. A method for producing processed plant milk according to claim 1, wherein the plant-based milk is the peanut milk, and α-amylase, β-amylase, glucoamylase, and / or hemicellulase are used in combination with the protein deamidation enzyme.

13. Use of a protein deamidase for the production of a dispersion stability enhancer for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk.

14. Use of a protein deamidase for the production of a solubility enhancer for plant-based milks selected from the group consisting of oat milk, walnut milk, and peanut milk.

15. Use of protein deamidation enzymes in the production of texture enhancers for peanut milk.

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