Enzyme agent for reducing pink feeling of vegetable food and drink or vegetable food and drink material

Abstract

Provided is a vegetable milk having a reduced pink feeling and a changed taste. The present technology provides: an enzyme agent for reducing the pink feeling of a plant food or beverage or a plant food or beverage material, which contains a cell wall polysaccharide-degrading enzyme; and a plant food or beverage or a plant food or beverage material, which uses the enzyme agent for reducing the pink feeling. Also provided are: a method for producing a plant food or beverage or a plant food or beverage material, said method comprising a step for causing a cell wall polysaccharide-decomposing enzyme to act on a plant raw material; and a method for reducing the pink feeling of a plant food or beverage or beverage material.

Description

Technical Field

[0001] The present technology relates to an enzyme for reducing the powdery feel of a plant-based food or beverage or a plant-based food or beverage material. More specifically, the present technology relates to an enzyme for reducing the powdery feel of a plant-based food or beverage or a plant-based food or beverage material, a plant-based food or beverage or a plant-based food or beverage material using the enzyme for reducing the powdery feel, a method for manufacturing a plant-based food or beverage or a plant-based food or beverage material, and a method for reducing the powdery feel of a plant-based food or beverage or a plant-based food or beverage material. Background Art

[0002] In recent years, due to the rise of health awareness, plant-based food and beverages have attracted much attention. Unlike animal milk, which is a secretion, plant-based milk is prepared by dispersing crushed plant tissues in water. Therefore, as a basic characteristic of a substitute for animal milk, it is required to have a smooth taste. In addition, when making plant-based milk, the preferred flavor from the raw materials is sometimes damaged, and sometimes the overall flavor is adversely affected. Therefore, in plant-based milk, it is required to improve the taste and flavor.

[0003] Various technologies have been studied for the purpose of giving plant milk properties closer to animal milk. For example, Patent Document 1 describes a technology for producing a cereal liquefied product that combines low viscosity and suppressed free sugar content by allowing 4-α-glucanotransferase, cellulase and peptidase to act on oats. Patent Document 2 describes a technology for producing an oat beverage with moderate sweetness by using one or more amylases to decompose the starch of an oat material. Patent Document 3 describes a technology for changing the nutritional components by allowing an enzyme to act on oat bran, and improving the taste of a food composition by a micronization process. Patent Document 4 describes a rice dispersion treated with protease, cellulase, pullulanase and α-amylase to produce a rice liquefied product that is not sweet and rich in oligosaccharides, and the taste of the rice liquefied product becomes smooth by further homogenization.

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: Japanese Patent Application Publication No. 2021-40553

[0007] Patent Document 2: Japanese Patent Application Publication No. 2018-075029

[0008] Patent Document 3: Japanese Patent Application Publication No. 2015-84687

[0009] Patent Document 4: Japanese Patent Application Publication No. 2004-121135 Summary of the invention

[0010] The compositions of plant-based milk and animal-based milk are different, so there is room for improvement to obtain satisfactory properties. For example, in order to achieve a smooth texture, special devices such as micronization treatment and homogenization treatment are required, so a simpler method is desired. In addition, regarding flavor, due to the diversification of demands and the trend towards health, a method of changing the flavor without adding unnecessary raw materials is also desired.

[0011] Therefore, in the present technology, the main object is to provide a plant-based milk with reduced powdery feeling and changed flavor.

[0012] The inventors of the present application conducted in-depth research and found that the powdery feeling can be reduced by allowing cell wall polysaccharide-degrading enzymes to act on plant-based milk. The present technology is based on this insight and has been completed through further repeated research.

[0013] That is, in the present technology, first, an enzyme agent for reducing the powdery feeling of plant-based food or plant-based food material containing cell wall polysaccharide-degrading enzyme is provided.

[0014] In the present technology, a plant-based food or plant-based food material using the enzyme agent for reducing the powdery feeling according to the present technology is also provided.

[0015] The plant-based food of the present technology can be oat milk or rice milk.

[0016] In the present technology, next, a manufacturing method of a plant-based food or plant-based food material including a step of allowing a cell wall polysaccharide-degrading enzyme to act on a plant-based raw material is provided.

[0017] In the present technology, a method for reducing the powdery feeling of a plant-based food or plant-based food material including a step of allowing a cell wall polysaccharide-degrading enzyme to act on a plant-based raw material is further provided.

[0018] According to the present technology, a plant-based food or plant-based food material with reduced powdery feeling and changed flavor is provided. Detailed Embodiments

[0019] Hereinafter, preferred modes for implementing the present technology will be described. In addition, the embodiments described below show an example of representative embodiments of the present technology, but the scope of the present technology will not be construed narrowly thereby.

[0020] 1. Enzyme Agent for Reducing Powdery Feeling

[0021] The enzyme agent for reducing the powdery feeling according to the present technology contains cell wall polysaccharide-degrading enzyme as an active ingredient. Carbohydrates are composed of saccharides containing polysaccharides such as starch, oligosaccharides, monosaccharides, disaccharides, etc. and dietary fibers containing cell wall polysaccharides, etc. Cell wall polysaccharides include cellulose, hemicellulose, pectin, etc., and the enzyme that decomposes these cell wall polysaccharides is cell wall polysaccharide-degrading enzyme.

[0022] As the cell wall polysaccharide-degrading enzyme that can be used in the present technology, as long as it is an enzyme having the function of degrading cell wall polysaccharides, there is no particular limitation, and one or more cell wall polysaccharide-degrading enzymes that can be used in the production of food and beverages can be freely combined without impairing the effects of the present technology. As the cell wall polysaccharide-degrading enzyme, for example, cellulase, hemicellulase, pectinase, etc. can be cited. Among them, cellulase is particularly preferably used as the enzyme agent for reducing the powdery feeling in the present technology. Hereinafter, the enzyme and the like that can be used for the enzyme agent for reducing the powdery feeling related to the present technology will be described in detail.

[0023] (1) Cellulase

[0024] The cellulase that can be used in the present technology is an enzyme that hydrolyzes the glycosidic bond of β-1,4-glucan and decomposes cellulose, which is one of the cell wall polysaccharides. As the cellulase that can be used in the present technology, as long as it has cellulase activity, it can also be an enzyme having other functions. In the present technology, by allowing cellulase to act on plant raw materials, the powdery feeling in the produced plant-based food and beverages can be reduced.

[0025] The source of the cellulase that can be used in the present technology is not particularly limited. For example, it can be cited from microorganisms belonging to the genus Aspergillus such as Aspergillus niger, Aspergillus oryzae, Aspergillus sojae, Aspergillus saitoi, Aspergillus awamori, Aspergillus flavus; the genus Trichoderma such as Trichoderma reesei, Trichoderma viride; the genus Acremonium such as Acremonium cellulolyticus, etc. These cellulases can be used alone or in combination of multiple kinds. Among them, from the viewpoint of reducing the powdery feeling of plant raw materials, the cellulase is more preferably a cellulase from microorganisms of the genus Trichoderma, particularly Trichoderma viride.

[0026] "Cellulase from Trichoderma viride" here means cellulase produced by a microorganism classified as Trichoderma viride (which can be a wild strain or a mutant strain), or cellulase obtained by genetic engineering methods using a cellulase gene. Therefore, a recombinant produced by a host microorganism into which a cellulase gene (or a gene obtained by modifying the gene) derived from Trichoderma viride has been introduced is also equivalent to "cellulase from Trichoderma viride".

[0027] There is no particular limitation on the type of cellulase used. For example, it can be any one of endoglucanase, exoglucanase, or a mixture thereof. As the cellulase used in the present invention, an enzyme preparation containing both endoglucanase and exoglucanase is preferably selected.

[0028] The cellulase used in this technology can be prepared from the culture broth of the microorganism that is the source of the above cellulase. As a specific preparation method, a method of recovering cellulase from the culture broth or cells of the above microorganism can be cited. For example, in the case of using a cellulase-secreting microorganism, after recovering the cells from the culture broth by pre-filtration, centrifugation, etc. as needed, the enzyme can be separated and / or purified. In addition, in the case of using a cellulase non-secreting microorganism, after recovering the cells from the culture broth in advance as needed, the cells are disrupted by pressure treatment, ultrasonic treatment, etc. to expose the enzyme, and then the enzyme can be separated and / or purified. As the method for separating and / or purifying the enzyme, publicly known protein separation and / or purification methods can be used without particular limitation. For example, centrifugation, UF concentration, salting out, various chromatography methods using ion exchange resins, etc. can be cited. The separated and / or purified enzyme can be powdered by drying methods such as freeze-drying and vacuum drying. In addition, appropriate excipients and / or drying aids can also be used in this drying method for powdering. In addition, the separated and / or purified enzyme can also be made into a liquid state by adding appropriate additives and performing filtration sterilization.

[0029] In this technology, commercially available products can also be used as the cellulase. As an example of a preferred commercially available product, cellulase from Trichoderma viride manufactured by Amano Enzyme Inc. can be cited.

[0030] Without impairing the technical effects of the present technology, the content of cellulase in the enzyme agent for reducing powdery feeling involved in the present technology can be freely set. With respect to 1 g of plant raw materials used in the manufactured plant-based food or plant-based food materials, the content of cellulase can be set, for example, to 0.01 U or more, and from the viewpoint of further improving the powdery feeling reduction effect, it can preferably be set to 0.05 U or more, more preferably to 0.1 U or more, further preferably to 0.3 U or more, and even more preferably to 0.5 U or more.

[0031] Without impairing the technical effects of the present technology, the upper limit of the content of cellulase is not particularly limited, and with respect to 1 g of plant raw materials used in the manufactured plant-based food or plant-based food materials, it can be set, for example, to 500 U or less, 200 U or less, 100 U or less, 50 U or less, 30 U or less, 20 U or less, or 10 U or less.

[0032] In addition, in the present technology, the activity of cellulase is a value defined by the following method.

[0033] [Definition of the activity of cellulase]

[0034] Regarding the activity of cellulase, the amount of enzyme that increases the reducing power equivalent to 1 μmole of glucose in 1 minute is defined as 1 unit (1 U). In the present technology, the activity of cellulase is a value measured by the cellulase activity measurement method described in the following examples.

[0035] (2) Hemicellulase

[0036] The hemicellulase that can be used in the present technology is an enzyme that hydrolyzes hemicellulose, which is one of the cell wall polysaccharides. Hemicellulose is a heteropolysaccharide composed of various constituent sugars. Generally, for the main chain from which it gets its name, other constituent sugars have a branched structure that forms side chains. As a specific example of hemicellulose, for example, mannan, β-1,3-1,4-glucan, glucomannan, xylan, xyloglucan, glucuronoxylan, etc. can be cited. The enzyme that decomposes these hemicelluloses is hemicellulase.

[0037] As specific examples of hemicellulase, for example, xylanase, galactosidase, mannanase, galactomannanase, arabinase, β-glucanase, etc. can be cited. These can be used alone or in combination of two or more. As the hemicellulase that can be used in the present technology, as long as it has hemicellulase activity, it can also be an enzyme that has other functions. In the present technology, by allowing hemicellulase to act on plant raw materials, the powdery feeling in the manufactured plant-based food can be reduced.

[0038] The source of the hemicellulase that can be used in the present technology is not particularly limited. For example, it can be derived from microorganisms such as basidiomycetes (genus Corticium, genus Pycnoporus), filamentous fungi (genus Aspergillus, genus Humicola, genus Penicillium, genus Trichoderma), actinomycetes (genus Streptomyces), or bacteria (genus Bacillus), etc., or it can also be a synthetic enzyme. Additionally, they can be used individually or in combination of two or more. Among them, in the present technology, those derived from filamentous fungi are preferred, those derived from microorganisms of the genus Aspergillus are more preferred, and hemicellulase derived from Aspergillus niger is further preferred.

[0039] The hemicellulase used in the present technology can be prepared from the culture broth of the microorganisms that are the source of the above hemicellulase. As a specific preparation method, a method of recovering hemicellulase from the culture broth or cells of the above microorganisms can be cited. For example, in the case of using hemicellulase-secreting microorganisms, after recovering the cells from the culture broth by pre-filtration, centrifugation, etc. as needed, the enzyme can be separated and / or purified. Additionally, in the case of using hemicellulase-non-secreting microorganisms, after recovering the cells from the culture broth in advance as needed, the cells are disrupted by pressure treatment, ultrasonic treatment, etc. to expose the enzyme, and then the enzyme can be separated and / or purified. As the method for separating and / or purifying the enzyme, publicly known protein separation and / or purification methods can be used without particular limitation. For example, centrifugation, UF concentration, salting out, various chromatography methods using ion exchange resins, etc. can be cited. The separated and / or purified enzyme can be powdered by drying methods such as freeze-drying and vacuum drying. Additionally, appropriate excipients and / or drying aids can also be used in this drying method for powdering. Additionally, the separated and / or purified enzyme can also be made into a liquid state by adding appropriate additives and performing filtration sterilization.

[0040] In the present technology, commercially available products can also be used as hemicellulase. As an example of a preferred commercially available product, hemicellulase derived from Aspergillus niger manufactured by Amano Enzyme Inc. can be cited.

[0041] Without impairing the technical effects of the present technology, the content of hemicellulase in the enzyme preparation for reducing powdery feeling involved in the present technology can be freely set. With respect to 1 g of plant raw material used in the manufactured plant-based food or plant-based food material, the content of hemicellulase can be set, for example, to 1 U or more. From the viewpoint of further improving the effect of reducing powdery feeling, it can preferably be set to 10 U or more, more preferably to 50 U or more, further preferably to 100 U or more, and even more preferably to 200 U or more.

[0042] Without impairing the technical effects of the present technology, the upper limit of the content of hemicellulase is not particularly limited either. With respect to 1 g of plant raw material used in the manufactured plant-based food or plant-based food material, it can be set, for example, to 100,000 U or less, 50,000 U or less, 10,000 U or less, 5,000 U or less, 2,000 U or less, or 1,000 U or less.

[0043] In addition, in the present technology, the activity of hemicellulase is a value defined by the following method.

[0044] [Definition of the activity of hemicellulase]

[0045] The amount of enzyme that generates reducing sugars equivalent to 1 mg of xylose in 1 minute using xylan as a substrate is taken as 100 units (100 U).

[0046] (3) Pectinase

[0047] The pectinase that can be used in the present technology is an enzyme that hydrolyzes pectin, which is one of the cell wall polysaccharides. Pectin is a complex polysaccharide mainly composed of polygalacturonic acid in which galacturonic acid is α-1,4-bonded. Specific examples of pectinase include, for example, polygalacturonase, pectin lyase, pectin methyl esterase, etc., and polygalacturonase is preferably cited. As the pectinase that can be used in the present technology, as long as it has pectinase activity, it can also be an enzyme that has other functions.

[0048] In the present technology, by allowing pectinase to act on plant raw materials, the powdery feeling in the manufactured plant-based food can be reduced.

[0049] The source of pectinase that can be used in the present technology is not particularly limited. For example, it can be derived from basidiomycetes (genus Corticium), filamentous fungi (genus Aspergillus, genus Rhizopus, and genus Trichoderma), yeasts (genus Geotrichum and genus Trichosporon), actinomycetes (genus Streptomyces), or bacteria (genus Bacillus) and other microorganisms, or can be a synthetic enzyme. Additionally, they can be used alone or in combination of two or more. Among them, in the present technology, pectinase derived from filamentous fungi is preferred, and pectinase derived from the genus Aspergillus is more preferred.

[0050] The pectinase used in the present technology can be prepared from the culture broth of the microorganisms that are the source of the above pectinase. As a specific preparation method, a method of recovering pectinase from the culture broth or cells of the above microorganisms can be cited. For example, in the case of using pectinase-secreting microorganisms, after recovering the cells from the culture broth by pre-filtration, centrifugation, etc. as needed, the enzyme can be separated and / or purified. Additionally, in the case of using pectinase-non-secreting microorganisms, after recovering the cells from the culture broth in advance as needed, the cells are disrupted by pressure treatment, ultrasonic treatment, etc. to expose the enzyme, and then the enzyme can be separated and / or purified. As the method for separating and / or purifying the enzyme, publicly known protein separation and / or purification methods can be used without particular limitation, such as centrifugation, UF concentration, salting out, various chromatography methods using ion exchange resins, etc. The separated and / or purified enzyme can be powdered by drying methods such as freeze-drying and vacuum drying. Additionally, appropriate excipients and / or drying aids can also be used in this drying method for powdering. Additionally, the separated and / or purified enzyme can also be liquefied by adding appropriate additives and performing filtration sterilization.

[0051] In the present technology, commercially available products can also be used as pectinase. As an example of a preferred commercially available product, pectinase derived from the genus Aspergillus manufactured by Amano Enzyme Inc. can be cited.

[0052] Without impairing the effects of the present technology, the content of pectinase in the enzyme agent for reducing powdery feeling according to the present technology can be freely set. Relative to 1 g of plant raw material used in the manufactured plant-based food or plant-based food material, the content of pectinase can be set, for example, to 0.01 U or more. From the perspective of further improving the effect of reducing powdery feeling, it can preferably be set to 0.1 U or more, more preferably to 0.5 U or more, further preferably to 1 U or more, and even more preferably to 2 U or more.

[0053] Without impairing the technical effects of the present invention, the upper limit of the content of pectinase is not particularly limited, and can be set, for example, to 2000 U or less, 500 U or less, 100 U or less, 50 U or less, 40 U or less, or 30 U or less per 1 g of vegetable raw material used in the produced vegetable food or vegetable food material.

[0054] In addition, in the present technology, the activity of pectinase is a value defined by the following method.

[0055] [Definition of the activity of pectinase]

[0056] The amount of enzyme that reduces the viscosity by 50% in 1 minute using low-methoxyl (LM) pectin as a substrate is defined as 1 unit (1 U).

[0057] (4) Glucoamylase

[0058] Glucoamylase may also be contained in the enzyme agent for reducing powdery feeling according to the present technology. The glucoamylase that can be used in the present technology is an enzyme having the activity of hydrolyzing the α-1,4-glycosidic bond of carbohydrates such as starch in units of glucose from the non-reducing end. As the glucoamylase that can be used in the present technology, as long as it has glucoamylase activity, it may also be an enzyme having other functions.

[0059] In the present technology, by allowing glucoamylase to act on vegetable raw materials, the sweetness of the produced vegetable food or vegetable food material can be enhanced.

[0060] The source of the glucoamylase that can be used in the present technology is not particularly limited, and examples include glucoamylases derived from the genus Aspergillus and the genus Rhizopus. These glucoamylases can be used alone or in combination. Among these glucoamylases, glucoamylases derived from Rhizopus, and more preferably glucoamylases derived from Rhizopus oryzae, are also exemplified.

[0061] Here, "glucoamylase derived from Rhizopus oryzae" means glucoamylase produced by a microorganism classified as Rhizopus oryzae (which may be a wild strain or a mutant strain), or glucoamylase obtained by genetic engineering using a glucoamylase gene. Therefore, a recombinant produced by a host microorganism into which a glucoamylase gene (or a gene obtained by modifying the gene) derived from Rhizopus oryzae has been introduced is also equivalent to "glucoamylase derived from Rhizopus oryzae".

[0062] The glucoamylase that can be used in the present technology can be prepared from the culture broth of the microorganism that is the source of the above glucoamylase. As a specific preparation method, it can be easily prepared by culturing glucoamylase-producing bacteria and separating glucoamylase by publicly known means, as well as by methods using transgenic technology, etc.

[0063] In the present technology, as the glucoamylase, commercially available products can also be used. As an example of a preferred commercially available product, glucoamylase from Rhizopus oryzae manufactured by Amano Enzyme Inc. can be cited.

[0064] Without impairing the effects of the present technology, the content of glucoamylase in the enzyme agent for reducing powdery feeling according to the present technology can be freely set. With respect to 1 g of plant raw material used in the manufactured plant-based food or plant-based food material, the content of glucoamylase can be set, for example, to 0.01 U or more, preferably 0.1 U or more, more preferably 1 U or more, further preferably 3 U or more, and even more preferably 5 U or more.

[0065] Without impairing the effects of the present technology, the upper limit of the content of glucoamylase is not particularly limited either. With respect to 1 g of plant raw material used in the manufactured plant-based food or plant-based food material, it can be set, for example, to 5000 U or less, 1000 U or less, 500 U or less, 200 U or less, 100 U or less, or 50 U or less.

[0066] In addition, in the present technology, the activity of glucoamylase is a value defined by the following method.

[0067] [Definition of the activity of glucoamylase]

[0068] The amount of enzyme that increases the reducing power equivalent to 1 mg of glucose in 1 minute using potato starch as the substrate is defined as 1 unit (1 U).

[0069] (5) α-Amylase

[0070] α-Amylase can also be used in the enzyme agent for reducing powdery feeling according to the present technology. The α-amylase that can be used in the present technology is an enzyme that acts on starch and mainly hydrolyzes α-1,4 glycosidic bonds. As the α-amylase that can be used in the present technology, as long as it has α-amylase activity, it can also be an enzyme that has other functions.

[0071] In the present technology, by allowing α-amylase to act on plant raw materials, the starch in the plant raw materials used in plant-based foods or plant-based food materials can be liquefied to improve solubility.

[0072] The source of the α-amylase that can be used in the present technology is not particularly limited. For example, α-amylases of biological origin belonging to the genus Aspergillus (e.g., Aspergillus oryzae, Aspergillus niger, etc.), the genus Bacillus (e.g., Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, etc.) can be cited. Preferred are α-amylases from the genus Bacillus or α-amylases from Aspergillus, and more preferred is the α-amylase from Bacillus amyloliquefaciens.

[0073] The α-amylase that can be used in the present technology can be prepared from the culture broth of the microorganism that is the source of the above α-amylase. As a specific preparation method, it can be easily prepared by culturing α-amylase-producing bacteria and separating the α-amylase using publicly known means, as well as methods using transgenic technology.

[0074] In the present technology, commercially available products can also be used as the α-amylase. As an example of a preferred commercially available product, the α-amylase (from Bacillus amyloliquefaciens) manufactured by Amano Enzyme Inc. can be cited.

[0075] Without impairing the effects of the present technology, the content of the α-amylase in the enzyme agent for reducing powderiness according to the present technology can be freely set. With respect to 1 g of plant raw materials used in the manufactured plant-based food or plant-based food material, the content of the α-amylase can be set, for example, to 0.01 U or more. From the viewpoint of further enhancing the effect of improving starch solubility, it can be preferably set to 0.1 U or more, more preferably set to 1 U or more, further preferably set to 3 U or more, and even more preferably set to 5 U or more.

[0076] Without impairing the effects of the present technology, the upper limit of the content of the α-amylase is also not particularly limited. With respect to 1 g of plant raw materials used in the manufactured plant-based food or plant-based food material, it can be set, for example, to 10000 U or less, 2000 U or less, 500 U or less, preferably set to 300 U or less, more preferably set to 100 U or less, and further preferably set to 50 U or less.

[0077] In the present technology, the activity of the α-amylase is a value defined by the following method.

[0078] [Definition of the activity of α - amylase]

[0079] It is measured by the iodine reaction using potato starch as the substrate. That is, the activity of amylase is determined by an enzymatic reaction using potato starch as the substrate by a conventional method, and the amount of enzyme that reduces the color development caused by the iodine reaction by 10% within 1 minute is defined as 1 unit (1U).

[0080] (6) Other enzymes

[0081] The enzyme agent for reducing powdery feeling involved in this technology can further contain β - amylase and / or α - glucosidase.

[0082] In this technology, by allowing β - amylase and / or α - glucosidase to act on plant - based raw materials, the powdery feeling of the produced plant - based food or plant - based food materials can be further reduced.

[0083] The source of β - amylase that can be used in this technology is not particularly limited. For example, β - amylase from plants (wheat, soybean) and from the genus Bacillus can be cited. Preferably, β - amylase from the genus Bacillus is selected, and more preferably, β - amylase from Bacillus Flexus is selected.

[0084] The source of α - glucosidase that can be used in this technology is not particularly limited. Preferably, α - glucosidase from the genus Aspergillus is selected, and more preferably, α - glucosidase from Aspergillus niger is selected.

[0085] The β - amylase and α - glucosidase that can be used in this technology can be prepared from the culture broth of the microorganisms that are the above - mentioned sources. As a specific preparation method, it can be easily prepared by culturing each producing bacterium and separating it by publicly known means, as well as by methods using transgenic technology, etc.

[0086] In this technology, as β - amylase and α - glucosidase, commercially available products can also be used. As examples of preferred commercially available products, β - amylase from Bacillus Flexus manufactured by Amano Enzyme Inc. and α - glucosidase from Aspergillus niger manufactured by Amano Enzyme Inc. can be cited.

[0087] Without impairing the technical effects of the present technology, the content of β-amylase in the enzyme preparation for reducing powdery feeling involved in the present technology can be freely set. With respect to 1 g of vegetable raw material used in the manufactured vegetable-based food or vegetable-based food material, the content of β-amylase can be set, for example, to 0.01 U or more, and from the viewpoint of further enhancing the effect of reducing powdery feeling, it is preferably set to 0.05 U or more, more preferably set to 0.1 U or more, further preferably set to 0.5 U or more, and still more preferably set to 1 U or more.

[0088] Without impairing the technical effects of the present technology, the upper limit of the content of β-amylase is not particularly limited either. With respect to 1 g of vegetable raw material used in the manufactured vegetable-based food or vegetable-based food material, it can be set, for example, to 1000 U or less, 200 U or less, 100 U or less, 50 U or less, 30 U or less, or 20 U or less.

[0089] In addition, in the present technology, the activity of β-amylase is a value defined by the following method.

[0090] [Definition of the activity of β-amylase]

[0091] The amount of enzyme that increases the reducing power equivalent to 1 mg of glucose within 1 minute using potato starch as a substrate is defined as 1 unit (1 U).

[0092] Without impairing the technical effects of the present technology, the content of α-glucosidase in the enzyme preparation for reducing powdery feeling involved in the present technology can be freely set. With respect to 1 g of vegetable raw material used in the manufactured vegetable-based food or vegetable-based food material, the content of α-glucosidase can be set, for example, to 1 U or more, and from the viewpoint of further enhancing the effect of reducing powdery feeling, it is preferably set to 10 U or more, more preferably set to 100 U or more, further preferably set to 300 U or more, and still more preferably set to 500 U or more, 1000 U or more.

[0093] Without impairing the technical effects of the present technology, the upper limit of the content of α-glucosidase is not particularly limited either. For 1 g of vegetable raw material used in the manufactured vegetable-based food or vegetable-based food material, it can be set, for example, to 300000 U or less, 100000 U or less, 20000 U or less, 10000 U or less, 6000 U or less, or 3000 U or less.

[0094] In addition, in the present technology, the activity of α-glucosidase is a value defined by the following method.

[0095] [Definition of the activity of α-glucosidase]

[0096] The activity of generating 1 mg of glucose from soluble starch in 60 minutes at 40°C is defined as 1 unit (1 U).

[0097] (7) Others

[0098] The enzyme preparation for reducing powdery feeling according to the present technology only needs to contain the above-mentioned cell wall polysaccharide-degrading enzyme, and may further contain one or more enzymes selected from the above-mentioned glucoamylase, α-amylase, β-amylase, and α-glucosidase. Without impairing the effects of the present technology, it is also possible to freely select to contain one or more other components. As other components, for example, components such as excipients, pH regulators, colorants, flavoring agents, disintegrants, lubricants, and stabilizers used in conventional formulation can be used. Moreover, functional components that are publicly available or discovered in the future can also be appropriately used in combination according to the purpose.

[0099] 2. Plant-based food or plant-based food materials

[0100] The plant-based food or plant-based food materials according to the present technology are plant-based foods or plant-based food materials produced using the above-mentioned enzyme preparation for reducing powdery feeling. Specific examples of plant-based beverages include oat beverages and rice beverages, and more specifically, oat milk (also known as "oat milk") and rice milk. Specific examples of plant-based foods include plant-based yogurt.

[0101] In addition to reducing the powdery feeling, the plant-based food or plant-based food materials according to the present technology can also be materials with enhanced sweetness and depth. In addition to treating with cell wall polysaccharide-degrading enzymes to reduce the powdery feeling, the solubility of plant-based raw materials is increased, and the depth of the taste can be enhanced. In addition, sweetness can be enhanced by treating with α-amylase and glucoamylase.

[0102] 3. Method for manufacturing plant-based food or plant-based food materials, method for reducing powdery feeling of plant-based food or plant-based food materials

[0103] The manufacturing method of the plant-based food or plant-based food material involved in the present technology is a method including a step of allowing a cell wall polysaccharide-degrading enzyme to act on a plant-based raw material (hereinafter, also referred to as "the step of allowing a cell wall polysaccharide-degrading enzyme to act"). The step of allowing a cell wall polysaccharide-degrading enzyme to act can also be carried out in the liquefaction step of the plant-based raw material. In addition, depending on the type of the plant-based food or plant-based food material, etc., the usual food manufacturing steps can also be carried out before and after each step or simultaneously with each step within the range that does not impair the effects of the present technology. In the above-mentioned manufacturing method of the plant-based food or plant-based food material and the method for reducing the powdery feeling of the plant-based food or plant-based food material, a liquefaction step of starch (α-amylase action step), glucoamylase action step, β-amylase action step, α-glucosidase action step, and recovery step, etc. can be further implemented.

[0104] (1) Plant-based raw material

[0105] The plant-based raw materials that can be used in the present technology are not particularly limited in terms of the origin, type, etc. of the plant-based raw material without impairing the effects of the present technology, and can be freely selected according to the target plant-based food. For example, soybeans, green peas, lentils, chickpeas, black beans, empty beans, mung beans, lupins, string beans, etc. can be cited as legumes; wheat, barley, oats, rice, rye, buckwheat, barnyard millet, millet, teff, etc. can be cited as cereals; almonds, coconuts, peanuts, cashews, hazelnuts, pecans, macadamia nuts, pistachios, walnuts, brazil nuts, pili nuts, chestnuts, sesame seeds, pinecones, etc. can be cited as nuts; chia seeds, quinoa, amaranthus, hypericum seeds, flax seeds, etc. In the present technology, they can be used alone or in combination of two or more. Among these raw materials, cereals are preferably selected, and oats and rice are more preferably selected.

[0106] In the present technology, without impairing the effects of the present technology, the properties of the plant-based raw material during various enzyme treatments are not particularly limited, and powdery (suspension), liquid, slurry, and paste are preferably selected. The powdery feeling is more easily felt when using powdery raw materials. In the present technology, by carrying out the step of allowing a cell wall polysaccharide-degrading enzyme to act, the powdery feeling of the plant-based food or plant-based food material can be reduced. Therefore, as a preferred raw material of the present invention, a liquid in which a powdery raw material is suspended can be cited.

[0107] The plant-based raw materials applicable to the present technology contain saccharides and cell wall polysaccharides. The content of the saccharides contained in the plant-based raw materials (based on the weight in the dried state of the plant-based raw materials) is not particularly limited. For example, 20% by weight or more can be cited. Preferably, 30% by weight or more is selected, more preferably 40% by weight or more, and further preferably 50% by weight or more. The upper limit of this content range is not particularly limited. For example, 90% by weight or less, 80% by weight or less, 70% by weight or less can be cited.

[0108] The content of the cell wall polysaccharides contained in the plant-based raw materials applicable to the present technology is not particularly limited. For example, 0.1% by weight or more is contained. Preferably, 0.3% by weight or more, 0.5% by weight or more, 3% by weight or more, 5% by weight or more, 7% by weight or more, 9% by weight or more are selected. The upper limit of the content is also not particularly limited. For example, 90% by weight or less, 70% by weight or less, 50% by weight or less, 30% by weight or less, 20% by weight or less, 15% by weight or less, 10% by weight or less, 5% by weight or less, 3% by weight or less can be cited. If specific contents are exemplified, the general content of rice (polished rice) is about 0.3 - 1%, and the general content of oats (rolled oats) is about 5 - 15%.

[0109] (2) Cell wall polysaccharide-degrading enzyme action step

[0110] The cell wall polysaccharide-degrading enzyme action step is a step of allowing a cell wall polysaccharide-degrading enzyme to act on a plant-based raw material. Without impairing the effects of the present technology, the addition amount of the cell wall polysaccharide-degrading enzyme in the cell wall polysaccharide-degrading enzyme action step can be freely set. Since the specific addition amount of the cell wall polysaccharide-degrading enzyme in the present technology is the same as the content of the cell wall polysaccharide-degrading enzyme in the above-mentioned enzyme agent for reducing powdery feeling, the description is omitted here.

[0111] Without impairing the effects of the present technology, various conditions of the cell wall polysaccharide-degrading enzyme action step can be freely set. For example, according to the physicochemical properties such as the optimum pH, stable pH range, optimum temperature, and temperature stability of the cell wall polysaccharide-degrading enzyme used, pH, temperature, action time, etc. can be set. The pH can be set to, for example, pH 5.0 - 8.0, preferably set to pH 5.5 - 7.5, and more preferably set to pH 5.5 - 7.0. The temperature can be set to, for example, 30°C - 80°C, preferably set to 40°C - 75°C, and more preferably set to 50°C - 70°C. The action time can be set to, for example, 1 minute - 12 hours, preferably set to 3 minutes - 3 hours, and more preferably set to 5 minutes - 1 hour. In addition, the optimal reaction conditions can be determined through preliminary experiments.

[0112] (3) α-Amylase action step

[0113] The α - amylase action step is the step of allowing α - amylase to act on vegetable raw materials. The α - amylase action step is mainly carried out for the purpose of liquefying starch, but can also be carried out for the purpose of improving taste. Without impairing the technical effects of the present technology, the addition amount of α - amylase in the α - amylase action step can be freely set. Since the specific addition amount of α - amylase in the present technology is the same as the content of α - amylase in the above - mentioned enzyme agent for reducing powdery feeling, the description is omitted here.

[0114] Without impairing the technical effects of the present technology, various conditions of the α - amylase action step can be freely set. For example, according to the physicochemical properties such as the optimum pH, stable pH range, optimum temperature, and temperature stability of the used α - amylase, pH, temperature, action time, etc. can be set. The pH can be set, for example, to pH 5.0 - 8.0, preferably set to pH 5.5 - 7.5, and more preferably set to pH 5.5 - 7.0. The temperature can be set, for example, to 30°C - 80°C, preferably set to 40°C - 75°C, and more preferably set to 50°C - 70°C. The action time can be set, for example, to 1 minute - 12 hours, preferably set to 3 minutes - 3 hours, and more preferably set to 5 minutes - 1 hour. In addition, the optimal reaction conditions can be determined through preliminary experiments.

[0115] (4)Glucoamylase action step

[0116] The glucoamylase action step is the step of allowing glucoamylase to act on vegetable raw materials. Without impairing the technical effects of the present technology, the addition amount of glucoamylase in the glucoamylase action step can be freely set. Since the specific addition amount of glucoamylase in the present technology is the same as the content of glucoamylase in the above - mentioned enzyme agent for reducing powdery feeling, the description is omitted here.

[0117] Without impairing the technical effects of the present technology, various conditions of the glucoamylase action step can be freely set. For example, according to the physicochemical properties such as the optimum pH, stable pH range, optimum temperature, and temperature stability of the used glucoamylase, pH, temperature, action time, etc. can be set. The pH can be set, for example, to pH 5.0 - 8.0, preferably set to pH 5.5 - 7.5, and more preferably set to pH 5.5 - 7.0. The temperature can be set, for example, to 30°C - 80°C, preferably set to 40°C - 75°C, and more preferably set to 50°C - 70°C. The action time can be set, for example, to 1 minute - 12 hours, preferably set to 3 minutes - 3 hours, and more preferably set to 5 minutes - 1 hour. In addition, the optimal reaction conditions can be determined through preliminary experiments.

[0118] (5)β - amylase action step

[0119] The β-amylase action step is a step of allowing β-amylase to act on a plant-based raw material. Without impairing the technical effects of the present technology, the addition amount of β-amylase in the β-amylase action step can be freely set. Since the specific addition amount of β-amylase in the present technology is the same as the content of β-amylase in the above-mentioned enzyme agent for reducing powdery feeling, the description thereof is omitted here.

[0120] Without impairing the technical effects of the present technology, various conditions of the β-amylase action step can be freely set. For example, according to the physicochemical properties such as the optimum pH, stable pH range, optimum temperature, and temperature stability of the β-amylase used, pH, temperature, action time, etc. can be set. The pH can be set, for example, to pH 5.0 to 8.0, preferably to pH 5.5 to 7.5, and more preferably to pH 5.5 to 7.0. The temperature can be set, for example, to 30°C to 80°C, preferably to 40°C to 75°C, and more preferably to 50°C to 70°C. The action time can be set, for example, to 1 minute to 12 hours, preferably to 3 minutes to 3 hours, and more preferably to 5 minutes to 1 hour. In addition, the optimum reaction conditions can be determined through preliminary experiments.

[0121] (6) α-Glucosidase action step

[0122] The α-glucosidase action step is a step of allowing α-glucosidase to act on a plant-based raw material. Without impairing the technical effects of the present technology, the addition amount of α-glucosidase in the α-glucosidase action step can be freely set. Since the specific addition amount of α-glucosidase in the present technology is the same as the content of α-glucosidase in the above-mentioned enzyme agent for reducing powdery feeling, the description thereof is omitted here.

[0123] Without impairing the technical effects of the present technology, various conditions of the α-glucosidase action step can be freely set. For example, according to the physicochemical properties such as the optimum pH, stable pH range, optimum temperature, and temperature stability of the α-glucosidase used, pH, temperature, action time, etc. can be set. The pH can be set, for example, to pH 5.0 to 8.0, preferably to pH 5.5 to 7.5, and more preferably to pH 5.5 to 7.0. The temperature can be set, for example, to 30°C to 80°C, preferably to 40°C to 75°C, and more preferably to 50°C to 70°C. The action time can be set, for example, to 1 minute to 12 hours, preferably to 3 minutes to 3 hours, and more preferably to 5 minutes to 1 hour. In addition, the optimum reaction conditions can be determined through preliminary experiments.

[0124] (7) Recovery step

[0125] The recovery process is a process for recovering plant-based food or plant-based food materials that have undergone the cell wall polysaccharide-degrading enzyme treatment process. In addition, in the case of performing the above liquefaction process (α-amylase treatment process), glucoamylase treatment process, β-amylase treatment process, α-glucosidase treatment process, etc., in the recovery process, plant-based food or plant-based food materials that have undergone one or more processes selected from the cell wall polysaccharide-degrading enzyme treatment process, liquefaction process (α-amylase treatment process), glucoamylase treatment process, β-amylase treatment process, and α-glucosidase treatment process are recovered. Specific recovery methods can be freely combined and used according to the types of plant-based food or plant-based food materials to be manufactured, using one or more than two common recovery methods in the manufacture of plant-based food or plant-based food materials.

[0126] The plant-based food or plant-based food materials that have undergone the above cell wall polysaccharide-degrading enzyme treatment process are characterized in that the powdery feeling is reduced and / or the insoluble components are decreased. In addition, the plant-based food or plant-based food materials that have undergone the α-amylase treatment process and / or the glucoamylase treatment process are characterized in that the sweetness is enhanced. Furthermore, the plant-based food or plant-based food materials that have undergone the β-amylase treatment process and / or the α-glucosidase treatment process are characterized in that the powdery feeling is further reduced and / or the insoluble components are further decreased.

[0127] In summary, in the recovery process of the present technology, plant-based food or plant-based food materials with a reduced powdery feeling and / or plant-based food or plant-based food materials with decreased insoluble components can be recovered. In addition, plant-based food or plant-based food materials with enhanced sweetness can also be recovered.

[0128] One mode of the method for manufacturing a plant-based food or plant-based food material and the method for reducing the powdery feeling of the present technology includes the following steps (1) and (2). In addition, an enzyme inactivation step can also be added after step (2).

[0129] (1) Step of preparing a plant-based raw material containing cell wall polysaccharides

[0130] (2) Step of treating the prepared raw material with a cell wall polysaccharide-degrading enzyme

[0131] One mode of the method for manufacturing a plant-based food or plant-based food material and the method for reducing the powdery feeling of the present technology includes the following steps (1) to (3). In addition, an enzyme inactivation step can also be added after step (2).

[0132] (1) Step of preparing a plant-based raw material containing cell wall polysaccharides

[0133] (2) A step of treating the prepared raw material with α-amylase and glucoamylase in addition to the cell wall polysaccharide-degrading enzyme

[0134] (3) A step of recovering a plant-based food or a plant-based food material with reduced powdery texture

[0135] One aspect of the method for producing a plant-based food or a plant-based food material and the method for reducing powdery texture of the present technology includes the following steps (1) to (3). In addition, an enzyme inactivation step may be added after step (2).

[0136] (1) A step of preparing a plant-based raw material containing cell wall polysaccharides

[0137] (2) A step of treating the prepared raw material with any one or more enzymes selected from α-amylase, glucoamylase, β-amylase, and α-glucosidase in addition to the cell wall polysaccharide-degrading enzyme

[0138] (3) A step of recovering a plant-based food or a plant-based food material with reduced powdery texture

[0139] In addition, the enzymes used in step (2) can be treated simultaneously or separately. The order in the case of separate treatment is not particularly limited. Simultaneous treatment is preferred. In addition, in the case of separate treatment, an enzyme inactivation step may be added as needed between each enzyme treatment.

[0140] Examples

[0141] Hereinafter, the present invention will be described in more detail based on examples. In addition, the examples described below show an example of a representative embodiment of the present invention, and the scope of the present invention is not construed narrowly thereby.

[0142] 1. Plant-based raw materials

[0143] The plant-based raw materials used in the examples are shown in Table 1 below.

[0144] [Table 1]

[0145] Raw materials Source Shape Product name Manufacturer Oat 1 Oat Powder Oatmeal (There is) OFCO Oat 2 Oat Powder Oat milk powder (Ltd.) Tomizawa Shoten Rice 1 Rice Powder Rice flour Kyoritsu Foods Co., Ltd. Rice 2 Rice Powder Pre-gelatinized rice flour (Ltd.) Yoshiyoshiya Taniguchi Store Rice 3 Rice Powder Pre-gelatinized japonica rice flour (There is) OFCO

[0146] 2. Enzymes used

[0147] The various enzymes used in the examples are shown in Table 2 below.

[0148] [Table 2]

[0149] Enzyme Source Manufacturer Cellulase Trichoderma viride Amano Enzyme Inc. Hemicellulase Aspergillus niger Amano Enzyme Inc. Pectinase Aspergillus pulvelentus Amano Enzyme Inc. α-Amylase 1 Bacillus amyloliquefaciens Amano Enzyme Inc. Glucoamylase 1 Rhizopus oryzae Amano Enzyme Inc. Glucoamylase 2 Aspergillus niger Amano Enzyme Inc. α-Amylase 2 Aspergillus oryzae Amano Enzyme Inc. β-Amylase Bacillus flexus Amano Enzyme Inc. α-Glucosidase Aspergillus niger Amano Enzyme Inc.

[0150] 3. Method for measuring enzyme activity

[0151] [Method for Measuring the Activity of Cellulase]

[0152] For the activity of cellulase, it is measured by a method based on the cellulase activity test method in the 9th Edition of the Official Monographs of Food Additives.

[0153] Precisely weigh approximately 1 g of sodium carboxymethyl cellulose (alias: CMC) (etherification degree 0.62 - 0.68) in advance, dry it at 105 °C for 4 hours and measure its weight loss. Accurately weigh sodium carboxymethyl cellulose corresponding to 0.625 g of its dried product, put it into a 100 mL Erlenmeyer flask, add 50 mL of water, heat and dissolve it. After cooling, a solution made by adding 10 mL of 1 mol / L acetic acid - sodium acetate buffer (pH 4.5) and water to make it exactly 100 mL is used as the substrate solution. Accurately measure 4 mL of the substrate solution, put it into a 50 mL Nessler tube, let it stand at 37 ± 0.5 °C for more than 10 minutes, then accurately measure 1 mL of the sample solution and immediately shake and mix it. Keep this liquid accurately at 37 ± 0.5 °C for 30 minutes, accurately add 2 mL of alkaline copper test solution (Somogyi test solution (I)) and shake and mix it. Plug the Nessler tube with a stopper and accurately heat it in a water bath for 30 minutes. After cooling with water, accurately add 2 mL of Nelson's solution and shake and mix it well. Further accurately add 3 mL of 0.5 mol / L sodium hydroxide test solution, shake and mix to dissolve the precipitate. After standing for 20 minutes, accurately add 13 mL of 1 mol / L acetic acid - sodium acetate buffer (pH 4.5). Accurately measure 1 mL of this liquid, accurately add 9 mL of 1 mol / L acetic acid - sodium acetate buffer (pH 4.5), and shake and mix it well. For this liquid, measure the absorbance (A1) at a wavelength of 750 nm with water as the control. In addition, accurately measure 1 mL of the sample solution, put it into a 50 mL Nessler tube, accurately add 2 mL of alkaline copper test solution (Somogyi test solution (I)) and shake and mix it. Next, accurately measure and add 4 mL of the substrate solution and shake and mix it. Then perform the following operations in the same way to measure the absorbance (A2). Under these conditions, the amount of enzyme that increases the reducing power equivalent to 1 μmole of glucose in 1 minute for the cellulase activity is defined as 1 unit, and the cellulase activity per 1 g of the sample or per 1 mL (U / g, U / mL) is calculated according to the following formula.

[0154] [Mathematical Formula 1]

[0155] Cellulase activity (U / g, U / mL) = G × (1 / 30) × (1 / 0.180) × n

[0156] G: The amount of glucose produced (mg) obtained from the (A1 - A2 / a) value according to the glucose standard curve

[0157] 1 / 30: Conversion factor per minute

[0158] 1 / 0.180: 1 μmole of glucose = 0.180 mg

[0159] a: Slope of the glucose standard curve

[0160] n: Dilution factor per 1 g of the sample or per 1 mL

[0161] [Method for determining the activity of hemicellulase]

[0162] For the activity of hemicellulase, it is determined by a method based on the hemicellulase activity test method in the 9th Edition of the Japanese Pharmacopoeia for Food Additives.

[0163] Measure an appropriate amount of the enzyme, dissolve it in water or disperse it uniformly, and use the appropriately diluted liquid as the test solution. Measure 0.50 g of xylan, add about 30 mL of water, heat it while stirring, and boil for 3 minutes after starting to boil. After cooling, add water to this liquid to make a 50 mL solution as the substrate solution. Measure 1 mL of the substrate solution into a test tube, add 3 mL of 0.1 mol / L acetic acid buffer (pH 4.5), heat it at 40 °C for 10 minutes, then add 1 mL of the test solution and mix by shaking, and heat it at 40 °C for 30 minutes. Add 2 mL of Somogyi reagent (III) to this liquid and mix, stopper the test tube, heat it in a boiling water bath for 20 minutes, and immediately cool it. After cooling, add 1 mL of Nelson reagent to this liquid, stir well until the red precipitate completely dissolves, let it stand at room temperature for about 20 minutes, and then add water to 25 mL. Centrifuge this liquid at 3000 revolutions per minute at 25 °C for 10 minutes, and use the supernatant as the test solution. Additionally, measure 1 mL of the substrate solution into a test tube, add 3 mL of 0.1 mol / L acetic acid buffer (pH 4.5) and 2 mL of Somogyi reagent (III), mix by shaking, then add 1 mL of the test solution, stopper the test tube, heat it in a boiling water bath for 20 minutes, and immediately cool it. Perform the following operations in the same manner as the preparation of the test solution to obtain the comparison solution. For the test solution and the comparison solution, measure the absorbance at a wavelength of 500 nm with water as the control. Under these conditions, the amount of enzyme that produces reducing sugar equivalent to 1 mg of xylose within 1 minute is defined as 100 units, and the enzyme activity is calculated.

[0164] [Method for determining the activity of pectinase]

[0165] The measurement is carried out by the following method based on the official document. Specifically, weigh 0.700 g of LM pectin and put it into about 70 mL of water preheated to 70 - 90 °C. After dissolution, cool it. After cooling, adjust the pH to 3.50 with 0.1 mol / L citric acid test solution or 0.2 mol / L disodium hydrogen phosphate test solution, and add 10 mL of McIlvaine buffer (pH 3.5) and water to make 100 mL. Weigh a specified amount of the sample, add the sample dilution solution to dissolve it to a specified dilution factor. In the case of powder, weigh it into a mortar, add a small amount (4 - 10 mL) of the sample dilution solution, and stir and dissolve it with a pestle.

[0166] <Measurement operation method based on viscometer>

[0167] Gently add 6 mL of LM pectin solution (pH 3.5) and 6 mL of McIlvaine buffer (pH 3.5) from tube 1 of the viscometer, vertically set it in a constant temperature water bath at 40 ± 0.5 °C, and after standing for 10 - 15 minutes, add 2 mL of the sample solution to start the reaction. Immediately after the addition, seal tube 3 with a finger, and blow air from tube 2 with the mouth to mix the content liquid.

[0168] Seal tube 3 with a finger. To prevent air bubbles from entering tube 2, gently suck from tube 2, suck the liquid level to the center of sphere C, then stop sucking, open the mouth of tube 3, and immediately measure the time ti seconds required for the liquid level to flow from the upper calibration line of sphere B to the lower calibration line. Repeat this operation 5 times. In addition, perform the same operation using 6 mL of LM pectin solution (pH 3.5), 6 mL of McIlvaine buffer (pH 3.5), and 2 mL of water, and measure the flow-down time t0 seconds. In addition, perform the same operation using 14 mL of water, and measure the flow-down time tw seconds.

[0169] Under these conditions, the amount of enzyme that reduces the viscosity by 50% within 1 minute is taken as 1 unit and calculated according to the following formula.

[0170] [Mathematical formula 2]

[0171] Viscosity reduction rate (V 50 ) = (t0 - ti) / (t0 - tw) × 100

[0172] Endo PGase activity (u / g, u / mL) = 60 / V 50 × n

[0173] t0: Flow-down time (seconds) of 6 mL of substrate + 6 mL of buffer + 2 mL of water

[0174] ti: Flow-down time (seconds) of 14 mL of reaction solution

[0175] tw: Flow-down time (seconds) of 14 mL of water

[0176] 60: Unit conversion factor (1 minute = 60 seconds)

[0177] n: Dilution factor per 1 g or 1 mL of the sample

[0178] V 50 : Time to achieve a 50% viscosity reduction rate [Ti + ti / 2 seconds]

[0179] Ti is the elapsed time from the start of the reaction to the start of the i-th viscosity measurement

[0180] Take the viscosity reduction rate on the vertical axis and time [Ti + ti / 2 seconds] on the horizontal axis, plot a curve, and read the time when the viscosity reduction rate reaches 50%.

[0181] [Method for measuring the activity of α-amylase]

[0182] After heating 10 mL of a 1% potato starch substrate solution (0.1 mol / L acetic acid (pH 5.0)) at 37°C for 10 minutes, add 1 mL of the sample solution containing α-amylase and immediately mix by shaking. After leaving this liquid at 37°C for 10 minutes, add 1 mL of this liquid to 10 mL of a 0.1 mol / L hydrochloric acid test solution and immediately mix by shaking. Next, measure 0.5 mL of this liquid, add 10 mL of a 0.0002 mol / L iodine test solution (Japanese Pharmacopoeia), mix by shaking, and measure the absorbance (AT) at a wavelength of 660 nm using water as a control. In addition, add 1 mL of water in place of the sample solution and perform the same operation to measure the absorbance (AB). Furthermore, the 0.0002 mol / L iodine test solution (Japanese Pharmacopoeia) is prepared as follows: Add 10 mL of water to 12.7 g of iodine and 25 g of potassium iodide, stir well, add water to make 100 mL, and then dilute 2500 times with water. Calculate the α-amylase activity using the following mathematical formula. Consider the amount of enzyme that reduces the color development caused by iodine in potato starch by 10% within 1 minute as 1 unit (1 U).

[0183] [Mathematical formula 3]

[0184] Activity of α-amylase (U / g, U / mL) = {(AB - AT) / AB} × 1 / W

[0185] AT: Absorbance of the reaction solution

[0186] AB: Absorbance of the blank solution

[0187] W: Amount of sample in 1 mL of the sample solution (g or mL)

[0188] [Method for measuring the activity of glucoamylase]

[0189] The titer (activity value) of glucoamylase is determined by the following method.

[0190] <Glucoamylase (GA) activity>

[0191] It is determined by the following method in accordance with Method 4 of the Glucoamylase Activity Test Method in the 9th Edition of the Japanese Pharmacopoeia for Food Additives.

[0192] Measure 0.50 g of the enzyme sample, and use the solution diluted with water to an appropriate concentration as the sample solution. Dry potato starch in advance at 105 °C for 2 hours, measure 1.0 g of the dried product, add 20 mL of water, and slowly add while stirring 5 mL of sodium hydroxide test solution (2 mol / L) to make a paste. After heating the paste in a water bath for 3 minutes while stirring, add 25 mL of water, cool, add hydrochloric acid test solution (2 mol / L) and hydrochloric acid test solution (0.1 mol / L) for neutralization, add 10 mL of 1 mol / L acetic acid-sodium acetate buffer solution (pH 4.5), and further add water to make a 100 mL solution as the substrate solution.

[0193] Measure 10 mL of the substrate solution, heat it at 37 °C for 10 minutes, add 1 mL of the sample solution and immediately mix well by shaking, after heating at 37 °C for 10 minutes, add 4 mL of Fehling's solution and mix gently by shaking, after heating in a water bath for 15 minutes, cool to below 25 °C, add 2 mL of potassium iodide test solution and 2 mL of sulfuric acid (dilute 1 volume part of sulfuric acid with 6 volume parts of water) to make the test solution. Additionally, use 10 mL of water instead of the substrate solution and perform the operation in the same manner as the preparation of the test solution to make the comparison solution. For the test solution and the comparison solution, titrate the free iodine with 0.05 mol / L sodium thiosulfate solution. The end point is when the blue color produced by adding 1 - 2 drops of soluble starch test solution disappears near the end of the titration. Under these conditions, the amount of enzyme that increases the reducing power equivalent to 1 mg of glucose within 1 minute is defined as 1 unit (1 U), and the glucoamylase activity is calculated according to the following formula.

[0194] [Mathematical formula 4]

[0195] Activity of glucoamylase (U / g) = Amount of glucose (mg) × 1 / 10 × 1 / M

[0196] Amount of glucose (mg) = (b - a) × 1.6 × f

[0197] a: Titration value of the test solution (mL)

[0198] b: Titration value of the comparison solution (mL)

[0199] 1.6: 1 mL of 0.05 mol / L sodium thiosulfate solution is equivalent to 1.6 mg of glucose

[0200] 1 / 10: Unit conversion factor for reaction time (minutes)

[0201] M: Amount of enzyme sample in 1 mL of sample solution (g or mL)

[0202] f: Factor of 0.05 mol / L sodium thiosulfate solution

[0203] [Method for measuring the activity of β - amylase]

[0204] The measurement is carried out according to the method described in the Food Additive Codex (9th Edition). Specifically, potato starch is used as the substrate, which is pre - dried at 105 °C for 2 hours. Weigh 1.0 g of its dried product, add 20 mL of water, and slowly add 5 mL of sodium hydroxide test solution (2 mol / L) while stirring to make a paste. Next, heat in a water bath for 3 minutes while stirring, and then add 25 mL of water. After cooling, add hydrochloric acid test solution (2 mol / L) and hydrochloric acid test solution (0.1 mol / L) for neutralization, add 10 mL of 1 mol / L acetic acid - sodium acetate buffer solution (pH 5.0), and further add water to make a 100 - mL solution as the substrate solution.

[0205] Measure 10 mL of the substrate solution, heat it at 37 °C for 10 minutes, add 1 mL of the sample solution and immediately mix by shaking. After heating at this temperature for 10 minutes or 30 minutes, further add 4 mL of Fehling's solution and gently mix by shaking. After heating in a water bath for 15 minutes, cool it to below 25 °C, add 2 mL of 30% potassium iodide solution and 2 mL of sulfuric acid (1→6) to make the test solution. In addition, Fehling's solution is prepared by mixing 34.66 g of fine crystals of copper(II) sulfate pentahydrate dissolved in water to make 500 mL of copper solution and 173 g of (+) - potassium sodium tartrate tetrahydrate and 50 g of sodium hydroxide dissolved in water to make 500 mL of alkaline tartrate solution in a ratio of 1 volume of copper solution to 1 volume of alkaline tartrate solution, and it is prepared when in use. Also, use 10 mL of water instead of the substrate solution and perform the operation in the same way as the preparation of the test solution as the comparison solution. For the test solution and the comparison solution, titrate the free iodine with 0.05 mol / L sodium thiosulfate solution. The end point is when the blue color produced by adding 1 - 2 drops of soluble starch test solution disappears when the titration is approaching the end point.

[0206] The amount of enzyme that increases the reducing power equivalent to 1 mg of glucose in 1 minute is defined as 1 unit (1 U) and is calculated according to the following formula.

[0207] [Mathematical formula 5]

[0208] Activity of β - amylase (U / g, U / mL) = Amount of glucose (mg) × 1 / 10 × 1 / M Amount of glucose (mg)=(b - a)×1.6×f

[0209] a: Titration value of enzyme reaction solution (mL) b: Titration value of blank solution (mL)

[0210] 1.6: 1 mL of 0.05 mol / L sodium thiosulfate solution is equivalent to 1.6 mg of glucose

[0211] 1 / 10: Unit conversion coefficient of reaction time (minutes)

[0212] M: Amount of sample in 1 mL of sample solution (g or mL)

[0213] f: Factor of 0.05 mol / L sodium thiosulfate solution (for quantification)

[0214] [Method for determining the activity of α-glucosidase]

[0215] The activity of α-glucosidase is determined by the second method of the α-glucosidase activity test method in the 9th edition of the Japanese Pharmacopoeia of Food Additives. Under these conditions, the amount of enzyme that produces 1 μg of glucose in 2.5 mL of the reaction solution within 60 minutes is defined as 1 unit.

[0216] [Mathematical formula 6]

[0217] α-glucosidase activity (U / g) = (E60 - E0) × G × 2.5 / 0.1 × n / 0.5

[0218] E60: Absorbance of the test solution

[0219] E0: Absorbance of the comparison solution

[0220] G: Amount of glucose (μg) when the absorbance difference is 1.000 (calculated according to the glucose standard curve)

[0221] 2.5: Volume of the reaction system solution (mL)

[0222] 0.1: Volume of the reaction solution collected (mL)

[0223] 0.5: Volume of the sample solution collected (mL)

[0224] n: Dilution factor of the sample

[0225] 4. Experimental examples

[0226] <Experimental example 1>

[0227] In Experimental example 1, the effect of reducing the powdery feeling produced by cell wall polysaccharide-degrading enzymes was investigated.

[0228] (1) Experimental method

[0229] Add an enzyme agent to 3.6 g of the above-mentioned vegetable raw material in an amount shown in Tables 3 and 4 below, add 30 mL of hot water at 60 °C, and use the liquid obtained by standing at 60 °C for 30 minutes as vegetable milk. In addition, a solution obtained by heating the powder of the above-mentioned vegetable raw material with hot water at 60 °C for 30 minutes without enzyme treatment is used as the vegetable milk of the reference example.

[0230] (2) Measurement and evaluation

[0231] [Measurement of Brix value]

[0232] Measure the Brix value after heating at 60 °C for 30 minutes using a Brix meter.

[0233] [Measurement of solubility]

[0234] To evaluate the liquefaction of the powder caused by the enzyme in the vegetable milk, collect 10 mL of the vegetable milk after heating at 60 °C for 30 minutes and centrifuge it at 10,000 rpm for 15 minutes. Calculate the solubility of the enzyme in the vegetable protein material based on the weight of the precipitate after centrifugation.

[0235] [Sensory evaluation]

[0236] Three panelists evaluate the change in the taste of the vegetable milk based on the enzyme. Regarding "powdery feeling", "sweetness", and "flavor of the material", evaluation is carried out in three grades according to the following evaluation criteria. In addition, for examples using oats 2 and rice 1-3 as raw materials, regarding "depth (complexity)", evaluation is also carried out in three grades according to the following evaluation criteria. In addition, the "flavor of the material" refers to the unique flavor of oats if it is oats, and the unique flavor of rice if it is rice.

[0237] (a) Powdery feeling

[0238] 3 points: Decreased compared to the reference example

[0239] 2 points: Slightly decreased compared to the reference example

[0240] 1 point: No change compared to the reference example

[0241] (b) Sweetness

[0242] 3 points: Sweeter compared to the reference example

[0243] 2 points: Slightly sweeter compared to the reference example

[0244] 1 point: No change compared to the reference example

[0245] (c) Flavor of the material

[0246] 3 points: The flavor of the material is enhanced compared to the reference example

[0247] 2 points: The flavor of the material is slightly enhanced compared with the reference example.

[0248] 1 point: No change compared with the reference example.

[0249] (d) Depth

[0250] 3 points: The depth increases compared with the reference example (becomes a complex flavor).

[0251] 2 points: The depth slightly increases compared with the reference example.

[0252] 1 point: No change compared with the reference example.

[0253] (3) Results

[0254] The results are shown in Tables 3 and 4 below.

[0255] [Table 3]

[0256]

[0257] [Table 4]

[0258]

[0259] (4) Discussion

[0260] As shown in Examples 1 to 15 of Tables 3 and 4, by treating with cell wall polysaccharide-degrading enzymes, an increase in Brix, a decrease in sediment, a reduction in powdery feeling, an enhancement of sweetness, and an effect of change in taste properties were confirmed in all raw materials. In addition, as shown in Examples 16 to 18 of Table 4, it was also confirmed that by further treating with β-amylase, α-glucosidase, and α-amylase 2, the powdery feeling was further improved.

Claims

1. An enzyme preparation for reducing the powdery feeling of a plant-based food or a plant-based food material, which contains a cell wall polysaccharide-degrading enzyme.

2. A plant-based food or a plant-based food material, which uses the enzyme preparation for reducing the powdery feeling according to claim 1.

3. The plant-based food and drink product according to claim 2, wherein, The plant-based food is oat milk or rice milk.

4. A method for manufacturing a plant-based food or a plant-based food material, which includes a step of allowing a cell wall polysaccharide-degrading enzyme to act on a plant-based raw material.

5. A method for reducing the powdery feeling of a plant-based food or a plant-based food material, which includes a step of allowing a cell wall polysaccharide-degrading enzyme to act on a plant-based raw material.

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