Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
  • A23L11/60—Drinks from legumes, e.g. lupine drinks
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF
  • A23C11/00—Milk substitutes, e.g. coffee whitener compositions
  • A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
  • A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
  • A23L2/38—Other non-alcoholic beverages
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
  • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
  • A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
  • A23L29/06—Enzymes
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
  • A23L7/10—Cereal-derived products
  • A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
  • A23L7/107—Addition or treatment with enzymes not combined with fermentation with microorganisms
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
  • C12N9/14—Hydrolases (3)
  • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
  • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
  • C12N9/2405—Glucanases
  • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
  • C12N9/14—Hydrolases (3)
  • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
  • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
  • C12N9/2405—Glucanases
  • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
  • C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)

Definitions

• the present technology relates to an enzyme agent for reducing the powdery texture of plant-based food and drink products or plant-based food and drink materials. More specifically, the present technology provides an enzyme agent for reducing the powdery texture of a plant-based food/beverage product or a plant-based food/drink material, a plant-based food/beverage product or a plant-based food/drink material using the enzyme agent for reducing the powdery texture, and , a method for producing a plant-based food/beverage product or a plant-based food/drink material, and a method for reducing the flouriness of a plant-based food/drink or food/drink material.
• Patent Document 1 describes a technology for producing a liquefied grain product that has both low viscosity and suppressed free sugar content by allowing 4- ⁇ -glucanotransferase, cellulase, and peptidase to act on oats.
• Patent Document 2 describes a technique for producing an oat drink with moderate sweetness by decomposing starch of an oat material using one or more amylases.
• Patent Document 3 describes that the nutritional components are changed by allowing phytase to act on oat bran, and that the mouthfeel of the food composition is improved by the pulverization process.
• Patent Document 4 states that by treating a rice dispersion with protease, cellulase, pullulanase, and ⁇ -amylase, it becomes a rice liquefied product that does not exhibit sweetness and is rich in oligosaccharides, and that by further homogenizing the rice, it becomes It is stated that the texture of the liquefied product is smooth.
• the main purpose of the present technology is to provide a plant-based milk with reduced flouriness and changed taste.
• the present technology first provides an enzyme agent for reducing the powdery texture of a plant-based food/beverage product or a plant-based food/beverage material that contains a cell wall polysaccharide degrading enzyme.
• the present technology also provides a plant-based food/beverage product or a plant-based food/drink material using the powdery texture reducing enzyme agent according to the present technology.
• the plant-based food/beverage product according to the present technology can be oat milk or rice milk.
• the present technology provides a method for producing a plant-based food/beverage product or a plant-based food/drink material, which includes a step of causing a cell wall polysaccharide-degrading enzyme to act on a plant-based raw material.
• the present technology further provides a method for reducing the flouriness of a plant-based food/beverage product or a plant-based food/drink material, which includes a step of causing a cell wall polysaccharide-degrading enzyme to act on a plant-based material.
• a plant-based food/beverage product or a plant-based food/beverage material with reduced flouriness and changed taste properties is provided.
• the enzyme preparation for reducing powdery texture contains a cell wall polysaccharide degrading enzyme as an active ingredient.
• Carbohydrates consist of carbohydrates that include polysaccharides such as starch, oligosaccharides, monosaccharides, disaccharides, etc., and dietary fibers that include cell wall polysaccharides.
• Cell wall polysaccharides include cellulose, hemicellulose, pectin, etc., and enzymes that decompose these cell wall polysaccharides are cell wall polysaccharide degrading enzymes.
• the cell wall polysaccharide degrading enzyme that can be used in this technology is not particularly limited as long as it has the ability to decompose cell wall polysaccharides; One type or two or more types can be used in any combination as long as the effects of the present technology are not impaired.
• Examples of cell wall polysaccharide degrading enzymes include cellulases, hemicellulases, and pectinases. Among these, it is particularly preferable to use cellulase in the enzyme agent for reducing powdery texture according to the present technology.
• enzymes and the like that can be used in the enzyme agent for reducing powdery texture according to the present technology will be explained in detail.
• Cellulase Cellulase that can be used in this technology is an enzyme that hydrolyzes the glycosidic bonds of ⁇ -1,4-glucan, and decomposes cellulose, which is one of the cell wall polysaccharides.
• the cellulase that can be used in the present technology may be an enzyme that additionally has other functions as long as it has cellulase activity. With this technology, by allowing cellulase to act on plant raw materials, it is possible to reduce the powdery texture of manufactured plant-based food and drink products.
• the origin of the cellulase that can be used in the present technology is not particularly limited, but examples include Aspergillus niger, Aspergillus oryzae, Aspergillus sojae, Aspergillus saitoi, Aspergillus genus such as Aspergillus awamori and Aspergillus flavus; Trichoderma genus such as Trichoderma reesei and Trichoderma viride; Acremonium cellulose Examples include microorganisms belonging to the genus Acremonium, such as Acremonium cellulolyticus. These cellulases may be used alone or in combination. Among these, from the viewpoint of reducing the flouriness of the plant material, the cellulase is preferably a cellulase derived from a microorganism of the genus Trichoderma, particularly Trichoderma viride.
• cellulase derived from Trichoderma viride refers to cellulase produced by a microorganism (which may be a wild strain or a mutant strain) classified as Trichoderma viride, or This means that the cellulase is obtained by genetic engineering using the cellulase gene. Therefore, a recombinant product produced by a host microorganism into which a cellulase gene obtained from Trichoderma viride (or a gene modified from the gene) is also classified as “cellulase derived from Trichoderma viride.” .
• the type of cellulase used is not particularly limited, and may be, for example, endoglucanase, exoglucanase, or a mixture thereof.
• the cellulase used in the present invention preferably includes an enzyme preparation containing both endoglucanase and exoglucanase.
• the cellulase used in the present technology can be prepared from the culture solution of the microorganism from which the cellulase described above is derived.
• a specific preparation method includes a method of recovering cellulase from the culture solution or cells of the above-mentioned microorganism.
• the enzyme when using a cellulase-secreting microorganism, the enzyme can be isolated and/or purified after the bacterial cells are recovered from the culture fluid by filtration, centrifugation, etc., as necessary.
• cellulase non-secreting microorganisms if necessary, after collecting the bacterial cells from the culture solution in advance, crushing the bacterial cells by pressure treatment, ultrasonication, etc. to expose the enzyme.
• the isolated and/or purified enzyme can be powdered by a drying method such as freeze drying or vacuum drying, and may be powdered using an appropriate excipient and/or drying aid in the drying method. You can also do that. Furthermore, the isolated and/or purified enzyme can be liquefied by adding appropriate additives and sterilizing it by filtration.
• a commercial product can also be used as the cellulase, and examples of preferred commercial products include cellulase derived from Trichoderma viride manufactured by Amano Enzyme Co., Ltd.
• the content of cellulase in the powdery texture reducing enzyme preparation according to the present technology can be freely set as long as it does not impair the effects of the present technology.
• the content of cellulase can be set to, for example, 0.01 U or more per 1 g of the plant raw material used for the plant-based food/beverage product or plant-based food/drink material to be produced, from the viewpoint of further enhancing the powdery texture reduction effect. From this, it can be set to preferably 0.05U or more, more preferably 0.1U or more, still more preferably 0.3U or more, even more preferably 0.5U or more.
• the upper limit of the content of cellulase is not particularly limited as long as it does not impair the effects of the present technology, but it may be, for example, 500 U or less, 200 U or less, 100 U or less per gram of the plant raw material used in the plant-based food/drink product or plant-based food/drink material to be produced. Below, it can be set to 50U or less, 30U or less, 20U or less, or 10U or less.
• the activity of cellulase is a value defined by the following method.
• 1 unit (1U) is the amount of enzyme that causes an increase in reducing power equivalent to 1 ⁇ mole of glucose per minute.
• the cellulase activity is a value measured by the cellulase activity measuring method described in the Examples below.
• Hemicellulase that can be used in this technology is an enzyme that hydrolyzes hemicellulose, which is one of cell wall polysaccharides.
• Hemicellulose is a heteropolysaccharide, composed of multiple constituent sugars, and generally has a branched structure in which side chains are formed by other constituent sugars to the main chain from which the name derives.
• Specific examples of hemicellulose include mannan, ⁇ -1,3-1,4-glucan, glucomannan, xylan, xyloglucan, glucuronoxylan, and the like.
• Hemicellulases are enzymes that degrade these hemicelluloses.
• hemicellulase examples include xylanase, galactanase, mannanase, galactomannanase, arabinase, ⁇ -glucanase, etc., and these may be used alone or in combination of two or more. You can.
• the hemicellulase that can be used in the present technology may be an enzyme that also has other functions as long as it has hemicellulase activity. With this technology, by allowing hemicellulase to act on plant raw materials, it is possible to reduce the powdery texture of manufactured plant-based foods and drinks.
• the origin of the hemicellulase that can be used in this technology is not particularly limited, but examples include basidiomycetes (genus Corticium, genus Pycnoporus), filamentous fungi (genus Aspergillus, genus Humicola), , Penicillium spp., Trichoderma spp.), actinomycetes (Streptomyces spp.) or bacteria (Bacillus spp.), but are artificially synthesized enzymes. It may be. Moreover, these may be used individually by 1 type, and may use 2 or more types together.
• hemicellulases derived from filamentous fungi are preferred, those derived from microorganisms of the genus Aspergillus are more preferred, and hemicellulases derived from Aspergillus niger are even more preferred.
• the hemicellulase used in the present technology can be prepared from the culture solution of the microorganism from which the hemicellulase described above is derived.
• a specific preparation method includes a method of recovering hemicellulase from the culture solution or cells of the above-mentioned microorganism.
• the enzyme can be isolated and/or purified after the bacterial cells are recovered from the culture fluid by filtration, centrifugation, etc., if necessary.
• Enzymes can be separated and/or purified.
• any known protein separation and/or purification method can be used without particular limitation, such as centrifugation method, UF concentration method, salting out method, ion exchange resin, etc. Examples include various chromatography methods using .
• the isolated and/or purified enzyme can be powdered by a drying method such as freeze drying or vacuum drying, and may be powdered using an appropriate excipient and/or drying aid in the drying method. You can also do that.
• the isolated and/or purified enzyme can be liquefied by adding appropriate additives and sterilizing it by filtration.
• hemicellulase a commercially available product can also be used as the hemicellulase, and examples of preferred commercially available products include hemicellulase derived from Aspergillus niger manufactured by Amano Enzyme Co., Ltd.
• the content of hemicellulase in the powdery texture reducing enzyme preparation according to the present technology can be freely set as long as it does not impair the effects of the present technology.
• the content of hemicellulase can be set to, for example, 1 U or more per 1 g of the plant raw material used for the plant-based food/beverage product or plant-based food/drink material to be produced, from the viewpoint of further increasing the powdery texture reduction effect. , preferably 10 U or more, more preferably 50 U or more, still more preferably 100 U or more, even more preferably 200 U or more.
• the upper limit of the content of hemicellulase is not particularly limited as long as it does not impair the effects of the present technology, but for example, 100,000 U or less, 50,000 U or less, It can be set to 10000U or less, 5000U or less, 2000U or less, or 1000U or less.
• the activity of hemicellulase is a value defined by the following method. [Definition of hemicellulase activity] Using xylan as a substrate, the amount of enzyme that produces reducing sugar equivalent to 1 mg of xylose per minute is 100 units (100 U).
• Pectinase that can be used in this technology is an enzyme that hydrolyzes pectin, which is one of cell wall polysaccharides.
• Pectin is a complex polysaccharide whose main component is polygalacturonic acid in which galacturonic acid is ⁇ -1,4-linked.
• Specific examples of pectinase include polygalacturonase, pectin lyase, pectin methylesterase, and the like, with polygalacturonase being preferred.
• the pectinase that can be used in the present technology may be an enzyme that also has other functions as long as it has pectinase activity.
• the origin of the pectinase that can be used in this technology is not particularly limited, but examples include basidiomycetes (genus Corticium), filamentous fungi (genus Aspergillus, genus Rhizopus, and genus Trichoderma). , yeast (genus Geotrichum and genus Trichosporon), actinomycetes (genus Streptomyces), or bacteria (genus Bacillus), even if they are artificially synthesized. It may also be an enzyme that has been Moreover, these may be used individually by 1 type, and may use 2 or more types together. Among these, in the present technology, pectinases derived from filamentous fungi are preferred, and pectinases derived from the genus Aspergillus are more preferred.
• the pectinase used in this technology can be prepared from the culture solution of the microorganism from which the above pectinase is derived.
• a specific preparation method includes a method of recovering pectinase from the culture solution or cells of the above-mentioned microorganism.
• the enzyme when using a pectinase-secreting microorganism, the enzyme can be isolated and/or purified after the bacterial cells have been recovered from the culture solution by filtration, centrifugation, etc., if necessary.
• pectinase non-secreting microorganisms if necessary, after collecting the bacterial cells from the culture solution, crush the bacterial cells by pressure treatment, ultrasonication, etc.
• any known protein separation and/or purification method can be used without particular limitation, such as centrifugation method, UF concentration method, salting out method, ion exchange resin, etc. Examples include various chromatography methods using .
• the isolated and/or purified enzyme can be powdered by a drying method such as freeze drying or vacuum drying, and may be powdered using an appropriate excipient and/or drying aid in the drying method. You can also do that.
• the isolated and/or purified enzyme can be liquefied by adding appropriate additives and sterilizing it by filtration.
• pectinase a commercially available product can also be used as the pectinase, and examples of preferred commercially available products include pectinase derived from the genus Aspergillus manufactured by Amano Enzyme Co., Ltd.
• the content of pectinase in the enzyme agent for reducing powdery texture according to the present technology can be freely set as long as it does not impair the effects of the present technology.
• the content of pectinase can be set to, for example, 0.01 U or more per 1 g of the plant raw material used for the plant-based food/beverage product or plant-based food/drink material to be produced, from the viewpoint of further enhancing the powdery texture reduction effect. Therefore, it can be set to preferably 0.1 U or more, more preferably 0.5 U or more, still more preferably 1 U or more, and even more preferably 2 U or more.
• the upper limit of the content of pectinase is also not particularly limited as long as it does not impair the effects of the present technology, but for example, 2000 U or less, 500 U or less, 100 U per gram of the plant raw material used in the plant-based food/drink product or plant-based food/drink material to be manufactured. Below, it can be set to 50U or less, 40U or less, or 30U or less.
• the activity of pectinase is a value defined by the following method. [Definition of pectinase activity] Using low methoxyl (LM) pectin as a substrate, the amount of enzyme that reduces the viscosity by 50% in 1 minute is 1 unit (1 U).
• LM low methoxyl
• the enzyme agent for reducing powdery texture according to the present technology can also contain glucoamylase.
• Glucoamylase that can be used in the present technology is an enzyme that has the activity of hydrolyzing ⁇ -1,4-glucoside bonds of carbohydrates such as starch into glucose units from the non-reducing end.
• the glucoamylase that can be used in the present technology may be an enzyme that additionally has other functions as long as it has glucoamylase activity.
• glucoamylase that can be used in the present technology is not particularly limited, but examples thereof include glucoamylase derived from the genus Aspergillus and the genus Rhizopus. These glucoamylases may be used alone or in combination. Among these glucoamylases, glucoamylase derived from the genus Rhizopus is preferred, and glucoamylase derived from Rhizopus oryzae is more preferred.
• Glucoamylase derived from Rhizopus oryzae here refers to glucoamylase produced by a microorganism (which may be a wild strain or a mutant strain) classified as Rhizopus oryzae. , or glucoamylase obtained by genetic engineering using the glucoamylase gene. Therefore, a recombinant produced by a host microorganism into which a glucoamylase gene obtained from Rhizopus oryzae (or a gene modified from the gene) is also referred to as "Glucoamylase derived from Rhizopus oryzae.” Applicable.
• Glucoamylase that can be used in the present technology can be prepared from the culture solution of the microorganism from which the glucoamylase is derived. As specific preparation methods, it can be easily prepared by culturing glucoamylase-producing bacteria and separating glucoamylase using known means, or by using genetic recombination technology.
• glucoamylase derived from Rhizopus oryzae manufactured by Amano Enzyme Co., Ltd.
• the content of glucoamylase in the powdery texture reducing enzyme agent according to the present technology can be freely set as long as it does not impair the effects of the present technology.
• the content of glucoamylase can be set to, for example, 0.01 U or more, preferably 0.1 U or more, per 1 g of the vegetable raw material used for the plant-based food/drink product or plant-based food/drink material to be produced. It can be set to preferably 1U or more, more preferably 3U or more, even more preferably 5U or more.
• the upper limit of the content of glucoamylase is not particularly limited as long as it does not impair the effects of the present technology, but it may be, for example, 5000 U or less, 1000 U or less, It can be set to 500U or less, 200U or less, 100U or less, or 50U or less.
• the activity of glucoamylase is a value defined by the following method. [Definition of glucoamylase activity] Using potato starch as a substrate, the amount of enzyme that causes an increase in reducing power equivalent to 1 mg of glucose per minute is defined as 1 unit (1 U).
• ⁇ -Amylase ⁇ -amylase can also be used in the enzyme agent for reducing powdery texture according to the present technology.
• ⁇ -amylase that can be used in the present technology is an enzyme that acts on starch and mainly hydrolyzes ⁇ -1,4 glycosidic bonds.
• the ⁇ -amylase that can be used in the present technology may be an enzyme that also has other functions as long as it has ⁇ -amylase activity.
• ⁇ -amylase that can be used in the present technology is not particularly limited, but examples include Aspergillus genus (e.g., Aspergillus oryzae, Aspergillus niger, etc.), Bacillus Examples include ⁇ -amylases derived from organisms of the genus Bacillus (for example, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, etc.), preferably Bacillus ( Examples include ⁇ -amylase derived from the genus Bacillus or ⁇ -amylase derived from Aspergillus, and more preferably ⁇ -amylase derived from Bacillus amyloliquefaciens.
• Aspergillus genus e.g., Aspergillus oryzae, Aspergillus niger, etc.
• Bacillus Examples include ⁇ -amylases derived from organisms of the genus Bacillus (for example, Bacill
• the ⁇ -amylase that can be used in the present technology can be prepared from the culture solution of the microorganism from which the ⁇ -amylase is derived. As specific preparation methods, it can be easily prepared by culturing ⁇ -amylase-producing bacteria and separating ⁇ -amylase using known means, or by using genetic recombination technology.
• ⁇ -amylase derived from Bacillus amyloliquefaciens manufactured by Amano Enzyme Co., Ltd.
• the content of ⁇ -amylase in the powdery texture reducing enzyme agent according to the present technology can be freely set as long as it does not impair the effects of the present technology.
• the content of ⁇ -amylase can be set to, for example, 0.01 U or more per 1 g of the plant raw material used in the plant-based food/beverage products or plant-based food/drink materials to be produced, thereby further enhancing the starch solubility improvement effect. From the viewpoint of increasing the density, it can be set to preferably 0.1 U or more, more preferably 1 U or more, still more preferably 3 U or more, even more preferably 5 U or more.
• the upper limit of the content of ⁇ -amylase is not particularly limited as long as it does not impair the effects of the present technology, but for example, 10,000 U or less per 1 g of the plant raw material used in the plant-based food and drink product or the plant-based food and drink material to be manufactured, It can be set to 2000U or less, 500U or less, preferably 300U or less, more preferably 100U or less, even more preferably 50U or less.
• the ⁇ -amylase activity is a value defined by the method below.
• [Definition of ⁇ -amylase activity] It is measured by iodine reaction using potato starch as a substrate. That is, the activity of amylase is determined by performing an enzymatic reaction using potato starch as a substrate in a conventional manner, and determining the amount of enzyme that reduces the coloring caused by the iodine reaction by 10% in 1 minute as 1 unit (1 U).
• the enzyme agent for reducing powdery texture according to the present technology may further contain ⁇ -amylase and/or ⁇ -glucosidase.
• ⁇ -amylase that can be used in the present technology is not particularly limited, but examples thereof include ⁇ -amylase derived from plants (wheat, soybean) and the genus Bacillus. Preferred examples include ⁇ -amylase derived from the genus Bacillus, and more preferred ⁇ -amylase derived from Bacillus Flexus.
• ⁇ -glucosidase that can be used in the present technology is not particularly limited, but ⁇ -glucosidase derived from the genus Aspergillus is preferred, and ⁇ -glucosidase derived from Aspergillus niger is more preferred. Can be mentioned.
• ⁇ -amylase and ⁇ -glucosidase that can be used in the present technology can be prepared from the culture solution of the microorganism from which they are derived. As specific preparation methods, it can be easily prepared by culturing each producing bacteria and separating it using known means, or by using genetic recombination technology.
• commercially available products can also be used as ⁇ -amylase and ⁇ -glucosidase.
• preferred commercially available products include ⁇ -amylase derived from the genus Bacillus Flexus, manufactured by Amano Enzyme Co., Ltd., and ⁇ -glucosidase derived from Aspergillus niger, manufactured by Amano Enzyme Co., Ltd.
• the content of ⁇ -amylase in the powdery texture reducing enzyme agent according to the present technology can be freely set as long as it does not impair the effects of the present technology.
• the content of ⁇ -amylase can be set to, for example, 0.01 U or more per 1 g of the plant raw material used in the plant-based food/beverage product or plant-based food/drink material to be produced, thereby further enhancing the powdery texture reduction effect. From the viewpoint of increasing the amount of heat, it can be set to preferably 0.05 U or more, more preferably 0.1 U or more, still more preferably 0.5 U or more, even more preferably 1 U or more.
• the upper limit of the content of ⁇ -amylase is not particularly limited as long as it does not impair the effects of the present technology, but it is, for example, 1000 U or less, 200 U or less per 1 g of the vegetable raw material used in the plant-based food/drink product or plant-based food/drink material to be manufactured. , 100U or less, 50U or less, 30U or less, or 20U or less.
• the activity of ⁇ -amylase is a value defined by the method below. [Definition of ⁇ -amylase activity] Using potato starch as a substrate, the amount of enzyme that causes an increase in reducing power equivalent to 1 mg of glucose per minute is defined as 1 unit (1 U).
• the content of ⁇ -glucosidase in the powdery texture reducing enzyme preparation according to the present technology can be freely set as long as it does not impair the effects of the present technology.
• the content of ⁇ -glucosidase can be set to, for example, 1 U or more per 1 g of the plant raw material used for the plant-based food/beverage products or plant-based food/drink materials to be produced, from the viewpoint of further enhancing the powdery texture reduction effect. , preferably 10 U or more, more preferably 100 U or more, still more preferably 300 U or more, still more preferably 500 U or more, and even more preferably 1000 U or more.
• the upper limit of the content of ⁇ -glucosidase is not particularly limited as long as it does not impair the effects of the present technology, but it is, for example, 300,000 U or less, 100,000 U or less per 1 g of the plant raw material used in the plant-based food/drink product or plant-based food/drink material to be manufactured. , 20000U or less, 10000U or less, 6000U or less, or 3000U or less.
• the ⁇ -glucosidase activity is a value defined by the method below. [Definition of ⁇ -glucosidase activity]
• One unit (1U) is the activity of producing 1mg of glucose from soluble starch in 60 minutes at 40°C.
• the enzyme preparation for reducing powdery texture according to the present technology only needs to contain the above-mentioned cell wall polysaccharide-degrading enzyme, and is further selected from the above-mentioned glucoamylase, ⁇ -amylase, ⁇ -amylase, and ⁇ -glucosidase. may further contain one or more enzymes.
• One or more other components can be freely selected and included as long as the effects of the present technology are not impaired.
• Other components that can be used include, for example, excipients, pH adjusters, coloring agents, flavoring agents, disintegrants, lubricants, stabilizers, and the like that are commonly used in formulations.
• components having functions that are known or that will be discovered in the future may be used in combination depending on the purpose.
• Plant-based food/beverage products or plant-based food/drink materials are the plant-based food/drinks or plant-based food/drink materials manufactured using the aforementioned powdery texture reducing enzyme agent. It is.
• plant-based beverages include oat drinks and rice drinks, and more specifically include oat milk (also referred to as "oat milk") and rice milk.
• oat milk also referred to as "oat milk”
• rice milk A specific example of a plant-based food is plant-based yogurt.
• the plant-based food/beverage products or plant-based food/drink materials according to the present technology include those that have enhanced sweetness and depth in addition to reduced flouriness.
• Treatment with a cell wall polysaccharide-degrading enzyme not only reduces the powdery texture but also improves the solubility of the vegetable raw material and enhances the depth of flavor.
• sweetness can be enhanced by treatment with ⁇ -amylase or glucoamylase.
• Method for producing plant-based food/beverage products or plant-based food/drink materials method for reducing flouriness of plant-based food/drinks or plant-based food/drink materials
• the method for producing plant-based food/drinks or plant-based food/drink materials according to the present technology includes a step of causing a cell wall polysaccharide degrading enzyme to act on a raw material (hereinafter also referred to as a "cell wall polysaccharide degrading enzyme action step").
• the cell wall polysaccharide degrading enzyme action step can also be carried out in the liquefaction step of plant raw materials.
• the method further includes a starch liquefaction step ( ⁇ -amylase action step) and a glucoamylase action step.
• ⁇ -amylase action step ⁇ -glucosidase action step, recovery step, etc. can also be carried out.
• Plant-based raw materials The origin, type, etc. of the plant-based raw materials that can be used in this technology are not particularly limited, as long as they do not impair the effects of this technology, and may vary depending on the desired plant-based food or drink. , can be freely selected.
• legumes such as soy beans, green peas, lentils, chickpeas, black beans, fava beans, mung beans, lupin beans, kidney beans; wheat, barley Grains such as oats, rice, rye, buckwheat, millet, millet, and teff; almonds, coconuts, peanuts, cashews, hazelnuts, pecans, macadamia nuts, pistachios, walnuts, Brazil nuts, pili nuts, chestnuts, sesame seeds, Nuts such as pine nuts; examples include hemp seeds (industrial hemp), chia seeds, quinoa, amaranthus, canary seeds, and flaxseeds. In the present technology, these may be used alone or in combination of two or more. Among these raw materials, cereals are preferred, and oats and rice are more preferred.
• the properties of the vegetable raw materials when subjected to various enzyme treatments are not particularly limited as long as they do not impair the effects of the present technology, but are preferably powder (suspension), liquid, slurry, Examples include paste form.
• powder suspension
• liquid slurry
• paste form The feeling of powder becomes more noticeable when powdered raw materials are used.
• by performing the cell wall polysaccharide degrading enzyme action step it is possible to reduce the powdery texture of plant-based food and drink products or plant-based food and drink materials. Examples include suspensions of .
• Plant raw materials that can be used in this technology contain carbohydrates and cell wall polysaccharides.
• the content of carbohydrates contained in the plant material is not particularly limited, but may be, for example, 20% by weight or more. Preferably it is 30% by weight or more, more preferably 40% by weight or more, still more preferably 50% by weight or more.
• the upper limit of the content range is not particularly limited, but includes, for example, 90% by weight or less, 80% by weight or less, and 70% by weight or less.
• the content of cell wall polysaccharides contained in the plant raw materials that can be used in the present technology is not particularly limited, and is contained, for example, at 0.1% by weight or more.
• Preferred examples include 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, and 9% by weight or more.
• the upper limit of the content is also not particularly limited, and 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.
• the following examples include 3% by weight or less.
• the general content of rice is about 0.3 to 1%
• the general content of oats (oatmeal) is about 5 to 15%.
• the cell wall polysaccharide degrading enzyme action step is a step in which a cell wall polysaccharide degrading enzyme is allowed to act on a plant material.
• the amount of the cell wall polysaccharide degrading enzyme added in the cell wall polysaccharide degrading enzyme action step can be freely set as long as it does not impair the effects of the present technology.
• the specific amount of the cell wall polysaccharide degrading enzyme added in the present technology is the same as the content of the cell wall polysaccharide degrading enzyme in the above-mentioned powdery texture reducing enzyme agent, so a description thereof will be omitted here.
• the pH, temperature, action time, etc. can be set depending on the physicochemical properties such as the optimum pH, stable pH range, optimum temperature, and temperature stability of the cell wall polysaccharide degrading enzyme used.
• the pH can be set, for example, to pH 5.0 to 8.0, preferably pH 5.5 to 7.5, more preferably pH 5.5 to 7.0.
• the temperature can be set to, for example, 30°C to 80°C, preferably 40°C to 75°C, more preferably 50°C to 70°C.
• the action time can be set, for example, from 1 minute to 12 hours, preferably from 3 minutes to 3 hours, and more preferably from 5 minutes to 1 hour. Note that the optimal reaction conditions can be determined through preliminary experiments.
• the ⁇ -amylase action step is a step in which ⁇ -amylase is allowed to act on vegetable raw materials.
• the ⁇ -amylase action step is mainly carried out for the purpose of liquefying starch, but it may also be carried out for the purpose of improving taste.
• the amount of ⁇ -amylase added in the ⁇ -amylase action step can be freely set as long as it does not impair the effects of the present technology.
• the specific amount of ⁇ -amylase added in the present technology is the same as the content of ⁇ -amylase in the above-mentioned enzyme agent for reducing powdery texture, so a description thereof will be omitted here.
• the pH, temperature, action time, etc. can be set depending on the physicochemical properties of the ⁇ -amylase used, such as the optimum pH, stable pH range, optimum temperature, and temperature stability.
• the pH can be set, for example, to pH 5.0 to 8.0, preferably pH 5.5 to 7.5, more preferably pH 5.5 to 7.0.
• the temperature can be set to, for example, 30°C to 80°C, preferably 40°C to 75°C, more preferably 50°C to 70°C.
• the action time can be set, for example, from 1 minute to 12 hours, preferably from 3 minutes to 3 hours, and more preferably from 5 minutes to 1 hour. Note that the optimal reaction conditions can be determined through preliminary experiments.
• the glucoamylase action step is a step in which glucoamylase is allowed to act on vegetable raw materials.
• the amount of glucoamylase added in the glucoamylase action step can be freely set as long as it does not impair the effects of the present technology.
• the specific amount of glucoamylase added in the present technology is the same as the content of glucoamylase in the powdery texture reducing enzyme agent described above, so a description thereof will be omitted here.
• the pH, temperature, action time, etc. can be set depending on the physicochemical properties of the glucoamylase used, such as the optimum pH, stable pH range, optimum temperature, and temperature stability.
• the pH can be set, for example, to pH 5.0 to 8.0, preferably pH 5.5 to 7.5, more preferably pH 5.5 to 7.0.
• the temperature can be set to, for example, 30°C to 80°C, preferably 40°C to 75°C, more preferably 50°C to 70°C.
• the action time can be set, for example, from 1 minute to 12 hours, preferably from 3 minutes to 3 hours, and more preferably from 5 minutes to 1 hour. Note that the optimal reaction conditions can be determined through preliminary experiments.
• the ⁇ -amylase action step is a step in which ⁇ -amylase is allowed to act on vegetable raw materials.
• the amount of ⁇ -amylase added in the ⁇ -amylase action step can be freely set as long as it does not impair the effects of the present technology.
• the specific amount of ⁇ -amylase added in the present technology is the same as the content of ⁇ -amylase in the powdery texture reducing enzyme agent described above, so a description thereof will be omitted here.
• the pH, temperature, action time, etc. can be set depending on the physicochemical properties of the ⁇ -amylase used, such as the optimum pH, stable pH range, optimum temperature, and temperature stability.
• the pH can be set, for example, to pH 5.0 to 8.0, preferably pH 5.5 to 7.5, more preferably pH 5.5 to 7.0.
• the temperature can be set to, for example, 30°C to 80°C, preferably 40°C to 75°C, more preferably 50°C to 70°C.
• the action time can be set, for example, from 1 minute to 12 hours, preferably from 3 minutes to 3 hours, and more preferably from 5 minutes to 1 hour. Note that the optimal reaction conditions can be determined through preliminary experiments.
• the ⁇ -glucosidase action step is a step in which ⁇ -glucosidase is allowed to act on vegetable raw materials.
• the amount of ⁇ -glucosidase added in the ⁇ -glucosidase action step can be freely set as long as the effect of the present technology is not impaired.
• the specific amount of ⁇ -glucosidase added in the present technology is the same as the content of ⁇ -glucosidase in the above-mentioned enzyme agent for reducing powdery texture, so a description thereof will be omitted here.
• the pH, temperature, action time, etc. can be set depending on the physicochemical properties of the ⁇ -glucosidase used, such as the optimum pH, stable pH range, optimum temperature, and temperature stability.
• the pH can be set, for example, to pH 5.0 to 8.0, preferably pH 5.5 to 7.5, more preferably pH 5.5 to 7.0.
• the temperature can be set to, for example, 30°C to 80°C, preferably 40°C to 75°C, more preferably 50°C to 70°C.
• the action time can be set, for example, from 1 minute to 12 hours, preferably from 3 minutes to 3 hours, and more preferably from 5 minutes to 1 hour. Note that the optimal reaction conditions can be determined through preliminary experiments.
• the recovery process is a process of recovering the plant food/drink or food/beverage material that has undergone the cell wall polysaccharide degrading enzyme action process.
• the recovery step when performing the above-mentioned liquefaction step ( ⁇ -amylase action step), glucoamylase action step, ⁇ -amylase action step, ⁇ -glucosidase action step, etc., in the recovery step, the cell wall polysaccharide degrading enzyme action step, and A plant food/drink or food/drink material that has undergone one or more steps selected from a liquefaction step ( ⁇ -amylase action step), a glucoamylase action step, a ⁇ -amylase action step, and an ⁇ -glucosidase action step is recovered.
• the specific recovery method may be one or two types of recovery methods used in the production of general plant-based food/drinks or plant-based food/drink materials. The above can be used in any combination.
• the plant-based food and drink or plant-based food and drink ingredients that have undergone the cell wall polysaccharide degrading enzyme action step are characterized by a reduced powdery feel and/or a reduced amount of insoluble components.
• the plant-based food and drink or plant-based food and drink ingredients that have undergone the ⁇ -amylase action step and/or the glucoamylase action step are characterized by enhanced sweetness.
• the plant-based food and drink or plant-based food and drink ingredients that have undergone the ⁇ -amylase action step and/or the ⁇ -glucosidase action step are characterized by a further reduced powdery feel and/or a further reduced amount of insoluble components.
• plant-based food/drinks or plant-based food/drink materials with reduced flouriness and/or plant-based food/drinks or plant-based food/drink materials with reduced insoluble components are recovered. It is also possible to recover plant-based food and drink products or plant-based food and drink materials with enhanced sweetness.
• One embodiment of the method for producing a plant-based food/beverage product or a plant-based food/drink material and the method for reducing powdery texture of the present technology includes the following steps (1) and (2). Note that an enzyme deactivation step may be added after step (2).
• Step of preparing plant raw materials containing cell wall polysaccharides (2) Step of treating the prepared raw materials with cell wall polysaccharide-degrading enzymes
• One embodiment of the method for producing a plant-based food/beverage product or a plant-based food/drink material and the method for reducing powdery texture of the present technology includes the following steps (1) to (3). Note that an enzyme deactivation step may be added after step (2).
• Step of preparing plant raw materials containing cell wall polysaccharides (2) Step of adding prepared raw materials to cell wall polysaccharide degrading enzyme and treating with ⁇ -amylase and glucoamylase (3) Plants with reduced powdery texture The process of collecting plant-based food and drink materials or plant-based food and drink materials
• One embodiment of the method for producing a plant-based food/beverage product or a plant-based food/drink material and the method for reducing powdery texture of the present technology includes the following steps (1) to (3).
• an enzyme deactivation step may be added after step (2).
• Step of preparing a vegetable raw material containing cell wall polysaccharide (2) Adding the prepared raw material to a cell wall polysaccharide degrading enzyme, an enzyme selected from ⁇ -amylase, glucoamylase, ⁇ -amylase, and ⁇ -glucosidase (3) Process of recovering plant-based food/drinks or plant-based food/drink materials with reduced flouriness.
• the enzymes used in step (2) may be treated at the same time or separately. may be processed. The order in which they are processed separately is not particularly limited. Preferably, simultaneous processing is performed.
• an enzyme deactivation step may be added between each enzyme treatment as necessary.
• Plant-based raw materials The plant-based raw materials used in the examples are shown in Table 1 below.
• Enzyme activity measurement method Cellulase activity measurement method
• Cellulase activity was measured by a method based on the cellulase activity test method of the 9th edition of the Food Additives Official Standards.
• carmellose sodium also known as sodium carboxymethylcellulose
• degree of etherification 0.62 to 0.68 was weighed accurately in advance, dried at 105° C. for 4 hours, and its weight loss was measured.
• Accurately weigh carmellose sodium corresponding to 0.625 g of the dried product put it in a 100 mL Erlenmeyer flask, add 50 mL of water and dissolve by heating. After cooling, 1 mol/L acetic acid/sodium acetate buffer (pH 4.
• the hemicellulase activity was measured by a method based on the hemicellulase activity test method of the 9th edition of the Food Additives Official Standards. An appropriate amount of enzyme was weighed, water was added to dissolve or uniformly disperse the enzyme, and the sample solution was diluted appropriately. Weighed out 0.50 g of xylan, added about 30 mL of water, heated it while stirring, and boiled for 3 minutes after it started to boil. After cooling, water was added to this liquid to make 50 mL, which was used as a substrate solution.
• tube 3 was closed with a finger, and air was blown through tube 2 with the mouth to mix the contents. Close the tube 3 with your fingers, and suck the liquid level from the tube 2 weakly to prevent air bubbles from entering the tube 2 until the liquid level reaches the center of the bulb C. Then, stop the suction and close the tube opening of the tube 3. Immediately after opening, the time ti seconds required for the liquid level to flow down from the marked line above ball B to the marked line below was measured. This operation was repeated 5 times.
• Glucoamylase activity was measured by the following method. ⁇ Glucoamylase (GA) activity> It was measured by the following method according to Method 4 of the Glucoamylase Activity Test Method of the Food Additives Official Standards, 9th Edition.
• a sample solution was prepared by weighing 0.50 g of an enzyme sample and diluting it with water to an appropriate concentration. Potato starch was previously dried at 105° C. for 2 hours, 1.0 g of the dried product was weighed, 20 mL of water was added, and 5 mL of sodium hydroxide test solution (2 mol/L) was gradually added with stirring to form a paste.
• hydrochloric acid test solution (2 mol/L) and hydrochloric acid test solution (0.1 mol/L) to neutralize, add 1 mol/L acetic acid/sodium acetate buffer (pH 5.0) 10 mL, and further add water.
• the 100 mL solution was used as a substrate solution. Measure out 10 mL of the substrate solution, warm it at 37°C for 10 minutes, add 1 mL of the sample solution, shake it immediately, warm it at the same temperature for 10 minutes or 30 minutes, then add 4 mL of Fehling's test solution and shake it gently. After heating in a water bath for 15 minutes, the mixture was cooled to 25° C.
• a comparative solution was prepared in the same manner as in the preparation of the test solution using 10 mL of water instead of the substrate solution.
• liberated iodine was titrated with a 0.05 mol/L sodium thiosulfate solution.
• the end point was defined as when the titration was near the end point, 1 to 2 drops of soluble starch sample solution was added, and the resulting blue color disappeared.
• the amount of enzyme that causes an increase in reducing power equivalent to 1 mg of glucose per minute was defined as 1 unit (1 U), and was calculated from the following formula.
• Powdery feel 3 points Reduced compared to the reference example 2 points: Slightly reduced compared to the reference example 1 point: No change compared to the reference example

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