Method for treating enzyme of yeast material

Abstract

Provided are a method for producing a yeast-enzyme-treated product for a milk substitute food having exceptional dispersibility, a treated product obtained thereby, and a milk substitute food / beverage containing said treated product.　The present invention provides: a method for producing a yeast-enzyme-treated product for a milk substitute food / beverage, the method being characterized by comprising (a) treating a yeast extract residue with glucanase and (b) recovering the treated product obtained in (a), and furthermore being characterized in that the glucanase is of at least two kinds, i.e., endo-acting glucanase and exo-acting glucanase; a treated product obtained thereby; and a milk substitute food / beverage containing said treated product. The yeast-enzyme-treated product according to the present invention has better dispersibility than prior-art products and is suitable for blending into a milk substitute food / beverage.

Description

Enzyme treatment method for yeast material

[0001] The present invention relates to a method for producing a yeast enzyme-treated product for use in a milk substitute food having excellent dispersibility, the treated product, and a milk substitute food or drink containing the treated product.

[0002] In recent years, due to the increasing demand for low-fat or fat-free foods due to dietary and health-conscious trends, as well as the increasing demand for vegan diets, attention has been increasingly focused on dairy substitute foods and beverages that use plant materials such as oats, soybeans, and almonds instead of dairy ingredients in order to avoid dairy-derived allergens. However, dairy substitute foods and beverages that use these plant materials are usually more expensive than cow's milk and are known to contain allergens.

[0003] In addition to such milk substitute foods and beverages using plant materials, milk substitute foods and beverages derived from yeast materials have also been reported. For example, the present applicant has reported the use of a composition containing a cell wall-lytic enzyme hydrolyzed product of the residue (yeast extract cell residue) after yeast extract production as a milk substitute (see, for example, Patent Document 1).

[0004] Japanese Patent Application Laid-Open No. 2022-135934

[0005] However, conventional milk substitutes using yeast materials have the drawback of forming a precipitate that tends to stick when left as a suspension. As such, dairy substitute foods and beverages using yeast materials for the purpose of protein supplementation have the problem that they are not suitable for direct consumption because they tend to form a precipitate and are difficult to redisperse.

[0006] As a result of extensive research to solve the above problems, the inventors discovered that by treating yeast material with at least two types of glucanases having different activities, particularly when the total amount added is less than a specific amount, a yeast enzyme-treated product can be obtained that has better dispersibility than conventional products and is suitable for incorporation into milk substitute foods and beverages, and thus completed the present invention.

[0007] The gist of the present invention is as follows: [1] A method for producing an enzyme-treated yeast product for use in a milk substitute food or beverage, comprising: (a) treating yeast extract cell residue with glucanase; and (b) recovering the treated product obtained in (a), wherein the glucanase is at least two types of glucanase: a glucanase having endoactivity and a glucanase having exoactivity. [2] The production method according to [1] above, wherein the glucanase having endoactivity is a glucanase belonging to the enzyme code EC 3.2.1.6 or EC 3.2.1.39. [3] The production method according to [1] or [2] above, wherein the glucanase having endoactivity is a glucanase derived from the genus Streptomyces and having β-1,3 activity. [4] The production method according to any of [1] to [3] above, wherein the amount of glucanase having endoactivity added is 0.1% by mass or more relative to the dry mass of the yeast extract cell residue. [5] The production method according to any one of [1] to [4] above, wherein the glucanase having exoactivity is a glucanase belonging to the enzyme code EC 3.2.1.58. [6] The production method according to any one of [1] to [5] above, wherein the glucanase having exoactivity is a glucanase derived from the genera Trichoderma, Talanomyces, and / or Aspergillus and having β-1,3, β-1,4, and / or β-1,6 activity. [7] The production method according to any one of [1] to [6] above, wherein the amount of glucanase having exoactivity added is less than 1% by mass relative to the dry mass of the yeast extract cell residue. [8] The production method according to any one of [1] to [7] above, wherein the yeast extract cell residue is a water-insoluble fraction obtained after hot water extraction of yeast or autolysis of yeast. [9] An enzyme-treated yeast product obtained by the production method according to any one of [1] to [8] above.

[10] A milk substitute food or drink containing a yeast enzyme-treated product obtained by the production method according to any one of [1] to [8] above.

[11] The milk substitute food or drink according to

[10] above, further containing 0.1 to 30 mass% of fats and oils.

[12] Use of a glucanase-treated product of yeast extract cell residue as a milk substitute food or drink, wherein the glucanase is at least two types of glucanases, namely, a glucanase having endoactivity and a glucanase having exoactivity.

[0008] According to the present invention, a milk substitute food or beverage can be provided using inexpensive and easily available yeast material, particularly yeast extract cell residue, as a raw material. In the production method of the present invention, by treating the yeast with at least two types of glucanase with different activities, the dispersibility of the resulting yeast enzyme-treated product is improved compared to conventional products, especially when the total amount added is less than a specific amount. Therefore, milk substitute food or beverages using this product are less likely to precipitate, and even if precipitate does occur, the redispersion is easy, making them suitable for consumption. Improvements in flavor as milk substitute food or beverages can also be expected. Therefore, milk substitute food or beverages using the yeast material obtained by the production method of the present invention can more easily ingest yeast-derived proteins, dietary fiber, and other nutrients.

[0009] 1 is a radar chart showing the results (average values) of sensory evaluation by five expert panelists for suspensions prepared at 10% by mass of each of the compositions obtained in Comparative Examples 1 and 3 and Examples 1 to 6 in Test Example 1.

[0010] In one embodiment of the present invention, there is provided a method for producing a yeast enzyme-treated product for use in a milk substitute food or beverage, comprising: (a) treating yeast extract cell residue with glucanase; and (b) recovering the treated product obtained in (a), wherein the glucanase is at least two types of glucanase, namely, a glucanase having endoactivity and a glucanase having exoactivity.

[0011] In the present invention, the term "dairy substitute food and beverage" refers to a composition that can be used in place of milk (typically cow's milk) consumed by humans. Human milk is a cloudy liquid secreted from the mammary glands of mammals that contains nutrients such as water, protein, lipids, carbohydrates, vitamins, and minerals, and has a unique milky flavor. Milk substitute food and beverage refers to a liquid consumed that possesses one or more of these characteristics. Milk substitute food and beverages are generally made from plant ingredients such as soybeans, almonds, oats, rice, and coconut, and contain plant-derived proteins and lipids, as well as dietary fiber that is rarely found in milk. Products made from these plant ingredients may be used alone, in combination with multiple ingredients, or mixed with animal-derived milk. The milk substitute food and beverage of the present invention can also be used in place of milk substitute food and beverages made from such known plant ingredients.

[0012] The "yeast extract cell residue" in (a) above is not particularly limited as long as it contains yeast cell walls and is the residue (insoluble fraction) of yeast cells obtained after subjecting yeast to extraction and removing the extract. Specifically, it refers to yeast cells that are generated as a residue after subjecting yeast to a known extraction treatment such as autolysis (protease treatment), hot water treatment, acid treatment, alkali treatment, and / or mechanical disruption, and removing the supernatant (soluble fraction (yeast extract)) separated by centrifugation or the like. The yeast extract cell residue is preferably the residue (also referred to as the water-insoluble fraction or heavy liquid) of yeast cells after hot water extraction or autolysis of yeast.

[0013] As used herein, the term "dry mass of yeast extract cell residue" refers to the mass of the yeast extract cell residue after drying the yeast extract cell residue by a known method. The drying method is not particularly limited, and known drying methods can be used, such as atmospheric heat drying, vacuum drying, spray drying, and freeze drying. For example, the dry mass can be obtained by drying a sample in an atmospheric heat dryer at 105°C for 5 hours and then weighing the residue.

[0014] Yeast extract cell residue is composed of proteins, lipids, ash, dietary fiber, and the like. The protein content is, for example, 20% to 60% by mass based on the dry mass of the yeast extract cell residue. The lipid content is, for example, 1% to 10% by mass based on the dry mass of the yeast extract cell residue. The ash content is, for example, 0% to 10% by mass based on the dry mass of the yeast extract cell residue. The dietary fiber content is, for example, 10% to 60% by mass based on the dry mass of the yeast extract cell residue. In addition to general nutritional components, yeast extract cell residue also contains components such as β-glucan and α-mannan, which have been reported to have physiological functions. β-glucan is a polysaccharide formed by the polymerization of D-glucose via β-1,3 and β-1,6 bonds, and is classified as dietary fiber because it is a resistant component in food that cannot be digested by human digestive enzymes. α-Mannan is a polysaccharide formed by the polymerization of D-mannose via α-1,6, α-1,2, or α-1,3 bonds. It is classified as dietary fiber because it is an indigestible food component that cannot be digested by human digestive enzymes. The β-glucan content is, for example, 10% to 40% by mass relative to the dry mass of the yeast extract cell residue. The α-mannan content is, for example, 10% to 40% by mass relative to the dry mass of the yeast extract cell residue. The content of each component can be measured by known analytical methods. The protein content can be determined, for example, by measuring the nitrogen content using the Kjeldahl method and multiplying the nitrogen content by a conversion factor of 6.25. The lipid content can be quantified, for example, by acid hydrolysis. The ash content can be quantified by direct ashing. The dietary fiber content can be quantified, for example, by the Prosky method (enzyme-gravimetric method) or high-performance liquid chromatography (enzyme-HPLC method). The α-mannan content can be measured, for example, by quantifying the amount of mannose produced by hydrolyzing mannan, and the β-glucan content can be measured, for example, using a (1-3), (1-4)-β-glucan measurement kit (Megazyme).

[0015] The yeast is not particularly limited as long as it is applicable to the food industry, and examples thereof include yeast for beer brewing, yeast for bread brewing, and yeast for sake brewing. Alternatively, the yeast is not limited to these, and examples thereof include those belonging to genera such as Saccharomyces, Saccharomycodes, Rhodotorula, Endomycopsis, Nematospora, Brettanomyces, Candida, and Torulopsis. Among these, Saccharomyces cerevisiae, Saccharomyces pastorianus, Saccharomyces bayanus, and Candida utilis are preferred, with Saccharomyces pastorianus (beer yeast) being particularly preferred. These may be used alone or in combination of two or more types.

[0016] The glucanase in (a) above may be, for example, a naturally occurring product, a commercially available product, or a product obtained by a method using genetic recombination technology or the like.

[0017] The production method of the present invention is characterized by using at least two types of glucanases: a glucanase having endoactivity and a glucanase having exoactivity. The total amount of the at least two types of glucanases added is, for example, 1.5% by mass or less, preferably 1.2% by mass or less, more preferably 1.0% by mass or less, and particularly preferably 0.8% by mass or less, based on the dry mass of the yeast extract cell residue. The blending ratio (by mass) of the glucanase having endoactivity to the glucanase having exoactivity is appropriately set depending on the enzymes used and is not particularly limited, but is, for example, 1:20 to 20:1, preferably 1:10 to 10:1, more preferably 1:5 to 5:1, and particularly preferably 1:3 to 3:1.

[0018] Examples of glucanases having endo activity include endo-β-1,3(4)-glucanases belonging to the enzyme number EC 3.2.1.6, endo-β-1,3-glucanases belonging to the enzyme number EC 3.2.1.39, licheninase belonging to the enzyme number EC 3.2.1.73, and endo-β-1,6-glucanases belonging to the enzyme number EC 3.2.1.75, preferably endo-1,3(4)-β-glucanases belonging to the enzyme number EC 3.2.1.6, and endo-β-1,3-glucanases belonging to the enzyme number EC 3.2.1.39. Examples of glucanases having endo activity include glucanases derived from the genus Streptomyces, preferably glucanases derived from the genus Streptomyces having β-1,3 activity (e.g., glucanases belonging to the enzyme number EC 3.2.1.6 or EC 3.2.1.39). Such a glucanase is available, for example, from Nagasevita Co., Ltd. under the name "Denazyme GEL-L1 / R." Furthermore, glucanases having endo activity are preferably those having only endo activity, and more preferably those having low, almost no, or no protease activity (i.e., no protease activity). The amount of glucanase having endo activity to be added is determined appropriately depending on the type of yeast extract cell residue and glucanase, but is preferably 0.01% by mass or more, more preferably 0.1% by mass or more, and preferably less than 1% by mass, more preferably 0.5% by mass or less, and particularly preferably 0.3% by mass or less, based on the dry mass of the yeast extract cell residue.

[0019] Examples of glucanases having exoactivity include β-glucosidases and glucan β-1,6-glucosidases belonging to the enzyme number EC 3.2.1.21, and glucan β-1,3-glucosidases belonging to the enzyme number EC 3.2.1.58, preferably glucan β-1,3-glucosidases, and glucanases derived from the genus Trichoderma, Talaromyces, and / or Aspergillus, preferably glucanases derived from the genus Trichoderma or Talaromyces and Aspergillus having β-1,3, β-1,4, and / or β-1,6 activity (e.g., glucanases derived from the genus EC 3.2.1.58). Such glucanases are available, for example, as "Sumiteam TG" from Shin-Nihon Chemical Industry Co., Ltd., "Sumiteam C" from Shin-Nihon Chemical Industry Co., Ltd., and "Filtrase Smart" from DSM Japan Co., Ltd. Furthermore, the glucanase having exo activity may be any glucanase as long as it has exo activity as its primary activity, and may also have endoactivity and / or protease activity. The amount of glucanase having exo activity added is appropriately determined depending on the type of yeast extract cell residue and glucanase, but is preferably 0.01% by mass or more, more preferably 0.1% by mass or more, and preferably less than 1% by mass, more preferably 0.75% by mass or less, even more preferably 0.6% by mass or less, and particularly preferably 0.5% by mass or less, relative to the dry mass of the yeast extract cell residue.

[0020] As used herein, the term "yeast enzyme-treated product" refers to a product obtained by decomposing yeast extract cell residue with at least two types of glucanases, specifically, a product obtained by decomposing yeast cell walls and the like contained in the yeast extract cell residue with glucanases having endoactivity and exoactivity.

[0021] The yeast enzyme-treated product is composed of proteins, lipids, ash, dietary fiber, and the like. The protein content is, for example, 20% to 60% by mass based on the dry mass of the yeast enzyme-treated product. The lipid content is, for example, 1% to 10% by mass based on the dry mass of the yeast enzyme-treated product. The ash content is, for example, 0% to 10% by mass based on the dry mass of the yeast enzyme-treated product. The dietary fiber content is, for example, 10% to 60% by mass based on the dry mass of the yeast enzyme-treated product. In addition to general nutritional components, the yeast enzyme-treated product also contains components such as β-glucan and α-mannan, which have been reported to have physiological functions. β-glucan is a polysaccharide formed by the polymerization of D-glucose via β-1,3 and β-1,6 bonds, and is classified as dietary fiber because it is a difficult-to-digest component in food that is not digested by human digestive enzymes. α-Mannan is a polysaccharide formed by polymerizing D-mannose via α-1,6, α-1,2, or α-1,3 bonds, and is classified as dietary fiber because it is an indigestible component in food that is not digested by human digestive enzymes. The β-glucan content varies due to degradation by the exoactivity of glucanase, but is, for example, 0% to 40% by mass relative to the dry mass of the yeast enzyme-treated product. The α-mannan content is, for example, 10% to 40% by mass relative to the dry mass of the yeast enzyme-treated product. The content of each component can be measured by known analytical methods, as described above. The yeast enzyme-treated product obtained by the production method of the present invention also contains protein, lipids, ash, dietary fiber, etc., but has improved dispersibility.

[0022] The "treatment" in (a) above is not particularly limited as long as the conditions are such that glucanase can decompose yeast cell walls and the like contained in the yeast extract cell residue, and can be appropriately changed depending on the origin of the yeast cell walls, the type and / or amount of glucanase, the desired properties, and the like. The treatment is usually carried out in a desired solvent (e.g., water). For example, the treatment is carried out using a suspension of yeast extract cell residue in a solvent (e.g., water). The suspension may be subjected to a sterilization treatment (e.g., heat sterilization, filtration sterilization, etc.) as necessary. Treatment of yeast extract cell residue with glucanase can be carried out, for example, at a temperature above 0°C but below about 100°C (preferably about 10 to about 70°C, more preferably about 25 to about 65°C, and even more preferably about 40 to about 60°C), for about 0.5 to about 120 hours (preferably about 0.5 to about 60 hours, more preferably about 1 to about 24 hours, even more preferably about 3 to about 24 hours, and still more preferably about 12 to about 24 hours), at a pH of about 1 to about 12 (preferably about 2 to about 10, more preferably about 3 to about 8, and even more preferably about 4 to about 6). After the above treatment, the glucanase may be inactivated, if necessary, by autoclaving, high-temperature treatment, acid or alkali treatment, or the like.

[0023] In (b) above, the treated product obtained in (a) may be used as is as a "yeast enzyme-treated product" in its entirety, including the residue (insoluble fraction), or, if necessary, may be further purified (e.g., HPLC, ultrafiltration, etc.), concentrated (e.g., air-drying, vacuum filtration, etc.), sterilized (e.g., heat sterilization, filtration sterilization, etc.), dried (e.g., air-drying, heating, vacuum, spray-drying, freeze-drying, etc.) and used as a "yeast enzyme-treated product." The conditions for these steps can be adjusted as appropriate by those skilled in the art.

[0024] The processed product obtained in (a) in (b) above, including the residue (insoluble fraction), may be used as is in its entirety as the milk substitute food or beverage of the present invention. Alternatively, the milk substitute food or beverage containing the yeast enzyme-treated product of the present invention may be in the form of a cloudy liquid or paste containing the processed product in an appropriate medium (preferably water) at an appropriate concentration, for example, about 0.001 to about 30% by mass, preferably about 5 to about 20% by mass, based on the dry mass. By using the milk substitute food or beverage of the present invention, nutrients such as yeast-derived protein and dietary fiber can be ingested.

[0025] The yeast enzyme-treated product described above can be used in place of milk in foods and beverages containing milk (cow's milk). For example, the "yeast enzyme-treated product" of the present invention may be incorporated into foods and beverages, or may be mixed with foods and beverages before, during, and / or after cooking.

[0026] In one embodiment of the present invention, a milk substitute food or beverage is provided, comprising a yeast enzyme-treated product obtained by the present invention and fats and / or carbohydrates. The amount of the yeast enzyme-treated product in the milk substitute food or beverage of the present invention varies depending on the form of the food or beverage, and is not particularly limited. However, the dry mass of the yeast enzyme-treated product can be approximately 1 to 30% by mass relative to the mass of the milk substitute food or beverage. The milk substitute food or beverage of the present invention may be consumed as yeast-based milk or used as a milk substitute during cooking. The milk substitute food or beverage of the present invention is a new milk substitute food or beverage made primarily from yeast, which has a protein-based main component, a moderate amount of fats and oils, and a milk-like appearance. If no animal-derived ingredients are used as ingredients, the food or beverage is suitable for vegetarians and vegans who avoid consuming animal products. If fats and oils produced using microorganisms are used as ingredients, the new milk substitute food or beverage is made using neither animals nor plants. The amount of yeast enzyme-treated product to be blended when used as yeast milk for drinking is not particularly limited, but can be approximately 1 to 30% by mass, preferably 2 to 20% by mass, and more preferably 5 to 10% by mass, of the dry mass of the yeast enzyme-treated product relative to the mass of the yeast milk. Blending a larger amount of yeast enzyme-treated product in yeast milk allows for more efficient intake of yeast-derived proteins, dietary fiber, and other nutrients. On the other hand, increasing the blending amount may increase the viscosity of the yeast milk and impair its dispersibility (redispersibility). However, if the blending amount of yeast enzyme-treated product is within the above range, a beverage can be provided that allows for efficient intake of yeast-derived proteins, dietary fiber, and other nutrients while maintaining the dispersibility and favorable mouthfeel of yeast milk. When used as a milk substitute during cooking, it can be used in foods such as gratin and confectionery.

[0027] "Oils and fats" as used herein includes, but is not limited to, vegetable oils and fats, animal oils and fats, processed oils and fats, such as edible safflower oil, edible grape oil, edible soybean oil, edible sunflower oil, edible corn oil, edible cottonseed oil, sesame oil, edible rapeseed oil, edible rice bran oil, edible peanut oil, edible olive oil, edible palm oil, edible palm olein, edible palm stearin, edible palm kernel oil, edible coconut oil, edible blended oils, flavored edible oils, beef tallow, lard, chicken oil, fish oil, dairy fat, hydrogenated oils and fats, and oils and fats produced by microorganisms such as yeast.

[0028] The amount of fat or oil contained in the milk substitute food or beverage of the present invention is not particularly limited, as it varies depending on the form of the food or beverage, but is generally 0.1 to 30% by mass relative to the mass of the milk substitute food or beverage. The amount of fat or oil when used as yeast milk is generally 0.1 to 30% by mass, preferably 1 to 10% by mass, and more preferably 2 to 5% by mass relative to the mass of the yeast milk. Incorporating a larger amount of fat or oil into yeast milk can provide richer flavor, a more satisfying taste, and a smoother drinking experience. On the other hand, excessively large amounts can have undesirable effects, such as a decrease in the drinkability of the yeast milk and an increase in the amount of energy ingested. However, if the amount of fat or oil is within the above range, it can provide richness, a satisfying taste, and a smoother drinking experience while also providing the yeast milk with an easy-to-drink beverage that provides adequate energy intake. When used as yeast milk, an appropriate medium (preferably water) can be incorporated in addition to the yeast enzyme-treated product and fat or oil. The amount of medium (preferably water) in the yeast milk is generally 30 to 97% by mass, preferably 40 to 95% by mass, and more preferably 45 to 93% by mass, relative to the mass of the yeast milk. When the amount of medium (preferably water) in the yeast milk is within the above range, the yeast enzyme-treated product is homogeneously dispersed in the medium, resulting in a drink with a pleasant texture.

[0029] The term "carbohydrate" as used herein includes, for example, monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, polysaccharides, starch hydrolysates, reduced starch hydrolysates, etc., and these may be used alone or in combination of two or more kinds. Examples of carbohydrates include, but are not limited to, ketotriose, aldotriose, erythrulose, ribulose, xylulose, lyxose, deoxyribose, psicose, fructose, sorbose, tagatose, allose, altrose, glucose, mannose, gulose, idose, galactose, talose, fucose, fuculose, rhamnose, sedoheptulose, sucrose, lactose, maltose, trehalose, turanose, cellobiose, raffinose, maltotriose, acarbose, stachyose, fructooligosaccharides, galactooligosaccharides, mannanoligosaccharides, glycogen, starch, cellulose, dextrin, corn syrup, powdered sugar, and glucan, which may be used alone or in combination of two or more.

[0030] The method for producing the milk substitute food or drink of the present invention is not particularly limited, and can be carried out by mixing the yeast enzyme-treated product with optional ingredients such as fats and / or carbohydrates, as needed, by any method. For example, the method can be carried out by mixing and emulsifying predetermined amounts of the yeast enzyme-treated product, fats and / or water using an appropriate stirrer (e.g., a homomixer, a homogenizer, etc.) to obtain a base liquid (emulsion), and then adding optional ingredients such as carbohydrates.

[0031] The milk substitute food and drink of the present invention may further contain additives such as, but not limited to, excipients, lubricants, binders, disintegrants, pH adjusters, solvents, solubilizers, suspending agents, buffers, preservatives, antioxidants, colorants, sweeteners, surfactants, flavorings, etc. These additives may be, for example, known additives for foods or pharmaceuticals, and the amount used may be adjusted as appropriate by those skilled in the art depending on the purpose.

[0032] Alternatively, foods and beverages to which the "yeast enzyme-treated product" obtained by the present invention can be applied or which contain the same include, but are not limited to, beverages and foods that contain or are intended to contain milk (e.g., cow's milk, soy milk, almond milk, all-milk, coconut milk, etc.) or milk powder, such as soups, dressings, semi-solid dressings, mayonnaise, confectioneries, chocolate, margarine, fat spreads, ice cream, lacto ice cream, ice milk, fresh cream, bread, retort foods, extract seasonings, processed oils and fats, prepared oils and fats, cheese, cheese food, processed cheese, pudding, and other processed foods.

[0033] The amount of the "yeast enzyme-treated product" obtained in the present invention to be applied is not particularly limited as long as the object of the present invention can be achieved. The amount of the yeast enzyme-treated product (dry mass) can be, for example, about 0.001 to about 30% by mass, preferably about 0.01 to about 20% by mass, and more preferably about 0.1 to about 15% by mass, based on the total mass of the food, drink, etc. or processed products thereof.

[0034] In another embodiment of the present invention, there is provided a use of a glucanase hydrolyzed product of yeast extract cell residue as a milk substitute food or beverage, wherein the glucanases are at least two types of glucanases, one having endoactivity and one having exoactivity. The explanation given above regarding the embodiment of the method for producing an enzyme-treated yeast product can be similarly applied to these embodiments of the invention.

[0035] The present invention will be described in more detail below using examples, but these examples are not intended to limit the scope of the present invention in any way.

[0036] Preparation Example 1 Brewer's yeast (manufactured by Asahi Group Foods Co., Ltd.) was extracted with hot water, and the heavy liquid (insoluble fraction) obtained by separating the yeast extract using a nozzle-type continuous centrifuge was sterilized at 125°C for 40 seconds and spray-dried to obtain a yeast extract cell residue as the composition of Preparation Example 1.

[0037] Comparative Example 1 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solid content 16%), which was sterilized in an autoclave (121°C, 15 minutes), and then a glucanase having endoactivity of 0.2% by mass relative to the dry mass of the yeast extract cell residue (Denazyme GEL-L1 / R, manufactured by Nagasevita Co., Ltd.) was added at 50°C and pH 5.4, followed by treatment at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize the product (121°C, 15 minutes), and freeze-dried to obtain a yeast enzyme-treated product as the composition of Comparative Example 1.

[0038] Comparative Example 2 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solid content 16%), which was sterilized in an autoclave (121°C, 15 minutes), and then glucanase (Shin-Nihon Chemical Industry Co., Ltd., Sumiteam TG) having exoactivity in an amount of 0.1% by mass relative to the dry mass of the yeast extract cell residue was added at 50°C and pH 5.4, followed by treatment at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize it (121°C, 15 minutes), and freeze-dried to obtain a yeast enzyme-treated product as the composition of Comparative Example 2.

[0039] Comparative Example 3 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solid content 16%), which was sterilized in an autoclave (121°C, 15 minutes), and a glucanase (Shin-Nihon Chemical Industry Co., Ltd., Sumiteam TG) having exoactivity in an amount of 0.75% by mass relative to the dry mass of the yeast extract cell residue was added at 50°C and pH 5.4, followed by treatment at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize it (121°C, 15 minutes), and freeze-dried to obtain a yeast enzyme-treated product as the composition of Comparative Example 3.

[0040] Example 1 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solids content 16%), which was sterilized in an autoclave (121°C, 15 minutes). A glucanase having endoactivity (Denazyme GEL-L1 / R, manufactured by Nagasevita Co., Ltd.) and a glucanase having exoactivity (Shin-Nihon Chemical Industry Co., Ltd., Sumizyme TG) were added at 0.25% by mass relative to the dry mass of the yeast extract cell residue at 50°C and pH 5.4, and the mixture was treated at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize the product (121°C, 15 minutes), followed by freeze-drying to obtain the yeast enzyme-treated product of Example 1.

[0041] Example 2 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solids content 16%), which was sterilized in an autoclave (121°C, 15 minutes). A glucanase having endoactivity (Denazyme GEL-L1 / R, manufactured by Nagasevita Co., Ltd.) and a glucanase having exoactivity (Shin-Nihon Chemical Industry Co., Ltd., Sumizyme TG) were added at 0.25% by mass relative to the dry mass of the yeast extract cell residue at 50°C and pH 5.4, and the mixture was treated at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize the product (121°C, 15 minutes), followed by freeze-drying to obtain the yeast enzyme-treated product of Example 2.

[0042] Example 3 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solid content 16%), which was sterilized in an autoclave (121°C, 15 minutes). A glucanase having 0.25% by mass of endoactivity (Denazyme GEL-L1 / R, manufactured by Nagasevita Co., Ltd.) and 0.2% by mass of exoactivity (Filtrase Smart, manufactured by DSM Japan Co., Ltd.) were added at 50°C and pH 5.4, relative to the dry mass of the yeast extract cell residue, and the mixture was treated at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize the product (121°C, 15 minutes), yielding the yeast enzyme-treated product of Example 3.

[0043] Example 4 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solids content 16%), which was sterilized in an autoclave (121°C, 15 minutes). A glucanase having endoactivity (Denazyme GEL-L1 / R, manufactured by Nagasevita Co., Ltd.) and a glucanase having exoactivity (Shin-Nihon Chemical Industry Co., Ltd., Sumizyme C) were added in an amount of 0.25% by mass relative to the dry mass of the yeast extract cell residue at 50°C and pH 5.4, and the mixture was treated at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize the product (121°C, 15 minutes), yielding the yeast enzyme-treated product described in Example 4.

[0044] Example 5 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solids content 16%), which was sterilized in an autoclave (121°C, 15 minutes), and then a glucanase having 0.25% by mass of endoactivity (Denazyme GEL-L1 / R, manufactured by Nagasevita Co., Ltd.) and a glucanase having 1.0% by mass of exoactivity (Shin-Nihon Chemical Industry Co., Ltd., Sumizyme TG) were added at 50°C and pH 5.4, relative to the dry mass of the yeast extract cell residue, and the mixture was treated at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize the product (121°C, 15 minutes), and freeze-dried to obtain the yeast enzyme-treated product of Example 5.

[0045] Example 6 The yeast extract cell residue obtained in Preparation Example 1 was suspended in deionized water to prepare a slurry (solids content 16%), which was sterilized in an autoclave (121°C, 15 minutes), and then a glucanase having endoactivity (Denazyme GEL-L1 / R, manufactured by Nagasevita Co., Ltd.) and a glucanase having exoactivity (Shin-Nihon Chemical Industry Co., Ltd., Sumizyme TG) were added in an amount of 0.1% by mass relative to the dry mass of the yeast extract cell residue at 50°C and pH 5.4, followed by treatment at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize the product (121°C, 15 minutes), and freeze-dried to obtain the yeast enzyme-treated product of Example 6.

[0046] Example 7: A slurry (10% solids content) of yeast extract cell residue (manufactured by Asahi Group Foods Co., Ltd., product name "Yeast Cell Wall") obtained by extracting an extract from brewer's yeast by autolysis was suspended in deionized water and sterilized in an autoclave (121°C, 15 minutes). A glucanase having 0.25% by mass of endoactivity (manufactured by Nagasevita Co., Ltd., Denatzyme GEL-L1 / R) and a glucanase having 0.1% by mass of exoactivity (manufactured by Shin-Nippon Chemical Industry Co., Ltd., Sumitzyme TG) were added at 50°C and pH 5.4, relative to the dry mass of the yeast extract cell residue. The mixture was then treated at 50°C for 18 hours. The treated product was then heated in an autoclave to inactivate the glucanase and sterilize it (121°C, 15 minutes), and freeze-dried to obtain the yeast enzyme-treated product of Example 7.

[0047] [Preparation Example 2-10] The raw materials were mixed according to the composition shown in Table 1, and then pre-emulsified (4000 rpm, 5 minutes) using a homogenizer (Labo-Lusion, Primix Corporation), followed by emulsification using a homogenizer (Homogenizer LAB1000, SMT Corporation: 20 MPa, 3 times) and sterilization in an autoclave (121°C, 40 minutes) to obtain a milk substitute food or beverage.

[0048]

[0049] Test Example 1 Sensory Evaluation of Yeast Enzyme-Treated Products Sensory evaluation was performed on each of the compositions obtained in Comparative Examples 1 and 3, and Examples 1-6, as follows. First, each treated product was added to room temperature water to a solids content of 10% by mass, and the mixture was thoroughly stirred to obtain a suspension. Then, sensory evaluation was performed on each suspension by five expert panelists. The composition of Comparative Example 1 was assigned a score of 0 as a control, and the flavor intensity of each sample was evaluated based on the following evaluation criteria. Note that negative values ​​(weaker than the control) are preferred for "sourness," "roasted flavor," and "astringency," and positive values ​​are preferred for "milk flavor" and "smoothness." Furthermore, "smoothness" refers to a mild taste with little unpleasant flavor, and "milk flavor" was evaluated based on a subtle sweet aroma after swallowing, the absence of off-flavors, and plainness, to determine whether the flavor was similar to animal milk overall.

[0050] [Evaluation criteria] -3 points: Very weak -2 points: Weak -1 point: Slightly weak 0 point: Control +1 point: Slightly strong +2 points: Strong +3 points: Very strong

[0051] [Results] The sensory evaluation results are shown as average values ​​in Figure 1. As a result, compared to the composition of Comparative Example 1, the compositions of Examples 1-4 and 6 had the same or reduced "sourness," "roasted feeling," and "astringency," and had increased "mellowness" and "milky flavor." With regard to "milky flavor," all of the compositions of Examples 1-4 and 6 received scores of 1.3 or higher, indicating favorable sensory characteristics. Among them, all of the compositions of Examples 1-3 and 6 received scores of 1.5 or higher, indicating particularly favorable sensory characteristics. These results demonstrated that treatment with at least two types of glucanases having different activities, particularly treatment with at least two types of glucanases whose total addition amount is less than a specific amount, can produce processed products exhibiting favorable sensory characteristics. Conversely, compared to the composition of Comparative Example 1, Comparative Example 3 had increased "sourness," "roasted feeling," and "astringency," and no significant differences were observed in "mellowness" and "milky flavor." These results demonstrated that treatment with a single type of glucanase can produce processed products exhibiting unfavorable sensory characteristics. It was also shown that treatment with an excessive amount of glucanase resulted in a treated product exhibiting undesirable organoleptic properties.

[0052] Test Example 2 Sensory Evaluation of Milk Substitute Food and Drink Preparation Examples 2-7, 9, and 10 were subjected to sensory evaluation by five expert panelists in the same manner as in Test Example 1. For the evaluation items of "mellowness," "acidity," "astringency," "roasted feeling," and "milk-like flavor," the sample of Preparation Example 9 (including the composition of Comparative Example 1) was assigned a score of 0 as a control, and the strength of the flavor of each sample was evaluated based on the following evaluation criteria.

[0053] [Results] Compared with the sample of Preparation Example 9 (including the composition of Comparative Example 1), the samples of Preparation Examples 2-5 and 7 (including Examples 1-4 and 6) had the same or reduced "sourness," "roasted feeling," and "astringency," and "mellowness" and "milk-like flavor" were increased. Conversely, compared with the sample of Preparation Example 9 (including the composition of Comparative Example 1), the sample of Preparation Example 10 (including the composition of Comparative Example 3) had increased "sourness," "roasted feeling," and "astringency," and no significant difference was observed in "mellowness" and "milk-like flavor." These results indicate that the evaluation results of the yeast enzyme-treated product obtained in Test Example 1 show similar sensory properties even in the form of a milk substitute food or beverage.

[0054] Test Example 3 Evaluation of Dispersibility of Precipitate of Yeast Enzyme-Treated Product Using the samples of Preparation Example 2-9, the re-dispersibility of precipitate was defined as follows and evaluated. 40 ml of each of the milk substitute foods and beverages of Preparation Example 2-9 was poured into a 50 ml round-bottom centrifuge tube (manufactured by BM Kiki Co., Ltd.) and centrifuged (2000 × g, 2 minutes) to precipitate, and 1 ml of the supernatant not containing the precipitate was collected and its mass (g) before drying was measured. Next, the supernatant was heated at 105°C for 5 hours to dry, and then its mass (g) after drying was measured, and the solids concentration (S ０ ) (%) was calculated according to the following formula:

[0055]

[0056] Thereafter, the mixture was stirred at 45 rpm for 5 minutes using a tube rotator (MX-RD-E, DLAB Scientific Co. Ltd.) to redisperse the precipitate, and 1 ml of the supernatant was similarly recovered and the solid content was measured in the same manner as above (S １ Finally, the solid content was measured after the precipitate was completely dispersed. (Total) The change in the solid content ratio in the precipitate before and after stirring was defined as the residual precipitate rate, which was calculated using the following formula:

[0057]

[0058] The results of each preparation example are shown in Table 2.

[0059]

[0060] [Results] The samples of Preparation Example 9 (including the composition of Comparative Example 1) were barely dispersed and the residual precipitate rate exceeded 80%, whereas the samples of Preparation Examples 2 to 8 (including Examples 1 to 7) all had residual precipitate rates below 20%, indicating improved redispersibility. These results demonstrate that treatment with a glucanase having exo activity in addition to a glucanase having endo activity can yield a treated product with improved precipitate dispersibility.

[0061] Test Example 4 Evaluation of Precipitate Volume of Enzyme-Treated Yeast Product The volume of precipitate that could not be decomposed by enzyme treatment was evaluated for each of the compositions obtained in Comparative Examples 1-3 and Examples 1-7 as follows. First, each treated product was added to room temperature water to a concentration of 10% by mass and stirred thoroughly to obtain a suspension. Next, 14 ml of this suspension was placed in a 15 ml Falcon tube and centrifuged (3000 × g, 5 minutes) to precipitate the precipitate. The volume of the precipitate was measured using the scale, and the proportion of the precipitate in the solution was calculated. Similar evaluations were also performed on the yeast extract cell residue (hot water treatment) obtained in Preparation Example 1 and the yeast extract cell residue (product name "yeast cell wall"; autolysis) used in Example 7, both of which were enzyme-untreated, as controls. The results for each Comparative Example, Example, and Control Example are shown in Table 3.

[0062]

[0063] [Results] In all of Comparative Examples 1-3, which were treated with one type of glucanase, the proportion of precipitate exceeded 20%. In particular, in Comparative Example 2, which was treated with 0.1% by mass of Sumizyme TG, the volume of precipitate was almost unchanged compared to Preparation Example 1, which was not treated with the enzyme, and the proportion of the volume occupied by precipitate was large. On the other hand, in all of Examples 1-7, the proportion of the volume occupied by precipitate was reduced to 20% or less. These results demonstrate that treatment with at least two types of glucanases having different activities can produce a treated product with a reduced precipitate volume. The results in Tables 2 and 3 demonstrate that treatment with at least two types of glucanases having different activities can produce a treated product with a reduced amount of precipitate that tends to stick when left as a suspension of a milk substitute using a yeast material.

[0064] As described above, in the production method of the present invention, by treating the yeast material with at least two types of glucanases having different activities, the dispersibility of the resulting yeast enzyme-treated product is improved compared to conventional products. Therefore, milk substitute foods and beverages using this product are less likely to precipitate, and even if precipitate does occur, it is easy to redisperse, making them suitable for consumption. Improvements in flavor as milk substitute foods and beverages were also demonstrated. Therefore, milk substitute foods and beverages using the yeast material obtained by the production method of the present invention make it easier to ingest yeast-derived proteins, dietary fiber, and other nutrients.

Claims

1. A method for producing a yeast enzyme-treated product for use in a dairy substitute food or beverage, comprising: (a) treating yeast extract cell residue with glucanase; and (b) recovering the treated product obtained in (a), wherein the glucanase is at least two types of glucanase: a glucanase having endoactivity and a glucanase having exoactivity.

2. The production method according to claim 1, wherein the glucanase having endoactivity is a glucanase belonging to enzyme number EC 3.2.1.6 or EC 3.2.1.

39.

3. The method according to claim 1, wherein the glucanase having endo activity is a glucanase derived from the genus Streptomyces and having β-1,3 activity.

4. The production method described in claim 1, wherein the amount of glucanase having endo activity added is 0.1 mass% or more based on the dry mass of the yeast extract cell residue.

5. The production method according to claim 1, wherein the glucanase having exoactivity is a glucanase belonging to the enzyme number EC 3.2.1.

58.

6. The production method according to claim 1, wherein the glucanase having exo activity is a glucanase having β-1,3, β-1,4, and / or β-1,6 activity derived from the genera Trichoderma, Talanomyces, and / or Aspergillus.

7. The production method according to claim 1, wherein the amount of glucanase having exoactivity added is less than 1% by mass relative to the dry mass of the yeast extract cell residue.

8. The production method according to claim 1, wherein the yeast extract cell residue is a water-insoluble fraction obtained after hot water extraction of yeast or autolysis of yeast.

9. An enzyme-treated yeast product obtained by the production method described in claim 1.

10. A milk substitute food or beverage containing a yeast enzyme-treated product obtained by the production method described in claim 1.

11. The milk substitute food or beverage according to claim 10, further comprising 0.1 to 30% by mass of fat or oil.

12. Use of a glucanase-treated product of yeast extract cell residue as a milk substitute food or beverage, wherein the glucanase is at least two types of glucanase, namely, a glucanase having endoactivity and a glucanase having exoactivity.

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