Classifications

• • 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
  • A23C20/00—Cheese substitutes
  • A23C20/02—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates
• • 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
  • A23C20/00—Cheese substitutes
  • A23C20/02—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates
  • A23C20/025—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates mainly containing proteins from pulses or oilseeds
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J3/00—Working-up of proteins for foodstuffs
  • A23J3/14—Vegetable proteins
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J3/00—Working-up of proteins for foodstuffs
  • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
  • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
  • A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
  • A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
• • 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
  • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
  • C12N9/2411—Amylases
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Y—ENZYMES
  • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)

Definitions

• the present invention relates to a method for producing a stretchable cheese substitute. More specifically, the present invention relates to a method for producing a heat-stretchable stretchable cheese substitute made from vegetable protein.
• the present invention provides a cheese substitute manufacturing technology that can impart improved stretchability to a cheese substitute containing a vegetable protein and 0.6 parts by weight or more of starch with respect to 1 part by weight of the vegetable protein. intended to provide
• the present inventors added a protease to a material composition having a specific composition containing vegetable protein and 0.6 parts by weight or more of starch with respect to 1 part by weight of the vegetable protein, cheese obtained by It was found that the stretchability of the substitute was improved. Furthermore, in the case where amylase is used in combination with protease, when amylase is added to the material composition together with protease, the starch refinement power of amylase allowed for the material composition is limited to a predetermined condition (that is, a condition that satisfies the relationship of 8 U or less of starch refinement power per 1 g of starch with respect to 100,000 U of protease activity per 1 g of vegetable protein).
• Section 1 A step of treating a material composition containing a vegetable protein and 0.6 parts by weight or more of starch per 1 part by weight of the vegetable protein with an enzymatic agent containing a protease and optionally containing an amylase; A substitute for stretchy cheese, wherein the enzymatic agent is used so that the starch refinement power of the amylase per 1 g of the starch is 8 U or less with respect to the protease activity of 100,000 U of the protease per 1 g of the vegetable protein. A method of making things.
• Item 3. Item 3. The production method according to Item 1 or 2, wherein the protease is a bacterial protease. Section 4.
• Item 4. The production method according to any one of Items 1 to 3, wherein the protease is derived from the genus Bacillus and/or Geobacillus.
• Item 6. The production method according to any one of Items 1 to 5, wherein the enzyme agent is used so that the protease activity is 10 to 500 U per 1 g of the vegetable protein.
• Item 7. Item 7.
• Item 8. Item 8. The production method according to any one of Items 1 to 7, wherein the vegetable protein is pea protein, broad bean protein, chickpea protein, and/or lentil protein.
• Item 9. Item 9. The production method according to any one of Items 1 to 8, wherein the content of the vegetable protein in the material composition is 1% by weight or more and less than 15% by weight.
• Item 10. Item 10.
• Item 11. The production method according to any one of Items 1 to 10, wherein the starch content per 1 part by weight of the vegetable protein is 5 parts by weight or less.
• Item 12. A stretchability improver for a stretchable cheese substitute, comprising an enzymatic agent containing protease and optionally amylase, and containing 0.6 parts by weight or more of starch per 1 part by weight of the vegetable protein. and A stretchability improver used so that the starch refinement power of the amylase per 1 g of the starch is 8 U or less for the protease activity of the protease of 100,000 U per 1 g of the vegetable protein.
• Item 13. Item 13.
• the method for producing a stretchable cheese substitute of the present invention comprises preparing a material composition comprising a vegetable protein and 0.6 parts by weight or more of starch per 1 part by weight of the vegetable protein, A step of treating with an enzymatic agent containing protease and optionally amylase (hereinafter also referred to as a "protease treatment step"), wherein the enzymatic agent is treated with the protease of the protease per 1 g of the vegetable protein It is characterized in that the amylase is used so that the starch gelatinization power of the amylase per 1 g of the starch is 8 U or less with respect to an activity of 100,000 U.
• the stretchability of the obtained cheese substitute is improved, or in addition to the improvement of stretchability, the heat meltability improvement effect and / or the hydrophobic peptide reduction effect (hydrophobic peptide reduction effect means bitterness It refers to the effect of degrading a presenting hydrophobic peptide and replacing it with a hydrophobic amino acid.) can be further imparted.
• Material composition containing vegetable protein and starch The plant from which the vegetable protein is derived is not particularly limited, but for example, beans such as peas, soybeans, broad beans, chickpeas, and lentils; Cereals such as rice, buckwheat, millet and millet; nuts such as almonds, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, peanuts and coconuts.
• beans such as peas, soybeans, broad beans, chickpeas, and lentils
• Cereals such as rice, buckwheat, millet and millet
• nuts such as almonds, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, peanuts and coconuts.
• vegetable proteins derived from these plants one type may be used alone, or two or more types having different origins may be used in combination.
• bean proteins are preferable.
• the content of vegetable protein in the material composition is not particularly limited, but is, for example, 1 to 30% by weight, preferably 1% to 15% by weight, more preferably 3 to 12% by weight, still more preferably 4% by weight. ⁇ 11 wt%, more preferably 4-9 wt%, 4-8 wt%, 4-7 wt%, 4-6 wt%, 6-11 wt%, 7-11 wt%, 8-11 wt%, Or 9 to 11% by weight.
• the plant from which starch is derived is not particularly limited as long as it can impart stretchability to the cheese substitute, but examples include cassava, potato, sweet potato, and kudzu.
• starch derived from these plants one type may be used alone, or two or more types having different origins may be used in combination.
• cassava starch (tapioca starch) is preferable from the viewpoint of further improving stretchability, or from the viewpoint of further imparting the effect of improving heat meltability and / or the effect of reducing hydrophobic peptides in addition to this viewpoint. mentioned.
• the content of starch per 1 part by weight of vegetable protein is 0.6 parts by weight or more. Since the production method of the present invention is excellent in the effect of improving stretchability, when an enzymatic agent containing amylase is used, the ratio of starch to vegetable protein is adversely affected by the starch gelatinization power. Even so, the stretchability can be effectively improved.
• a suitable example of the content of starch per 1 part by weight of vegetable protein is preferably 0.7 parts by weight or more, more preferably 0.8 parts by weight per 1 part by weight of vegetable protein. More preferably 1 part by weight or more, still more preferably 1.5 parts by weight or more, even more preferably 2 parts by weight or more, and particularly preferably 2.5 parts by weight or more.
• the upper limit of the content range of starch per 1 part by weight of vegetable protein is not particularly limited, but from the viewpoint of appropriately blending a predetermined amount of vegetable protein, for example, it is 5 parts by weight or less. Moreover, since the production method of the present invention is excellent in the effect of improving the stretchability, the stretchability can be effectively improved even when the content ratio of starch to the vegetable protein is relatively small. From such a viewpoint, the upper limit of the starch content range per 1 part by weight of vegetable protein is preferably 3 parts by weight or less, more preferably 2 parts by weight or less, and still more preferably 1.2 parts by weight. part or less, more preferably 0.9 part by weight or less.
• the upper limit of the content range of starch per part by weight of vegetable protein is preferably 3 parts by weight or less, more preferably 2 parts by weight or less, even more preferably 1.2 parts by weight or less, and even more preferably 0.9 parts by weight or less.
• the content of starch in the material composition is not particularly limited as long as stretchability can be imparted, but examples include 4% by weight or more. From the viewpoint of further improving the stretchability, or in addition to this viewpoint, from the viewpoint of further imparting the effect of improving the heat meltability and / or the effect of reducing the hydrophobic peptide, the content of starch in the material composition is preferably 5% by weight or more, more preferably 6% by weight or more, still more preferably 7% by weight or more, even more preferably 8% by weight or more, 9% by weight or more, 10% by weight or more, 11% by weight or more, 12% by weight or more, or 13% by weight or more.
• the upper limit of the content range of starch in the material composition is not particularly limited, but from the viewpoint of appropriately blending a predetermined amount of vegetable protein, for example, 20% by weight or less can be mentioned.
• the production method of the present invention is excellent in the effect of improving the stretchability, the stretchability can be effectively improved even when the starch content is relatively small.
• the upper limit of the starch content range in the material composition is preferably 17% by weight or less, more preferably 15% by weight or less, still more preferably 13% by weight or less, and even more preferably 11% by weight. Below, more preferably 9% by weight or less is mentioned.
• the material composition can contain any material component (hereinafter also referred to as "other material component") used for cheese substitutes as a component other than vegetable protein and starch.
• other material components include vegetable oils and fats, polysaccharide thickeners, water, and salt.
• Vegetable oils and fats are not particularly limited, but examples include canola oil (rapeseed oil), coconut oil, corn oil, olive oil, soybean oil, peanut oil, walnut oil, almond oil, sesame oil, cottonseed oil, sunflower seed oil, safflower oil, and flax. Seed oil, palm oil, palm kernel oil, palm fruit oil, babassu oil, shea butter, mango butter, cocoa butter, wheat germ oil, rice bran oil and the like. These vegetable oils and fats may be used singly or in combination of two or more. Canola oil (rapeseed oil) and coconut oil are preferred from the viewpoint of further improving stretchability, or from the viewpoint of further imparting the effect of improving heat meltability and / or the effect of reducing hydrophobic peptides in addition to this viewpoint.
• canola oil (rapeseed oil) and coconut oil are preferred from the viewpoint of further improving stretchability, or from the viewpoint of further imparting the effect of improving heat meltability and / or the effect of reducing hydrophobic peptides
• the content of the vegetable oil in the material composition is not particularly limited. Or from the viewpoint of further imparting a hydrophobic peptide reduction effect, for example, 5 to 30% by weight, preferably 8 to 25% by weight, more preferably 10 to 20% by weight, even more preferably 10 to 17% by weight, 12 to 17% by weight %, 12-14% by weight, or 14-17% by weight.
• the content ratio of vegetable protein and vegetable oil is determined by the content of each component described above.
• Polysaccharide thickeners are not particularly limited, but examples include locust bean gum, guar gum, carrageenan, xanthan gum, tragacanth gum, tamarind seed gum, pectin, gum arabic, curdlan, tara gum, gellan gum, gati gum, CMC (carboxymethylcellulose), and alginic acid. Examples include sodium and pullulan, preferably carrageenan. These polysaccharide thickeners may be used alone or in combination of two or more. Carrageenan is preferred from the viewpoint of further improving stretchability, or from the viewpoint of further imparting an effect of improving heat meltability and/or an effect of reducing hydrophobic peptides in addition to the above viewpoint.
• the content of the polysaccharide thickener in the material composition is not particularly limited, but from the viewpoint of further improving stretchability, or in addition to the viewpoint, heat meltability From the viewpoint of further imparting an improvement effect and/or a hydrophobic peptide reduction effect, it is, for example, 0.3 to 1.8% by weight, preferably 0.8 to 1.2% by weight.
• the content ratio of the vegetable protein and the polysaccharide thickener is determined by the content of each of the components described above. 3 parts by weight, preferably 0.08 to 0.12 parts by weight.
• the water content is not particularly limited, but from the viewpoint of further improving stretchability, or in addition to this viewpoint, the effect of improving heat meltability and / or the effect of reducing hydrophobic peptides. From the viewpoint of further imparting, for example, 50 to 72% by weight, preferably 55 to 70% by weight, more preferably 62 to 68% by weight.
• the content ratio of vegetable protein and water is determined by the content of each component described above. parts, more preferably 6 to 13 parts by weight, 6 to 9 parts by weight, or 9 to 13 parts by weight.
• the content of salt is not particularly limited, but the viewpoint of further improving stretchability, or in addition to this viewpoint, the effect of improving heat meltability and / or the effect of reducing hydrophobic peptides. From the viewpoint of further imparting, it is, for example, 0.1 to 1% by weight, more preferably 0.3 to 0.5% by weight.
• the content ratio of vegetable protein and salt is determined by the content of each component described above. 0.01 to 0.12 parts by weight, 0.02 to 0.12 parts by weight, 0.02 to 0.1 parts by weight, 0.02 to 0.09 parts by weight, more preferably 0.03 to 0.09 parts by weight parts by weight, 0.03 to 0.06 parts by weight, or 0.06 to 0.09 parts by weight.
• the enzymatic agent used for treating the above material composition contains at least protease and may contain amylase.
• the amylase is not limited to being added to the protease preparation, being blended in the protease preparation, or being used together with the protease in any other manner.
• protease refers to endo-peptidase.
• the origin of the protease is not particularly limited, for example, proteases derived from bacteria such as Bacillus, Geobacillus, etc.; Aspergillus, Mucor, Neurospora, Proteases from fungi of the genus Penicillium, Rhizomucor, Rhizopus, Sclerotinia; yeasts of the genus Saccharomyces; Actinomyces of the genus Streptomyces Bacteria-derived proteases and the like can be used.
• proteases may be used alone, or two or more of them may be used in combination.
• proteases derived from bacteria are preferred from the viewpoint of further enhancing stretchability, or from the viewpoint of further imparting an effect of improving heat melting property and/or an effect of reducing hydrophobic peptides in addition to this viewpoint.
• proteases derived from Bacillus and/or Geobacillus more preferably Bacillus stearothermophilus, Bacillus licheniformis and proteases derived from these Geobacillus.
• Bacillus stearothermophilus and Geobacillus stearothermophilus and particularly preferably Geobacillus stearothermophilus.
• the enzymatic agent can be used so that the protease activity of the protease is, for example, 10 to 500 U per 1 g of vegetable protein.
• the enzyme agent has a protease activity of protease per 1 g of vegetable protein. It can be used in an amount of preferably 30 to 500 U, more preferably 50 to 500 U, and even more preferably 80 to 500 U. Since the production method of the present invention is excellent in the stretchability-improving effect, it is possible to effectively obtain the stretchability-improving effect even with a relatively small amount of protease.
• the enzymatic agent may be used so that the protease activity of the protease is, for example, 10 to 400 U, 10 to 300 U, 10 to 200 U, 10 to 150 U, or 10 to 100 U per 1 g of vegetable protein. .
• amylase means ⁇ -amylase.
• the origin of the amylase is not particularly limited. .licheniformis), etc.), bacteria such as Geobacillus; fungi such as the genus Neurospora, the genus Penicillium, the genus Rhizomucor, the genus Rhizopus, the genus Sclerotinia; and the actinomycetes of the genus Streptomyces.
• bacteria such as Geobacillus
• fungi such as the genus Neurospora, the genus Penicillium, the genus Rhizomucor, the genus Rhizopus, the genus Sclerotinia
• actinomycetes of the genus Streptomyces may be used singly or in combination.
• the enzymatic agent is used so that the starch gelatinization power of amylase per 1 g of starch is 8 U or less for the protease activity of 100,000 U per 1 g of vegetable protein. From the viewpoint of further increasing the stretchability, or in addition to this viewpoint, from the viewpoint of further imparting the effect of improving heat meltability and / or the effect of reducing hydrophobic peptides, 1 g of starch for 100,000 U of protease activity per 1 g of vegetable protein.
• the starch paste refinement power per unit is preferably 7 U or less, more preferably 6.5 U or less, still more preferably 5.5 U or less, still more preferably 4.5 U or less, still more preferably 3.5 U or less or 2 U or less. is mentioned.
• the lower limit of the range of starch refinement power per 1 g of starch for protease activity of 100,000 U per 1 g of vegetable protein is 0 U or more.
• the starch gelatinization power per 1 g of starch is 0 U with respect to the protease activity of 100,000 U per 1 g of vegetable protein.
• the starch gelatinizing power per 1 g of starch with respect to 100,000 U of protease activity per 1 g of vegetable protein is more than 0 U, preferably 0.5 U or more, more preferably. 1 U or more, more preferably 1.5 U or more, 2 U or more, or 3 U or more.
• the protease activity is measured by the Folin method using casein as a substrate. Specifically, an enzymatic reaction is performed using casein as a substrate by a conventional method, and a Folin's reagent solution coloring substance equivalent to 1 ⁇ g of tyrosine per minute is obtained. 1 unit (1 U) is the amount of enzyme that causes an increase in the enzyme activity.
• the starch refinement power is an enzymatic activity in which the amount of enzyme that reduces the coloration of potato starch due to iodine by 10% per minute is defined as 1 unit (1U).
• peptidase treatment step in addition to the protease treatment step, it is treated with peptidase. It is preferable to further include a step (hereinafter also referred to as "peptidase treatment step").
• the peptidase treatment step may be performed simultaneously with the protease treatment step, or may be performed after the protease treatment step.
• a material composition containing vegetable protein and starch may be treated with both the protease and the peptidase at the same time, or a material composition containing vegetable protein and starch may be treated with Alternatively, after the treatment with protease, the treatment with peptidase may be further performed.
• peptidase refers to exo-type peptidase.
• the origin of the peptidase is not particularly limited, for example, peptidases derived from fungi such as the genus Rhizopus and the genus Aspergillus; peptidases derived from actinomycetes of the genus Streptomyces; the genus Bacillus and Geobacillus. (Geobacillus), Lactobacillus (Lactobacillus), Lactococcus (Lactococcus) bacteria-derived peptidases can be used, more specifically, fungi such as Rhizopus (Rhizopus), Aspergillus (Aspergillus), etc. Peptidases derived from Rhizopus oryzae and Aspergillus oryzae can be used more specifically. One of these peptidases may be used alone, or two or more of them may be used in combination.
• Rhizopus-derived peptidases are preferable, Peptidases derived from Rhizopus oryzae are more preferred.
• the peptidase can be used so that the peptidase activity per 1 g of vegetable protein is, for example, 0.001 to 1 U. From the viewpoint of further enhancing stretchability, or from the viewpoint of further imparting the effect of improving heat meltability and/or the effect of reducing hydrophobic peptides in addition to the above viewpoint, the peptidase has a peptidase activity per 1 g of vegetable protein of, for example, 0.5.
• 002 to 0.8 U preferably 0.0025 to 0.7 U, more preferably 0.003 to 0.6 U, still more preferably 0.0035 to 0.4 U, still more preferably 0.0035 to 0.3 U, more More preferably 0.004-0.25U, 0.004-0.02U, 0.004-0.01U, 0.004-0.008U, 0.004-0.006U, 0.01-0.25U , 0.02-0.25 U, 0.03-0.25 U, 0.04-0.25 U, 0.05-0.25 U, 0.1-0.25 U, or 0.2-0.25 U can be used as
• the peptidase activity shall be measured using L-leucyl-glycyl-glycine as a substrate and by a method based on the 9th edition of the Japanese Code of Food Additives. Specifically, L-leucyl-glycyl-glycine as a substrate
• the enzymatic activity is defined as 1 unit (1 U) of the amount of enzyme that causes an increase in ninhydrin coloring substance corresponding to 1 ⁇ mol of leucine per minute when an enzymatic reaction is performed by a conventional method.
• protease treatment step such as treatment conditions and the optional peptidase treatment step are not particularly limited as long as the substance to be treated with the enzyme is in contact with the enzyme.
• a material composition may be prepared and then protease (or protease and amylase) may be added, or the constituent materials of the material composition and protease (or protease and amylase) may be mixed at the same time.
• the material composition may be prepared, and then the protease (or protease and amylase) and peptidase may be added simultaneously or sequentially, or the constituent materials of the material composition may be added. and protease (or protease and amylase) and peptidase may be mixed simultaneously.
• the temperature in the protease treatment step and, if necessary, the peptidase treatment step is not particularly limited, and can be appropriately determined by those skilled in the art according to the optimum temperature of each enzyme used. is mentioned. Also, in the present invention, the treatment temperature can be changed stepwise.
• heating condition 1 is 45 to 70°C, preferably 45 to 60°C, more preferably 45 to 55°C
• heating condition 2 is 70 to 90°C, preferably 80 to 90°C.
• the treatment under the heating condition 2 can be performed.
• the time required for these treatment steps is not particularly limited, and may be appropriately determined according to the preparation scale of the enzyme-treated object, and is, for example, 10 minutes or longer, preferably 15 minutes or longer.
• the upper limit of the enzymatic treatment reaction time range is not particularly limited, but includes, for example, 6 hours or less, 3 hours or less, 1 hour or less, or 30 minutes or less.
• the treatment under the heating condition 1 is performed for 10 to 30 minutes, and then the treatment under the heating condition 2 is performed for 5 to 10 minutes.
• the treated material composition can be filled into a container and cooled as needed. This gives a stretchy cheese substitute.
• the enzymatic agent containing protease and optionally amylase contains vegetable protein and 0.6 parts by weight or more of starch per 1 part by weight of vegetable protein.
• the stretchability can be improved when used so that the starch refinement power per 1g of starch is 8U or less with respect to the protease activity of 100,000U per 1g of vegetable protein. Therefore, the present invention provides a stretchable cheese alternative comprising a vegetable protein, which comprises an enzymatic agent containing protease and optionally amylase, and 0.6 parts by weight or more of starch per 1 part by weight of the vegetable protein.
• the starch refinement power of the amylase per 1 g of the starch is 8 U or less for the protease activity of the protease of 100,000 U per 1 g of the vegetable protein.
• a stretchability improver is also provided. From the viewpoint of further improving the stretchability, the stretchability improver of the stretchable cheese substitute preferably further contains a peptidase.
• the amount of enzyme that causes an increase in Folin's test solution coloring substance equivalent to 1 ⁇ g of tyrosine per minute was defined as 1 unit (1 U).
• One unit (1 U) was defined as the amount of enzyme that reduces the iodine-induced coloration of potato starch by 10% per minute.
• 0.1 mL of the sample liquid was weighed into a stoppered test tube and heated in a boiling water bath for 5 minutes. After cooling, 1 mL of the substrate solution was added and mixed, heated at 37° C. for 5 minutes, and then cooled to room temperature. 2 mL of ninhydrin/2-methoxyethanol/citrate buffer test solution and 0.1 mL of tin (II) chloride test solution were added to this solution, and the solution was stoppered and heated in a boiling water bath for 20 minutes. After cooling, 10 mL of 1-propanol (1 ⁇ 2) was added and shaken to prepare a comparative solution.
• the absorbance of the test solution is greater than that of the comparative solution.
• centrifugation was performed and the supernatant was measured.
• One unit (1 U) was defined as the amount of the enzyme that increased the ninhydrin coloring substance corresponding to 1 ⁇ mol of leucine per minute.
• the percent value of the stretch length in each example was derived as a stretchability improvement evaluation index when the stretch length of the comparative example using no enzymatic agent was taken as 100%.
• the stretchability improvement evaluation index exceeds 100%, it is evaluated that improved stretchability is imparted.
• the larger the stretchability improvement evaluation index the higher the effect of improving the stretchability. Table 3 shows the results.
• Test Example 2 Pure water (RO water), pea protein material, tapioca starch, canola oil, coconut oil, salt, and enzyme agents shown in Tables 4 to 6 were added in the amounts shown in Tables 4 to 6 Test examples except that A cheese substitute was prepared in the same manner as in 1 and evaluated for stretchability. The results are shown in Tables 4-6. In Table 4, the results of Comparative Example 1 and Example 1 of Test Example 1 are transcribed.
• starch is 0.6 parts by weight or more per 1 part by weight of vegetable protein (Examples 3 to 5, Comparative Examples 2 and 3, Examples 7 to 9, Comparative Example 5), the enzyme agent
• the protease activity ([A] value in the table) per 1 g of vegetable protein is 100,000 U
• the starch gelatinizing power ([B] value in the table) per 1 g of starch is 8 U or less.
• the material composition has a specific composition in which the amount of starch is 0.6 parts by weight or more per 1 part by weight of vegetable protein (Examples 3 to 5, Comparative Examples 2 and 3, Examples 7 to 9, Comparative Although it is not clear that only Example 5) is accompanied by a specific restriction on the [B] value for the [A] value of 100,000 U, it is considered as follows.
• protein has the property of absorbing moisture, and a material composition that does not have the specific composition (such as used in Reference Examples 2 to 5, the amount of starch blended is 0 per part by weight of vegetable protein.
• the amount of vegetable protein relative to the amount of starch is large, so the protein can absorb water relative to the amount of starch.
• the amount of starch used in the material composition of a specific composition (Examples 3-5, Comparative Examples 2, 3, Examples 7-9, Comparative Example 5, 0.6 parts by weight or more per 1 part by weight of natural protein, hereinafter referred to as “material composition (2)), since the relative amount of vegetable protein to the amount of starch is small, starch The amount of protein that can absorb water is small relative to the amount.
• starch is reduced by excessive starch sizing power (starch sizing power where the [B] value exceeds 8 U with respect to the [A] value of 100,000 U). Even if the protein is decomposed and becomes less viscous, the protein absorbs enough water to avoid a significant decrease in viscosity, masking the effect of starch gelatinization and completely losing the effect of improving stretchability. (Reference Examples 3 to 5).
• Heat-meltability evaluation was performed using the prepared cheese substitute.
• Commercial frozen pizza dough (7 inches) was cut into pieces and coated with commercial pizza sauce.
• the prepared cheese substitute was placed thereon and cooked in a steam oven at 110° C. for 30 minutes.
• the thermal meltability of the cheese substitute after cooking was evaluated according to the following criteria. Table 7 shows the results.
• ++ Some form of cheese fragment remains.
• +++ The shape of the cheese fragment does not remain.
• the cheese substitute produced using protease As is clear from Table 7, compared to the stretchability of the cheese substitute produced without using protease (Comparative Example 7), the cheese substitute produced using protease (Examples 10 and 11) has improved stretchability. In particular, a more excellent effect of improving the stretchability was observed by using a protease in combination with a peptidase (Example 11). Regarding the heat meltability, the heat meltability was improved in the cheese substitutes produced using protease (Examples 10 and 11) compared to the cheese substitute produced without using protease (Comparative Example 7). By using a peptidase in combination with the protease, even better heat-meltability was exhibited (Example 11).
• the cheese substitutes produced using protease were more hydrophobic than the amount of hydrophobic amino acids in the cheese substitute produced without using protease (Comparative Example 7).
• the amount of amino acids was improved, and in particular, a more excellent effect of increasing the amount of hydrophobic amino acids was observed by using protease in combination with peptidase (Example 11).
• Example 11 bitterness was felt in the cheese substitute of Comparative Example 7, while bitterness was suppressed in the cheese substitute of Example 10. It was confirmed that the cheese substitute of Example 11 lost its bitterness and had a good taste.
• Test Example 4 Same as Test Example 1, except that pure water (RO water), broad bean protein material, tapioca starch, coconut oil, ⁇ -carrageenan, salt, and the enzymatic agent shown in Table 8 were added in the amounts shown in Table 8.
• a cheese substitute was prepared by
• the cheese substitute produced using protease As is clear from Table 8, compared to the stretchability of the cheese substitute produced without using protease (Comparative Example 8), the cheese substitute produced using protease (Example 12.13) has improved stretchability. Among them, a more excellent effect of improving the stretchability was observed by using a protease in combination with a peptidase (Example 13).
• the amount of hydrophobic amino acids compared to the amount of hydrophobic amino acids in the cheese substitute produced without using protease (Comparative Example 8), the cheese substitute produced using protease (Examples 12 and 13) was hydrophobic. The amount of amino acids was improved, and in particular, a more excellent effect of increasing the amount of hydrophobic amino acids was observed by using protease in combination with peptidase (Example 13). It was suggested that
• Test Example 5 Same as Test Example 1, except that pure water (RO water), chickpea protein material, tapioca starch, coconut oil, ⁇ -carrageenan, salt, and the enzymatic agents shown in Table 9 were added in the amounts shown in Table 9.
• a cheese substitute was prepared by
• the cheese substitute produced using protease As is clear from Table 9, compared to the stretchability of the cheese substitute produced without using protease (Comparative Example 9), the cheese substitute produced using protease (Examples 14 and 15) has improved stretchability. In particular, the use of protease in combination with peptidase was found to have an even more excellent effect of improving the stretchability (Example 15).
• the cheese substitute produced using protease Examples 14 and 15 was more hydrophobic than the amount of hydrophobic amino acids in the cheese substitute produced without using protease (Comparative Example 9). The amount of amino acids was improved, and in particular, a more excellent effect of increasing the amount of hydrophobic amino acids was observed by using protease in combination with peptidase (Example 15). It was suggested that
• Test Example 6 Same as Test Example 1, except that pure water (RO water), lentil protein material, tapioca starch, coconut oil, ⁇ -carrageenan, salt, and the enzymatic agent shown in Table 10 were added in the amounts shown in Table 10.
• a cheese substitute was prepared by
• the cheese substitute produced using protease As is clear from Table 10, compared to the stretchability of the cheese substitute produced without using protease (Comparative Example 10), the cheese substitute produced using protease (Examples 16 and 17) has improved stretchability. In particular, the use of protease in combination with peptidase was found to have an even more excellent stretchability-enhancing effect (Example 17). Regarding heat melting property, the cheese substitute produced using protease (Example 16) and the cheese substitute produced using protease in combination with peptidase compared to the cheese substitute produced without using protease (Comparative Example 10). In the product (Example 17), it showed excellent thermal meltability.
• the cheese substitute produced using protease was more hydrophobic than the amount of hydrophobic amino acids in the cheese substitute produced without using protease (Comparative Example 10).
• the amount of amino acids was improved, and in particular, a more excellent effect of increasing the amount of hydrophobic amino acids was observed by using protease in combination with peptidase (Example 17). It was suggested that

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