Method for producing stretchy cheese substitute

Abstract

The purpose of the present invention is to provide a cheese alternative producing technique with which it is possible to add improved stretchability to a cheese alternative containing vegetable protein and starch. A stretchable cheese alternative producing method including a step for treating a material composition containing vegetable protein and starch with protease provides a cheese alternative having improved stretchability.

Description

[Technical Field]

[0001] The present invention relates to a method for producing a stretchable cheese substitute, more specifically, a method for producing a stretchable cheese substitute made from vegetable protein and having heat stretchability. [Background technology]

[0002] Due to various reasons, such as the recent health boom, addressing allergy issues, and religious reasons, plant-based protein foods have become increasingly popular as substitutes for animal-based protein foods.

[0003] Because vegetable protein materials are significantly different from animal protein materials, various processing techniques have been investigated in the creation of vegetable protein foods to bring the flavor, texture, etc. closer to those of animal protein foods.

[0004] Cheese-like foods made from vegetable proteins, so-called plant-based cheese, have been studied as potential substitutes for cheese made from animal milk. For example, Patent Document 1 describes that a cream cheese-like food, which is obtained by oxidizing an emulsion containing soy protein hydrolysate and fats and oils through the action of protease in a neutral to alkaline range, has a smooth texture and a good flavor similar to cream cheese. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2006 / 135089 Summary of the Invention [Problem to be solved by the invention]

[0006] Among the properties of cheese made from animal milk, one that is particularly distinctive is the ability to stretch when heated (hereinafter, this property will also be referred to as "stretchability"). This property is related to the casein contained in animal milk, and is one of the reasons why cheese is so appetizing. On the other hand, plant-based cheese (hereinafter, also referred to as "cheese substitute"), which has a completely different protein composition, does not inherently have stretchability.

[0007] Addition of starch is one way to impart stretchability to cheese alternatives, but there is a limit to the effect of imparting stretchability to cheese alternatives by adding starch alone, so a technology that can further enhance stretchability is desired.

[0008] Therefore, an object of the present invention is to provide a technology for producing a cheese substitute that can impart improved stretchability to a cheese substitute containing vegetable protein and starch. [Means for solving the problem]

[0009] The present inventors have found that adding a protease to a material composition containing a vegetable protein and starch improves the stretchability of the resulting cheese alternative. Based on this finding, the present invention was completed through further investigation.

[0010] That is, the present invention provides the following aspects. Item 1. A method for producing a stretchable cheese substitute, comprising a step of treating a material composition containing vegetable protein and starch with a protease. Item 2. The method according to Item 1, wherein the content of the starch per part by weight of the vegetable protein is 0.1 part by weight or more and less than 0.6 parts by weight. Item 3. The production method according to Item 1 or 2, wherein the protease is a bacterial protease. 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 5. The production method according to any one of Items 1 to 4, wherein the protease is selected from the group consisting of proteases derived from Bacillus stearothermophilus, Bacillus licheniformis, Bacillus amyloliquefaciens, and these Geobacillus species. Item 6. The production method according to any one of Items 1 to 5, wherein the protease has a protease activity of 10 to 500 U per 1 g of the vegetable protein. Item 7. The production method according to any one of Items 1 to 6, further comprising a step of treating with peptidase. Item 8. The production method according to any one of Items 1 to 7, wherein the vegetable protein is pea protein, fava bean protein, chickpea protein, and / or lentil protein. Item 9. The method according to any one of Items 1 to 8, wherein the content of the vegetable protein in the material composition is 15 to 30 wt %. Item 10. The method according to any one of Items 1 to 9, wherein the starch is tapioca starch. Item 11. A stretchability improver for a stretchable cheese substitute containing vegetable protein and starch, which improves stretchability and contains a protease. Item 12. The stretchability improving agent according to Item 11, further comprising a peptidase. [Effects of the Invention]

[0011] According to the present invention, a technology for producing a cheese substitute is provided that can impart improved stretchability to a cheese substitute containing vegetable protein and starch. DETAILED DESCRIPTION OF THE INVENTION

[0012] 1. Method for producing stretchable cheese substitute The method for producing a stretchable cheese substitute of the present invention is characterized by including a step of treating a material composition containing vegetable protein and starch with a protease (hereinafter also referred to as a "protease treatment step"). The method for producing a stretchable cheese substitute of the present invention will be described in detail below. The present invention can improve the stretchability of the resulting cheese substitute, or in addition to improving the stretchability, can further impart an effect of improving thermal melting properties and / or an effect of reducing hydrophobic peptides (the hydrophobic peptide reduction effect refers to the effect of degrading bitter hydrophobic peptides and replacing them with hydrophobic amino acids).

[0013] 1-1. Material composition containing vegetable protein and starch The plant from which the vegetable protein originates is not particularly limited, and examples thereof include beans such as peas, soybeans, broad beans, chickpeas, and lentils; grains such as barley, wheat, oats, rice, buckwheat, barnyard millet, and foxtail millet; and nuts such as almonds, cashew nuts, hazelnuts, pecan nuts, macadamia nuts, pistachios, walnuts, Brazil nuts, peanuts, and coconuts. One type of vegetable protein derived from these plants may be used alone, or two or more types from different origins may be used in combination.

[0014] Among these, from the viewpoint of further improving stretchability, or in addition to that viewpoint, from the viewpoint of further imparting a thermal melting property improving effect and / or a hydrophobic peptide reducing effect, bean proteins are preferred, and pea, fava bean, chickpea, and lentil proteins are more preferred.

[0015] The vegetable protein content in the material composition is not particularly limited, but may be, for example, 1 to 30% by weight. From the viewpoint of further improving stretchability, or from the viewpoint of further imparting a thermal melting property improving effect and / or a hydrophobic peptide reducing effect in addition to the above, the vegetable protein content in the material composition is preferably 4 to 25% by weight, 4 to 22% by weight, more preferably 9 to 21% by weight, 9 to 20% by weight, even more preferably 14 to 20% by weight, even more preferably 16 to 19% by weight, and even more preferably 17 to 18% by weight.

[0016] The plant from which the starch originates is not particularly limited as long as it can impart stretchability to the cheese alternative, and examples include cassava, potato, sweet potato, kudzu, etc. Starches derived from these plants may be used singly or in combination of two or more species from different origins.

[0017] Among these, cassava starch (tapioca starch) is preferred from the viewpoint of further improving stretchability, or in addition to that viewpoint, further imparting the effect of improving thermal melting property and / or the effect of reducing hydrophobic peptides.

[0018] The starch content in the material composition is not particularly limited as long as it can impart stretchability, but can be, for example, 4% by weight or more. From the perspective of further improving stretchability, or from the perspective of further imparting a thermal melting property improving effect and / or a hydrophobic peptide reducing effect in addition to the above, the starch content in the material composition is preferably 5% by weight or more, more preferably 6% by weight or more, even more preferably 7% by weight or more, still 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.

[0019] The upper limit of the starch content in the material composition is not particularly limited, but can be, for example, 20% by weight or less, from the viewpoint of appropriately blending a predetermined amount of vegetable protein. Furthermore, because the manufacturing method of the present invention is excellent in improving stretchability, stretchability can be effectively improved even when the starch content is relatively low. From this viewpoint, the upper limit of the starch content in the material composition is preferably 17% by weight or less, more preferably 15% by weight or less, even more preferably 13% by weight or less, even more preferably 11% by weight or less, and even more preferably 9% by weight or less.

[0020] The starch content per part by weight of the vegetable protein in the material composition is determined by the content of each of the above-mentioned components, but may be, for example, 0.1 to 5 parts by weight. From the viewpoint of further enhancing the stretchability-improving effect, or from the viewpoint of further imparting the thermal melting property-improving effect and / or the hydrophobic peptide-reducing effect in addition to the above-mentioned effect, the starch content is preferably 0.1 to 2 parts by weight, more preferably 0.1 to 0.6 parts by weight, even more preferably 0.1 to 0.55 parts by weight, and even more preferably 0.1 to 0.5 parts by weight, 0.2 to 0.5 parts by weight, 0.35 to 0.5 parts by weight, or 0.4 to 0.5 parts by weight.

[0021] The material composition can contain any ingredient used in cheese substitutes (hereinafter also referred to as "other ingredient") as ingredients other than vegetable protein and starch. Examples of other ingredient ingredients include vegetable oils and fats, thickening polysaccharides, water, salt, etc.

[0022] The vegetable oil is not particularly limited, but examples thereof 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, flaxseed oil, palm oil, palm kernel oil, palm fruit oil, babassu oil, shea butter, mango butter, cocoa butter, wheat germ oil, and rice bran oil. These vegetable oils may be used alone or in combination of two or more. From the viewpoint of further improving stretchability, or from the viewpoint of further imparting a thermal melting property improving effect and / or a hydrophobic peptide reducing effect in addition to the above, canola oil (rapeseed oil) and coconut oil are preferred.

[0023] When the material composition contains vegetable oil, the vegetable oil content in the material composition is not particularly limited, but from the viewpoint of further improving stretchability, or from the viewpoint of further imparting the effects of improving thermal melting and / or reducing hydrophobic peptides in addition to the above viewpoint, for example, 5 to 30 wt%, preferably 8 to 25 wt%, more preferably 10 to 20 wt%, even more preferably 10 to 17 wt%, 12 to 17 wt%, 12 to 14 wt%, or 14 to 17 wt%. The vegetable protein to vegetable oil content ratio is determined by the content of each of the above components, but the vegetable oil content per part by weight of vegetable protein is, for example, 0.3 to 5 wt%, preferably 0.5 to 4.5 wt%, more preferably 0.7 to 4 wt%, 0.7 to 3.5 wt%, even more preferably 0.9 to 3.5 wt%, even more preferably 1.1 to 3.3 wt%, 1.1 to 3.2 wt%, or 1.2 to 3 wt%.

[0024] The thickening polysaccharide is not particularly limited, but examples thereof include locust bean gum, guar gum, carrageenan, xanthan gum, tragacanth gum, tamarind seed gum, pectin, gum arabic, curdlan, tara gum, gellan gum, gum ghatti, CMC (carboxymethylcellulose), sodium alginate, pullulan, etc., with carrageenan being preferred. These thickening polysaccharides may be used alone or in combination of two or more. From the viewpoint of further improving stretchability, or from the viewpoint of further imparting a thermal melting property improving effect and / or a hydrophobic peptide reducing effect in addition to the above, carrageenan is preferred.

[0025] When the material composition contains a thickening polysaccharide, the content of the thickening polysaccharide in the material composition is not particularly limited, but from the viewpoint of further improving stretchability, or from the viewpoint of further imparting the effect of improving thermal melting property and / or the effect of reducing hydrophobic peptides in addition to the above viewpoint, for example, 0.3 to 1.8 wt%, preferably 0.8 to 1.2 wt%. The content ratio of the vegetable protein to the thickening polysaccharide is determined by the content of each of the above components, but the content of the thickening polysaccharide per part by weight of the vegetable protein is, for example, 0.1 to 0.3 parts by weight, preferably 0.04 to 0.12 parts by weight.

[0026] When the material composition contains water, the water content is not particularly limited, but from the viewpoint of further improving stretchability, or from the viewpoint of further imparting the effect of improving thermal melting property and / or the effect of reducing hydrophobic peptides in addition to the above viewpoint, for example, 30 to 72 wt%, 35 to 72 wt%, preferably 40 to 72 wt%, 45 to 72 wt%, 50 to 72 wt%, 55 to 70 wt%, more preferably 62 to 68 wt%. The vegetable protein to water content ratio is determined by the content of each of the above components, but the water content per part by weight of vegetable protein is, for example, 1 to 17 wt%, preferably 2 to 15 wt%, 3 to 15 wt%, more preferably 6 to 13 wt%, 6 to 9 wt%, or 9 to 13 wt%.

[0027] When the material composition contains salt, the salt content is not particularly limited, but can be, for example, 0.1 to 1 wt %, more preferably 0.3 to 0.5 wt %, from the viewpoint of further improving stretchability, or, in addition to that, from the viewpoint of further imparting the effect of improving thermal melting property and / or the effect of reducing hydrophobic peptides. The ratio of vegetable protein to salt is determined by the content of each of the above components, and can be, for example, 0.008 to 0.15 parts by weight, preferably 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, 0.03 to 0.06 parts by weight, or 0.06 to 0.09 parts by weight.

[0028] 1-2.Protease In the present invention, protease refers to an endo-type peptidase. The origin of the protease used to treat the material composition is not particularly limited. For example, proteases derived from bacteria such as Bacillus and Geobacillus; proteases derived from fungi such as Aspergillus, Mucor, Neurospora, Penicillium, Rhizomucor, Rhizopus, and Sclerotinia; proteases derived from yeasts of the Saccharomyces genus; and proteases derived from actinomycetes of the Streptomyces genus. These proteases may be used alone or in combination.

[0029] Among these proteases, from the viewpoint of further enhancing stretchability, or, in addition to that viewpoint, further imparting a thermal melting property improving effect and / or a hydrophobic peptide reducing effect, proteases derived from bacteria are preferred, more preferably proteases derived from the genus Bacillus and / or Geobacillus are preferred, even more preferably Bacillus stearothermophilus, Bacillus licheniformis, Bacillus amyloliquefaciens and proteases derived from these Geobacillus species are even more preferred, Bacillus stearothermophilus, Geobacillus stearothermophilus and Bacillus amyloliquefaciens are even more preferred, and Geobacillus stearothermophilus is particularly preferred.

[0030] The protease can be used so that the protease activity per 1 g of vegetable protein is, for example, 10 to 500 U. From the viewpoint of further enhancing stretchability, or from the viewpoint of further imparting a thermal melting property improving effect and / or a hydrophobic peptide reducing effect in addition to the above, the protease can be used so that the protease activity per 1 g of vegetable protein is preferably 30 to 500 U, more preferably 50 to 500 U, and even more preferably 80 to 500 U. Because the production method of the present invention is excellent in improving stretchability, even a relatively small amount of protease can effectively achieve the effect of improving stretchability. From this viewpoint, the protease may be used so that the protease activity per 1 g of vegetable protein is, for example, 10 to 400 U, 10 to 300 U, 10 to 200 U, 10 to 150 U, or 10 to 100 U.

[0031] Protease activity is measured by the Folin method using casein as a substrate. Specifically, the amount of enzyme that causes an increase in the Folin test solution color substance equivalent to 1 μg of tyrosine per minute when an enzymatic reaction is carried out using casein as a substrate in a standard manner is defined as 1 unit (1 U).

[0032] 1-3. Peptidase From the viewpoint of further improving stretchability, or in addition to that viewpoint, from the viewpoint of further imparting the effect of improving thermal melting property and / or the effect of reducing hydrophobic peptides, it is preferable that the present invention further includes a step of treating with peptidase (hereinafter also referred to as a "peptidase treatment step") in addition to the protease treatment step.

[0033] The peptidase treatment step may be carried out simultaneously with the protease treatment step or after the protease treatment step. That is, a material composition containing a vegetable protein and starch may be treated with both the protease and peptidase at the same time, or a material composition containing a vegetable protein and starch may be treated with a protease and then treated with a peptidase.

[0034] In the present invention, peptidase refers to exo-type peptidase. The origin of the peptidase is not particularly limited, but for example, peptidases derived from fungi such as Rhizopus and Aspergillus; peptidases derived from actinomycetes such as Streptomyces; peptidases derived from bacteria such as Bacillus, Geobacillus, Lactobacillus, and Lactococcus can be used. More specifically, peptidases derived from fungi such as Rhizopus and Aspergillus can be used. Even more specifically, peptidases derived from Rhizopus oryzae and Aspergillus oryzae can be used. These peptidases may be used singly or in combination of two or more kinds.

[0035] Among these peptidases, from the viewpoint of further enhancing stretchability, or in addition to that viewpoint, further imparting the effect of improving thermal melting property and / or the effect of reducing hydrophobic peptides, peptidases derived from Rhizopus are preferred, and peptidases derived from Rhizopus oryzae are more preferred.

[0036] 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 in addition to that viewpoint, from the viewpoint of further imparting the effect of improving thermal melting property and / or the effect of reducing hydrophobic peptides, the peptidase can be used so that the peptidase activity per 1 g of vegetable protein is, for example, 0.002 to 0.8 U, preferably 0.0025 to 0.7 U, more preferably 0.003 to 0.6 U, even more preferably 0.0035 to 0.4 U, and even more preferably It can be used at a concentration of 0.0035 to 0.3 U, more preferably 0.004 to 0.25 U, 0.004 to 0.02 U, 0.004 to 0.01 U, 0.004 to 0.008 U, 0.004 to 0.006 U, 0.01 to 0.25 U, 0.02 to 0.25 U, 0.03 to 0.25 U, 0.04 to 0.25 U, 0.05 to 0.25 U, 0.1 to 0.25 U, or 0.2 to 0.25 U.

[0037] Peptidase activity is measured using L-leucyl-glycyl-glycine as a substrate according to the method of the 9th edition of the Japanese Standards for Food Additives. Specifically, one unit (1U) of enzyme activity is defined as the amount of enzyme that causes an increase in ninhydrin color-reactive substance equivalent to 1 μmol of leucine per minute when an enzymatic reaction is carried out using L-leucyl-glycyl-glycine as a substrate in a conventional manner.

[0038] 1-4. Processing conditions, etc. The specific procedures for the protease treatment step and the optional peptidase treatment step are not particularly limited as long as the enzyme contacts the target material. For example, in the protease treatment step, a material composition may be prepared and then the protease may be added, or the constituent materials of the material composition and the protease may be simultaneously mixed. Furthermore, in a combination of the protease treatment step and the peptidase treatment step, a material composition may be prepared and then the protease and peptidase may be simultaneously or sequentially added, or the constituent materials of the material composition may be simultaneously mixed with the protease and peptidase.

[0039] The temperatures in the protease treatment step and the optional peptidase treatment step are not particularly limited and can be appropriately determined by those skilled in the art depending on the optimum temperature of each enzyme used, and examples include 45 to 90°C. In the present invention, the treatment temperature can also be changed stepwise. For example, heating condition 1 can be a combination of 45°C or higher but lower than 70°C, preferably 45 to 60°C, and more preferably 45 to 55°C, with heating condition 2 being 70 to 90°C, preferably 80 to 90°C. Preferably, these treatment steps can be performed under heating condition 1, followed by heating under heating condition 2.

[0040] The time required for these treatment steps is not particularly limited and may be determined appropriately depending on the preparation scale of the material to be treated with the enzyme, but may be, for example, 10 minutes or more, preferably 15 minutes or more. The upper limit of the range of the enzyme treatment reaction time is not particularly limited, but may be, for example, 6 hours or less, 3 hours or less, 1 hour or less, or 30 minutes or less. Preferably, in these treatment steps, treatment under the heating condition 1 above is carried out for 10 to 30 minutes, and then treatment under the heating condition 2 above is carried out for 5 to 10 minutes.

[0041] After the necessary processing steps are completed, the processed material composition can be filled into containers as needed and cooled to provide a stretchable cheese replica.

[0042] 2. Cheese substitute stretch improver As described above, protease can improve the stretchability in the production of a stretchable cheese substitute containing vegetable protein and starch. Therefore, the present invention also provides a stretchability improver for a stretchable cheese substitute containing vegetable protein and starch, which includes a protease. From the viewpoint of further improving the stretchability, it is preferable that the stretchability improver for the stretchable cheese substitute further includes a peptidase.

[0043] The types and amounts of components used in the stretchability improver are as shown in the section "1. Method for producing stretchable cheese substitute." [Example]

[0044] The present invention will be specifically described below with reference to examples, but the present invention should not be construed as being limited to the following examples.

[0045] [Enzyme used] The commercially available enzymes shown in the table below were used. [Table 1]

[0046] [Protease activity measurement method] Five mL of 0.6% (w / v) casein solution (0.05 mol / L sodium hydrogen phosphate, pH 8.0) was heated at 37°C for 10 minutes, after which 1 mL of the protease-containing sample solution was added and immediately shaken. After allowing this solution to stand at 37°C for 10 minutes, 5 mL of trichloroacetic acid TS containing 1.8% trichloroacetic acid, 1.8% sodium acetate, and 0.33 mol / L acetic acid was added and shaken. The solution was then allowed to stand at 37°C for another 30 minutes and filtered. The first 3 mL of filtrate was discarded, and 2 mL of the next filtrate was measured. 5 mL of 0.55 mol / L sodium carbonate TS and 1 mL of Folin's TS (1→3) were added, shaken well, and allowed to stand at 37°C for 30 minutes. The absorbance (AT) of this solution (enzyme reaction solution) at 660 nm was measured, using water as a control.

[0047] Separately, 1 mL of sample solution containing protease was measured, 5 mL of trichloroacetic acid test solution containing 1.8% trichloroacetic acid, 1.8% sodium acetate, and 0.33 mol / L acetic acid was added, and the mixture was shaken. 5 mL of 0.6% (w / v) casein solution was then added, and the mixture was immediately shaken and left to stand at 37°C for 30 minutes. The absorbance AB of this solution (blank) was measured in the same manner as the enzyme reaction solution described above.

[0048] The amount of enzyme that causes an increase in the colored substance in Folin's test solution equivalent to 1 μg of tyrosine per minute was defined as 1 unit (1 U).

[0049] 1 mL, 2 mL, 3 mL, and 4 mL of a 1 mg / mL tyrosine standard stock solution (0.2 mol / L hydrochloric acid) were measured, and 0.2 mol / L hydrochloric acid TS was added to each to make a 100 mL solution. 2 mL of each solution was measured, and 5 mL of 0.55 mol / L sodium carbonate TS and 1 mL of Folin's TS (1→3) were added. The mixture was immediately shaken and left at 37°C for 30 minutes. For each of these solutions, 2 mL of 0.2 mol / L hydrochloric acid TS was measured and the resulting solution was used as a control. Absorbances A1, A2, A3, and A4 at a wavelength of 660 nm were measured. A calibration curve was created, plotting absorbances A1, A2, A3, and A4 on the vertical axis and the amount of tyrosine (μg) in 2 mL of each solution on the horizontal axis, and the amount of tyrosine (μg) per 1 mL of absorbance difference was calculated.

[0050]

number

[0051] [Peptidase activity measurement method] An appropriate amount of enzyme was weighed and dissolved or dispersed uniformly in water, pH 7.0 potassium phosphate buffer (0.005 mol / L), or potassium phosphate buffer (0.005 mol / L, pH 7.0, containing zinc sulfate) to make a 50 mL solution, or this was further diluted 10-fold, 100-fold, or 1000-fold with water or the same buffer to prepare the sample solution.

[0052] 30 mg of L-leucyl-glycyl-glycine was weighed out and dissolved in pH 7.0 potassium phosphate buffer (0.05 mol / L) to make a 50 mL solution. This solution was diluted 10-fold with pH 7.0 potassium phosphate buffer (0.05 mol / L) to prepare the substrate solution. This substrate solution was prepared immediately before use.

[0053] 1 mL of substrate solution was measured into a stoppered test tube and heated at 37°C for 5 minutes. 0.1 mL of sample solution was added and mixed. The tube was then heated at 37°C for 60 minutes, then heated in a boiling water bath for 5 minutes, and cooled to room temperature. To this solution, 2 mL of ninhydrin-2-methoxyethanol-citric acid buffer solution and 0.1 mL of tin(II) chloride TS were added, the tube was stoppered, and heated in a boiling water bath for 20 minutes. After cooling, 10 mL of 1-propanol (1→2) was added, shaken, and used as the test solution. Separately, 0.1 mL of sample solution was measured into a stoppered test tube and heated in a boiling water bath for 5 minutes. After cooling, 1 mL of substrate solution was added, mixed, heated at 37°C for 5 minutes, and cooled to room temperature. 2 mL of ninhydrin-2-methoxyethanol-citric acid buffer solution and 0.1 mL of tin(II) chloride TS were added, the tube 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 comparison solution. When the absorbance of the test solution and comparison solution was measured at a wavelength of 570 nm within 5 to 30 minutes after preparation, the absorbance of the test solution was greater than that of the comparison solution. If the test solution or comparison solution for which absorbance was to be measured was turbid, the solution was centrifuged and the supernatant was measured. The amount of enzyme that caused an increase in the amount of ninhydrin color-reactive substance equivalent to 1 μmol of leucine per minute was defined as 1 unit (1 U).

[0054]

number

[0055] [Materials used] The materials used were as shown in the table below. [Table 2]

[0056] [Test Example 1] (1) Production of stretchable cheese substitute Purified water (RO water) was placed in a Thermomix mixer, and while stirring at 50°C and speed 3, the pea protein material, tapioca starch, canola oil, coconut oil, salt, and the enzyme agent shown in Table 3 were added in the amounts shown in Table 3. After stirring at speed 3 for 15 minutes at 50°C, the mixture was heated to 85°C and stirred at speed 3 for 7 minutes, and then filled into three aluminum containers (bottom inner diameter: 5 cm) at 100 g per container, covered, cooled to 4°C, and stored. In this way, cheese substitutes (three for each Example / Comparative Example) were obtained.

[0057] (2) Stretchability evaluation The cheese substitute sample filled in an aluminum container was heated for 30 minutes in a steam oven set to 110°C (to eliminate the effect of water evaporation during heating). The sample was then removed from the steam oven, and after confirming that the internal temperature of the sample had reached 70°C, it was stirred with a fork. After confirming that the sample was covered with the fork, the tip of the fork was lifted at a rate of 5 cm / sec, as if scooping up the sample. The distance between the start point of lifting the tip of the fork and the point at which the stretch of the sample ceased (stretch length (mm)) was measured. The stretch length was calculated as the average value obtained by similarly testing three samples prepared for each Example / Comparative Example. In addition, when no stretchability was observed, the stretch length was recorded as "<10." Furthermore, the relative value of the stretch length in each Example, when the stretch length of the Comparative Example without an enzyme agent was set to 1, was calculated as a stretchability improvement evaluation index. For Comparative Examples where the stretch length was "<10," the stretch length was assumed to be 1 cm for convenience. A stretchability improvement evaluation index greater than 1 is evaluated as having improved stretchability. The larger the stretchability improvement evaluation index, the greater the effect of improving stretchability. The results are shown in Table 3.

[0058] [Table 3]

[0059] As is clear from Table 3, the cheese substitutes produced using protease (Examples 1-9) had improved stretchability compared to the cheese substitutes produced without protease (Comparative Examples 1-4). Furthermore, in the cheese substitutes produced without protease (Comparative Examples 1-4), the stretchability was impaired as the protein content increased, whereas in the cheese substitutes produced with protease (Examples 1-9), the stretchability improvement evaluation index increased as the protein content increased, indicating a tendency for the stretchability improvement effect to be enhanced. Furthermore, in light of the results of Example 1 compared to Example 2, Example 4 compared to Examples 5 and 6, and Example 7 compared to Examples 8 and 9, the cheese substitutes (Examples 1, 3, 4, and 7) produced using a protease derived from Geobacillus stearothermophilus (Thermoase GL30) exhibited an even more excellent effect in improving stretchability.

[0060] [Test Example 2] Cheese replicas were prepared in the same manner as in Test Example 1, except that purified water (RO water), pea protein material, tapioca starch, coconut oil, canola oil, nutritional yeast, κ-carrageenan, salt, and the enzyme agents shown in Table 4 were added in the amounts shown in Table 4.

[0061] (1) Stretchability evaluation The stretchability was evaluated in the same manner as in Test Example 1. The results are shown in Table 4.

[0062] (2) Thermal melting evaluation The prepared cheese substitutes were used to evaluate their thermal meltability. Commercially available frozen pizza dough (7 inches) was cut into pieces and spread with commercially available pizza sauce. The prepared cheese substitutes were placed on top and cooked in a steam oven at 110°C for 30 minutes. The thermal meltability of the cheese substitutes after cooking was evaluated according to the following criteria. The results are shown in Table 4. -: No melting of cheese pieces was observed. +: Melted but the cheese pieces still retain their shape. ++: The shape of the cheese pieces remains slightly. +++: No trace of cheese remains.

[0063] (3) Evaluation of bitter peptide reduction (hydrophobic peptide degradation, i.e., increase in hydrophobic amino acids) The prepared cheese substitute was used to examine the increase in hydrophobic amino acids to evaluate the reduction of bitter peptides. 1 ml of water was added to 1 g of cheese substitute and homogenized using a vortex mixer. The mixture was centrifuged at 13,000 rpm for 5 minutes, and the supernatant was collected. The collected supernatant was filtered through a syringe filter and used as a sample for HPLC analysis. Analysis was performed using HPLC with a post-column reactor using the ninhydrin reaction, and the total amount of Gly, Ala, Val, Met, Ile, Leu, Phe, and Pro (calculated as the amount (mg) per 1 g of cheese substitute) was measured as the hydrophobic amino acid content. Furthermore, the relative value, where the amount of hydrophobic amino acids in the corresponding comparative example (i.e., an example prepared under the same conditions except without enzyme treatment) was set to 1, was calculated as the hydrophobic amino acid content increase rate. A higher hydrophobic amino acid content increase rate indicates that more bitter hydrophobic peptides have been decomposed into amino acids, i.e., the bitterness has been reduced more. The results are shown in Table 4. Analytical column: TSKgel Aminopak Mobile phase: HITACHI AMINO ACID ANALYSIS Buffer pH1-4

[0064] [Table 4]

[0065] As is clear from Table 4, the cheese substitutes produced using protease (Examples 10 to 13) had improved stretchability compared to the cheese substitutes produced without protease (Comparative Examples 5 and 6), and in particular, as shown in Example 11, which is compared to Example 10, and Example 13, which is compared to Example 12, the use of peptidase in combination with protease showed an even more excellent effect of improving stretchability (Examples 11 and 13). With regard to heat-meltability, the cheese substitutes produced using protease or a combination of protease and peptidase showed excellent heat-meltability compared to the cheese substitutes produced without protease (Comparative Examples 5 and 6) (Examples 10 to 13). The amount of hydrophobic amino acids was increased in the cheese substitutes produced using protease (Examples 10-13) compared to the amount of hydrophobic amino acids in the cheese substitutes produced without protease (Comparative Examples 5 and 6). In particular, as shown in Example 11 (compared to Example 10) and Example 13 (compared to Example 12), the combined use of protease and peptidase demonstrated an even greater effect of increasing the amount of hydrophobic amino acids (Examples 11 and 13). The increase in the amount of hydrophobic amino acids correlated with the degradation of bitter-tasting hydrophobic peptides, suggesting a reduction in bitterness. In fact, sensory tests were performed on Comparative Example 5, Example 10, and Example 11, confirming that the cheese substitute of Comparative Example 5 tasted bitter, while the bitterness was suppressed in the cheese substitute of Example 10, and the bitterness disappeared in the cheese substitute of Example 11, resulting in a favorable taste.

[0066] [Test Example 3] A cheese replica was prepared in the same manner as in Test Example 1, except that purified water (RO water), fava bean protein material, tapioca starch, coconut oil, κ-carrageenan, salt, and the enzyme agent shown in Table 5 were added in the amounts shown in Table 5.

[0067] (1) Stretchability evaluation The stretchability was evaluated in the same manner as in Test Example 1. The results are shown in Table 5.

[0068] (2) Thermal melting evaluation For some of the comparative examples and examples, the thermal melting properties were evaluated in the same manner as in Test Example 2. The results are shown in Table 5.

[0069] (3) Evaluation of bitter peptide reduction (hydrophobic peptide degradation, i.e., increase in hydrophobic amino acids) The increase in hydrophobic amino acids was examined to evaluate the reduction in bitter peptides in the same manner as in Test Example 2. The results are shown in Table 5.

[0070] [Table 5]

[0071] As is clear from Table 5, the cheese substitutes produced using protease (Examples 14-19) had improved stretchability compared to the cheese substitutes produced without protease (Comparative Examples 7-9), and in particular, as shown in Examples 15, 17, and 19, which are compared with Examples 14, 16, and 18, respectively, the use of peptidase in combination with protease demonstrated an even more excellent effect in improving stretchability (Examples 15, 17, and 19).With regard to thermal meltability, the cheese substitutes produced using protease or a combination of protease and peptidase exhibited excellent thermal meltability compared to the cheese substitutes produced without protease (Comparative Examples 8 and 9) (Examples 16-19). With regard to the amount of hydrophobic amino acids, the amount of hydrophobic amino acids was improved in the cheese alternatives produced using protease (Examples 14 to 19) compared to the amount of hydrophobic amino acids in the cheese alternatives produced without protease (Comparative Examples 7 to 9). In particular, as shown in Examples 15, 17, and 19, which are compared with Examples 14, 16, and 18, respectively, an even greater effect of increasing the amount of hydrophobic amino acids was observed by using peptidase in combination with protease (Examples 15, 17, and 19), suggesting that the hydrophobic peptides that cause bitterness were reduced.

[0072] [Test Example 4] Cheese replicas were prepared in the same manner as in Test Example 1, except that purified water (RO water), chickpea protein material, tapioca starch, coconut oil, κ-carrageenan, salt, and the enzyme agents shown in Table 6 were added in the amounts shown in Table 6.

[0073] (1) Stretchability evaluation The stretchability was evaluated in the same manner as in Test Example 1. The results are shown in Table 6.

[0074] (2) Evaluation of bitter peptide reduction (hydrophobic peptide degradation, i.e., increase in hydrophobic amino acids) The increase in hydrophobic amino acids was examined to evaluate the reduction in bitter peptides in the same manner as in Test Example 2. The results are shown in Table 6.

[0075] [Table 6]

[0076] As is clear from Table 6, the stretchability of the cheese substitutes produced using protease (Examples 20 to 25) was improved compared to the stretchability of the cheese substitutes produced without using protease (Comparative Examples 10 to 12). In particular, as shown in Examples 21, 23, and 25, which are compared with Examples 20, 22, and 24, an even more excellent effect of improving stretchability was observed by using peptidase in combination with protease (Examples 21, 23, and 25). With regard to the amount of hydrophobic amino acids, the amount of hydrophobic amino acids was increased in the cheese alternatives produced using protease (Examples 20 to 25) compared to the amount of hydrophobic amino acids in the cheese alternatives produced without protease (Comparative Examples 10 to 12). In particular, as shown in Examples 21, 23, and 25, which are compared with Examples 20, 22, and 24, an even greater effect of increasing the amount of hydrophobic amino acids was observed by using peptidase in combination with protease (Examples 21, 23, and 25), suggesting that the hydrophobic peptides that cause bitterness were reduced.

[0077] [Test Example 5] Cheese replicas were prepared in the same manner as in Test Example 1, except that purified water (RO water), lentil protein material, tapioca starch, coconut oil, κ-carrageenan, salt, and the enzyme agents shown in Table 7 were added in the amounts shown in Table 7.

[0078] (1) Stretchability evaluation The stretchability was evaluated in the same manner as in Test Example 1. The results are shown in Table 7.

[0079] (2) Thermal melting evaluation The heat melting property was evaluated in the same manner as in Test Example 2. The results are shown in Table 7.

[0080] (3) Evaluation of bitter peptide reduction (hydrophobic peptide degradation, i.e., increase in hydrophobic amino acids) The increase in hydrophobic amino acids was examined to evaluate the reduction in bitter peptides in the same manner as in Test Example 2. The results are shown in Table 7.

[0081] [Table 7]

[0082] As is clear from Table 7, the cheese substitutes produced using protease (Examples 26 to 29) had improved stretchability compared to the cheese substitutes produced without protease (Comparative Examples 13 and 14), and in particular, as shown in Example 27, which is compared with Example 26, and Example 29, which is compared with Example 28, the use of peptidase in combination with protease demonstrated an even more excellent effect of improving stretchability (Examples 27 and 29). With regard to heat-meltability, the cheese substitutes produced using protease or a combination of protease and peptidase exhibited excellent heat-meltability compared to the cheese substitutes produced without protease (Comparative Examples 13 and 14) (Examples 26 to 29). With regard to the amount of hydrophobic amino acids, the amount of hydrophobic amino acids was increased in the cheese alternatives produced using protease (Examples 26 to 29) compared to the amount of hydrophobic amino acids in the cheese alternatives produced without protease (Comparative Examples 13 and 14). In particular, as shown in Example 27, which is compared with Example 26, and Example 29, which is compared with Example 28, the use of peptidase in combination with protease showed an even greater effect of increasing the amount of hydrophobic amino acids (Examples 27 and 29), suggesting that the hydrophobic peptides that cause bitterness were reduced.

Claims

1. The method comprises treating a material composition containing a vegetable protein and a starch with a protease, the vegetable protein is selected from the group consisting of pea protein, fava bean protein, chickpea protein, and lentil protein; A method for producing a stretchable cheese substitute, wherein the protease is an endopeptidase.

2. The method according to claim 1, wherein the content of the starch per 1 part by weight of the vegetable protein is 0.1 part by weight or more and less than 0.6 part by weight.

3. The method according to claim 1 or 2, wherein the protease is a bacterial protease.

4. The method according to any one of claims 1 to 3, wherein the protease is derived from the genus Bacillus and / or Geobacillus.

5. The production method according to any one of claims 1 to 4, wherein the protease is selected from the group consisting of proteases derived from Bacillus stearothermophilus, Bacillus licheniformis, Bacillus amyloliquefaciens, and these proteases derived from Geobacillus genus.

6. The method according to any one of claims 1 to 5, wherein the protease has a protease activity of 10 to 500 U per 1 g of the vegetable protein.

7. A manufacturing method described in any one of claims 1 to 6, further comprising a step of treating with an exopeptidase.

8. The method according to any one of claims 1 to 7, wherein the content of the vegetable protein in the material composition is 15 to 30 wt%.

9. The method according to any one of claims 1 to 8, wherein the starch is tapioca starch.

10. A stretchability improver for a stretchable cheese substitute comprising vegetable protein and starch, comprising a protease; the protease is an endopeptidase, The stretchability improving agent, wherein the vegetable protein is selected from the group consisting of pea protein, fava bean protein, chickpea protein, and lentil protein.

11. A stretchability improver as described in claim 10, further containing an exopeptidase.

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