WO2021167062A1 - Method for meat modification, composition for modification, and food product containing modified meat - Google Patents
Method for meat modification, composition for modification, and food product containing modified meat Download PDFInfo
- Publication number
- WO2021167062A1 WO2021167062A1 PCT/JP2021/006333 JP2021006333W WO2021167062A1 WO 2021167062 A1 WO2021167062 A1 WO 2021167062A1 JP 2021006333 W JP2021006333 W JP 2021006333W WO 2021167062 A1 WO2021167062 A1 WO 2021167062A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- meat
- thermolysin
- neutral protease
- bacillus amyloliquefaciens
- present
- Prior art date
Links
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- 108090001109 Thermolysin Proteins 0.000 claims abstract description 85
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/70—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/70—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor
- A23L13/72—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions
- A23L13/74—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions using microorganisms or enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L35/00—Food or foodstuffs not provided for in groups A23L5/00 – A23L33/00; Preparation or treatment thereof
Definitions
- the present invention relates to a method for modifying meat that can easily impart looseness to meat in a short time and easily, a composition for modifying meat, and a food containing meat to which ease of loosening has been imparted. ..
- Tenderize treatment that cuts fibers short while keeping, complicated pretreatment such as tumbling treatment that makes it easier for fibers to loosen at the same time as physically infiltrating the seasoning liquid, long-term cooking such as vacuum low temperature cooking, pressure cooking In many cases, complicated cooking such as is performed. Therefore, there is a demand for a technique that can easily give the chunky meat easy to loosen in a short time without performing such processing or cooking.
- the texture peculiar to stewed dishes that loosens in the mouth is preferred, but in order to realize the texture without complicated operations, it is necessary to stew for a long time. The need for shortening the cooking time is very high.
- proteases include a method of allowing protease and transglutaminase to act on chicken and animal meat (Patent Document 1), and a modification containing protease and sucrose fatty acid ester.
- Patent Document 2 A pawnbroker (Patent Document 2), a method of contacting meat with a heat-resistant protease (Patent Document 3), a neutral protease derived from Bacillus amyloliquefaciens and having specific properties, and trisodium citrate or
- Patent Document 4 A method for producing a processed ground meat food in which arginine and arginine act on the ground meat
- Patent Document 1 and Patent Document 2 the type of protease is not limited, and there is room for consideration in the superiority of the meat softening effect.
- Patent Document 3 although a heat-resistant protease is used and a certain superior effect is shown, there are still problems in terms of work complexity and length of processing time. It had been.
- Patent Document 4 describes that a processed meat product having excellent juiciness and texture can be produced by a neutral protease derived from Bacillus amyloliquefaciens, but meat other than minced meat, For example, its effect on chunky meat has not been investigated.
- Patent Document 5 describes a method for modifying hard, streaky, low-quality meat by combining treatment with one or more types of protease and high-pressure treatment under a pressure of 100 kg / cm 2 to 4000 kg / cm 2. Is disclosed. Patent Document 6 describes a method of softening meat and fish and shellfish with an enzyme-treated solution controlled to a specific viscosity, and Patent Document 7 describes a method of softening meat and seafood with an enzyme-treated solution containing protease, starch and curdran. , A method of softening meat and seafood is disclosed. Although thermolysin is mentioned as a protease in these patent documents, no detailed study using thermolysin has been made, and no mention is made of the ease of loosening of meat in stewed dishes.
- Patent Document 8 As a method of softening meat in a short time in stewed dishes, a method of adding glutinous rice (Patent Document 8), a method of adding one or more of potassium chloride, trisodium citrate, sorbitol, and trehalose (Patent Document 8). Although 9) are disclosed, there remains a problem in modifying the lumpy meat in a short time with the techniques described in these.
- Japanese Patent No. 3235278 Japanese Unexamined Patent Publication No. 7-313108 Special Table 2003-508804 Japanese Patent No. 6398198 Japanese Unexamined Patent Publication No. 5-7476 Japanese Unexamined Patent Publication No. 2014-30410 Japanese Patent No. 4850943 Japanese Unexamined Patent Publication No. 8-131122 Japanese Patent No. 4809297
- an object of the present invention is to provide a method for modifying meat that can soften meat in a short time and easily to impart easy loosening to the meat, and to provide meat in a short time and easily. It is an object of the present invention to provide a composition for modifying meat, which can be softened to impart looseness to the meat.
- the present inventor uses a neutral protease derived from Bacillus amyloliquefaciens, which has specific characteristics, and a thermolysin. As a result, it has been found that the meat can be softened and easily loosened in a short time and easily, and the present invention has been completed. That is, the present invention is as follows.
- EDTA ethylenediaminetetraacetic acid
- the amount of neutral protease derived from Bacillus amyloliquefaciens to be brought into contact with meat is 10 U or more and 5,000,000 U or less per 100 g of meat, and the amount of thermolysin to be brought into contact with meat is 100 g of meat.
- the method according to [1], wherein the amount is 10 U or more and 5,000,000 U or less per unit.
- a method for producing a stewed dish which comprises using the meat modified by the method according to [1] or [2].
- a composition for modifying meat which is a neutral protease derived from Bacillus amyloliquefaciens and contains a neutral protease having the following characteristics (a) to (e) and thermolysin.
- thermolysin An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
- B The optimum pH is 7,
- D The optimum temperature is 50 ° C to 55 ° C.
- E A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
- EDTA ethylenediaminetetraacetic acid
- B) The optimum pH is 7,
- C) Stable at pH 5-7,
- D) Optimal temperature 50 ° C to 55 ° C,
- E A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
- the meat can be softened and the meat can be easily loosened in a short time and easily. Further, according to the present invention, it is possible to provide a composition for modifying meat that can soften meat and impart easy loosening to the meat in a short time and easily. Further, according to the present invention, it is possible to provide a food containing meat that has been softened and imparted with ease of loosening.
- the present invention provides a method for modifying meat (hereinafter, also referred to as "method for modifying the present invention” in the present specification).
- method for modifying the present invention refers to softening the meat and imparting ease of loosening to the meat.
- the modification method of the present invention comprises contacting meat with a neutral protease from Bacillus amyloliquefaciens and thermolysin.
- the neutral protease derived from Bacillus amyloliquefaciens used in the modification method of the present invention is a protease produced by Bacillus amyloliquefaciens, and the proteases produced by Bacillus amyloliquefaciens are described below (a) to (a). Those having the characteristics of e) are used.
- A An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
- B) The optimum pH is 7,
- the optimum temperature is 50 ° C to 55 ° C.
- E A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
- the neutral protease for example, Bacillus amyloliquefaciens is cultured, and the culture solution is collected, concentrated, and dried. Ion exchange column chromatography, gel filtration column chromatography, or the like is used from the culture solution. It is possible to use a product produced by purification by purification or a product produced by a gene recombination method, but a commercially available enzyme preparation can also be used. Examples of commercially available enzyme preparations include "Protin SD-NY10" and "Protin NY100" commercially available from Amano Enzyme Co., Ltd.
- the neutral protease can be used in various forms such as powder, granular, granular, liquid, and paste. In the present invention, the above-mentioned neutral protease derived from Bacillus amyloliquefaciens can be used alone or in combination of two or more.
- thermolysin used in the modification method of the present invention is a thermolysin, which is also called thermolysin, thermolysin, etc., and is a neutral metalloproteinase produced by Bacillus thermoproteolitics. It is an enzyme that selectively hydrolyzes peptide bonds containing hydrophobic amino acids in proteins, and has a CAS Registry Number of 9073-78-3.
- other basic enzymes such as Geobacillus stearothermophilus may be used as long as they have the above-mentioned characteristics, and a modifying enzyme or a recombinant enzyme by amino acid substitution or the like may be used. can.
- thermolysin manufactured by Fujifilm Wako Pure Chemical Industries, Ltd. and "thermolysin” manufactured by Promega Co., Ltd.
- the thermolysin can be used in various forms such as powder, granular, granular, liquid, and paste. In the present invention, the thermolysin can be used alone or in combination of two or more.
- meat refers to meat to be used for food, that is, meat derived from livestock, poultry, and other wild animals, which is used for food.
- Livestock include pigs, cows, sheep, goats, horses, camels, rabbits, alpaca, etc.
- Poultry includes chickens, pigeons, ducks, ducks, sardines, etc.
- wild animals include deer, boars, etc. , Bears, etc.
- the meat meat of various parts derived from the above animals is used, and it can be appropriately cut into a desired size and shape and used, and can be used as chunky block meat or sliced meat (thick sliced meat).
- the reforming method of the present invention is suitable for reforming chunky meat, which was difficult to soften in a short time by the conventional softening treatment method, and hard, low-quality meat, and is used for stewed dishes. It is particularly suitable for modifying meat.
- the neutral protease derived from Bacillus amyloliquefaciens and the means or mode of contact between the thermolysin and the meat are such that the neutral protease and the thermolysin act on the meat for a certain period of time.
- the enzyme is added to the meat, such as by sprinkling the enzyme in the form of powder or granules, or the enzyme in the form of a liquid or paste is applied to the meat, or a solution or dispersion of the enzyme.
- the suspension may be injected into the meat, or the meat may be immersed in the enzyme solution or dispersion or suspension.
- a method of sprinkling a powdered or granular enzyme and contacting the meat, or a method of immersing the meat in a solution, dispersion or suspension of the enzyme and contacting the meat is preferable.
- the neutral protease derived from Bacillus amyloliquefaciens and thermolysin may be contacted in a state of being prepared as a composition for modifying meat described later.
- the contact between the enzyme and the meat can be appropriately performed at the stage of processing and cooking the meat, and the raw meat may be brought into contact with the enzyme and then cooked, or the heated meat may be cooked.
- the meat may be cooked again after being brought into contact with the enzyme, but from the viewpoint of convenience of treatment, it is preferably brought into contact with raw meat.
- the order in which the neutral protease derived from Bacillus amyloliquefaciens and the thermolysin are brought into contact with the meat is not particularly limited, and the neutral protease derived from Bacillus amyloliquefaciens and the thermolysin derived from Bacillus amyloliquefaciens are not particularly limited. They may be brought into contact first or both may be brought into contact at the same time, but from the viewpoint of simplicity of the reforming treatment, it is preferable to bring them into contact at the same time.
- the amount of neutral protease derived from Bacillus amyloliquefaciens to be brought into contact with meat may be 10 units (U) or more and 5,000,000 U or less per 100 g of meat. It is preferably 50 U or more and 3,000,000 U or less, and more preferably 100 U or more and 1,000,000 U or less.
- the amount of the enzyme that causes an increase in the folin test solution color-forming substance corresponding to 1 ⁇ g of tyrosine per minute using casein as a substrate was defined as 1U.
- the amount of thermolysin to be brought into contact with meat is preferably 10 U or more and 5,000,000 U or less, and more preferably 50 U or more and 3,000,000 U or less with respect to 100 g of meat. It is more preferably 100 U or more and 1,000,000 U or less.
- the enzyme activity of thermolysin was defined in the same way as the above-mentioned neutral protease.
- the enzymatic activities of various proteases used in the test examples described later are all defined in the same manner as the above-mentioned neutral proteases.
- the ratio of the neutral protease amount to the thermolysin amount when contacting the meat with a neutral protease derived from Bacillus amyloliquefaciens and thermolysin is preferably 1: 0.000002 to 1: 500,000, more preferably 1: 0.00002 to 1: 50,000, and 1: 0 in terms of the activity ratio of these enzymes. It is more preferably .0001 to 1: 10,000.
- the neutral protease derived from Bacillus amyloliquefaciens and the time for contacting the thermolysin with the meat are sufficient for the neutral protease and the thermolysin to be a substrate substance, that is, a protein contained in the meat.
- the time is not particularly limited as long as it can act on, but specifically, 1 minute to 24 hours (for example, 1 minute to 12 hours, 1 minute to 8 hours, 1 minute to 4 hours, 1 minute to 2 hours, 1 minute to 2 hours). Minutes to 1 hour, etc.) is preferable.
- the present invention is characterized in that by using a neutral protease and thermolysin in combination, the meat can be softened and easily loosened in a short time and easily, and the meat can be easily loosened for about 1 minute to 1 hour.
- the utility value of the present invention is further improved by setting the action time to be short.
- the neutral protease derived from Bacillus amyloliquefaciens and the temperature at which the thermolysin is brought into contact with the meat are also within the range in which the neutral protease and the thermolysin maintain their activities.
- ° C to 80 ° C for example, 0 ° C to 75 ° C, 0 ° C to 70 ° C, 0 ° C to 65 ° C, 0 ° C to 60 ° C, 0 ° C to 55 ° C, 0).
- ° C. to 50 ° C., 0 ° C. to 45 ° C., etc. is preferable. That is, they can be sufficiently allowed to act by contacting the enzymes under normal meat processing / cooking conditions.
- the meat can be softened and the meat can be easily loosened in a short time and easily. Therefore, the modification method of the present invention is effective in a stewed dish using meat, particularly a stewed dish using chunky meat.
- the present invention also provides a method for producing a stewed dish, which comprises using the meat modified by the modification method of the present invention.
- the “simmered dish” refers to a dish in which meat, particularly chunky meat, is cooked in a state of being immersed in a liquid. , Soup, etc.
- the method for producing a stewed dish of the present invention is modified by a normal process adopted in the production of a stewed dish, for example, a processing step such as shredding other ingredients, boiling underneath, or the above-mentioned modification method of the present invention.
- a step of mixing the meat and other ingredients, a step of cooking the meat and other ingredients by heating (frying, baking, boiling, steaming, etc.), a seasoning step, and the like can be further included.
- the method for producing a stewed dish of the present invention includes a step of cooling the manufactured stewed dish and storing it by refrigerating, freezing, etc., a sterilization process such as high-pressure heat sterilization, and a container such as a can, a bottle, and a retort pouch. The process of filling and packaging can also be included.
- Each of the above steps is carried out by a general method under the conditions normally carried out in the production of food.
- the present invention also provides a composition for modifying meat (hereinafter, also referred to as “composition of the present invention” in the present specification).
- composition of the present invention refers to a composition used to soften meat and impart ease of loosening to the meat.
- the composition of the present invention contains a neutral protease derived from Bacillus amyloliquefaciens, and a thermolysin.
- the neutral protease and thermolysin contained in the composition of the present invention are as described above in the modification method of the present invention, and each content thereof in the composition of the present invention is the modification method of the present invention.
- the above-mentioned amounts of the neutral protease and thermolysin are set as an amount that can be brought into contact with each other.
- the content ratio of the neutral protease to the thermolysin in the composition of the present invention is preferably 1: 0.000002 to 1: 500 in terms of the enzyme activity (U) ratio. It is 000, more preferably 1: 0.00002 to 1: 50,000, and even more preferably 1: 0.0001 to 1: 10,000.
- the composition of the present invention is a mixture of a neutral protease derived from Bacillus amyloliquefaciens and thermolysin, and if necessary, general food additives are added to make a powdery, granular, granular composition. It can be in various forms such as solids such as granules and tablets; semi-solids such as gels, pastes and creams; liquids such as solutions, suspensions and dispersed liquids. Further, it can be provided in a solid form such as powder, granule, or granule, and can be used as a time-prepared type composition which is dissolved, dispersed or suspended in a solvent such as water at the time of use.
- the food additive examples include water, solvents such as polyhydric alcohol; bases such as polyethylene glycol; excipients such as dextrin and lactose; gelatin, pregelatinized starch, partially pregelatinized starch, cellulose and derivatives thereof (crystalline cellulose). , Hydroxypropyl cellulose, etc.); Disintegrants such as crospovidone, povidone, crystalline cellulose, etc .; Starch, magnesium stearate, etc.
- Emulsifiers such as lecithin, sodium caseinate; seasonings such as salt, amino acids and salts thereof, nucleic acids and salts thereof, yeast extract, livestock meat extract; proteins such as plant protein, gluten, egg white, gelatin, casein; protein hydrolyzate; protein Partial decomposition products; pH adjusters such as organic acid salts (fumarate etc.); chelating agents such as polymerized phosphates; reducing agents such as glutathione and cysteine; thickening polysaccharides such as alginic acid and xanthan gum; casein; pigments; Acidulants; fragrances and the like can be mentioned.
- the modifying composition of the present invention is a formulation well known in the field of formulation by adding the above-mentioned food additives to the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, if necessary. It can be manufactured according to the means for making it, for example, the method described in each article of the 17th revised Japanese Pharmacopoeia General Regulations for Pharmaceuticals [3] Pharmaceuticals.
- the composition of the present invention is used for modifying meat by contacting it with meat by the means or embodiment described above in the modification method of the present invention.
- the contact amount of the neutral protease derived from Bacillus amyloliquefaciens and the thermolysin is the same amount as described above in the modification method of the present invention with respect to 100 g of meat, respectively. , Used.
- composition of the present invention can easily soften the meat in a short time and impart the meat to be easily loosened. Therefore, the composition of the present invention is preferably used in meat, especially stewed dishes using chunky meat.
- the present invention provides a food containing meat that has been softened and imparted with ease of loosening (hereinafter, also referred to as "food of the present invention” in the present specification).
- the food product of the present invention contains a neutral protease derived from Bacillus amyloliquefaciens, and meat on which thermolysin is acted.
- Neutral protease derived from Bacillus amyloliquefaciens and meat on which thermolysin acts refers to neutral protease derived from Bacillus amyloliquefaciens and modified contact with thermolysin. Meat.
- the neutral protease derived from Bacillus amyloliquefaciens used for preparing the meat contained in the food of the present invention and the thermolysin are as described above in the modification method of the present invention. Further, in the preparation of the meat contained in the food of the present invention, the amount of the neutral protease and thermolysin to be brought into contact with the meat (contact amount with respect to 100 g of meat), the method of contacting the neutral protease and the thermolysin, the contact time, The temperature at the time of contact is also the same as described above in the reforming method of the present invention.
- the food product of the present invention can be prepared by a cooking means commonly used in the field of food products, using a neutral protease derived from Bacillus amyloliquefaciens and meat on which thermolysin is acted, for example, as described above. If necessary, other ingredients and food additives such as preservatives and seasonings are added to the meat, and the meat is prepared by ordinary cooking means such as roasting, baking, boiling, and steaming. Processed foods such as meat that are further used for cooking as ingredients, and processed foods such as stew, curry, potov, boiled, roast beef, roast pork, roast chicken, roasted meat, roasted pork, roasted chicken, steak, cutlet, etc. Can be provided as. Further, the storage form of the food of the present invention is not particularly limited, and it can be stored in various forms such as room temperature storage, refrigerated storage, and frozen storage, and may be prepared as a retort pouch food.
- the food product of the present invention contains meat that has been softened and imparted with ease of loosening, and exhibits a good texture. Therefore, the food product of the present invention can be particularly preferably provided as a stewed dish using chunky meat.
- the sensory evaluation was performed on a panel of 4 people by a scoring method of -5 to 5 points with the additive-free group (test group 1) as 0 points for softness and ease of loosening.
- the grades were given to the first decimal place, and the evaluation results were shown by the numerical values up to the first decimal place, rounded off from the average value of the four people.
- "softness” means the chewing force required to chew the whole meat
- "easiness to loosen” means the feeling of loosening like a white fish when chewing the meat.
- the ease of loosening is a texture that is particularly preferred in stewed dishes, and is generally improved by boiling for a long time.
- the ease of unraveling is the texture that is felt when the myofibrils of the meat collapse as a bundle to some extent. Even with a soft texture such as liver or foie gras, where the whole meat is crushed evenly, the ease of loosening is not felt so much.
- the scores for softness and ease of loosening were defined as follows by comparison with the additive-free group.
- thermolysin improved the ease of loosening most (Test Group 4).
- the softness was most improved by adding papain, but the softness given was a liver-like uniform softness, and the ease of loosening was not felt so much (Test Group 2).
- a high softness-imparting effect is obtained in the neutral protease-added group (test group 8) derived from Aspergillus oryzae and the acidic protease-added group (test group 5) derived from Aspergillus sp. Although it was observed, there was not much improvement in the ease of loosening.
- thermolysin which showed the effect of improving the looseness most, may be used in combination with other proteases to further improve the looseness.
- Test Example 2 In Test Example 1, thermolysin, which had the highest effect of imparting looseness to meat, was examined for its combined effect with other proteases.
- test Example 2 In the same manner as in Test Example 1, domestic young chicken breast meat was cut into 20 g pieces, placed in a vinyl bag, and the proteases of each test group shown in Table 2 were added and shaken. In Table 2, "-" in the enzyme 1 column and "-" in the enzyme 2 column indicate that each enzyme is not added. Test group 2 was a group to which thermolysin was added alone, and test groups 3 to 11 were groups to which thermolysin was added in combination with other proteases. As the thermolysin and other proteases shown in Table 2, the same ones used in Test Example 1 were used. The amount of each protease added was 10,000 U per 20 g of chicken breast meat (50,000 U per 100 g of chicken breast meat).
- thermolysin was based on the evaluation results of the test group to which only thermolysin was added (test group 2) and the test group to which a protease other than thermolysin was added alone (test groups 2 to 11 of test example 1).
- the theoretical value of the evaluation of the test group added in combination with other proteases was calculated as the sum of the evaluation results (average value of the scores) in the test group added individually. For example, for the test group 3 in Table 2, the theoretical value for the evaluation of looseness is "2" for the test group to which only thermolysin was added (test group 2 in Table 2), and only papain.
- the difference between the theoretical value calculated in this way and the actual sensory evaluation result was obtained.
- Test Group 7 the difference between the score of ease of loosening obtained by sensory evaluation and the theoretical value was 1.1 points, and a neutral protease derived from Bacillus amyloliquefaciens was used in combination with thermolysin. It was shown that a synergistic effect of improving the ease of loosening can be obtained. However, in Test Group 9, the difference between the score of ease of loosening obtained by the sensory evaluation and the theoretical value was -0.7 points, and no synergistic improvement in ease of loosening was observed. In addition, the combined use of papain and thermolysin greatly improved the softness, but the texture became uniform like a lever, and the ease of loosening decreased on the contrary (Test Group 3).
- thermolysin As described above, the action on meat tissue differs depending on the type of protease used in combination with thermolysin, but the effect of improving the ease of loosening by using a neutral protease derived from Bacillus amyloliquefaciens in combination with thermolysin is It was found to be clearly larger and synergistic than the combination of thermolysin and other proteases. In particular, in stewed dishes in which the ease of loosening of meat is preferred, the effect of imparting ease of loosening based on the findings is extremely useful.
- Examples 1 to 15 Modification of chicken breast meat and preparation of cream stew
- domestic young chicken breast meat is cut into 20 g pieces, placed in a vinyl bag, and Bacillus amilo.
- Neutral protease and thermolysin derived from Bacillus amyloliquefaciens were added in the amounts shown in Table 3 and shaken, allowed to stand at 20 ° C. for 15 minutes, and the chicken breast meat pieces were brought into contact with the protease.
- the chicken breast was modified.
- Table 3 "-" in the neutral protease column and the thermolysin column derived from Bacillus amyloliquefaciens indicates that each enzyme is not added.
- Chicken breast meat was taken out from the obtained cream stew of chicken breast meat and used as a sample for sensory evaluation.
- all of the above proteases were treated in the same manner without addition, cooked to prepare a cream stew, and chicken breast meat was taken out and used as a control.
- the sensory evaluation was performed on a panel of 4 people by a scoring method of -5 to 5 points with a control of 0 points for softness and ease of loosening, and the score was calculated by the same method as in Test Example 1.
- the evaluation result is shown by the average value.
- " ⁇ " was given to samples with an average score of 3 points or more
- " ⁇ " was given to samples with 2 points or more and less than 3 points
- samples with less than 2 points were given.
- the evaluation results are also shown in Table 3.
- the neutral protease and thermolysin derived from Bacillus amyloliquefaciens are brought into contact with the meat in an amount of 10 U to 5,000,000 U per 100 g of the meat, respectively, and then boiled. It was suggested that the meat can be given the ease of loosening, which is a texture that is particularly preferred in cooking.
- Example 16 Beef thigh curry A US beef thigh is die-cut into 2 cm squares, placed in a vinyl bag, and a neutral protease and thermolysin derived from Bacillus amyloliquefaciens are added. It was shaken and allowed to stand at 20 ° C. for 15 minutes for contact. As the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, the same proteases used in Test Example 1 were used, and the amount of each protease added was 50, per 100 g of beef thigh meat. It was set to 000U.
- the curry was prepared by cooking as follows in a pan. That is, the surface of the beef thigh meat was lightly fried, then water was added, and after boiling, it was simmered for 15 minutes, then a commercially available roux (manufactured by House Foods Co., Ltd.) was dissolved, and the meat was simmered for another 10 minutes. The total boiling time was 25 minutes.
- a curry was prepared using beef thigh meat treated in the same manner without contacting the above protease, and used as Comparative Example 1.
- Example 17 Kakuni pork belly Die-cut domestic pork belly into 3 cm squares, put them in a vinyl bag, and add a neutral protease and thermolysin derived from Bacillus amyloliquefaciens. It was shaken and allowed to stand at 20 ° C. for 15 minutes for contact. As the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, the same proteases used in Test Example 1 were used, and the amount of each protease added was 50, per 100 g of pork belly. It was set to 000U.
- the beef thighs and pork ribs were taken out from the curry of each beef thigh of Example 16 and Comparative Example 1 and the pork ribs of Example 17 and Comparative Example 2, respectively, and subjected to sensory evaluation.
- the sensory evaluation was carried out focusing on the ease of loosening of the meat, and was evaluated by a panel of four people according to the following evaluation criteria.
- the sensory evaluation results were decided through discussions among four people. For pork ribs, the ease of loosening in the meat layer was evaluated among the fat layer and the meat layer.
- the beef in the curry of Example 16 and the pork ribs in the kakuni of Example 17 are both easier to loosen than the beef in the curry of Comparative Example 1 and the pork ribs in Comparative Example 2.
- Beef thighs and pork ribs in curry and kakuni prepared using beef thighs and pork ribs on which neutral protease and thermolysin from Bacillus amyloliquefaciens acted, respectively. It was confirmed that the ribs were clearly given the ease of loosening.
- the present invention it is possible to provide a method for modifying meat that can easily soften meat in a short time and impart ease of loosening to the meat. Further, according to the present invention, it is possible to provide a composition for modifying meat that can easily soften meat in a short time and impart ease of loosening to the meat. Further, according to the present invention, it is possible to provide a food containing meat that has been softened and imparted with ease of loosening. The present invention is extremely useful in the food field, especially in stewed dishes in which the ease of loosening of meat is preferred as a texture.
Abstract
The present invention relates to: a neutral protease derived from Bacillus amyloliquefaciens, the neutral protease having properties (a) through (e); a method for meat modification, the method including bringing thermolysin into contact with the meat; and a composition for modification that contains the neutral protease and thermolysin. Property (a): being an enzyme that hydrolyzes a substrate having a peptide bond to free peptides.
Property (b): having an optimum pH of 7.
Property (c): being stable at a pH between 5 and 7.
Property (d): having an optimum temperature of 50°C-55°C.
Property (e): being a metalloprotease and inhibited by ethylenediaminetetraacetate (EDTA).
The present invention is capable of rapidly and easily tenderizing meat and imparting an easy-to-break-up property to the meat.
Description
本発明は、短時間でかつ簡便に、食肉にほぐれやすさを付与し得る食肉の改質方法、及び食肉の改質用組成物、ならびに、ほぐれやすさが付与された食肉を含有する食品に関する。
The present invention relates to a method for modifying meat that can easily impart looseness to meat in a short time and easily, a composition for modifying meat, and a food containing meat to which ease of loosening has been imparted. ..
食肉加工及び調理の分野においては、様々なニーズが存在する。例えば、ハンバーグにおける肉粒感とジューシー感、ソーセージにおけるプリプリ感、から揚げやステーキにおけるやわらかさとジューシー感、煮込み料理における肉のほぐれやすさと調理時間の短縮等が挙げられる。
とりわけ、煮込み料理等、塊状の肉を使用する調理においては、短時間での肉の改質が難しく、赤身の肉に脂質、タンパク質、食品添加物等を注入するインジェクション加工や、肉の原形を保ったまま繊維を短く切断するテンダライズ処理、調味液を物理的に肉に浸透させると同時に繊維同士をほぐれやすくするタンブリング処理等の煩雑な前処理、真空低温調理等の長時間にわたる調理、圧力調理等の煩雑な調理等が行われることが多い。それゆえ、かかる処理や調理を行うことなく、短時間で簡便に塊状の肉にほぐれやすさを付与できる技術が求められている。特に、煮込み料理に用いられる肉においては、口の中でホロホロとほぐれる煮込み料理に特有の食感が好まれるが、煩雑な操作を伴わずにかかる食感を実現するには、長時間煮込むことを要し、調理時間の短縮の必要性は非常に高い。 There are various needs in the fields of meat processing and cooking. For example, the feeling of meat grain and juiciness in hamburger steak, the feeling of pre-preparation in sausage, the softness and juiciness in fried chicken and steak, the ease of loosening of meat and shortening of cooking time in steaked dishes can be mentioned.
In particular, in cooking using chunky meat such as stewed dishes, it is difficult to modify the meat in a short time, so injection processing that injects lipids, proteins, food additives, etc. into lean meat and the original form of meat can be used. Tenderize treatment that cuts fibers short while keeping, complicated pretreatment such as tumbling treatment that makes it easier for fibers to loosen at the same time as physically infiltrating the seasoning liquid, long-term cooking such as vacuum low temperature cooking, pressure cooking In many cases, complicated cooking such as is performed. Therefore, there is a demand for a technique that can easily give the chunky meat easy to loosen in a short time without performing such processing or cooking. In particular, for meat used in stewed dishes, the texture peculiar to stewed dishes that loosens in the mouth is preferred, but in order to realize the texture without complicated operations, it is necessary to stew for a long time. The need for shortening the cooking time is very high.
とりわけ、煮込み料理等、塊状の肉を使用する調理においては、短時間での肉の改質が難しく、赤身の肉に脂質、タンパク質、食品添加物等を注入するインジェクション加工や、肉の原形を保ったまま繊維を短く切断するテンダライズ処理、調味液を物理的に肉に浸透させると同時に繊維同士をほぐれやすくするタンブリング処理等の煩雑な前処理、真空低温調理等の長時間にわたる調理、圧力調理等の煩雑な調理等が行われることが多い。それゆえ、かかる処理や調理を行うことなく、短時間で簡便に塊状の肉にほぐれやすさを付与できる技術が求められている。特に、煮込み料理に用いられる肉においては、口の中でホロホロとほぐれる煮込み料理に特有の食感が好まれるが、煩雑な操作を伴わずにかかる食感を実現するには、長時間煮込むことを要し、調理時間の短縮の必要性は非常に高い。 There are various needs in the fields of meat processing and cooking. For example, the feeling of meat grain and juiciness in hamburger steak, the feeling of pre-preparation in sausage, the softness and juiciness in fried chicken and steak, the ease of loosening of meat and shortening of cooking time in steaked dishes can be mentioned.
In particular, in cooking using chunky meat such as stewed dishes, it is difficult to modify the meat in a short time, so injection processing that injects lipids, proteins, food additives, etc. into lean meat and the original form of meat can be used. Tenderize treatment that cuts fibers short while keeping, complicated pretreatment such as tumbling treatment that makes it easier for fibers to loosen at the same time as physically infiltrating the seasoning liquid, long-term cooking such as vacuum low temperature cooking, pressure cooking In many cases, complicated cooking such as is performed. Therefore, there is a demand for a technique that can easily give the chunky meat easy to loosen in a short time without performing such processing or cooking. In particular, for meat used in stewed dishes, the texture peculiar to stewed dishes that loosens in the mouth is preferred, but in order to realize the texture without complicated operations, it is necessary to stew for a long time. The need for shortening the cooking time is very high.
肉を軟化させる技術としては多くの報告があるが、そのうち、プロテアーゼを用いる方法としては、プロテアーゼとトランスグルタミナーゼを鳥獣肉に作用させる方法（特許文献１）、プロテアーゼとショ糖脂肪酸エステルを含有する改質剤（特許文献２）、肉を不耐熱性プロテアーゼと接触させる方法（特許文献３）、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来で特定の特性を有する中性プロテアーゼと、クエン酸三ナトリウムまたはアルギニンとを挽き肉に作用させる挽き肉加工食品の製造方法（特許文献４）等が開示されている。
特許文献１及び特許文献２に記載された技術においては、プロテアーゼの種類は限定されておらず、肉の軟化効果の優位性において検討の余地が残されていた。また、特許文献３に記載された技術においては、不耐熱性プロテアーゼを使用し、一定の優位性ある効果が示されているものの、作業の煩雑さや処理時間の長さという点で、課題が残されていた。さらに、特許文献４には、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼにより、ジューシー感と食感に優れた挽き肉加工食品を製造できることが記載されているが、挽き肉以外の肉、たとえば塊状の肉に対する効果は検討されていない。 There are many reports on techniques for softening meat, and among them, methods using proteases include a method of allowing protease and transglutaminase to act on chicken and animal meat (Patent Document 1), and a modification containing protease and sucrose fatty acid ester. A pawnbroker (Patent Document 2), a method of contacting meat with a heat-resistant protease (Patent Document 3), a neutral protease derived from Bacillus amyloliquefaciens and having specific properties, and trisodium citrate or A method for producing a processed ground meat food in which arginine and arginine act on the ground meat (Patent Document 4) and the like are disclosed.
In the techniques described in Patent Document 1 and Patent Document 2, the type of protease is not limited, and there is room for consideration in the superiority of the meat softening effect. Further, in the technique described in Patent Document 3, although a heat-resistant protease is used and a certain superior effect is shown, there are still problems in terms of work complexity and length of processing time. It had been. Further, Patent Document 4 describes that a processed meat product having excellent juiciness and texture can be produced by a neutral protease derived from Bacillus amyloliquefaciens, but meat other than minced meat, For example, its effect on chunky meat has not been investigated.
特許文献１及び特許文献２に記載された技術においては、プロテアーゼの種類は限定されておらず、肉の軟化効果の優位性において検討の余地が残されていた。また、特許文献３に記載された技術においては、不耐熱性プロテアーゼを使用し、一定の優位性ある効果が示されているものの、作業の煩雑さや処理時間の長さという点で、課題が残されていた。さらに、特許文献４には、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼにより、ジューシー感と食感に優れた挽き肉加工食品を製造できることが記載されているが、挽き肉以外の肉、たとえば塊状の肉に対する効果は検討されていない。 There are many reports on techniques for softening meat, and among them, methods using proteases include a method of allowing protease and transglutaminase to act on chicken and animal meat (Patent Document 1), and a modification containing protease and sucrose fatty acid ester. A pawnbroker (Patent Document 2), a method of contacting meat with a heat-resistant protease (Patent Document 3), a neutral protease derived from Bacillus amyloliquefaciens and having specific properties, and trisodium citrate or A method for producing a processed ground meat food in which arginine and arginine act on the ground meat (Patent Document 4) and the like are disclosed.
In the techniques described in Patent Document 1 and Patent Document 2, the type of protease is not limited, and there is room for consideration in the superiority of the meat softening effect. Further, in the technique described in Patent Document 3, although a heat-resistant protease is used and a certain superior effect is shown, there are still problems in terms of work complexity and length of processing time. It had been. Further, Patent Document 4 describes that a processed meat product having excellent juiciness and texture can be produced by a neutral protease derived from Bacillus amyloliquefaciens, but meat other than minced meat, For example, its effect on chunky meat has not been investigated.
また、特許文献５には、１種類以上のプロテアーゼによる処理と１００ｋｇ／ｃｍ２～４０００ｋｇ／ｃｍ２の圧力下での高圧処理の併用により、硬くてスジのある低品質な肉を改質する方法が開示されている。特許文献６には、特定の粘度にコントロールされた酵素処理液により、食肉や魚介類を軟質化する方法が記載され、特許文献７には、プロテアーゼと澱粉及びカードランを含有する酵素処理液により、食肉や魚介類を軟質化する方法が開示されている。
これらの特許文献において、プロテアーゼとしてサーモリシンが挙げられているが、いずれにおいても、サーモリシンを用いた詳細な検討はなされておらず、煮込み料理における肉のほぐれやすさに関しては言及されていない。 Further, Patent Document 5 describes a method for modifying hard, streaky, low-quality meat by combining treatment with one or more types of protease and high-pressure treatment under a pressure of 100 kg / cm 2 to 4000 kg / cm 2. Is disclosed. Patent Document 6 describes a method of softening meat and fish and shellfish with an enzyme-treated solution controlled to a specific viscosity, and Patent Document 7 describes a method of softening meat and seafood with an enzyme-treated solution containing protease, starch and curdran. , A method of softening meat and seafood is disclosed.
Although thermolysin is mentioned as a protease in these patent documents, no detailed study using thermolysin has been made, and no mention is made of the ease of loosening of meat in stewed dishes.
これらの特許文献において、プロテアーゼとしてサーモリシンが挙げられているが、いずれにおいても、サーモリシンを用いた詳細な検討はなされておらず、煮込み料理における肉のほぐれやすさに関しては言及されていない。 Further, Patent Document 5 describes a method for modifying hard, streaky, low-quality meat by combining treatment with one or more types of protease and high-pressure treatment under a pressure of 100 kg / cm 2 to 4000 kg / cm 2. Is disclosed. Patent Document 6 describes a method of softening meat and fish and shellfish with an enzyme-treated solution controlled to a specific viscosity, and Patent Document 7 describes a method of softening meat and seafood with an enzyme-treated solution containing protease, starch and curdran. , A method of softening meat and seafood is disclosed.
Although thermolysin is mentioned as a protease in these patent documents, no detailed study using thermolysin has been made, and no mention is made of the ease of loosening of meat in stewed dishes.
煮込み料理において肉を短時間で軟質化する方法としては、もち米を添加する方法（特許文献８）、塩化カリウム、クエン酸三ナトリウム、ソルビトール、トレハロースのうち１種以上を添加する方法（特許文献９）が開示されているが、これらに記載された技術では、短時間で塊状の肉を改質するには課題が残されていた。
As a method of softening meat in a short time in stewed dishes, a method of adding glutinous rice (Patent Document 8), a method of adding one or more of potassium chloride, trisodium citrate, sorbitol, and trehalose (Patent Document 8). Although 9) are disclosed, there remains a problem in modifying the lumpy meat in a short time with the techniques described in these.
それゆえ、たとえば塊状の食肉であっても、短時間でかつ簡便に軟質化することができ、食肉にほぐれやすさを付与することができる改質方法が、依然として求められている。
Therefore, for example, there is still a demand for a reforming method that can easily soften lumpy meat in a short time and can impart easiness to loosen the meat.
よって、本発明の目的は、短時間でかつ簡便に、食肉を軟質化して食肉にほぐれやすさを付与し得る食肉の改質方法を提供すること、及び、短時間でかつ簡便に、食肉を軟質化して食肉にほぐれやすさを付与することのできる、食肉の改質用組成物を提供することである。
Therefore, an object of the present invention is to provide a method for modifying meat that can soften meat in a short time and easily to impart easy loosening to the meat, and to provide meat in a short time and easily. It is an object of the present invention to provide a composition for modifying meat, which can be softened to impart looseness to the meat.
本発明者は、上記課題を解決すべく鋭意研究を行った結果、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、特定の特性を有する中性プロテアーゼ、及びサーモリシンを用いることにより、短時間でかつ簡便に、食肉を軟質化して食肉にほぐれやすさを付与し得ることを見出し、本発明を完成するに至った。すなわち、本発明は以下の通りである。
As a result of diligent research to solve the above problems, the present inventor uses a neutral protease derived from Bacillus amyloliquefaciens, which has specific characteristics, and a thermolysin. As a result, it has been found that the meat can be softened and easily loosened in a short time and easily, and the present invention has been completed. That is, the present invention is as follows.
［１］バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼ、及びサーモリシンを食肉と接触させることを含む、食肉の改質方法；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
［２］食肉に接触させるバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼの量が、食肉１００ｇあたり１０Ｕ以上５，０００，０００Ｕ以下であり、食肉に接触させるサーモリシンの量が、食肉１００ｇあたり１０Ｕ以上５，０００，０００Ｕ以下である、［１］に記載の方法。
［３］［１］または［２］に記載の方法により改質された食肉を用いることを含む、煮込み料理の製造方法。
［４］バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼ、及びサーモリシンを含有する、食肉の改質用組成物；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
［５］バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼと、サーモリシンとの含有量比（バチルス アミロリケファシエンス由来の中性プロテアーゼ：サーモリシン）が、酵素活性比にて１：０．０００００２～１：５００，０００である、［４］に記載の組成物。
［６］煮込み料理に用いる食肉の改質用である、［４］または［５］に記載の組成物。
［７］バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼおよび、サーモリシンが作用した食肉を含有する、食品；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度＝５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
［８］食肉１００ｇあたり、１０Ｕ以上５，０００，０００Ｕ以下のバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼと、１０Ｕ以上５，０００，０００Ｕ以下のサーモリシンが作用した食肉を含有する、［７］に記載の食品。
［９］煮込み料理として供される、［７］または［８］に記載の食品。 [1] A neutral protease derived from Bacillus amyloliquefaciens, which comprises contacting a neutral protease having the following characteristics (a) to (e) and a thermolysin with the meat. Modification method;
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
[2] The amount of neutral protease derived from Bacillus amyloliquefaciens to be brought into contact with meat is 10 U or more and 5,000,000 U or less per 100 g of meat, and the amount of thermolysin to be brought into contact with meat is 100 g of meat. The method according to [1], wherein the amount is 10 U or more and 5,000,000 U or less per unit.
[3] A method for producing a stewed dish, which comprises using the meat modified by the method according to [1] or [2].
[4] A composition for modifying meat, which is a neutral protease derived from Bacillus amyloliquefaciens and contains a neutral protease having the following characteristics (a) to (e) and thermolysin. ;
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
[5] The content ratio of the neutral protease derived from Bacillus amyloliquefaciens to thermolysin (neutral protease derived from Bacillus amyloliquefaciens: thermolysin) has an enzyme activity ratio of 1: 0. The composition according to [4], which is 000002-1: 500,000.
[6] The composition according to [4] or [5], which is used for modifying meat used in stewed dishes.
[7] A food product containing a neutral protease derived from Bacillus amyloliquefaciens, which has the following characteristics (a) to (e), and meat on which thermolysin acts.
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) Optimal temperature = 50 ° C to 55 ° C,
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
[8] Per 100 g of meat contains a neutral protease derived from Bacillus amyloliquefaciens of 10 U or more and 5,000,000 U or less and a meat on which thermolysin of 10 U or more and 5,000,000 U or less acts. The food product according to [7].
[9] The food according to [7] or [8], which is served as a stewed dish.
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
［２］食肉に接触させるバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼの量が、食肉１００ｇあたり１０Ｕ以上５，０００，０００Ｕ以下であり、食肉に接触させるサーモリシンの量が、食肉１００ｇあたり１０Ｕ以上５，０００，０００Ｕ以下である、［１］に記載の方法。
［３］［１］または［２］に記載の方法により改質された食肉を用いることを含む、煮込み料理の製造方法。
［４］バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼ、及びサーモリシンを含有する、食肉の改質用組成物；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
［５］バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼと、サーモリシンとの含有量比（バチルス アミロリケファシエンス由来の中性プロテアーゼ：サーモリシン）が、酵素活性比にて１：０．０００００２～１：５００，０００である、［４］に記載の組成物。
［６］煮込み料理に用いる食肉の改質用である、［４］または［５］に記載の組成物。
［７］バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼおよび、サーモリシンが作用した食肉を含有する、食品；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度＝５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
［８］食肉１００ｇあたり、１０Ｕ以上５，０００，０００Ｕ以下のバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼと、１０Ｕ以上５，０００，０００Ｕ以下のサーモリシンが作用した食肉を含有する、［７］に記載の食品。
［９］煮込み料理として供される、［７］または［８］に記載の食品。 [1] A neutral protease derived from Bacillus amyloliquefaciens, which comprises contacting a neutral protease having the following characteristics (a) to (e) and a thermolysin with the meat. Modification method;
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
[2] The amount of neutral protease derived from Bacillus amyloliquefaciens to be brought into contact with meat is 10 U or more and 5,000,000 U or less per 100 g of meat, and the amount of thermolysin to be brought into contact with meat is 100 g of meat. The method according to [1], wherein the amount is 10 U or more and 5,000,000 U or less per unit.
[3] A method for producing a stewed dish, which comprises using the meat modified by the method according to [1] or [2].
[4] A composition for modifying meat, which is a neutral protease derived from Bacillus amyloliquefaciens and contains a neutral protease having the following characteristics (a) to (e) and thermolysin. ;
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
[5] The content ratio of the neutral protease derived from Bacillus amyloliquefaciens to thermolysin (neutral protease derived from Bacillus amyloliquefaciens: thermolysin) has an enzyme activity ratio of 1: 0. The composition according to [4], which is 000002-1: 500,000.
[6] The composition according to [4] or [5], which is used for modifying meat used in stewed dishes.
[7] A food product containing a neutral protease derived from Bacillus amyloliquefaciens, which has the following characteristics (a) to (e), and meat on which thermolysin acts.
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) Optimal temperature = 50 ° C to 55 ° C,
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
[8] Per 100 g of meat contains a neutral protease derived from Bacillus amyloliquefaciens of 10 U or more and 5,000,000 U or less and a meat on which thermolysin of 10 U or more and 5,000,000 U or less acts. The food product according to [7].
[9] The food according to [7] or [8], which is served as a stewed dish.
本発明により、短時間でかつ簡便に、食肉を軟質化し、食肉にほぐれやすさを付与することができる。
また本発明により、短時間でかつ簡便に、食肉を軟質化し、食肉にほぐれやすさを付与し得る食肉の改質用組成物を提供することができる。
さらに本発明により、軟質化され、ほぐれやすさが付与された食肉を含有する食品を提供することができる。 According to the present invention, the meat can be softened and the meat can be easily loosened in a short time and easily.
Further, according to the present invention, it is possible to provide a composition for modifying meat that can soften meat and impart easy loosening to the meat in a short time and easily.
Further, according to the present invention, it is possible to provide a food containing meat that has been softened and imparted with ease of loosening.
また本発明により、短時間でかつ簡便に、食肉を軟質化し、食肉にほぐれやすさを付与し得る食肉の改質用組成物を提供することができる。
さらに本発明により、軟質化され、ほぐれやすさが付与された食肉を含有する食品を提供することができる。 According to the present invention, the meat can be softened and the meat can be easily loosened in a short time and easily.
Further, according to the present invention, it is possible to provide a composition for modifying meat that can soften meat and impart easy loosening to the meat in a short time and easily.
Further, according to the present invention, it is possible to provide a food containing meat that has been softened and imparted with ease of loosening.
本発明は、食肉の改質方法（以下、本明細書にて「本発明の改質方法」ともいう）を提供する。
本明細書において、「食肉の改質」とは、食肉を軟質化し、食肉にほぐれやすさを付与することをいう。
本発明の改質方法は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを食肉に接触させることを含む。 The present invention provides a method for modifying meat (hereinafter, also referred to as "method for modifying the present invention" in the present specification).
As used herein, the term "meat reforming" refers to softening the meat and imparting ease of loosening to the meat.
The modification method of the present invention comprises contacting meat with a neutral protease from Bacillus amyloliquefaciens and thermolysin.
本明細書において、「食肉の改質」とは、食肉を軟質化し、食肉にほぐれやすさを付与することをいう。
本発明の改質方法は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを食肉に接触させることを含む。 The present invention provides a method for modifying meat (hereinafter, also referred to as "method for modifying the present invention" in the present specification).
As used herein, the term "meat reforming" refers to softening the meat and imparting ease of loosening to the meat.
The modification method of the present invention comprises contacting meat with a neutral protease from Bacillus amyloliquefaciens and thermolysin.
本発明の改質方法において用いられるバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼとしては、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）により産生されるプロテアーゼであって、下記（ａ）～（ｅ）の特性を有するものが用いられる。
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。 The neutral protease derived from Bacillus amyloliquefaciens used in the modification method of the present invention is a protease produced by Bacillus amyloliquefaciens, and the proteases produced by Bacillus amyloliquefaciens are described below (a) to (a). Those having the characteristics of e) are used.
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。 The neutral protease derived from Bacillus amyloliquefaciens used in the modification method of the present invention is a protease produced by Bacillus amyloliquefaciens, and the proteases produced by Bacillus amyloliquefaciens are described below (a) to (a). Those having the characteristics of e) are used.
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
上記中性プロテアーゼとしては、たとえば、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）を培養し、培養液を回収して濃縮、乾燥する、培養液よりイオン交換カラムクロマトグラフィーやゲル濾過カラムクロマトグラフィー等を用いて精製する等して製造したものや、遺伝子組み換え法により製造したものを用いることが可能であるが、市販の酵素製剤を用いることもできる。
市販の酵素製剤の例として、天野エンザイム株式会社より市販されている「プロチンＳＤ－ＮＹ１０」、「プロチンＮＹ１００」等が挙げられる。
上記中性プロテアーゼは、粉末状、粒状、顆粒状、液状、ペースト状等、種々の形態で用いることができる。
本発明において、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の上記中性プロテアーゼは、１種を単独で、または２種以上を組み合わせて用いることができる。 As the neutral protease, for example, Bacillus amyloliquefaciens is cultured, and the culture solution is collected, concentrated, and dried. Ion exchange column chromatography, gel filtration column chromatography, or the like is used from the culture solution. It is possible to use a product produced by purification by purification or a product produced by a gene recombination method, but a commercially available enzyme preparation can also be used.
Examples of commercially available enzyme preparations include "Protin SD-NY10" and "Protin NY100" commercially available from Amano Enzyme Co., Ltd.
The neutral protease can be used in various forms such as powder, granular, granular, liquid, and paste.
In the present invention, the above-mentioned neutral protease derived from Bacillus amyloliquefaciens can be used alone or in combination of two or more.
市販の酵素製剤の例として、天野エンザイム株式会社より市販されている「プロチンＳＤ－ＮＹ１０」、「プロチンＮＹ１００」等が挙げられる。
上記中性プロテアーゼは、粉末状、粒状、顆粒状、液状、ペースト状等、種々の形態で用いることができる。
本発明において、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の上記中性プロテアーゼは、１種を単独で、または２種以上を組み合わせて用いることができる。 As the neutral protease, for example, Bacillus amyloliquefaciens is cultured, and the culture solution is collected, concentrated, and dried. Ion exchange column chromatography, gel filtration column chromatography, or the like is used from the culture solution. It is possible to use a product produced by purification by purification or a product produced by a gene recombination method, but a commercially available enzyme preparation can also be used.
Examples of commercially available enzyme preparations include "Protin SD-NY10" and "Protin NY100" commercially available from Amano Enzyme Co., Ltd.
The neutral protease can be used in various forms such as powder, granular, granular, liquid, and paste.
In the present invention, the above-mentioned neutral protease derived from Bacillus amyloliquefaciens can be used alone or in combination of two or more.
本発明の改質方法において用いられるサーモリシンは、サーモライシン、テルモリシンなどとも呼ばれる耐熱性プロテアーゼで、バチルス サーモプロテオリティクス（Ｂａｃｉｌｌｕｓ ｔｈｅｒｍｏｐｒｏｔｅｏｌｙｔｉｃｕｓ）により生産される中性の金属プロテアーゼである。タンパク質中、疎水性アミノ酸を含むペプチド結合を選択的に加水分解する酵素であり、ＣＡＳ登録番号は９０７３－７８－３である。
本発明においては、上記特性を有するものであれば、ゲオバチルス ステアロサーモフィルス（Ｇｅｏｂａｃｉｌｌｕｓ ｓｔｅａｒｏｔｈｅｒｍｏｐｈｉｌｕｓ）等、他の基原の酵素を用いてもよく、アミノ酸置換等による改変酵素や組み換え酵素を用いることもできる。また、化学的合成法、発酵法、菌体からの抽出、精製による方法等のいずれの方法で製造されたものでもよく、市販の酵素製剤を利用することもできる。
市販の酵素製剤としては、富士フイルム和光純薬株式会社製「サーモリシン」、プロメガ株式会社製「サーモリシン」等が挙げられる。
上記サーモリシンは、粉末状、粒状、顆粒状、液状、ペースト状等、種々の形態で用いることができる。
本発明において、上記サーモリシンは、１種を単独で、または２種以上を組み合わせて用いることができる。 The thermolysin used in the modification method of the present invention is a thermolysin, which is also called thermolysin, thermolysin, etc., and is a neutral metalloproteinase produced by Bacillus thermoproteolitics. It is an enzyme that selectively hydrolyzes peptide bonds containing hydrophobic amino acids in proteins, and has a CAS Registry Number of 9073-78-3.
In the present invention, other basic enzymes such as Geobacillus stearothermophilus may be used as long as they have the above-mentioned characteristics, and a modifying enzyme or a recombinant enzyme by amino acid substitution or the like may be used. can. Further, it may be produced by any method such as a chemical synthesis method, a fermentation method, an extraction from bacterial cells, a method by purification, or the like, and a commercially available enzyme preparation can also be used.
Examples of commercially available enzyme preparations include "thermolysin" manufactured by Fujifilm Wako Pure Chemical Industries, Ltd. and "thermolysin" manufactured by Promega Co., Ltd.
The thermolysin can be used in various forms such as powder, granular, granular, liquid, and paste.
In the present invention, the thermolysin can be used alone or in combination of two or more.
本発明においては、上記特性を有するものであれば、ゲオバチルス ステアロサーモフィルス（Ｇｅｏｂａｃｉｌｌｕｓ ｓｔｅａｒｏｔｈｅｒｍｏｐｈｉｌｕｓ）等、他の基原の酵素を用いてもよく、アミノ酸置換等による改変酵素や組み換え酵素を用いることもできる。また、化学的合成法、発酵法、菌体からの抽出、精製による方法等のいずれの方法で製造されたものでもよく、市販の酵素製剤を利用することもできる。
市販の酵素製剤としては、富士フイルム和光純薬株式会社製「サーモリシン」、プロメガ株式会社製「サーモリシン」等が挙げられる。
上記サーモリシンは、粉末状、粒状、顆粒状、液状、ペースト状等、種々の形態で用いることができる。
本発明において、上記サーモリシンは、１種を単独で、または２種以上を組み合わせて用いることができる。 The thermolysin used in the modification method of the present invention is a thermolysin, which is also called thermolysin, thermolysin, etc., and is a neutral metalloproteinase produced by Bacillus thermoproteolitics. It is an enzyme that selectively hydrolyzes peptide bonds containing hydrophobic amino acids in proteins, and has a CAS Registry Number of 9073-78-3.
In the present invention, other basic enzymes such as Geobacillus stearothermophilus may be used as long as they have the above-mentioned characteristics, and a modifying enzyme or a recombinant enzyme by amino acid substitution or the like may be used. can. Further, it may be produced by any method such as a chemical synthesis method, a fermentation method, an extraction from bacterial cells, a method by purification, or the like, and a commercially available enzyme preparation can also be used.
Examples of commercially available enzyme preparations include "thermolysin" manufactured by Fujifilm Wako Pure Chemical Industries, Ltd. and "thermolysin" manufactured by Promega Co., Ltd.
The thermolysin can be used in various forms such as powder, granular, granular, liquid, and paste.
In the present invention, the thermolysin can be used alone or in combination of two or more.
本発明の改質方法において、「食肉」とは、食用として供される肉、すなわち家畜、家禽、その他野生動物由来の肉であって、食用として利用される肉を指す。
家畜としては、豚、牛、羊、山羊、馬、らくだ、ウサギ、アルパカ等が挙げられ、家禽としては、鶏、鳩、アヒル、鴨、鶉等が挙げられ、野生動物としては、鹿、猪、熊等が挙げられる。
また、本発明において、食肉としては、上記動物由来の種々の部位の肉が用いられ、適宜所望の大きさ、形状にカットして用いることができ、塊状のブロック肉、スライス肉（厚切り肉、薄切り肉）、こま切れ肉等が例示される。挽肉を加工したハンバーグやソーセージ等は本発明の「食肉」には含まれないが、カット肉、屑肉等の肉の小片を結着剤で固め、形状を整えて製造される成形肉等は、本発明の「食肉」に含まれる。
本発明の改質方法は、従来の軟質化処理法では短時間での軟質化が困難であった塊状の食肉や、硬い低品質な食肉の改質に好適であり、煮込み料理に用いられる塊状の食肉の改質に特に好適である。 In the reforming method of the present invention, "meat" refers to meat to be used for food, that is, meat derived from livestock, poultry, and other wild animals, which is used for food.
Livestock include pigs, cows, sheep, goats, horses, camels, rabbits, alpaca, etc. Poultry includes chickens, pigeons, ducks, ducks, sardines, etc., and wild animals include deer, boars, etc. , Bears, etc.
Further, in the present invention, as the meat, meat of various parts derived from the above animals is used, and it can be appropriately cut into a desired size and shape and used, and can be used as chunky block meat or sliced meat (thick sliced meat). , Thinly sliced meat), chopped meat and the like. Processed minced meat such as hamburger steak and sausage is not included in the "meat" of the present invention. , Included in the "meat" of the present invention.
The reforming method of the present invention is suitable for reforming chunky meat, which was difficult to soften in a short time by the conventional softening treatment method, and hard, low-quality meat, and is used for stewed dishes. It is particularly suitable for modifying meat.
家畜としては、豚、牛、羊、山羊、馬、らくだ、ウサギ、アルパカ等が挙げられ、家禽としては、鶏、鳩、アヒル、鴨、鶉等が挙げられ、野生動物としては、鹿、猪、熊等が挙げられる。
また、本発明において、食肉としては、上記動物由来の種々の部位の肉が用いられ、適宜所望の大きさ、形状にカットして用いることができ、塊状のブロック肉、スライス肉（厚切り肉、薄切り肉）、こま切れ肉等が例示される。挽肉を加工したハンバーグやソーセージ等は本発明の「食肉」には含まれないが、カット肉、屑肉等の肉の小片を結着剤で固め、形状を整えて製造される成形肉等は、本発明の「食肉」に含まれる。
本発明の改質方法は、従来の軟質化処理法では短時間での軟質化が困難であった塊状の食肉や、硬い低品質な食肉の改質に好適であり、煮込み料理に用いられる塊状の食肉の改質に特に好適である。 In the reforming method of the present invention, "meat" refers to meat to be used for food, that is, meat derived from livestock, poultry, and other wild animals, which is used for food.
Livestock include pigs, cows, sheep, goats, horses, camels, rabbits, alpaca, etc. Poultry includes chickens, pigeons, ducks, ducks, sardines, etc., and wild animals include deer, boars, etc. , Bears, etc.
Further, in the present invention, as the meat, meat of various parts derived from the above animals is used, and it can be appropriately cut into a desired size and shape and used, and can be used as chunky block meat or sliced meat (thick sliced meat). , Thinly sliced meat), chopped meat and the like. Processed minced meat such as hamburger steak and sausage is not included in the "meat" of the present invention. , Included in the "meat" of the present invention.
The reforming method of the present invention is suitable for reforming chunky meat, which was difficult to soften in a short time by the conventional softening treatment method, and hard, low-quality meat, and is used for stewed dishes. It is particularly suitable for modifying meat.
本発明の改質方法において、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンと食肉との接触の手段または態様は、前記中性プロテアーゼ及びサーモリシンが、一定の時間食肉に作用する状態で行うことができれば、特に制限されない。前記酵素を食肉と接触させる手段としては、粉末状または顆粒状の酵素を振りかける等、前記酵素を食肉に添加する、液状またはペースト状の前記酵素を食肉に塗布する、前記酵素の溶液または分散液もしくは懸濁液を食肉に注入する、前記酵素の溶液または分散液もしくは懸濁液に食肉を浸漬する等が挙げられる。簡便かつ短時間で食肉を改質するという観点からは、粉末状または顆粒状の酵素を振りかけて接触させる方法、あるいは酵素の溶液または分散液もしくは懸濁液に浸漬して接触させる方法が好ましい。
バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンは、後述する食肉の改質用組成物として調製された状態で接触させてもよい。
また、前記酵素と食肉との接触は、食肉の加工、調理の段階にて適宜行わせることができ、生の食肉に酵素を接触させた後に加熱調理をしてもよいし、加熱した食肉に酵素を接触させた後に再度加熱調理をしてもよいが、処理の簡便性の観点からは、生の食肉に接触させることが好ましい。 In the modification method of the present invention, the neutral protease derived from Bacillus amyloliquefaciens and the means or mode of contact between the thermolysin and the meat are such that the neutral protease and the thermolysin act on the meat for a certain period of time. There is no particular limitation as long as it can be done in the state of doing. As a means for bringing the enzyme into contact with the meat, the enzyme is added to the meat, such as by sprinkling the enzyme in the form of powder or granules, or the enzyme in the form of a liquid or paste is applied to the meat, or a solution or dispersion of the enzyme. Alternatively, the suspension may be injected into the meat, or the meat may be immersed in the enzyme solution or dispersion or suspension. From the viewpoint of modifying the meat easily and in a short time, a method of sprinkling a powdered or granular enzyme and contacting the meat, or a method of immersing the meat in a solution, dispersion or suspension of the enzyme and contacting the meat is preferable.
The neutral protease derived from Bacillus amyloliquefaciens and thermolysin may be contacted in a state of being prepared as a composition for modifying meat described later.
Further, the contact between the enzyme and the meat can be appropriately performed at the stage of processing and cooking the meat, and the raw meat may be brought into contact with the enzyme and then cooked, or the heated meat may be cooked. The meat may be cooked again after being brought into contact with the enzyme, but from the viewpoint of convenience of treatment, it is preferably brought into contact with raw meat.
バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンは、後述する食肉の改質用組成物として調製された状態で接触させてもよい。
また、前記酵素と食肉との接触は、食肉の加工、調理の段階にて適宜行わせることができ、生の食肉に酵素を接触させた後に加熱調理をしてもよいし、加熱した食肉に酵素を接触させた後に再度加熱調理をしてもよいが、処理の簡便性の観点からは、生の食肉に接触させることが好ましい。 In the modification method of the present invention, the neutral protease derived from Bacillus amyloliquefaciens and the means or mode of contact between the thermolysin and the meat are such that the neutral protease and the thermolysin act on the meat for a certain period of time. There is no particular limitation as long as it can be done in the state of doing. As a means for bringing the enzyme into contact with the meat, the enzyme is added to the meat, such as by sprinkling the enzyme in the form of powder or granules, or the enzyme in the form of a liquid or paste is applied to the meat, or a solution or dispersion of the enzyme. Alternatively, the suspension may be injected into the meat, or the meat may be immersed in the enzyme solution or dispersion or suspension. From the viewpoint of modifying the meat easily and in a short time, a method of sprinkling a powdered or granular enzyme and contacting the meat, or a method of immersing the meat in a solution, dispersion or suspension of the enzyme and contacting the meat is preferable.
The neutral protease derived from Bacillus amyloliquefaciens and thermolysin may be contacted in a state of being prepared as a composition for modifying meat described later.
Further, the contact between the enzyme and the meat can be appropriately performed at the stage of processing and cooking the meat, and the raw meat may be brought into contact with the enzyme and then cooked, or the heated meat may be cooked. The meat may be cooked again after being brought into contact with the enzyme, but from the viewpoint of convenience of treatment, it is preferably brought into contact with raw meat.
食肉にバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを接触させる順序は、特に制限されず、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンのいずれを先に接触させてもよく、両者を同時に接触させてもよいが、改質処理の簡便性の観点からは、両者を同時に接触させることが好ましい。
The order in which the neutral protease derived from Bacillus amyloliquefaciens and the thermolysin are brought into contact with the meat is not particularly limited, and the neutral protease derived from Bacillus amyloliquefaciens and the thermolysin derived from Bacillus amyloliquefaciens are not particularly limited. They may be brought into contact first or both may be brought into contact at the same time, but from the viewpoint of simplicity of the reforming treatment, it is preferable to bring them into contact at the same time.
本発明の改質方法において、食肉に接触させるバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼの量は、食肉１００ｇに対し１０ユニット（Ｕ）以上５，０００，０００Ｕ以下とすることが好ましく、５０Ｕ以上３，０００，０００Ｕ以下とすることがより好ましく、１００Ｕ以上１，０００，０００Ｕ以下とすることがさらに好ましい。
なお、上記中性プロテアーゼの酵素活性については、カゼインを基質として、１分間にチロシン１μｇに相当するフォリン試液呈色物質の増加をもたらす酵素量を、１Ｕと定義した。 In the modification method of the present invention, the amount of neutral protease derived from Bacillus amyloliquefaciens to be brought into contact with meat may be 10 units (U) or more and 5,000,000 U or less per 100 g of meat. It is preferably 50 U or more and 3,000,000 U or less, and more preferably 100 U or more and 1,000,000 U or less.
Regarding the enzymatic activity of the neutral protease, the amount of the enzyme that causes an increase in the folin test solution color-forming substance corresponding to 1 μg of tyrosine per minute using casein as a substrate was defined as 1U.
なお、上記中性プロテアーゼの酵素活性については、カゼインを基質として、１分間にチロシン１μｇに相当するフォリン試液呈色物質の増加をもたらす酵素量を、１Ｕと定義した。 In the modification method of the present invention, the amount of neutral protease derived from Bacillus amyloliquefaciens to be brought into contact with meat may be 10 units (U) or more and 5,000,000 U or less per 100 g of meat. It is preferably 50 U or more and 3,000,000 U or less, and more preferably 100 U or more and 1,000,000 U or less.
Regarding the enzymatic activity of the neutral protease, the amount of the enzyme that causes an increase in the folin test solution color-forming substance corresponding to 1 μg of tyrosine per minute using casein as a substrate was defined as 1U.
本発明の改質方法において、食肉に接触させるサーモリシンの量は、食肉１００ｇに対し１０Ｕ以上５，０００，０００Ｕ以下とすることが好ましく、５０Ｕ以上３，０００，０００Ｕ以下とすることがより好ましく、１００Ｕ以上１，０００，０００Ｕ以下とすることがさらに好ましい。
なお、サーモリシンの酵素活性については、上記中性プロテアーゼと同様の定義とした。
また、後述する試験例において用いられる各種プロテアーゼの酵素活性についても、全て上記中性プロテアーゼと同様の定義とした。 In the modification method of the present invention, the amount of thermolysin to be brought into contact with meat is preferably 10 U or more and 5,000,000 U or less, and more preferably 50 U or more and 3,000,000 U or less with respect to 100 g of meat. It is more preferably 100 U or more and 1,000,000 U or less.
The enzyme activity of thermolysin was defined in the same way as the above-mentioned neutral protease.
In addition, the enzymatic activities of various proteases used in the test examples described later are all defined in the same manner as the above-mentioned neutral proteases.
なお、サーモリシンの酵素活性については、上記中性プロテアーゼと同様の定義とした。
また、後述する試験例において用いられる各種プロテアーゼの酵素活性についても、全て上記中性プロテアーゼと同様の定義とした。 In the modification method of the present invention, the amount of thermolysin to be brought into contact with meat is preferably 10 U or more and 5,000,000 U or less, and more preferably 50 U or more and 3,000,000 U or less with respect to 100 g of meat. It is more preferably 100 U or more and 1,000,000 U or less.
The enzyme activity of thermolysin was defined in the same way as the above-mentioned neutral protease.
In addition, the enzymatic activities of various proteases used in the test examples described later are all defined in the same manner as the above-mentioned neutral proteases.
本発明の改質方法において、食肉にバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを接触させる際の、前記中性プロテアーゼ量とサーモリシン量の比（前記中性プロテアーゼ量：サーモリシン量）は、これら酵素の活性比にて１：０．０００００２～１：５００，０００とすることが好ましく、１：０．００００２～１：５０，０００とすることがより好ましく、１：０．０００１～１：１０，０００とすることがさらに好ましい。
In the modification method of the present invention, the ratio of the neutral protease amount to the thermolysin amount when contacting the meat with a neutral protease derived from Bacillus amyloliquefaciens and thermolysin (the neutral protease amount: The amount of thermolysin) is preferably 1: 0.000002 to 1: 500,000, more preferably 1: 0.00002 to 1: 50,000, and 1: 0 in terms of the activity ratio of these enzymes. It is more preferably .0001 to 1: 10,000.
本発明の改質方法において、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを食肉に接触させる時間は、中性プロテアーゼ及びサーモリシンが基質物質、すなわち食肉に含まれるタンパク質に十分に作用し得る時間であれば特に制限されないが、具体的には１分～２４時間（例えば、１分～１２時間、１分～８時間、１分～４時間、１分～２時間、１分～１時間等）とすることが好ましい。
とりわけ本発明においては、中性プロテアーゼとサーモリシンを併用することにより、短時間でかつ簡便に、食肉を軟質化して食肉にほぐれやすさを付与し得ることが特徴であり、１分～１時間程度の短い作用時間とすることにより、本発明の利用価値がより向上する。
また、本発明の改質方法において、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを食肉に接触させる際の温度に関しても、前記中性プロテアーゼ及びサーモリシンが活性を保つ範囲であれば特に制限されないが、具体的には０℃～８０℃（例えば、０℃～７５℃、０℃～７０℃、０℃～６５℃、０℃～６０℃、０℃～５５℃、０℃～５０℃、０℃～４５℃等）とすることが好ましい。
すなわち、通常の食肉の加工・調理の条件下において前記酵素を接触させることで、十分にこれらを作用させることができる。 In the modification method of the present invention, the neutral protease derived from Bacillus amyloliquefaciens and the time for contacting the thermolysin with the meat are sufficient for the neutral protease and the thermolysin to be a substrate substance, that is, a protein contained in the meat. The time is not particularly limited as long as it can act on, but specifically, 1 minute to 24 hours (for example, 1 minute to 12 hours, 1 minute to 8 hours, 1 minute to 4 hours, 1 minute to 2 hours, 1 minute to 2 hours). Minutes to 1 hour, etc.) is preferable.
In particular, the present invention is characterized in that by using a neutral protease and thermolysin in combination, the meat can be softened and easily loosened in a short time and easily, and the meat can be easily loosened for about 1 minute to 1 hour. The utility value of the present invention is further improved by setting the action time to be short.
Further, in the modification method of the present invention, the neutral protease derived from Bacillus amyloliquefaciens and the temperature at which the thermolysin is brought into contact with the meat are also within the range in which the neutral protease and the thermolysin maintain their activities. If there is, it is not particularly limited, but specifically, it is 0 ° C to 80 ° C (for example, 0 ° C to 75 ° C, 0 ° C to 70 ° C, 0 ° C to 65 ° C, 0 ° C to 60 ° C, 0 ° C to 55 ° C, 0). ° C. to 50 ° C., 0 ° C. to 45 ° C., etc.) is preferable.
That is, they can be sufficiently allowed to act by contacting the enzymes under normal meat processing / cooking conditions.
とりわけ本発明においては、中性プロテアーゼとサーモリシンを併用することにより、短時間でかつ簡便に、食肉を軟質化して食肉にほぐれやすさを付与し得ることが特徴であり、１分～１時間程度の短い作用時間とすることにより、本発明の利用価値がより向上する。
また、本発明の改質方法において、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを食肉に接触させる際の温度に関しても、前記中性プロテアーゼ及びサーモリシンが活性を保つ範囲であれば特に制限されないが、具体的には０℃～８０℃（例えば、０℃～７５℃、０℃～７０℃、０℃～６５℃、０℃～６０℃、０℃～５５℃、０℃～５０℃、０℃～４５℃等）とすることが好ましい。
すなわち、通常の食肉の加工・調理の条件下において前記酵素を接触させることで、十分にこれらを作用させることができる。 In the modification method of the present invention, the neutral protease derived from Bacillus amyloliquefaciens and the time for contacting the thermolysin with the meat are sufficient for the neutral protease and the thermolysin to be a substrate substance, that is, a protein contained in the meat. The time is not particularly limited as long as it can act on, but specifically, 1 minute to 24 hours (for example, 1 minute to 12 hours, 1 minute to 8 hours, 1 minute to 4 hours, 1 minute to 2 hours, 1 minute to 2 hours). Minutes to 1 hour, etc.) is preferable.
In particular, the present invention is characterized in that by using a neutral protease and thermolysin in combination, the meat can be softened and easily loosened in a short time and easily, and the meat can be easily loosened for about 1 minute to 1 hour. The utility value of the present invention is further improved by setting the action time to be short.
Further, in the modification method of the present invention, the neutral protease derived from Bacillus amyloliquefaciens and the temperature at which the thermolysin is brought into contact with the meat are also within the range in which the neutral protease and the thermolysin maintain their activities. If there is, it is not particularly limited, but specifically, it is 0 ° C to 80 ° C (for example, 0 ° C to 75 ° C, 0 ° C to 70 ° C, 0 ° C to 65 ° C, 0 ° C to 60 ° C, 0 ° C to 55 ° C, 0). ° C. to 50 ° C., 0 ° C. to 45 ° C., etc.) is preferable.
That is, they can be sufficiently allowed to act by contacting the enzymes under normal meat processing / cooking conditions.
本発明の改質方法により、短時間でかつ簡便に、食肉を軟質化し、食肉にほぐれやすさを付与することができる。
従って、本発明の改質方法は、食肉を用いた煮込み料理、特に塊状の食肉を用いた煮込み料理において効果を発揮する。 According to the modification method of the present invention, the meat can be softened and the meat can be easily loosened in a short time and easily.
Therefore, the modification method of the present invention is effective in a stewed dish using meat, particularly a stewed dish using chunky meat.
従って、本発明の改質方法は、食肉を用いた煮込み料理、特に塊状の食肉を用いた煮込み料理において効果を発揮する。 According to the modification method of the present invention, the meat can be softened and the meat can be easily loosened in a short time and easily.
Therefore, the modification method of the present invention is effective in a stewed dish using meat, particularly a stewed dish using chunky meat.
それゆえ、本発明は、上記本発明の改質方法により改質された食肉を用いることを含む、煮込み料理の製造方法をも提供する。
なお、本明細書において「煮込み料理」とは、食肉、特に塊状の食肉が液体に浸漬された状態で加熱調理される料理を指し、例えば、シチュー、カレー、ポトフ、角煮、鍋料理、味噌汁、スープ等が挙げられる。
本発明の煮込み料理の製造方法は、煮込み料理の製造において採用される通常の工程、たとえば他の具材の細切、下茹で等の処理工程、上記本発明の改質方法により改質された食肉と他の具材とを混合する工程、前記食肉および他の具材を加熱調理する（炒める、焼く、煮る、蒸す等）工程、調味工程等をさらに含むことができる。
また、本発明の煮込み料理の製造方法には、製造された煮込み料理を冷却し、冷蔵、冷凍等して保存する工程、高圧加熱殺菌等の殺菌処理工程、缶、びん、レトルトパウチ等の容器に充填、包装する工程等も含まれ得る。
上記した各工程は、食品の製造に際して通常実施される条件にて、一般的な方法で行われる。 Therefore, the present invention also provides a method for producing a stewed dish, which comprises using the meat modified by the modification method of the present invention.
In the present specification, the “simmered dish” refers to a dish in which meat, particularly chunky meat, is cooked in a state of being immersed in a liquid. , Soup, etc.
The method for producing a stewed dish of the present invention is modified by a normal process adopted in the production of a stewed dish, for example, a processing step such as shredding other ingredients, boiling underneath, or the above-mentioned modification method of the present invention. Further, a step of mixing the meat and other ingredients, a step of cooking the meat and other ingredients by heating (frying, baking, boiling, steaming, etc.), a seasoning step, and the like can be further included.
Further, the method for producing a stewed dish of the present invention includes a step of cooling the manufactured stewed dish and storing it by refrigerating, freezing, etc., a sterilization process such as high-pressure heat sterilization, and a container such as a can, a bottle, and a retort pouch. The process of filling and packaging can also be included.
Each of the above steps is carried out by a general method under the conditions normally carried out in the production of food.
なお、本明細書において「煮込み料理」とは、食肉、特に塊状の食肉が液体に浸漬された状態で加熱調理される料理を指し、例えば、シチュー、カレー、ポトフ、角煮、鍋料理、味噌汁、スープ等が挙げられる。
本発明の煮込み料理の製造方法は、煮込み料理の製造において採用される通常の工程、たとえば他の具材の細切、下茹で等の処理工程、上記本発明の改質方法により改質された食肉と他の具材とを混合する工程、前記食肉および他の具材を加熱調理する（炒める、焼く、煮る、蒸す等）工程、調味工程等をさらに含むことができる。
また、本発明の煮込み料理の製造方法には、製造された煮込み料理を冷却し、冷蔵、冷凍等して保存する工程、高圧加熱殺菌等の殺菌処理工程、缶、びん、レトルトパウチ等の容器に充填、包装する工程等も含まれ得る。
上記した各工程は、食品の製造に際して通常実施される条件にて、一般的な方法で行われる。 Therefore, the present invention also provides a method for producing a stewed dish, which comprises using the meat modified by the modification method of the present invention.
In the present specification, the “simmered dish” refers to a dish in which meat, particularly chunky meat, is cooked in a state of being immersed in a liquid. , Soup, etc.
The method for producing a stewed dish of the present invention is modified by a normal process adopted in the production of a stewed dish, for example, a processing step such as shredding other ingredients, boiling underneath, or the above-mentioned modification method of the present invention. Further, a step of mixing the meat and other ingredients, a step of cooking the meat and other ingredients by heating (frying, baking, boiling, steaming, etc.), a seasoning step, and the like can be further included.
Further, the method for producing a stewed dish of the present invention includes a step of cooling the manufactured stewed dish and storing it by refrigerating, freezing, etc., a sterilization process such as high-pressure heat sterilization, and a container such as a can, a bottle, and a retort pouch. The process of filling and packaging can also be included.
Each of the above steps is carried out by a general method under the conditions normally carried out in the production of food.
本発明はまた、食肉の改質用組成物（以下、本明細書にて「本発明の組成物」ともいう）を提供する。
本明細書において、「食肉の改質用組成物」とは、食肉を軟質化し、食肉にほぐれやすさを付与するために用いられる組成物をいう。
本発明の組成物は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを含有する。
本発明の組成物に含有される前記中性プロテアーゼおよびサーモリシンについては、本発明の改質方法において上記した通りであり、本発明の組成物におけるそれらの各含有量は、本発明の改質方法において、食肉１００ｇに対し、上記した量の前記中性プロテアーゼおよびサーモリシンを、接触させ得る量として設定される。
また、本発明の組成物における前記中性プロテアーゼとサーモリシンの含有量比（前記中性プロテアーゼ：サーモリシン）は、これらの酵素活性（Ｕ）比にして、好ましくは１：０．０００００２～１：５００，０００であり、より好ましくは１：０．００００２～１：５０，０００であり、さらに好ましくは１：０．０００１～１：１０，０００である。 The present invention also provides a composition for modifying meat (hereinafter, also referred to as “composition of the present invention” in the present specification).
As used herein, the term "meat reforming composition" refers to a composition used to soften meat and impart ease of loosening to the meat.
The composition of the present invention contains a neutral protease derived from Bacillus amyloliquefaciens, and a thermolysin.
The neutral protease and thermolysin contained in the composition of the present invention are as described above in the modification method of the present invention, and each content thereof in the composition of the present invention is the modification method of the present invention. In 100 g of meat, the above-mentioned amounts of the neutral protease and thermolysin are set as an amount that can be brought into contact with each other.
The content ratio of the neutral protease to the thermolysin in the composition of the present invention (the neutral protease: thermolysin) is preferably 1: 0.000002 to 1: 500 in terms of the enzyme activity (U) ratio. It is 000, more preferably 1: 0.00002 to 1: 50,000, and even more preferably 1: 0.0001 to 1: 10,000.
本明細書において、「食肉の改質用組成物」とは、食肉を軟質化し、食肉にほぐれやすさを付与するために用いられる組成物をいう。
本発明の組成物は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを含有する。
本発明の組成物に含有される前記中性プロテアーゼおよびサーモリシンについては、本発明の改質方法において上記した通りであり、本発明の組成物におけるそれらの各含有量は、本発明の改質方法において、食肉１００ｇに対し、上記した量の前記中性プロテアーゼおよびサーモリシンを、接触させ得る量として設定される。
また、本発明の組成物における前記中性プロテアーゼとサーモリシンの含有量比（前記中性プロテアーゼ：サーモリシン）は、これらの酵素活性（Ｕ）比にして、好ましくは１：０．０００００２～１：５００，０００であり、より好ましくは１：０．００００２～１：５０，０００であり、さらに好ましくは１：０．０００１～１：１０，０００である。 The present invention also provides a composition for modifying meat (hereinafter, also referred to as “composition of the present invention” in the present specification).
As used herein, the term "meat reforming composition" refers to a composition used to soften meat and impart ease of loosening to the meat.
The composition of the present invention contains a neutral protease derived from Bacillus amyloliquefaciens, and a thermolysin.
The neutral protease and thermolysin contained in the composition of the present invention are as described above in the modification method of the present invention, and each content thereof in the composition of the present invention is the modification method of the present invention. In 100 g of meat, the above-mentioned amounts of the neutral protease and thermolysin are set as an amount that can be brought into contact with each other.
The content ratio of the neutral protease to the thermolysin in the composition of the present invention (the neutral protease: thermolysin) is preferably 1: 0.000002 to 1: 500 in terms of the enzyme activity (U) ratio. It is 000, more preferably 1: 0.00002 to 1: 50,000, and even more preferably 1: 0.0001 to 1: 10,000.
本発明の組成物は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼと、サーモリシンとを混合し、必要に応じて、一般的な食品添加物を添加して、粉末状、粒状、顆粒状、タブレット状等の固形状；ゲル状、ペースト状、クリーム状等の半固形状；溶液状、懸濁液状、分散液状等の液状等、種々の形態とすることができる。
また、粉末状、粒状、顆粒状等の固形状の形態で提供され、使用時に水等の溶媒に溶解しまたは分散もしくは懸濁して用いられる用時調製タイプの組成物とすることもできる。
上記食品添加物としては、水、多価アルコール等の溶剤；ポリエチレングリコール等の基剤；デキストリン、乳糖等の賦形剤；ゼラチン、アルファー化デンプン、部分アルファー化デンプン、セルロースおよびその誘導体（結晶セルロース、ヒドロキシプロピルセルロース等）等の結合剤；クロスポビドン、ポビドン、結晶セルロース等の崩壊剤；タルク、ステアリン酸マグネシウム等の滑沢剤；グリセリン脂肪酸エステル、ショ糖脂肪酸エステル、ポリグリセリン脂肪酸エステル、サポニン、レシチン、カゼインナトリウム等の乳化剤；食塩、アミノ酸およびその塩、核酸およびその塩、酵母エキス、畜肉エキス等の調味料；植物タンパク、グルテン、卵白、ゼラチン、カゼイン等のタンパク質；タンパク加水分解物；タンパク部分分解物；有機酸塩（フマル酸塩等）等のｐＨ調整剤；重合リン酸塩等のキレート剤；グルタチオン、システイン等の還元剤；アルギン酸、キサンタンガム等の増粘多糖類；かんすい；色素；酸味料；香料等が挙げられる。 The composition of the present invention is a mixture of a neutral protease derived from Bacillus amyloliquefaciens and thermolysin, and if necessary, general food additives are added to make a powdery, granular, granular composition. It can be in various forms such as solids such as granules and tablets; semi-solids such as gels, pastes and creams; liquids such as solutions, suspensions and dispersed liquids.
Further, it can be provided in a solid form such as powder, granule, or granule, and can be used as a time-prepared type composition which is dissolved, dispersed or suspended in a solvent such as water at the time of use.
Examples of the food additive include water, solvents such as polyhydric alcohol; bases such as polyethylene glycol; excipients such as dextrin and lactose; gelatin, pregelatinized starch, partially pregelatinized starch, cellulose and derivatives thereof (crystalline cellulose). , Hydroxypropyl cellulose, etc.); Disintegrants such as crospovidone, povidone, crystalline cellulose, etc .; Starch, magnesium stearate, etc. Emulsifiers such as lecithin, sodium caseinate; seasonings such as salt, amino acids and salts thereof, nucleic acids and salts thereof, yeast extract, livestock meat extract; proteins such as plant protein, gluten, egg white, gelatin, casein; protein hydrolyzate; protein Partial decomposition products; pH adjusters such as organic acid salts (fumarate etc.); chelating agents such as polymerized phosphates; reducing agents such as glutathione and cysteine; thickening polysaccharides such as alginic acid and xanthan gum; casein; pigments; Acidulants; fragrances and the like can be mentioned.
また、粉末状、粒状、顆粒状等の固形状の形態で提供され、使用時に水等の溶媒に溶解しまたは分散もしくは懸濁して用いられる用時調製タイプの組成物とすることもできる。
上記食品添加物としては、水、多価アルコール等の溶剤；ポリエチレングリコール等の基剤；デキストリン、乳糖等の賦形剤；ゼラチン、アルファー化デンプン、部分アルファー化デンプン、セルロースおよびその誘導体（結晶セルロース、ヒドロキシプロピルセルロース等）等の結合剤；クロスポビドン、ポビドン、結晶セルロース等の崩壊剤；タルク、ステアリン酸マグネシウム等の滑沢剤；グリセリン脂肪酸エステル、ショ糖脂肪酸エステル、ポリグリセリン脂肪酸エステル、サポニン、レシチン、カゼインナトリウム等の乳化剤；食塩、アミノ酸およびその塩、核酸およびその塩、酵母エキス、畜肉エキス等の調味料；植物タンパク、グルテン、卵白、ゼラチン、カゼイン等のタンパク質；タンパク加水分解物；タンパク部分分解物；有機酸塩（フマル酸塩等）等のｐＨ調整剤；重合リン酸塩等のキレート剤；グルタチオン、システイン等の還元剤；アルギン酸、キサンタンガム等の増粘多糖類；かんすい；色素；酸味料；香料等が挙げられる。 The composition of the present invention is a mixture of a neutral protease derived from Bacillus amyloliquefaciens and thermolysin, and if necessary, general food additives are added to make a powdery, granular, granular composition. It can be in various forms such as solids such as granules and tablets; semi-solids such as gels, pastes and creams; liquids such as solutions, suspensions and dispersed liquids.
Further, it can be provided in a solid form such as powder, granule, or granule, and can be used as a time-prepared type composition which is dissolved, dispersed or suspended in a solvent such as water at the time of use.
Examples of the food additive include water, solvents such as polyhydric alcohol; bases such as polyethylene glycol; excipients such as dextrin and lactose; gelatin, pregelatinized starch, partially pregelatinized starch, cellulose and derivatives thereof (crystalline cellulose). , Hydroxypropyl cellulose, etc.); Disintegrants such as crospovidone, povidone, crystalline cellulose, etc .; Starch, magnesium stearate, etc. Emulsifiers such as lecithin, sodium caseinate; seasonings such as salt, amino acids and salts thereof, nucleic acids and salts thereof, yeast extract, livestock meat extract; proteins such as plant protein, gluten, egg white, gelatin, casein; protein hydrolyzate; protein Partial decomposition products; pH adjusters such as organic acid salts (fumarate etc.); chelating agents such as polymerized phosphates; reducing agents such as glutathione and cysteine; thickening polysaccharides such as alginic acid and xanthan gum; casein; pigments; Acidulants; fragrances and the like can be mentioned.
本発明の改質用組成物は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンに、必要に応じて上記した食品添加物を添加して、製剤の分野で周知の製剤化手段、たとえば第十七改正日本薬局方製剤総則［３］製剤各条に記載された方法等に準じて、製造することができる。
The modifying composition of the present invention is a formulation well known in the field of formulation by adding the above-mentioned food additives to the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, if necessary. It can be manufactured according to the means for making it, for example, the method described in each article of the 17th revised Japanese Pharmacopoeia General Regulations for Pharmaceuticals [3] Pharmaceuticals.
本発明の組成物は、本発明の改質方法において上記した手段または態様にて、食肉に接触させて、食肉の改質に用いられる。
本発明の組成物は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンの各接触量が、それぞれ食肉１００ｇに対し、本発明の改質方法において上記した量となるように、用いられる。 The composition of the present invention is used for modifying meat by contacting it with meat by the means or embodiment described above in the modification method of the present invention.
In the composition of the present invention, the contact amount of the neutral protease derived from Bacillus amyloliquefaciens and the thermolysin is the same amount as described above in the modification method of the present invention with respect to 100 g of meat, respectively. , Used.
本発明の組成物は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンの各接触量が、それぞれ食肉１００ｇに対し、本発明の改質方法において上記した量となるように、用いられる。 The composition of the present invention is used for modifying meat by contacting it with meat by the means or embodiment described above in the modification method of the present invention.
In the composition of the present invention, the contact amount of the neutral protease derived from Bacillus amyloliquefaciens and the thermolysin is the same amount as described above in the modification method of the present invention with respect to 100 g of meat, respectively. , Used.
本発明の組成物は、短時間でかつ簡便に食肉を軟質化して、食肉にほぐれやすさを付与することができる。
従って、本発明の組成物は、食肉、特に塊状の食肉を用いた煮込み料理において、好適に用いられる。 The composition of the present invention can easily soften the meat in a short time and impart the meat to be easily loosened.
Therefore, the composition of the present invention is preferably used in meat, especially stewed dishes using chunky meat.
従って、本発明の組成物は、食肉、特に塊状の食肉を用いた煮込み料理において、好適に用いられる。 The composition of the present invention can easily soften the meat in a short time and impart the meat to be easily loosened.
Therefore, the composition of the present invention is preferably used in meat, especially stewed dishes using chunky meat.
さらに本発明は、軟質化され、ほぐれやすさが付与された食肉を含有する食品（以下、本明細書にて「本発明の食品」とも称する）を提供する。
本発明の食品は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンが作用した食肉を含有する。
「バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンが作用した食肉」とは、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを接触させて改質された食肉をいう。
本発明の食品に含有される食肉の調製に用いられるバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、およびサーモリシンについては、本発明の改質方法において上記した通りである。
また、本発明の食品に含有される食肉の調製に際し、食肉に接触させる前記中性プロテアーゼおよびサーモリシンの量（食肉１００ｇに対する接触量）、前記中性プロテアーゼおよびサーモリシンを接触させる方法、接触させる時間、接触させる際の温度についても、本発明の改質方法において上記した通りである。 Furthermore, the present invention provides a food containing meat that has been softened and imparted with ease of loosening (hereinafter, also referred to as "food of the present invention" in the present specification).
The food product of the present invention contains a neutral protease derived from Bacillus amyloliquefaciens, and meat on which thermolysin is acted.
"Neutral protease derived from Bacillus amyloliquefaciens and meat on which thermolysin acts" refers to neutral protease derived from Bacillus amyloliquefaciens and modified contact with thermolysin. Meat.
The neutral protease derived from Bacillus amyloliquefaciens used for preparing the meat contained in the food of the present invention and the thermolysin are as described above in the modification method of the present invention.
Further, in the preparation of the meat contained in the food of the present invention, the amount of the neutral protease and thermolysin to be brought into contact with the meat (contact amount with respect to 100 g of meat), the method of contacting the neutral protease and the thermolysin, the contact time, The temperature at the time of contact is also the same as described above in the reforming method of the present invention.
本発明の食品は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンが作用した食肉を含有する。
「バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンが作用した食肉」とは、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンを接触させて改質された食肉をいう。
本発明の食品に含有される食肉の調製に用いられるバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、およびサーモリシンについては、本発明の改質方法において上記した通りである。
また、本発明の食品に含有される食肉の調製に際し、食肉に接触させる前記中性プロテアーゼおよびサーモリシンの量（食肉１００ｇに対する接触量）、前記中性プロテアーゼおよびサーモリシンを接触させる方法、接触させる時間、接触させる際の温度についても、本発明の改質方法において上記した通りである。 Furthermore, the present invention provides a food containing meat that has been softened and imparted with ease of loosening (hereinafter, also referred to as "food of the present invention" in the present specification).
The food product of the present invention contains a neutral protease derived from Bacillus amyloliquefaciens, and meat on which thermolysin is acted.
"Neutral protease derived from Bacillus amyloliquefaciens and meat on which thermolysin acts" refers to neutral protease derived from Bacillus amyloliquefaciens and modified contact with thermolysin. Meat.
The neutral protease derived from Bacillus amyloliquefaciens used for preparing the meat contained in the food of the present invention and the thermolysin are as described above in the modification method of the present invention.
Further, in the preparation of the meat contained in the food of the present invention, the amount of the neutral protease and thermolysin to be brought into contact with the meat (contact amount with respect to 100 g of meat), the method of contacting the neutral protease and the thermolysin, the contact time, The temperature at the time of contact is also the same as described above in the reforming method of the present invention.
本発明の食品は、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、及びサーモリシンが作用した食肉を用いて、食品の分野で通常用いられる調理手段により調製することができ、たとえば、前記食肉に、必要に応じて他の具材や保存料、調味料等の食品添加物を添加し、炒める、焼く、煮る、蒸す等の通常の加熱調理手段により調製され、たとえば、水煮、蒸し肉等、食材としてさらに調理に供される加工食品や、シチュー、カレー、ポトフ、角煮、ローストビーフ、ローストポーク、ローストチキン、焼き肉、焼き豚、焼き鳥、ステーキ、カツ等、惣菜として利用される加工食品として提供され得る。
また、本発明の食品の保存形態は特に制限されず、常温保存、冷蔵保存、冷凍保存等、種々の形態で保存することができ、レトルトパウチ食品として調製されてもよい。 The food product of the present invention can be prepared by a cooking means commonly used in the field of food products, using a neutral protease derived from Bacillus amyloliquefaciens and meat on which thermolysin is acted, for example, as described above. If necessary, other ingredients and food additives such as preservatives and seasonings are added to the meat, and the meat is prepared by ordinary cooking means such as roasting, baking, boiling, and steaming. Processed foods such as meat that are further used for cooking as ingredients, and processed foods such as stew, curry, potov, boiled, roast beef, roast pork, roast chicken, roasted meat, roasted pork, roasted chicken, steak, cutlet, etc. Can be provided as.
Further, the storage form of the food of the present invention is not particularly limited, and it can be stored in various forms such as room temperature storage, refrigerated storage, and frozen storage, and may be prepared as a retort pouch food.
また、本発明の食品の保存形態は特に制限されず、常温保存、冷蔵保存、冷凍保存等、種々の形態で保存することができ、レトルトパウチ食品として調製されてもよい。 The food product of the present invention can be prepared by a cooking means commonly used in the field of food products, using a neutral protease derived from Bacillus amyloliquefaciens and meat on which thermolysin is acted, for example, as described above. If necessary, other ingredients and food additives such as preservatives and seasonings are added to the meat, and the meat is prepared by ordinary cooking means such as roasting, baking, boiling, and steaming. Processed foods such as meat that are further used for cooking as ingredients, and processed foods such as stew, curry, potov, boiled, roast beef, roast pork, roast chicken, roasted meat, roasted pork, roasted chicken, steak, cutlet, etc. Can be provided as.
Further, the storage form of the food of the present invention is not particularly limited, and it can be stored in various forms such as room temperature storage, refrigerated storage, and frozen storage, and may be prepared as a retort pouch food.
本発明の食品は、軟質化され、ほぐれやすさが付与された食肉を含有し、良好な食感を呈する。
従って、本発明の食品は、塊状の食肉を用いた煮込み料理として特に好適に提供され得る。 The food product of the present invention contains meat that has been softened and imparted with ease of loosening, and exhibits a good texture.
Therefore, the food product of the present invention can be particularly preferably provided as a stewed dish using chunky meat.
従って、本発明の食品は、塊状の食肉を用いた煮込み料理として特に好適に提供され得る。 The food product of the present invention contains meat that has been softened and imparted with ease of loosening, and exhibits a good texture.
Therefore, the food product of the present invention can be particularly preferably provided as a stewed dish using chunky meat.
以下に試験例および実施例を挙げ、本発明をさらに詳しく説明する。本発明は、これら実施例等により何ら限定されない。
The present invention will be described in more detail with reference to test examples and examples below. The present invention is not limited to these examples and the like.
［試験例１］
各種プロテアーゼについて、食肉に対するやわらかさ付与効果およびほぐれやすさ付与効果を、下記の通り検討した。 [Test Example 1]
For various proteases, the effect of imparting softness and the effect of imparting looseness to meat were examined as follows.
各種プロテアーゼについて、食肉に対するやわらかさ付与効果およびほぐれやすさ付与効果を、下記の通り検討した。 [Test Example 1]
For various proteases, the effect of imparting softness and the effect of imparting looseness to meat were examined as follows.
（ｉ）国産若鶏ムネ肉を２０ｇの小片にカットしてビニール製の袋に入れ、表１に示す各試験区のプロテアーゼを加えて振り混ぜた。なお、表１中、「－」は、プロテアーゼを添加しないことを示す。
バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼとしては、次の(ａ)～(ｅ)の特性を有するものを用いた。
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
鶏ムネ肉２０ｇに対する各プロテアーゼの添加量は、いずれも１０，０００Ｕ（鶏ムネ肉１００ｇあたり５０，０００Ｕ）とした。２０℃にて１５分間静置して各プロテアーゼを鶏ムネ肉に接触させた。
（ｉｉ）次いで、各プロテアーゼを接触させた鶏ムネ肉を用いて、鍋にて下記の通り調理し、クリームシチューを調製した。
＜クリームシチューの調製方法＞
鶏ムネ肉の表面を軽く炒めた後に水を入れ、沸騰後１５分間煮込んだ後に市販のルウ（ハウス食品株式会社製）を溶かし、５分間煮込み、次いで牛乳を加えて更に５分間煮込んだ。合計の煮込み時間は２５分間であった。水、牛乳、ルウの割合は、市販のルウ推奨の割合（水：牛乳：ルウ＝７００：１００：９０（重量比））とし、野菜を使用しない鶏ムネ肉のみのモデル系とした。
（ｉｉｉ）調製された鶏ムネ肉のクリームシチューから鶏ムネ肉を取り出し、官能評価に供した。
官能評価は、やわらかさとほぐれやすさについて、無添加区（試験区１）を０点とした－５点～５点の評点法にて、４名のパネルにて行った。評点は、小数点以下第一位までとし、４名の平均値を四捨五入した小数点以下第一位までの数値にて評価結果を示した。
なお、本官能評価において、「やわらかさ」とは、肉全体を噛み潰すために必要な咀嚼力を意味し、「ほぐれやすさ」とは、肉を噛み潰す時に白身魚のようにホロホロとほぐれる感覚を意味する。ほぐれやすさは、煮込み料理において特に好まれる食感で、一般的には長時間煮込むことで向上する。また、ほぐれやすさは、肉の筋原線維がある程度の束として崩れることで感じられる食感であり、肉全体が硬いと筋原線維が崩れないため、ほぐれやすさは感じられにくく、逆に肉全体が均一に潰れるレバーやフォアグラのようなやわらかい食感の場合も、ほぐれやすさはあまり感じられない。
やわらかさおよびほぐれやすさの評点については、無添加区との比較により、下記の通り定義した。
＜やわらかさの評点＞
－５点：極めて顕著に硬い
－４点：非常に硬い
－３点：硬い
－２点：やや硬い
－１点：わずかに硬い
０点：無添加区と差がない
１点：わずかにやわらかい
２点：やややわらかい
３点：やわらかい
４点：非常にやわらかい
５点：極めて顕著にやわらかい
＜ほぐれやすさの評点＞
－５点：極めて顕著にほぐれにくい
－４点：非常にほぐれにくい
－３点：ほぐれにくい
－２点：ややほぐれにくい
－１点：わずかにほぐれにくい
０点：無添加区と差がない
１点：わずかにほぐれやすい
２点：ややほぐれやすい
３点：ほぐれやすい
４点：非常にほぐれやすい
５点：極めて顕著にほぐれやすい
（ｉｖ）評価結果を表１に示す。 (I) Domestic young chicken breast meat was cut into 20 g pieces, placed in a vinyl bag, and the proteases of each test group shown in Table 1 were added and shaken. In Table 1, "-" indicates that protease is not added.
As the neutral protease derived from Bacillus amyloliquefaciens, those having the following characteristics (a) to (e) were used.
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
The amount of each protease added to 20 g of chicken breast meat was 10,000 U (50,000 U per 100 g of chicken breast meat). Each protease was brought into contact with chicken breast meat by allowing it to stand at 20 ° C. for 15 minutes.
(Ii) Next, using chicken breast meat contacted with each protease, it was cooked in a pan as follows to prepare a cream stew.
<How to prepare cream stew>
After lightly frying the surface of chicken breast meat, water was added, and after boiling, it was simmered for 15 minutes, then commercially available roux (manufactured by House Foods Co., Ltd.) was dissolved, simmered for 5 minutes, then milk was added and simmered for another 5 minutes. The total boiling time was 25 minutes. The ratio of water, milk, and roux was the recommended ratio of commercially available roux (water: milk: roux = 700: 100: 90 (weight ratio)), and a model system of chicken breast meat without vegetables was used.
(Iii) Chicken breast meat was taken out from the prepared chicken breast meat cream stew and subjected to sensory evaluation.
The sensory evaluation was performed on a panel of 4 people by a scoring method of -5 to 5 points with the additive-free group (test group 1) as 0 points for softness and ease of loosening. The grades were given to the first decimal place, and the evaluation results were shown by the numerical values up to the first decimal place, rounded off from the average value of the four people.
In this sensory evaluation, "softness" means the chewing force required to chew the whole meat, and "easiness to loosen" means the feeling of loosening like a white fish when chewing the meat. Means. The ease of loosening is a texture that is particularly preferred in stewed dishes, and is generally improved by boiling for a long time. In addition, the ease of unraveling is the texture that is felt when the myofibrils of the meat collapse as a bundle to some extent. Even with a soft texture such as liver or foie gras, where the whole meat is crushed evenly, the ease of loosening is not felt so much.
The scores for softness and ease of loosening were defined as follows by comparison with the additive-free group.
<Score of softness>
-5 points: Extremely hard -4 points: Very hard -3 points: Hard -2 points: Slightly hard -1 point: Slightly hard 0 points: No difference from the additive-free group 1 point: Slightly soft 2 Point: Slightly soft 3 points: Soft 4 points: Very soft 5 points: Extremely soft <Score of ease of loosening>
-5 points: Very noticeably hard to loosen -4 points: Very hard to loosen -3 points: Hard to loosen -2 points: Slightly hard to loosen -1 point: Slightly hard to loosen 0 points: No difference from additive-free group 1 point : Slightly easy to unravel 2 points: Slightly easy to unravel 3 points: Easy to unravel 4 points: Very easy to unravel 5 points: Very significantly easy to unravel (iv) The evaluation results are shown in Table 1.
バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼとしては、次の(ａ)～(ｅ)の特性を有するものを用いた。
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。
鶏ムネ肉２０ｇに対する各プロテアーゼの添加量は、いずれも１０，０００Ｕ（鶏ムネ肉１００ｇあたり５０，０００Ｕ）とした。２０℃にて１５分間静置して各プロテアーゼを鶏ムネ肉に接触させた。
（ｉｉ）次いで、各プロテアーゼを接触させた鶏ムネ肉を用いて、鍋にて下記の通り調理し、クリームシチューを調製した。
＜クリームシチューの調製方法＞
鶏ムネ肉の表面を軽く炒めた後に水を入れ、沸騰後１５分間煮込んだ後に市販のルウ（ハウス食品株式会社製）を溶かし、５分間煮込み、次いで牛乳を加えて更に５分間煮込んだ。合計の煮込み時間は２５分間であった。水、牛乳、ルウの割合は、市販のルウ推奨の割合（水：牛乳：ルウ＝７００：１００：９０（重量比））とし、野菜を使用しない鶏ムネ肉のみのモデル系とした。
（ｉｉｉ）調製された鶏ムネ肉のクリームシチューから鶏ムネ肉を取り出し、官能評価に供した。
官能評価は、やわらかさとほぐれやすさについて、無添加区（試験区１）を０点とした－５点～５点の評点法にて、４名のパネルにて行った。評点は、小数点以下第一位までとし、４名の平均値を四捨五入した小数点以下第一位までの数値にて評価結果を示した。
なお、本官能評価において、「やわらかさ」とは、肉全体を噛み潰すために必要な咀嚼力を意味し、「ほぐれやすさ」とは、肉を噛み潰す時に白身魚のようにホロホロとほぐれる感覚を意味する。ほぐれやすさは、煮込み料理において特に好まれる食感で、一般的には長時間煮込むことで向上する。また、ほぐれやすさは、肉の筋原線維がある程度の束として崩れることで感じられる食感であり、肉全体が硬いと筋原線維が崩れないため、ほぐれやすさは感じられにくく、逆に肉全体が均一に潰れるレバーやフォアグラのようなやわらかい食感の場合も、ほぐれやすさはあまり感じられない。
やわらかさおよびほぐれやすさの評点については、無添加区との比較により、下記の通り定義した。
＜やわらかさの評点＞
－５点：極めて顕著に硬い
－４点：非常に硬い
－３点：硬い
－２点：やや硬い
－１点：わずかに硬い
０点：無添加区と差がない
１点：わずかにやわらかい
２点：やややわらかい
３点：やわらかい
４点：非常にやわらかい
５点：極めて顕著にやわらかい
＜ほぐれやすさの評点＞
－５点：極めて顕著にほぐれにくい
－４点：非常にほぐれにくい
－３点：ほぐれにくい
－２点：ややほぐれにくい
－１点：わずかにほぐれにくい
０点：無添加区と差がない
１点：わずかにほぐれやすい
２点：ややほぐれやすい
３点：ほぐれやすい
４点：非常にほぐれやすい
５点：極めて顕著にほぐれやすい
（ｉｖ）評価結果を表１に示す。 (I) Domestic young chicken breast meat was cut into 20 g pieces, placed in a vinyl bag, and the proteases of each test group shown in Table 1 were added and shaken. In Table 1, "-" indicates that protease is not added.
As the neutral protease derived from Bacillus amyloliquefaciens, those having the following characteristics (a) to (e) were used.
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA).
The amount of each protease added to 20 g of chicken breast meat was 10,000 U (50,000 U per 100 g of chicken breast meat). Each protease was brought into contact with chicken breast meat by allowing it to stand at 20 ° C. for 15 minutes.
(Ii) Next, using chicken breast meat contacted with each protease, it was cooked in a pan as follows to prepare a cream stew.
<How to prepare cream stew>
After lightly frying the surface of chicken breast meat, water was added, and after boiling, it was simmered for 15 minutes, then commercially available roux (manufactured by House Foods Co., Ltd.) was dissolved, simmered for 5 minutes, then milk was added and simmered for another 5 minutes. The total boiling time was 25 minutes. The ratio of water, milk, and roux was the recommended ratio of commercially available roux (water: milk: roux = 700: 100: 90 (weight ratio)), and a model system of chicken breast meat without vegetables was used.
(Iii) Chicken breast meat was taken out from the prepared chicken breast meat cream stew and subjected to sensory evaluation.
The sensory evaluation was performed on a panel of 4 people by a scoring method of -5 to 5 points with the additive-free group (test group 1) as 0 points for softness and ease of loosening. The grades were given to the first decimal place, and the evaluation results were shown by the numerical values up to the first decimal place, rounded off from the average value of the four people.
In this sensory evaluation, "softness" means the chewing force required to chew the whole meat, and "easiness to loosen" means the feeling of loosening like a white fish when chewing the meat. Means. The ease of loosening is a texture that is particularly preferred in stewed dishes, and is generally improved by boiling for a long time. In addition, the ease of unraveling is the texture that is felt when the myofibrils of the meat collapse as a bundle to some extent. Even with a soft texture such as liver or foie gras, where the whole meat is crushed evenly, the ease of loosening is not felt so much.
The scores for softness and ease of loosening were defined as follows by comparison with the additive-free group.
<Score of softness>
-5 points: Extremely hard -4 points: Very hard -3 points: Hard -2 points: Slightly hard -1 point: Slightly hard 0 points: No difference from the additive-free group 1 point: Slightly soft 2 Point: Slightly soft 3 points: Soft 4 points: Very soft 5 points: Extremely soft <Score of ease of loosening>
-5 points: Very noticeably hard to loosen -4 points: Very hard to loosen -3 points: Hard to loosen -2 points: Slightly hard to loosen -1 point: Slightly hard to loosen 0 points: No difference from additive-free group 1 point : Slightly easy to unravel 2 points: Slightly easy to unravel 3 points: Easy to unravel 4 points: Very easy to unravel 5 points: Very significantly easy to unravel (iv) The evaluation results are shown in Table 1.
表１に示す通り、サーモリシンを添加することで、ほぐれやすさが最も向上した（試験区４）。
一方、やわらかさは、パパインを添加することで最も向上したが、付与されるやわらかさはレバー様の均一なやわらかさであり、ほぐれやすさはあまり感じられなかった（試験区２）。同様に、アスペルギルス オリザエ（Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ）由来の中性プロテアーゼ添加区（試験区８）や、アスペルギルス属（Ａｓｐｅｒｇｉｌｌｕｓ ｓｐ．）由来の酸性プロテアーゼ添加区（試験区５）においても、高いやわらかさ付与効果が認められたが、ほぐれやすさの向上はあまり認められなかった。 As shown in Table 1, the addition of thermolysin improved the ease of loosening most (Test Group 4).
On the other hand, the softness was most improved by adding papain, but the softness given was a liver-like uniform softness, and the ease of loosening was not felt so much (Test Group 2). Similarly, a high softness-imparting effect is obtained in the neutral protease-added group (test group 8) derived from Aspergillus oryzae and the acidic protease-added group (test group 5) derived from Aspergillus sp. Although it was observed, there was not much improvement in the ease of loosening.
一方、やわらかさは、パパインを添加することで最も向上したが、付与されるやわらかさはレバー様の均一なやわらかさであり、ほぐれやすさはあまり感じられなかった（試験区２）。同様に、アスペルギルス オリザエ（Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ）由来の中性プロテアーゼ添加区（試験区８）や、アスペルギルス属（Ａｓｐｅｒｇｉｌｌｕｓ ｓｐ．）由来の酸性プロテアーゼ添加区（試験区５）においても、高いやわらかさ付与効果が認められたが、ほぐれやすさの向上はあまり認められなかった。 As shown in Table 1, the addition of thermolysin improved the ease of loosening most (Test Group 4).
On the other hand, the softness was most improved by adding papain, but the softness given was a liver-like uniform softness, and the ease of loosening was not felt so much (Test Group 2). Similarly, a high softness-imparting effect is obtained in the neutral protease-added group (test group 8) derived from Aspergillus oryzae and the acidic protease-added group (test group 5) derived from Aspergillus sp. Although it was observed, there was not much improvement in the ease of loosening.
以上の結果より、プロテアーゼの種類によって食肉組織に対する作用は異なり、やわらかくする効果とほぐれやすさを付与する効果は別であることが明らかとなった。
従って、最もほぐれやすさを向上させる効果を示したサーモリシンを、他のプロテアーゼと併用することで、ほぐれやすさを更に向上させ得る可能性があることが示唆された。 From the above results, it was clarified that the action on the meat tissue differs depending on the type of protease, and the effect of softening and the effect of imparting looseness are different.
Therefore, it was suggested that the thermolysin, which showed the effect of improving the looseness most, may be used in combination with other proteases to further improve the looseness.
従って、最もほぐれやすさを向上させる効果を示したサーモリシンを、他のプロテアーゼと併用することで、ほぐれやすさを更に向上させ得る可能性があることが示唆された。 From the above results, it was clarified that the action on the meat tissue differs depending on the type of protease, and the effect of softening and the effect of imparting looseness are different.
Therefore, it was suggested that the thermolysin, which showed the effect of improving the looseness most, may be used in combination with other proteases to further improve the looseness.
［試験例２］
試験例１にて、食肉に対するほぐれやすさ付与効果が最も高かったサーモリシンについて、他のプロテアーゼとの併用効果の検討を実施した。 [Test Example 2]
In Test Example 1, thermolysin, which had the highest effect of imparting looseness to meat, was examined for its combined effect with other proteases.
試験例１にて、食肉に対するほぐれやすさ付与効果が最も高かったサーモリシンについて、他のプロテアーゼとの併用効果の検討を実施した。 [Test Example 2]
In Test Example 1, thermolysin, which had the highest effect of imparting looseness to meat, was examined for its combined effect with other proteases.
（ｉ）試験例１と同様の方法にて、国産若鶏ムネ肉を２０ｇの小片にカットしてビニール製の袋に入れ、表２に示す各試験区のプロテアーゼを加えて振り混ぜた。表２中、酵素１欄の「－」および酵素２欄の「－」は、それぞれ各酵素を添加しないことを示す。試験区２はサーモリシンの単独添加区、試験区３～１１は、サーモリシンと他のプロテアーゼとの併用添加区とした。
表２に示されたサーモリシンおよび他のプロテアーゼとしては、試験例１にて用いたものと同じものを用いた。各プロテアーゼの添加量は、いずれも鶏ムネ肉２０ｇに対し１０，０００Ｕ（鶏ムネ肉１００ｇあたり５０，０００Ｕ）とした。
（ｉｉ）２０℃にて１５分間静置して各プロテアーゼを接触させた後、各プロテアーゼを接触させた鶏ムネ肉を用いて鍋にて調理し、クリームシチューを調製した。クリームシチューの調理方法は、試験例１と同様とした。
（ｉｉｉ）調製された鶏ムネ肉のクリームシチューから鶏ムネ肉を取り出し、官能評価に供した。官能評価は、やわらかさとほぐれやすさについて、酵素無添加区（試験区１）を０点とした－５点～５点の評点法にて、４名のパネルにて行い、試験例１と同様の方法にて４名の評点の平均値を算出した。
また、ほぐれやすさについては、サーモリシンのみを添加した試験区（試験区２）、サーモリシン以外のプロテアーゼを単独で添加した試験区（試験例１の試験区２～１１）の評価結果に基づき、サーモリシンと他のプロテアーゼとを併用して添加した試験区についての評価の理論値を、それぞれ単独で添加した試験区における評価結果（評点の平均値）の和として算出した。
例えば、表２の試験区３について、ほぐれやすさの評価の理論値は、サーモリシンのみを添加した試験区（表２の試験区２）のほぐれやすさの評価結果が「２」、パパインのみを添加した試験区（表１の試験区２）のほぐれやすさの評価結果が「０．２」であり、これらの和により「２＋０．２＝２．２」と算出される。よって「２．２」が表２の試験区３についてのほぐれやすさの評価の理論値となる。このようにして算出した理論値と実際の官能評価結果との差を求めた。例えば、表２の試験区３については、官能評価結果が「０．１」、理論値が「２．２」であるため、その差は「０．１－２．２＝－２．１」と算出される。
上記のようにして求めた理論値と実際の官能評価結果との差がゼロより大きければ、単独で添加した場合の効果の相加効果として求められる理論値よりも良好な効果、すなわち相乗効果が得られたことを意味する。相乗効果の得られた試験区については、相乗効果欄に「○」を、相乗効果の得られなかった試験区については、相乗効果欄に「×」を付した。
（ｉｖ）試験例２の評価結果を表２に示す。 (I) In the same manner as in Test Example 1, domestic young chicken breast meat was cut into 20 g pieces, placed in a vinyl bag, and the proteases of each test group shown in Table 2 were added and shaken. In Table 2, "-" in the enzyme 1 column and "-" in the enzyme 2 column indicate that each enzyme is not added. Test group 2 was a group to which thermolysin was added alone, and test groups 3 to 11 were groups to which thermolysin was added in combination with other proteases.
As the thermolysin and other proteases shown in Table 2, the same ones used in Test Example 1 were used. The amount of each protease added was 10,000 U per 20 g of chicken breast meat (50,000 U per 100 g of chicken breast meat).
(Ii) After allowing to stand at 20 ° C. for 15 minutes to contact each protease, the chicken breast meat to which each protease was contacted was cooked in a pan to prepare a cream stew. The cooking method of the cream stew was the same as in Test Example 1.
(Iii) Chicken breast meat was taken out from the prepared chicken breast meat cream stew and subjected to sensory evaluation. The sensory evaluation was performed on a panel of 4 people with a scoring method of -5 to 5 points with the enzyme-free group (test group 1) as 0 points for softness and ease of loosening, and was the same as in Test Example 1. The average value of the scores of 4 people was calculated by the method of.
Regarding the ease of loosening, thermolysin was based on the evaluation results of the test group to which only thermolysin was added (test group 2) and the test group to which a protease other than thermolysin was added alone (test groups 2 to 11 of test example 1). The theoretical value of the evaluation of the test group added in combination with other proteases was calculated as the sum of the evaluation results (average value of the scores) in the test group added individually.
For example, for the test group 3 in Table 2, the theoretical value for the evaluation of looseness is "2" for the test group to which only thermolysin was added (test group 2 in Table 2), and only papain. The evaluation result of the looseness of the added test group (test group 2 in Table 1) is "0.2", and the sum of these is calculated as "2 + 0.2 = 2.2". Therefore, "2.2" is the theoretical value for the evaluation of the ease of loosening for the test group 3 in Table 2. The difference between the theoretical value calculated in this way and the actual sensory evaluation result was obtained. For example, for the test group 3 in Table 2, the sensory evaluation result is "0.1" and the theoretical value is "2.2", so the difference is "0.1-2.2 = -2.1". Is calculated.
If the difference between the theoretical value obtained as described above and the actual sensory evaluation result is larger than zero, an effect better than the theoretical value obtained as an additive effect of the effect when added alone, that is, a synergistic effect is obtained. It means that it was obtained. For the test plots where the synergistic effect was obtained, "○" was added to the synergistic effect column, and for the test plots where the synergistic effect was not obtained, "x" was added to the synergistic effect column.
(Iv) Table 2 shows the evaluation results of Test Example 2.
表２に示されたサーモリシンおよび他のプロテアーゼとしては、試験例１にて用いたものと同じものを用いた。各プロテアーゼの添加量は、いずれも鶏ムネ肉２０ｇに対し１０，０００Ｕ（鶏ムネ肉１００ｇあたり５０，０００Ｕ）とした。
（ｉｉ）２０℃にて１５分間静置して各プロテアーゼを接触させた後、各プロテアーゼを接触させた鶏ムネ肉を用いて鍋にて調理し、クリームシチューを調製した。クリームシチューの調理方法は、試験例１と同様とした。
（ｉｉｉ）調製された鶏ムネ肉のクリームシチューから鶏ムネ肉を取り出し、官能評価に供した。官能評価は、やわらかさとほぐれやすさについて、酵素無添加区（試験区１）を０点とした－５点～５点の評点法にて、４名のパネルにて行い、試験例１と同様の方法にて４名の評点の平均値を算出した。
また、ほぐれやすさについては、サーモリシンのみを添加した試験区（試験区２）、サーモリシン以外のプロテアーゼを単独で添加した試験区（試験例１の試験区２～１１）の評価結果に基づき、サーモリシンと他のプロテアーゼとを併用して添加した試験区についての評価の理論値を、それぞれ単独で添加した試験区における評価結果（評点の平均値）の和として算出した。
例えば、表２の試験区３について、ほぐれやすさの評価の理論値は、サーモリシンのみを添加した試験区（表２の試験区２）のほぐれやすさの評価結果が「２」、パパインのみを添加した試験区（表１の試験区２）のほぐれやすさの評価結果が「０．２」であり、これらの和により「２＋０．２＝２．２」と算出される。よって「２．２」が表２の試験区３についてのほぐれやすさの評価の理論値となる。このようにして算出した理論値と実際の官能評価結果との差を求めた。例えば、表２の試験区３については、官能評価結果が「０．１」、理論値が「２．２」であるため、その差は「０．１－２．２＝－２．１」と算出される。
上記のようにして求めた理論値と実際の官能評価結果との差がゼロより大きければ、単独で添加した場合の効果の相加効果として求められる理論値よりも良好な効果、すなわち相乗効果が得られたことを意味する。相乗効果の得られた試験区については、相乗効果欄に「○」を、相乗効果の得られなかった試験区については、相乗効果欄に「×」を付した。
（ｉｖ）試験例２の評価結果を表２に示す。 (I) In the same manner as in Test Example 1, domestic young chicken breast meat was cut into 20 g pieces, placed in a vinyl bag, and the proteases of each test group shown in Table 2 were added and shaken. In Table 2, "-" in the enzyme 1 column and "-" in the enzyme 2 column indicate that each enzyme is not added. Test group 2 was a group to which thermolysin was added alone, and test groups 3 to 11 were groups to which thermolysin was added in combination with other proteases.
As the thermolysin and other proteases shown in Table 2, the same ones used in Test Example 1 were used. The amount of each protease added was 10,000 U per 20 g of chicken breast meat (50,000 U per 100 g of chicken breast meat).
(Ii) After allowing to stand at 20 ° C. for 15 minutes to contact each protease, the chicken breast meat to which each protease was contacted was cooked in a pan to prepare a cream stew. The cooking method of the cream stew was the same as in Test Example 1.
(Iii) Chicken breast meat was taken out from the prepared chicken breast meat cream stew and subjected to sensory evaluation. The sensory evaluation was performed on a panel of 4 people with a scoring method of -5 to 5 points with the enzyme-free group (test group 1) as 0 points for softness and ease of loosening, and was the same as in Test Example 1. The average value of the scores of 4 people was calculated by the method of.
Regarding the ease of loosening, thermolysin was based on the evaluation results of the test group to which only thermolysin was added (test group 2) and the test group to which a protease other than thermolysin was added alone (test groups 2 to 11 of test example 1). The theoretical value of the evaluation of the test group added in combination with other proteases was calculated as the sum of the evaluation results (average value of the scores) in the test group added individually.
For example, for the test group 3 in Table 2, the theoretical value for the evaluation of looseness is "2" for the test group to which only thermolysin was added (test group 2 in Table 2), and only papain. The evaluation result of the looseness of the added test group (test group 2 in Table 1) is "0.2", and the sum of these is calculated as "2 + 0.2 = 2.2". Therefore, "2.2" is the theoretical value for the evaluation of the ease of loosening for the test group 3 in Table 2. The difference between the theoretical value calculated in this way and the actual sensory evaluation result was obtained. For example, for the test group 3 in Table 2, the sensory evaluation result is "0.1" and the theoretical value is "2.2", so the difference is "0.1-2.2 = -2.1". Is calculated.
If the difference between the theoretical value obtained as described above and the actual sensory evaluation result is larger than zero, an effect better than the theoretical value obtained as an additive effect of the effect when added alone, that is, a synergistic effect is obtained. It means that it was obtained. For the test plots where the synergistic effect was obtained, "○" was added to the synergistic effect column, and for the test plots where the synergistic effect was not obtained, "x" was added to the synergistic effect column.
(Iv) Table 2 shows the evaluation results of Test Example 2.
表２に示す通り、バチルス（Ｂａｃｉｌｌｕｓ）属由来の中性プロテアーゼとサーモリシンを併用することでほぐれやすさが大きく向上する傾向が確認された（試験区７及び９）。
試験区７では、官能評価により得られたほぐれやすさの評点と、理論値の差が１．１点であり、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼとサーモリシンを併用することにより、相乗的なほぐれやすさ向上効果が得られることが示された。
しかし、試験区９では、官能評価により得られたほぐれやすさの評点と、理論値の差が－０．７点であり、ほぐれやすさの相乗的な向上は認められなかった。
また、パパインとサーモリシンを併用することによりやわらかさが大きく向上したが、レバー様の均一な食感となり、ほぐれやすさは逆に低下した（試験区３）。 As shown in Table 2, it was confirmed that the combined use of a neutral protease derived from the genus Bacillus and thermolysin tends to greatly improve the looseness (test groups 7 and 9).
In Test Group 7, the difference between the score of ease of loosening obtained by sensory evaluation and the theoretical value was 1.1 points, and a neutral protease derived from Bacillus amyloliquefaciens was used in combination with thermolysin. It was shown that a synergistic effect of improving the ease of loosening can be obtained.
However, in Test Group 9, the difference between the score of ease of loosening obtained by the sensory evaluation and the theoretical value was -0.7 points, and no synergistic improvement in ease of loosening was observed.
In addition, the combined use of papain and thermolysin greatly improved the softness, but the texture became uniform like a lever, and the ease of loosening decreased on the contrary (Test Group 3).
試験区７では、官能評価により得られたほぐれやすさの評点と、理論値の差が１．１点であり、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼとサーモリシンを併用することにより、相乗的なほぐれやすさ向上効果が得られることが示された。
しかし、試験区９では、官能評価により得られたほぐれやすさの評点と、理論値の差が－０．７点であり、ほぐれやすさの相乗的な向上は認められなかった。
また、パパインとサーモリシンを併用することによりやわらかさが大きく向上したが、レバー様の均一な食感となり、ほぐれやすさは逆に低下した（試験区３）。 As shown in Table 2, it was confirmed that the combined use of a neutral protease derived from the genus Bacillus and thermolysin tends to greatly improve the looseness (test groups 7 and 9).
In Test Group 7, the difference between the score of ease of loosening obtained by sensory evaluation and the theoretical value was 1.1 points, and a neutral protease derived from Bacillus amyloliquefaciens was used in combination with thermolysin. It was shown that a synergistic effect of improving the ease of loosening can be obtained.
However, in Test Group 9, the difference between the score of ease of loosening obtained by the sensory evaluation and the theoretical value was -0.7 points, and no synergistic improvement in ease of loosening was observed.
In addition, the combined use of papain and thermolysin greatly improved the softness, but the texture became uniform like a lever, and the ease of loosening decreased on the contrary (Test Group 3).
以上のように、サーモリシンと併用するプロテアーゼの種類によって食肉組織に対する作用が異なるが、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼとサーモリシンを組み合わせて用いることによるほぐれやすさ向上効果は、サーモリシンと他のプロテアーゼとを組み合わせた場合に比べて明らかに大きく、かつ相乗的であることが認められた。特に、肉のほぐれやすさが好まれる煮込み料理においては、当該知見によるほぐれやすさ付与効果は非常に有用である。
As described above, the action on meat tissue differs depending on the type of protease used in combination with thermolysin, but the effect of improving the ease of loosening by using a neutral protease derived from Bacillus amyloliquefaciens in combination with thermolysin is It was found to be clearly larger and synergistic than the combination of thermolysin and other proteases. In particular, in stewed dishes in which the ease of loosening of meat is preferred, the effect of imparting ease of loosening based on the findings is extremely useful.
［実施例１～１５］鶏ムネ肉の改質およびクリームシチューの調製
試験例１と同様の方法にて、国産若鶏ムネ肉を２０ｇの小片にカットしてビニール製の袋に入れ、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンを、それぞれ表３に示す量を添加して振り混ぜ、２０℃にて１５分間静置し、鶏ムネ肉小片にプロテアーゼを接触させて、鶏ムネ肉の改質を行った。表３中、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ欄およびサーモリシン欄の「－」は、それぞれ各酵素を添加しないことを示す。
各実施例にて、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンとしては、試験例１にて用いたものと同じものを用いた。
次いで、上記各プロテアーゼを接触させて改質した鶏ムネ肉を用いて鍋にて調理し、クリームシチューを調製した。クリームシチューの調理方法は、試験例１と同様とした。 [Examples 1 to 15] Modification of chicken breast meat and preparation of cream stew By the same method as in Test Example 1, domestic young chicken breast meat is cut into 20 g pieces, placed in a vinyl bag, and Bacillus amilo. Neutral protease and thermolysin derived from Bacillus amyloliquefaciens were added in the amounts shown in Table 3 and shaken, allowed to stand at 20 ° C. for 15 minutes, and the chicken breast meat pieces were brought into contact with the protease. The chicken breast was modified. In Table 3, "-" in the neutral protease column and the thermolysin column derived from Bacillus amyloliquefaciens indicates that each enzyme is not added.
In each example, the same neutral protease and thermolysin derived from Bacillus amyloliquefaciens were used as those used in Test Example 1.
Next, a cream stew was prepared by cooking in a pan using chicken breast meat modified by contacting each of the above proteases. The cooking method of the cream stew was the same as in Test Example 1.
試験例１と同様の方法にて、国産若鶏ムネ肉を２０ｇの小片にカットしてビニール製の袋に入れ、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンを、それぞれ表３に示す量を添加して振り混ぜ、２０℃にて１５分間静置し、鶏ムネ肉小片にプロテアーゼを接触させて、鶏ムネ肉の改質を行った。表３中、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ欄およびサーモリシン欄の「－」は、それぞれ各酵素を添加しないことを示す。
各実施例にて、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンとしては、試験例１にて用いたものと同じものを用いた。
次いで、上記各プロテアーゼを接触させて改質した鶏ムネ肉を用いて鍋にて調理し、クリームシチューを調製した。クリームシチューの調理方法は、試験例１と同様とした。 [Examples 1 to 15] Modification of chicken breast meat and preparation of cream stew By the same method as in Test Example 1, domestic young chicken breast meat is cut into 20 g pieces, placed in a vinyl bag, and Bacillus amilo. Neutral protease and thermolysin derived from Bacillus amyloliquefaciens were added in the amounts shown in Table 3 and shaken, allowed to stand at 20 ° C. for 15 minutes, and the chicken breast meat pieces were brought into contact with the protease. The chicken breast was modified. In Table 3, "-" in the neutral protease column and the thermolysin column derived from Bacillus amyloliquefaciens indicates that each enzyme is not added.
In each example, the same neutral protease and thermolysin derived from Bacillus amyloliquefaciens were used as those used in Test Example 1.
Next, a cream stew was prepared by cooking in a pan using chicken breast meat modified by contacting each of the above proteases. The cooking method of the cream stew was the same as in Test Example 1.
得られた鶏ムネ肉のクリームシチューから鶏ムネ肉を取り出し、試料として官能評価に供した。なお、上記プロテアーゼのいずれも添加せずに同様に処理し、調理してクリームシチューを調製し、鶏ムネ肉を取り出して対照とした。
官能評価は、やわらかさとほぐれやすさについて、対照を０点とした－５点～５点の評点法にて、４名のパネルにて行い、試験例１と同様の方法にて算出した評点の平均値にて評価結果を示した。
また、ほぐれやすさについては、評点の平均値が３点以上であった試料については「○」、２点以上３点未満であった試料については「△」、２点未満であった試料については「×」と判定した。
評価結果を表３に併せて示した。 Chicken breast meat was taken out from the obtained cream stew of chicken breast meat and used as a sample for sensory evaluation. In addition, all of the above proteases were treated in the same manner without addition, cooked to prepare a cream stew, and chicken breast meat was taken out and used as a control.
The sensory evaluation was performed on a panel of 4 people by a scoring method of -5 to 5 points with a control of 0 points for softness and ease of loosening, and the score was calculated by the same method as in Test Example 1. The evaluation result is shown by the average value.
Regarding the ease of loosening, "○" was given to samples with an average score of 3 points or more, "△" was given to samples with 2 points or more and less than 3 points, and samples with less than 2 points were given. Was determined to be "x".
The evaluation results are also shown in Table 3.
官能評価は、やわらかさとほぐれやすさについて、対照を０点とした－５点～５点の評点法にて、４名のパネルにて行い、試験例１と同様の方法にて算出した評点の平均値にて評価結果を示した。
また、ほぐれやすさについては、評点の平均値が３点以上であった試料については「○」、２点以上３点未満であった試料については「△」、２点未満であった試料については「×」と判定した。
評価結果を表３に併せて示した。 Chicken breast meat was taken out from the obtained cream stew of chicken breast meat and used as a sample for sensory evaluation. In addition, all of the above proteases were treated in the same manner without addition, cooked to prepare a cream stew, and chicken breast meat was taken out and used as a control.
The sensory evaluation was performed on a panel of 4 people by a scoring method of -5 to 5 points with a control of 0 points for softness and ease of loosening, and the score was calculated by the same method as in Test Example 1. The evaluation result is shown by the average value.
Regarding the ease of loosening, "○" was given to samples with an average score of 3 points or more, "△" was given to samples with 2 points or more and less than 3 points, and samples with less than 2 points were given. Was determined to be "x".
The evaluation results are also shown in Table 3.
表３に示す通り、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼとサーモリシンを添加して接触させた鶏ムネ肉を用いて調理した場合、調理後の鶏ムネ肉のほぐれやすさの向上が確認された（実施例１～１５）。
かかる結果から、食肉１００ｇに対し、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、サーモリシンともに１０Ｕ～５，０００，０００Ｕの添加により、ほぐれやすさを付与し得ることが示唆された。また、鶏ムネ肉のほぐれやすさは、両酵素ともに１００Ｕ以上の添加により、より向上することが認められた。
上記実施例の結果から、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンを、それぞれ食肉１００ｇに対し１０Ｕ～５，０００，０００Ｕの添加量にて食肉に接触させることにより、煮込み料理において特に好まれる食感であるほぐれやすさを、食肉に付与できることが示唆された。 As shown in Table 3, when cooked using chicken breast meat that has been contacted by adding thermolysin to a neutral protease derived from Bacillus amyloliquefaciens, the ease of loosening of the chicken breast meat after cooking Improvement was confirmed (Examples 1 to 15).
From these results, it was suggested that the ease of loosening can be imparted to 100 g of meat by adding 10 U to 5,000,000 U of both the neutral protease and thermolysin derived from Bacillus amyloliquefaciens. In addition, it was found that the ease of loosening of chicken breast meat was further improved by adding 100 U or more of both enzymes.
From the results of the above examples, the neutral protease and thermolysin derived from Bacillus amyloliquefaciens are brought into contact with the meat in an amount of 10 U to 5,000,000 U per 100 g of the meat, respectively, and then boiled. It was suggested that the meat can be given the ease of loosening, which is a texture that is particularly preferred in cooking.
かかる結果から、食肉１００ｇに対し、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ、サーモリシンともに１０Ｕ～５，０００，０００Ｕの添加により、ほぐれやすさを付与し得ることが示唆された。また、鶏ムネ肉のほぐれやすさは、両酵素ともに１００Ｕ以上の添加により、より向上することが認められた。
上記実施例の結果から、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンを、それぞれ食肉１００ｇに対し１０Ｕ～５，０００，０００Ｕの添加量にて食肉に接触させることにより、煮込み料理において特に好まれる食感であるほぐれやすさを、食肉に付与できることが示唆された。 As shown in Table 3, when cooked using chicken breast meat that has been contacted by adding thermolysin to a neutral protease derived from Bacillus amyloliquefaciens, the ease of loosening of the chicken breast meat after cooking Improvement was confirmed (Examples 1 to 15).
From these results, it was suggested that the ease of loosening can be imparted to 100 g of meat by adding 10 U to 5,000,000 U of both the neutral protease and thermolysin derived from Bacillus amyloliquefaciens. In addition, it was found that the ease of loosening of chicken breast meat was further improved by adding 100 U or more of both enzymes.
From the results of the above examples, the neutral protease and thermolysin derived from Bacillus amyloliquefaciens are brought into contact with the meat in an amount of 10 U to 5,000,000 U per 100 g of the meat, respectively, and then boiled. It was suggested that the meat can be given the ease of loosening, which is a texture that is particularly preferred in cooking.
［実施例１６］牛モモ肉のカレー
米国産牛モモ肉を２ｃｍ角にダイスカットし、ビニール製の袋に入れ、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンを添加して振り混ぜ、２０℃にて１５分間静置して接触させた。バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンとしては、試験例１にて用いたものと同じものを用い、前記各プロテアーゼの添加量は、いずれも牛モモ肉１００ｇあたり５０，０００Ｕとした。
次いで、上記処理にて各プロテアーゼを接触させた牛肉（すなわち各プロテアーゼが作用した牛モモ肉）を用いて、鍋にて下記の通り調理し、カレーを調製した。
すなわち、上記牛モモ肉は表面を軽く炒めた後に水を入れ、沸騰後１５分間煮込んだ後に市販のルウ（ハウス食品株式会社製）を溶かし、更に１０分間煮込んだ。合計の煮込み時間は２５分間であった。水とルウの割合は、市販のルウ推奨の割合（水：ルウ＝８５０：１１５（重量比））とし、野菜を使用しない牛モモ肉のみのカレーを調製して、実施例１６とした。
なお、上記プロテアーゼを接触させずに同様に処理した牛モモ肉を用いてカレーを調製し、比較例１とした。 [Example 16] Beef thigh curry A US beef thigh is die-cut into 2 cm squares, placed in a vinyl bag, and a neutral protease and thermolysin derived from Bacillus amyloliquefaciens are added. It was shaken and allowed to stand at 20 ° C. for 15 minutes for contact. As the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, the same proteases used in Test Example 1 were used, and the amount of each protease added was 50, per 100 g of beef thigh meat. It was set to 000U.
Next, using the beef to which each protease was contacted in the above treatment (that is, beef thigh meat on which each protease acted), the curry was prepared by cooking as follows in a pan.
That is, the surface of the beef thigh meat was lightly fried, then water was added, and after boiling, it was simmered for 15 minutes, then a commercially available roux (manufactured by House Foods Co., Ltd.) was dissolved, and the meat was simmered for another 10 minutes. The total boiling time was 25 minutes. The ratio of water to roux was the recommended ratio of commercially available roux (water: roux = 850: 115 (weight ratio)), and a curry containing only beef thigh meat without vegetables was prepared and used as Example 16.
A curry was prepared using beef thigh meat treated in the same manner without contacting the above protease, and used as Comparative Example 1.
米国産牛モモ肉を２ｃｍ角にダイスカットし、ビニール製の袋に入れ、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンを添加して振り混ぜ、２０℃にて１５分間静置して接触させた。バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンとしては、試験例１にて用いたものと同じものを用い、前記各プロテアーゼの添加量は、いずれも牛モモ肉１００ｇあたり５０，０００Ｕとした。
次いで、上記処理にて各プロテアーゼを接触させた牛肉（すなわち各プロテアーゼが作用した牛モモ肉）を用いて、鍋にて下記の通り調理し、カレーを調製した。
すなわち、上記牛モモ肉は表面を軽く炒めた後に水を入れ、沸騰後１５分間煮込んだ後に市販のルウ（ハウス食品株式会社製）を溶かし、更に１０分間煮込んだ。合計の煮込み時間は２５分間であった。水とルウの割合は、市販のルウ推奨の割合（水：ルウ＝８５０：１１５（重量比））とし、野菜を使用しない牛モモ肉のみのカレーを調製して、実施例１６とした。
なお、上記プロテアーゼを接触させずに同様に処理した牛モモ肉を用いてカレーを調製し、比較例１とした。 [Example 16] Beef thigh curry A US beef thigh is die-cut into 2 cm squares, placed in a vinyl bag, and a neutral protease and thermolysin derived from Bacillus amyloliquefaciens are added. It was shaken and allowed to stand at 20 ° C. for 15 minutes for contact. As the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, the same proteases used in Test Example 1 were used, and the amount of each protease added was 50, per 100 g of beef thigh meat. It was set to 000U.
Next, using the beef to which each protease was contacted in the above treatment (that is, beef thigh meat on which each protease acted), the curry was prepared by cooking as follows in a pan.
That is, the surface of the beef thigh meat was lightly fried, then water was added, and after boiling, it was simmered for 15 minutes, then a commercially available roux (manufactured by House Foods Co., Ltd.) was dissolved, and the meat was simmered for another 10 minutes. The total boiling time was 25 minutes. The ratio of water to roux was the recommended ratio of commercially available roux (water: roux = 850: 115 (weight ratio)), and a curry containing only beef thigh meat without vegetables was prepared and used as Example 16.
A curry was prepared using beef thigh meat treated in the same manner without contacting the above protease, and used as Comparative Example 1.
［実施例１７］豚バラ肉の角煮
国産豚バラ肉を３ｃｍ角にダイスカットし、ビニール製の袋に入れ、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンを添加して振り混ぜ、２０℃にて１５分間静置して接触させた。バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンとしては、試験例１にて用いたものと同じものを用い、前記各プロテアーゼの添加量は、いずれも豚バラ肉１００ｇあたり５０，０００Ｕとした。
次いで、上記処理にて各プロテアーゼを接触させた豚バラ肉（すなわち各プロテアーゼが作用した豚バラ肉）を用いて、鍋にて下記の通り調理し、豚バラ肉の角煮を調製した。
すなわち、上記豚バラ肉を軽く下茹でし、水及び市販の角煮のたれ（モランボン株式会社製）を加えて４０分間フタをして煮込み、フタを取って更に１５分間煮込み、豚バラ肉の角煮を調製して、実施例１７とした。合計の煮込み時間は５５分間であった。水とたれの割合は、市販のたれ推奨の割合（水：たれ＝４００：１２０（重量比））とした。
なお、上記プロテアーゼを接触させずに同様に調理して豚の角煮を調製し、比較例２とした。 [Example 17] Kakuni pork belly Die-cut domestic pork belly into 3 cm squares, put them in a vinyl bag, and add a neutral protease and thermolysin derived from Bacillus amyloliquefaciens. It was shaken and allowed to stand at 20 ° C. for 15 minutes for contact. As the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, the same proteases used in Test Example 1 were used, and the amount of each protease added was 50, per 100 g of pork belly. It was set to 000U.
Next, using the pork belly to which each protease was brought into contact in the above treatment (that is, the pork belly on which each protease acted), the pork belly was cooked in a pan as follows to prepare a stewed pork belly.
That is, the above pork belly is lightly boiled, water and a commercially available kakuni sauce (manufactured by Moranborg Co., Ltd.) are added, and the pork belly is covered and simmered for 40 minutes. Kakuni was prepared and used as Example 17. The total boiling time was 55 minutes. The ratio of water to dripping was the recommended ratio of commercially available dripping (water: dripping = 400: 120 (weight ratio)).
In addition, the pork kakuni was prepared by cooking in the same manner without contacting the above protease, and used as Comparative Example 2.
国産豚バラ肉を３ｃｍ角にダイスカットし、ビニール製の袋に入れ、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンを添加して振り混ぜ、２０℃にて１５分間静置して接触させた。バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼおよびサーモリシンとしては、試験例１にて用いたものと同じものを用い、前記各プロテアーゼの添加量は、いずれも豚バラ肉１００ｇあたり５０，０００Ｕとした。
次いで、上記処理にて各プロテアーゼを接触させた豚バラ肉（すなわち各プロテアーゼが作用した豚バラ肉）を用いて、鍋にて下記の通り調理し、豚バラ肉の角煮を調製した。
すなわち、上記豚バラ肉を軽く下茹でし、水及び市販の角煮のたれ（モランボン株式会社製）を加えて４０分間フタをして煮込み、フタを取って更に１５分間煮込み、豚バラ肉の角煮を調製して、実施例１７とした。合計の煮込み時間は５５分間であった。水とたれの割合は、市販のたれ推奨の割合（水：たれ＝４００：１２０（重量比））とした。
なお、上記プロテアーゼを接触させずに同様に調理して豚の角煮を調製し、比較例２とした。 [Example 17] Kakuni pork belly Die-cut domestic pork belly into 3 cm squares, put them in a vinyl bag, and add a neutral protease and thermolysin derived from Bacillus amyloliquefaciens. It was shaken and allowed to stand at 20 ° C. for 15 minutes for contact. As the neutral protease and thermolysin derived from Bacillus amyloliquefaciens, the same proteases used in Test Example 1 were used, and the amount of each protease added was 50, per 100 g of pork belly. It was set to 000U.
Next, using the pork belly to which each protease was brought into contact in the above treatment (that is, the pork belly on which each protease acted), the pork belly was cooked in a pan as follows to prepare a stewed pork belly.
That is, the above pork belly is lightly boiled, water and a commercially available kakuni sauce (manufactured by Moranborg Co., Ltd.) are added, and the pork belly is covered and simmered for 40 minutes. Kakuni was prepared and used as Example 17. The total boiling time was 55 minutes. The ratio of water to dripping was the recommended ratio of commercially available dripping (water: dripping = 400: 120 (weight ratio)).
In addition, the pork kakuni was prepared by cooking in the same manner without contacting the above protease, and used as Comparative Example 2.
実施例１６および比較例１の各牛モモ肉のカレー、ならびに実施例１７および比較例２の各豚バラ肉の角煮から牛モモ肉および豚バラ肉をそれぞれ取り出し、官能評価に供した。
官能評価は、食肉のほぐれやすさに着目して実施し、下記評価基準にて、４名のパネルに評価させて行った。官能評価結果は、４名の協議により決した。なお、豚バラ肉については、脂の層と肉の層のうち、肉の層におけるほぐれやすさを評価した。
＜評価基準＞
〇：比較例に比べてほぐれやすい
△：比較例に比べてややほぐれやすい
×：比較例と同等のほぐれやすさであるか、比較例よりほぐれにくい
評価結果を表４に示す。 The beef thighs and pork ribs were taken out from the curry of each beef thigh of Example 16 and Comparative Example 1 and the pork ribs of Example 17 and Comparative Example 2, respectively, and subjected to sensory evaluation.
The sensory evaluation was carried out focusing on the ease of loosening of the meat, and was evaluated by a panel of four people according to the following evaluation criteria. The sensory evaluation results were decided through discussions among four people. For pork ribs, the ease of loosening in the meat layer was evaluated among the fat layer and the meat layer.
<Evaluation criteria>
〇: Easy to unravel compared to the comparative example Δ: Slightly unraveled compared to the comparative example ×: Evaluated as easy to unravel as the comparative example or harder to unravel than the comparative example is shown in Table 4.
官能評価は、食肉のほぐれやすさに着目して実施し、下記評価基準にて、４名のパネルに評価させて行った。官能評価結果は、４名の協議により決した。なお、豚バラ肉については、脂の層と肉の層のうち、肉の層におけるほぐれやすさを評価した。
＜評価基準＞
〇：比較例に比べてほぐれやすい
△：比較例に比べてややほぐれやすい
×：比較例と同等のほぐれやすさであるか、比較例よりほぐれにくい
評価結果を表４に示す。 The beef thighs and pork ribs were taken out from the curry of each beef thigh of Example 16 and Comparative Example 1 and the pork ribs of Example 17 and Comparative Example 2, respectively, and subjected to sensory evaluation.
The sensory evaluation was carried out focusing on the ease of loosening of the meat, and was evaluated by a panel of four people according to the following evaluation criteria. The sensory evaluation results were decided through discussions among four people. For pork ribs, the ease of loosening in the meat layer was evaluated among the fat layer and the meat layer.
<Evaluation criteria>
〇: Easy to unravel compared to the comparative example Δ: Slightly unraveled compared to the comparative example ×: Evaluated as easy to unravel as the comparative example or harder to unravel than the comparative example is shown in Table 4.
表４に示す通り、実施例１６のカレーにおける牛肉、および実施例１７の角煮における豚バラ肉は、いずれも比較例１のカレーにおける牛肉、および比較例２における豚バラ肉に比べてほぐれやすいと評価され、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンが作用した牛モモ肉および豚バラ肉をそれぞれ用いて調製されたカレーおよび角煮において、牛モモ肉および豚バラ肉に明確にほぐれやすさが付与されたことが確認された。
As shown in Table 4, the beef in the curry of Example 16 and the pork ribs in the kakuni of Example 17 are both easier to loosen than the beef in the curry of Comparative Example 1 and the pork ribs in Comparative Example 2. Beef thighs and pork ribs in curry and kakuni prepared using beef thighs and pork ribs on which neutral protease and thermolysin from Bacillus amyloliquefaciens acted, respectively. It was confirmed that the ribs were clearly given the ease of loosening.
上記実施例において示された結果より、バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼ及びサーモリシンを食肉に接触させることで、肉の種類や部位、調理メニューに関わらず、食肉にほぐれやすさを付与し得ることが示された。
かかるほぐれやすさ付与効果は、とりわけ、食肉のほぐれやすさが食感として好まれる煮込み料理において、有用である。 From the results shown in the above examples, by contacting the neutral protease and thermolysin derived from Bacillus amyloliquefaciens with the meat, it is easy to loosen the meat regardless of the type and part of the meat and the cooking menu. It has been shown that it can be given.
This effect of imparting looseness is particularly useful in stewed dishes in which the looseness of meat is preferred as a texture.
かかるほぐれやすさ付与効果は、とりわけ、食肉のほぐれやすさが食感として好まれる煮込み料理において、有用である。 From the results shown in the above examples, by contacting the neutral protease and thermolysin derived from Bacillus amyloliquefaciens with the meat, it is easy to loosen the meat regardless of the type and part of the meat and the cooking menu. It has been shown that it can be given.
This effect of imparting looseness is particularly useful in stewed dishes in which the looseness of meat is preferred as a texture.
以上詳述したように、本発明により、短時間でかつ簡便に食肉を軟質化し、食肉にほぐれやすさを付与し得る食肉の改質方法を提供することができる。
また本発明により、短時間でかつ簡便に食肉を軟質化して、食肉にほぐれやすさを付与し得る食肉の改質用組成物を提供することができる。
さらに本発明により、軟質化され、ほぐれやすさの付与された食肉を含有する食品を提供することができる。
本発明は、食品分野、特に食肉のほぐれやすさが食感として好まれる煮込み料理において極めて有用である。 As described in detail above, according to the present invention, it is possible to provide a method for modifying meat that can easily soften meat in a short time and impart ease of loosening to the meat.
Further, according to the present invention, it is possible to provide a composition for modifying meat that can easily soften meat in a short time and impart ease of loosening to the meat.
Further, according to the present invention, it is possible to provide a food containing meat that has been softened and imparted with ease of loosening.
The present invention is extremely useful in the food field, especially in stewed dishes in which the ease of loosening of meat is preferred as a texture.
また本発明により、短時間でかつ簡便に食肉を軟質化して、食肉にほぐれやすさを付与し得る食肉の改質用組成物を提供することができる。
さらに本発明により、軟質化され、ほぐれやすさの付与された食肉を含有する食品を提供することができる。
本発明は、食品分野、特に食肉のほぐれやすさが食感として好まれる煮込み料理において極めて有用である。 As described in detail above, according to the present invention, it is possible to provide a method for modifying meat that can easily soften meat in a short time and impart ease of loosening to the meat.
Further, according to the present invention, it is possible to provide a composition for modifying meat that can easily soften meat in a short time and impart ease of loosening to the meat.
Further, according to the present invention, it is possible to provide a food containing meat that has been softened and imparted with ease of loosening.
The present invention is extremely useful in the food field, especially in stewed dishes in which the ease of loosening of meat is preferred as a texture.
本願は、日本国で出願された特願２０２０－０２８８６１を基礎としており、その内容は本明細書にすべて包含されるものである。
This application is based on Japanese Patent Application No. 2020-028861 filed in Japan, the contents of which are all included in the present specification.
Claims (9)
- バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼ、及びサーモリシンを食肉と接触させることを含む、食肉の改質方法；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。 A method for modifying meat, which is a neutral protease derived from Bacillus amyloliquefaciens and which comprises contacting a neutral protease having the following characteristics (a) to (e) and thermolysin with meat. ;
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA). - 食肉に接触させるバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼの量が、食肉１００ｇあたり１０Ｕ以上５，０００，０００Ｕ以下であり、食肉に接触させるサーモリシンの量が、食肉１００ｇあたり１０Ｕ以上５，０００，０００Ｕ以下である、請求項１に記載の方法。 The amount of neutral protease derived from Bacillus amyloliquefaciens in contact with meat is 10 U or more and 5,000,000 U or less per 100 g of meat, and the amount of thermolysin in contact with meat is 10 U or more per 100 g of meat. The method according to claim 1, which is 5,000,000 U or less.
- 請求項１または２に記載の方法により改質された食肉を用いることを含む、煮込み料理の製造方法。 A method for producing a stewed dish, which comprises using the meat modified by the method according to claim 1 or 2.
- バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼ、及びサーモリシンを含有する、食肉の改質用組成物；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。 A composition for modifying meat, which is a neutral protease derived from Bacillus amyloliquefaciens and contains a neutral protease having the following characteristics (a) to (e) and thermolysin;
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA). - バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼと、サーモリシンとの含有量比（バチルス アミロリケファシエンス由来の中性プロテアーゼ：サーモリシン）が、酵素活性比にて１：０．０００００２～１：５００，０００である、請求項４に記載の組成物。 The content ratio of the neutral protease derived from Bacillus amyloliquefaciens to thermolysin (neutral protease derived from Bacillus amyloliquefaciens: thermolysin) has an enzyme activity ratio of 1: 0.000002 to 1. : The composition according to claim 4, which is 500,000.
- 煮込み料理に用いる食肉の改質用である、請求項４または５に記載の組成物。 The composition according to claim 4 or 5, which is used for modifying meat used in stewed dishes.
- バチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼであって、下記（ａ）～（ｅ）の特性を有する中性プロテアーゼおよび、サーモリシンが作用した食肉を含有する、食品；
（ａ）ペプチド結合を有する基質を加水分解して、ペプチドを遊離する酵素である、
（ｂ）至適ｐＨが７である、
（ｃ）ｐＨ＝５～７にて安定である、
（ｄ）至適温度が５０℃～５５℃である、
（ｅ）金属プロテアーゼであり、エチレンジアミン四酢酸（ＥＤＴＡ）によって阻害される。 A food product containing a neutral protease derived from Bacillus amyloliquefaciens, which has the following characteristics (a) to (e), and meat on which thermolysin has acted;
(A) An enzyme that hydrolyzes a substrate having a peptide bond to release a peptide.
(B) The optimum pH is 7,
(C) Stable at pH = 5-7,
(D) The optimum temperature is 50 ° C to 55 ° C.
(E) A metalloproteinase that is inhibited by ethylenediaminetetraacetic acid (EDTA). - 食肉１００ｇあたり、１０Ｕ以上５，０００，０００Ｕ以下のバチルス アミロリクエファシエンス（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）由来の中性プロテアーゼと、１０Ｕ以上５，０００，０００Ｕ以下のサーモリシンが作用した食肉を含有する、請求項７に記載の食品。 17. Foods listed in.
- 煮込み料理として供される、請求項７または８に記載の食品。 The food according to claim 7 or 8, which is served as a stewed dish.
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|---|---|---|---|---|
| JP2009219383A (en) * | 2008-03-13 | 2009-10-01 | Kagawa Univ | Psychrophilic bacterium producing collagen degrading enzyme, collagen degrading enzyme, method for producing the same, and method for producing tenderized meat using the enzyme |
| JP2014158463A (en) * | 2013-01-28 | 2014-09-04 | Ajinomoto Co Inc | Manufacturing method of processed meat product, and modifier for processed meat product |
| WO2017018503A1 (en) * | 2015-07-29 | 2017-02-02 | 味の素株式会社 | Meat modifying agent |
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| JP2009219383A (en) * | 2008-03-13 | 2009-10-01 | Kagawa Univ | Psychrophilic bacterium producing collagen degrading enzyme, collagen degrading enzyme, method for producing the same, and method for producing tenderized meat using the enzyme |
| JP2014158463A (en) * | 2013-01-28 | 2014-09-04 | Ajinomoto Co Inc | Manufacturing method of processed meat product, and modifier for processed meat product |
| WO2017018503A1 (en) * | 2015-07-29 | 2017-02-02 | 味の素株式会社 | Meat modifying agent |
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| Title |
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| OSAGO HARUMI, MIKIKO KOBAYASHI-MIURA, YOSHIFUMI HAMASAKI, NOBUMASA HARA, MINEYOSHI HIYOSHI, MIKAKO TSUCHIYA: "Complete solubilization of cartilage using the heat-stable protease thermolysin for comprehensive GAG analysis", ANALYTICAL BIOCHEMISTRY, vol. 548, 1 May 2018 (2018-05-01), pages 115 - 118, XP055856035, DOI: 10.1016/j.ab.2018.02.028 * |
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