Method and enzyme composition for improving the meat quality and flavor of meat of birds and beasts

Abstract

To provide a protease processing technique for bird and beast meat capable of improving texture so as to soften it and enhancing flavor.SOLUTION: Provided is a method for improving texture and flavor of bird and beast meat by processing the bird and beast meat with an enzyme composition containing protease, wherein alkaline protease derived from bacterium of genus Bacillus and the protease derived from Aspergillus fungus are used in combination.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a method and an enzyme composition for improving the meat quality and flavor of livestock and poultry meat. More specifically, the present invention relates to a method for improving the meat quality and flavor of livestock and poultry meat by treating the meat with an enzyme composition containing protease and the like.

Background Art

[0002] Conventionally, attempts have been made to improve the meat quality and flavor of livestock and poultry meat by treating the meat with an enzyme composition containing protease (protease treatment) (see Patent Document 1). For example, the meat of hens (parent birds) that have finished laying eggs and are used as minced meat because of their tough meat quality can be expected to have an expanded range of uses by changing the meat quality to be softer by protease treatment.

[0003] In addition, by increasing the amino acids contained in the meat by protease treatment, an improvement in flavor (especially umami) can also be expected. On the other hand, it is known that depending on the type of protease used, off-flavors and bitterness derived from the decomposition products may occur (see Patent Document 2).

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0005] The main object of the present invention is to provide a protease treatment technology for livestock and poultry meat that can improve the meat quality to be softer and also improve the flavor.

Means for Solving the Problems

[0006] In order to solve the above problems, the present invention provides the following [1]-[9]. [1] A method for improving the meat quality and flavor of livestock and poultry meat by treating the livestock and poultry meat with an enzyme composition containing protease, wherein the enzyme composition contains an alkaline protease derived from a Bacillus bacterium and a protease derived from a fungus of the genus Aspergillus, the protease derived from a fungus of the genus Aspergillus has the activities of both an endo-type protease and an exo-type protease under the conditions of pH 8 and 10 °C, the total protease activity is 5,000 U / g or more, and the activity of the exo-type protease in the total protease activity is 1.0% or more. [2] The method according to [1], wherein the protease derived from a fungus of the genus Aspergillus is a protease derived from Aspergillus oryzae. [3] The method according to [1] or [2], wherein the alkaline protease derived from a Bacillus bacterium is an alkaline protease derived from Bacillus subtilis. [4] A method for producing livestock and poultry meat with improved meat quality and flavor, comprising a step of treating the livestock and poultry meat with an enzyme composition containing protease, wherein the enzyme composition contains an alkaline protease derived from a Bacillus bacterium and a protease derived from a fungus of the genus Aspergillus, the protease derived from a fungus of the genus Aspergillus has the activities of both an endo-type protease and an exo-type protease under the conditions of pH 8 and 10 °C, the total protease activity is 5,000 U / g or more, and the activity of the exo-type protease in the total protease activity is 1.0% or more. [5] The production method according to [4], wherein the protease derived from a fungus of the genus Aspergillus is a protease derived from Aspergillus oryzae. [6] The production method according to [4] or [5], wherein the alkaline protease derived from a Bacillus bacterium is an alkaline protease derived from Bacillus subtilis.

[0007] [7] An enzyme composition for improving the meat quality and flavor of bird and animal meat, comprising protease, comprising an alkaline protease derived from a Bacillus bacterium and a protease derived from a fungus of the genus Aspergillus, wherein the protease derived from the fungus of the genus Aspergillus has the activities of both endo-type protease and exo-type protease under the conditions of pH 8 and 10 °C, the total protease activity is 5,000 U / g or more, and the activity of the exo-type protease in the total protease activity is 1.0% or more. [8] The enzyme composition according to [7], wherein the protease derived from the fungus of the genus Aspergillus is a protease derived from Aspergillus oryzae. [9] The enzyme composition according to [7] or [8], wherein the alkaline protease derived from a Bacillus bacterium is an alkaline protease derived from Bacillus subtilis.

[0008] In the present invention, "meat quality" means the quality of meat that affects palatability. "Meat quality" particularly includes "softness of meat parts", "ease of biting through the skin", "juiciness", "water retention", "appearance", etc.

[0009] "Flavor" means the aroma and taste of food and drink. "Flavor" particularly includes "umami" (richness), "saltiness", "meat aroma", "bitterness / off-flavors", "sweetness", "sourness", etc.

Advantages of the Invention

[0010] The present invention provides a protease treatment technology for bird and animal meat that can improve the meat quality to be soft and also improve the flavor.

Brief Description of the Drawings

[0011]

Figure 1

Mode for Carrying Out the Invention

[0012] Hereinafter, preferred embodiments for carrying out the present invention will be described. Note that the embodiments described below show an example of typical embodiments of the present invention, and the scope of the present invention is not construed narrowly thereby.

[0013] [Protease derived from Aspergillus fungus] The protease derived from Aspergillus fungus contained in the enzyme composition according to the present invention may be derived from any Aspergillus fungus. For example, proteases derived from Aspergillus fumigatus, Aspergillus melleus, Aspergillus niger, Aspergillus oryzae, and Aspergillus sojae can be mentioned. As the protease derived from Aspergillus fungus, one kind of protease may be used, or two or more kinds of proteases may be used in combination.

[0014] The protease derived from Aspergillus fungus is preferably a protease derived from Aspergillus oryzae.

[0015] The protease derived from Aspergillus fungus has the activities of both endo-type protease and exo-type protease under the conditions of pH 8 and 10°C, the total protease activity is 5,000 U / g or more, and the activity of exo-type protease (exo-type protease activity ratio) in the total protease activity is 1.0% or more. Such a protease is particularly preferred. More preferred proteases derived from Aspergillus fungi have a total protease activity of 7,000 U / g or more, 8,000 U / g or more, or 9,000 U / g or more, and an exo-type protease activity ratio of 1.2% or more, 1.5% or more, or 1.6% or more.

[0016] The measurement of the total protease activity can be carried out by a conventionally known method, for example, in accordance with the "Protease Activity Measurement Method" in the "Self-Specification of Existing Additives (4th Edition)", Report 1. The measurement of the exo-type protease activity (aminopeptidase activity) can also be carried out by a conventionally known method, for example, in accordance with the "Aminopeptidase Activity Measurement Method" in the "Self-Specification of Existing Additives (4th Edition)".

[0017] For the protease derived from Aspergillus fungi described above, a commercially available product may be used, or one appropriately produced and obtained may be used. Examples of commercially available enzymes include Kocuriazyme P from Mitsubishi Chemical Foods Co., Ltd.

[0018] [Alkaline protease derived from Bacillus bacteria] The alkaline protease derived from Bacillus bacteria contained in the enzyme composition according to the present invention may be derived from any Bacillus bacteria. Examples include alkaline proteases derived from Bacillus amyloliquefaciens, Bacillus cereus, Bacillus clausii, Bacillus intermedius, Bacillus lentus, Bacillus licheniformis, Bacillus stearothermophilus, Bacillus subtilis, and Bacillus thermoproteolyticus. The alkaline protease derived from Bacillus bacteria may use one kind of protease, or may use a combination of two or more kinds of proteases.

[0019] The alkaline protease derived from Bacillus bacteria is preferably an alkaline protease derived from Bacillus subtilis.

[0020] For the above-mentioned alkaline protease derived from Bacillus bacteria, a commercially available product may be used, or a product appropriately manufactured and obtained may be used. Examples of commercially available enzymes are as follows. Protine SD-AY10 (derived from Bacillus licheniformis; Amano Enzyme Inc.) Delvolase (derived from Bacillus licheniformis; DSM Japan Co., Ltd.) Esperase (derived from Bacillus sp.; Novozymes Japan Co., Ltd.) Savinase (derived from Bacillus sp.; Novozymes Japan Co., Ltd.) Ebalase (derived from Bacillus sp.; Novozymes Japan Co., Ltd.) Alcalase (derived from Bacillus licheniformis; Novozymes Japan Co., Ltd.) Bioprase OP (derived from Bacillus sp.; Nagase ChemteX Corporation) Bioprase SP-20FG (derived from Bacillus sp.; Nagase ChemteX Corporation) Orientase 22BF (derived from Bacillus subtilis; HBI Co., Ltd.)

[0021] When producing proteases derived from Aspergillus fungi and alkaline proteases derived from Bacillus bacteria, known methods can be utilized. Specifically, it can be produced by culturing microorganisms that produce proteases and recovering the proteases from the culture. The microorganisms that produce proteases may inherently produce proteases or may be modified to produce proteases. Microorganisms that produce proteases can be obtained, for example, by introducing a gene encoding a protease into a microorganism so that it can be expressed. The introduction of the gene can be achieved, for example, by introducing a vector carrying the gene into a microorganism or by introducing the gene onto the chromosome of the microorganism. The amino acid sequences of various proteases derived from Bacillus bacteria or Aspergillus fungi and the nucleotide sequences of the genes encoding them can be obtained from public databases such as NCBI (http: / / www.ncbi.nlm.nih.gov / ), etc. The culture conditions of the microorganisms are not particularly limited as long as the microorganisms can grow and proteases can be produced. The microorganisms can be cultured, for example, under the normal conditions for culturing microorganisms such as bacteria and fungi. The proteases may be used as purified products, or may be used as cultures of microorganisms that produce proteases, culture supernatants separated from the cultures, cells separated from the cultures, and processed products of the cells, etc. As the protease derived from Aspergillus fungi and the alkaline protease derived from Bacillus bacteria, homologs of the known proteases exemplified above may be used. The homologs are not particularly limited as long as they are found in Aspergillus fungi and Bacillus bacteria and have the desired protease activity. Also, as the protease derived from Aspergillus fungi and the alkaline protease derived from Bacillus bacteria, artificial variants of the known proteases or their homologs as exemplified above may be used. The artificial variants are not particularly limited as long as they have the desired protease activity.

[0022] [Other Enzymes] The enzyme composition according to the present invention may contain various proteases in addition to the alkaline protease derived from Bacillus bacteria and the protease derived from Aspergillus fungi.

[0023] As additional proteases, for example, as enteropeptidases, papain, bromelain, ficin, actinidin, sutilisin, thermolysin, trypsin, cathepsin, chymotrypsin, rennet, etc. can be mentioned. Also, as exopeptidases, leucine aminopeptidase, aminopeptidase M, carboxypeptidase A, Y, etc. can be mentioned.

[0024] The enzyme composition according to the present invention may contain various enzymes such as amylase, lipase, phospholipase, esterase, glutaminase, transglutaminase, oxidase, nuclease, etc. in addition to the protease. Also, the enzyme composition according to the present invention may contain additives usually blended in foods.

[0025] The enzyme composition according to the present invention can be dissolved in a solvent (such as a pH adjusting solution) or used in powder form.

[0026] [Livestock and poultry meat] Livestock and poultry meat is not particularly limited and may be, for example, livestock meat such as beef, pork, horse, sheep, wild boar, deer, etc., and poultry meat such as chicken, turkey, duck, goose, etc. The present invention can be suitably applied to livestock and poultry meat such as parent birds (adult chicken meat) with a long breeding period and tough meat quality, breeding hens, parent pigs, dairy cows, etc. The part of the meat is also not particularly limited, but parts containing a large amount of hard proteins such as shank, shoulder, neck, thigh, cheek, thigh, tail, leg, chest, wing, etc. and parts with a large reduction in moisture due to cooking are preferred because there is a high possibility of obtaining a large effect on improving the meat quality.

[0027] [Protease treatment] By treating livestock and poultry meat with an enzyme composition containing an alkaline protease derived from Bacillus bacteria and a protease derived from Aspergillus fungi, the meat quality (especially "softness") can be improved, and while suppressing "bitterness / off-flavors" compared to treatment with each protease alone, the "umami" can be significantly improved to obtain a synergistic flavor improvement effect.

[0028] The method of treating livestock and poultry meat with the enzyme composition is not particularly limited as long as the enzyme acts effectively on the meat from immediately after slaughter until the meat is consumed. Generally, methods such as immersing the meat in a solution of the enzyme composition, injecting the same solution into the meat, tumbling in the same solution, or sprinkling the powder or solution of the enzyme composition on the meat are employed. It is preferable that the alkaline protease derived from Bacillus bacteria and the protease derived from Aspergillus fungi act on the meat simultaneously, but there is a possibility of obtaining the desired effect even if they act separately.

[0029] The treatment with the enzyme composition is preferably carried out by maintaining the contact state between the meat and the enzyme for a certain period of time under the optimal conditions of the enzyme so that the enzyme reaction proceeds sufficiently and the quality of the meat is maintained. The temperature is 0 to 40°C, preferably 0 to 25°C. The pH is 5 to 10, preferably 7 to 10. The time is 0.5 to 48 hours, preferably 0.5 to 24 hours. However, conditions such as temperature, pH, and time can be appropriately set according to the type, shape, and size of the meat to be treated, and the desired meat quality and flavor. After the enzyme treatment, it is preferable to quickly perform heat treatment or freeze to stop the progress of the enzyme reaction.

[0030] The usage amount of the enzyme composition can also be appropriately set according to the type, shape, and size of the meat to be treated, and the desired meat quality and flavor. The amount of protease derived from Aspergillus fungi can be, for example, 0.001 - 1% (w / v), preferably 0.01 - 0.5% (w / v), more preferably 0.05 - 0.1% (w / v) when meat is immersed in the enzyme composition solution. The amount of alkaline protease derived from Bacillus bacteria can be, for example, 0.001 - 1% (w / v), preferably 0.005 - 0.5% (w / v), more preferably 0.01 - 0.05% (w / v) when meat is immersed in the enzyme composition solution.

Examples

[0031] [Test Example 1: Sensory Evaluation 1] Parent bird peach meat was treated with an enzyme composition, and a sensory evaluation of the meat quality and flavor was conducted.

[0032] (1) Enzymes As the alkaline protease derived from Bacillus bacteria, a commercially available enzyme AP (Oryza 22BF, HBI Co., Ltd.) derived from Bacillus subtilis was used. As the protease derived from Aspergillus fungi, commercially available enzymes A, B, C, D, F, G, and H were used. For enzyme A, Kokurase P of Mitsubishi Chemical Foods Co., Ltd., which is an alkaline protease derived from Aspergillus oryzae, was used. Also, as enzyme E, commercially available papain (purified papain, Mitsubishi Chemical Foods Co., Ltd.) was used.

[0033] (2) Activity Measurement The total protease activity and aminopeptidase activity (exo-type protease activity) of enzymes A - D, F - H were measured. The measurement of the total protease activity was carried out according to the "Protease Activity Measurement Method" in the "Self-Specified Standards for Existing Additives (4th Edition)", Report 1. The measurement of the aminopeptidase activity was carried out according to the "Aminopeptidase Activity Measurement Method" in the "Self-Specified Standards for Existing Additives (4th Edition)". The measurement conditions were a reaction temperature of 10°C and a pH of 8.0 (Atkins-Pantin buffer).

[0034] The measurement results are shown in Table 1. As shown in the table, for enzymes A, F, and G, the total protease activity was 5,000 U / g or more, and the activity of exo-type protease in the total protease activity was 1.0% or more.

[0035]

Table 1

[0036] (3) Enzyme treatment The thawed parent chicken momotaro meat was immersed in a pH adjuster solution with or without enzyme added and then sealed in a vacuum pack. The volume of the pH adjuster solution was 30% (w / w) of the parent chicken momotaro meat. As the pH adjuster solution, a solution prepared by dissolving the pH adjuster shown in Table 2 in water at 3.5% (w / v) was used. The added concentration of the enzyme to the pH adjuster solution was 0.05% (w / v) for enzyme AP and 0.1% (w / v) for enzymes A - H. Treatment was carried out at 4°C using a vacuum tumbling device (25 minutes of rotation (under vacuum conditions), 5 minutes of rest (vacuum released), 4 sets). Baking was carried out at 250°C (steam output 40%) for 8 minutes using a steam convection oven.

[0037]

Table 2

[0038] (4) Sensory evaluation The "softness" and "tastiness" of the samples treated with the pH adjuster solution with enzyme added were evaluated based on the samples treated with the pH adjuster solution without enzyme added. When it was better than the standard, it was marked as 〇, when it was equivalent, it was marked as △, and when it was inferior, it was marked as ×. The results are shown in Table 3.

[0039]

Table 3

[0040] In the sample treated with enzyme AP alone, the "softness" was better than the standard, but the "umami" was equivalent to the standard. Also, in the sample treated with enzyme A alone, both the "softness" and "umami" were equivalent to the standard. However, in the sample treated with a combination of enzyme AP and enzyme A, both the "softness" and "umami" were better than the standard. It was clarified that by using a combination of enzyme AP and enzyme A, an improvement effect on "umami" can be obtained compared to the use of each enzyme alone. Similarly, for enzymes F and G, it was confirmed that both the "softness" and "umami" were better than the standard when used in combination with enzyme AP.

[0041] For enzymes B, C, D, and H, no improvement in "umami" was observed when used in combination with enzyme AP compared to the treatment with enzyme AP alone. For enzymes H and E, the "umami" that was equivalent to the standard in the sample treated with enzyme AP alone was inferior to the standard in the treatment with the combination of enzyme AP, and there was a tendency for the flavor to decrease rather than increase when combined with enzyme AP.

[0042] [Test Example 2: Sensory Evaluation 2] The sensory evaluation of the meat quality and flavor of the parent chicken peach meat treated with the enzyme composition was entrusted to an external testing agency.

[0043] The parent chicken peach meat that had been subjected to enzyme treatment (enzyme AP alone or in combination with enzyme A) in the same manner as in Test Example 1 was cut into pieces approximately 3 cm square and used for evaluation. The "softness of the meat part", "umami", "ease of chewing the skin", "saltiness", "overall meat aroma", and "preference (overall evaluation)" of the sample treated with the pH adjuster solution with enzyme addition were evaluated based on the sample treated with the pH adjuster solution without enzyme addition. The evaluation was conducted by 10 panelists. For each evaluation item, the scores were assigned as follows: equal to the standard was 3, slightly better than the standard was 4, better than the standard was 5, slightly worse than the standard was 2, and worse than the standard was 1.

[0044] Table 4 shows the average value of the scores of 10 panelists.

[0045]

Table 4

[0046] In the sample treated with enzyme AP alone, the "softness of the meat part" and "umami" tended to improve compared to the standard. In the sample treated with a combination of enzyme AP and enzyme A, a further improvement in the "softness of the meat part" and "umami" was observed, and the "preference (overall evaluation)" also improved compared to the standard.

[0047] [Test Example 3: Taste Sensor Measurement] The taste sensor measurement of the parent bird peach meat treated with the enzyme composition was entrusted to an external testing institution.

[0048] In the same manner as in Test Example 1, 5 times the amount of water was added to the parent bird peach meat (baked meat) treated with enzyme treatment (enzyme AP alone or a combination of enzyme AP and enzyme A), and it was pulverized and used for measurement after removing the solid content.

[0049] The measurement results are shown in Figure 1 and Table 5. In Figure 1, the horizontal axis represents the response value of AT0 (bitter / strange taste / front taste), and the vertical axis represents the response value of AAE (umami / aftertaste). The response values were corrected so that the response value of the sample treated with the pH adjuster solution without enzyme addition became 0. A numerical difference of 1 in the response value corresponds to a 20% difference in the component concentration of "bitter / strange taste / front taste" or "umami / aftertaste", and it is considered sufficient to give different tastes to humans.

[0050]

Table 5

[0051] Regarding the treatment by combining enzyme AP, for which the improvement effect of "umami and aftertaste" has been confirmed by the sensory evaluation in Test Examples 1 and 2, the same effect was also shown by the response value of the sensor. The AAE response values in the treatments with enzyme AP alone and enzyme A alone were 0.65 and 0.08 respectively, and the AAE response value in the treatment by combining enzyme AP and enzyme A was 1.9. The AAE response value (1.9) in the treatment by combining enzyme AP and enzyme A was significantly higher than the sum (0.73) of the AAE response values in the treatments with enzyme AP alone and enzyme A alone, and was improved by 1 or more compared to the sum.

[0052] Furthermore, the treatment by combining enzyme AP and enzyme A also showed the effect of suppressing the "bitter taste / strange taste" generated in the treatment with enzyme AP alone. The AT0 response value in the treatment with enzyme AP alone was 0.98, while the AT0 response value in the treatment by combining enzyme AP and enzyme A was suppressed to 0.23. It was shown that by combining enzyme A with enzyme AP, the improvement effect of "umami and aftertaste" can be obtained, and at the same time, the suppression effect of "bitter taste / strange taste and pre-taste" can also be obtained.

[0053] On the other hand, regarding enzymes B, C, and D, no improvement in "umami" was observed when combined with enzyme AP. Regarding enzyme E, the "umami" was the same in the treatment with enzyme AP alone and the combined treatment with enzyme AP, but the "bitter taste / strange taste" rather increased in the combined treatment with enzyme AP.

Claims

1. A method for improving the meat quality and flavor of livestock and poultry meat by treating the livestock and poultry meat with an enzyme composition containing protease, wherein the enzyme composition contains an alkaline protease derived from Bacillus subtilis and a protease derived from a fungus of the genus Aspergillus, the protease derived from the fungus of the genus Aspergillus has the activities of both endo-type protease and exo-type protease under the conditions of pH 8 and 10°C, the total protease activity is 5,000 U / g or more, and the activity of the exo-type protease in the total protease activity is 1.0% or more.

2. An enzyme composition for improving the meat quality and flavor of livestock and poultry meat, containing protease, which contains an alkaline protease derived from Bacillus subtilis and a protease derived from a fungus of the genus Aspergillus, wherein the protease derived from the fungus of the genus Aspergillus has the activities of both endo-type protease and exo-type protease under the conditions of pH 8 and 10°C, the total protease activity is 5,000 U / g or more, and the activity of the exo-type protease in the total protease activity is 1.0% or more.

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