JP2022118303A - Method for improving texture and flavor of bird and beast meat and enzyme composition - Google Patents

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

The present invention relates to methods and enzyme compositions for improving meat quality and flavor of poultry meat. More particularly, it relates to a method for improving the meat quality and flavor of game meat by treating it with an enzyme composition containing a protease.

Conventionally, attempts have been made to improve the meat quality and flavor of game meat by treating game meat with an enzyme composition containing protease (protease treatment) (see Patent Document 1). For example, meat from chickens (parent birds) that have finished their egg-laying season, which is used as minced meat due to its hard meat, can be expected to expand its use by softening its meat quality with protease treatment.

In addition, protease treatment increases the amount of amino acids contained in meat, which can be expected to improve flavor (especially umami). On the other hand, it is known that depending on the type of protease used, an offensive odor or bitter taste derived from decomposition products may occur (see Patent Document 2).

JP-A-05-007476 Japanese Patent Application Laid-Open No. 2003-033161

The main object of the present invention is to provide a protease treatment technology for poultry and animal meat that can soften the meat quality and improve the flavor.

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 game meat by treating it with an enzyme composition containing a protease, comprising:
The enzyme composition contains an alkaline protease derived from a bacterium belonging to the genus Bacillus and a protease derived from a fungus belonging to the genus Aspergillus,
The protease derived from a fungus belonging to the genus Aspergillus has both endo-type protease and exo-type protease activities at pH 8 and 10° C., has a total protease activity of 5,000 U/g or more, and accounts for the total protease activity. A method, wherein the exo-protease activity is 1.0% or more.
[2] The method of [1], wherein the Aspergillus fungus-derived protease is Aspergillus oryzae-derived protease.
[3] The method of [1] or [2], wherein the Bacillus bacterium-derived alkaline protease is Bacillus subtilis-derived alkaline protease.
[4] A method for producing poultry meat with improved meat quality and flavor, comprising:
treating said poultry meat with an enzyme composition comprising a protease;
The enzyme composition contains an alkaline protease derived from a bacterium belonging to the genus Bacillus and a protease derived from a fungus belonging to the genus Aspergillus,
The protease derived from a fungus belonging to the genus Aspergillus has both endo-type protease and exo-type protease activities at pH 8 and 10°C, has a total protease activity of 5,000 U/g or more, and accounts for the total protease activity. A production method, wherein the exo-protease activity is 1.0% or more.
[5] The production method of [4], wherein the Aspergillus fungus-derived protease is Aspergillus oryzae-derived protease.
[6] The production method of [4] or [5], wherein the Bacillus bacterium-derived alkaline protease is Bacillus subtilis-derived alkaline protease.

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

In the present invention, "meat quality" means quality of meat that affects palatability. "Meat quality" particularly includes "softness of meat", "ease of biting skin", "juiciness", "water retention", "appearance (appearance)" and the like.

"Flavor" means the aroma and taste of food and drink. "Flavor" particularly includes "umami" (richness), "salty taste", "meat aroma", "bitterness/off-taste", "sweetness", "sourness" and the like.

INDUSTRIAL APPLICABILITY According to the present invention, a protease treatment technique for poultry and animal meat is provided, which makes it possible to soften meat quality and improve flavor.

The results of taste sensor measurement of parent chicken thigh meat treated with the enzyme composition are shown (Test Example 3). The horizontal axis indicates the response value of AT0 (bitter/off-taste/early taste), and the vertical axis indicates the response value of AAE (umami/aftertaste).

Preferred embodiments for carrying out the present invention will be described below. It should be noted that the embodiments described below are examples of representative embodiments of the present invention, and the scope of the present invention should not be construed narrowly.

[Protease Derived from Fungi of the Genus Aspergillus]
The Aspergillus fungus-derived protease contained in the enzyme composition according to the present invention may be derived from Aspergillus fumigatus, Aspergillus fumigatus, Aspergillus melleus, Aspergillus melleus, and Aspergillus fumigatus. Proteases from Aspergillus niger, Aspergillus oryzae, and Aspergillus sojae.
As the Aspergillus fungus-derived protease, one kind of protease may be used, or two or more kinds of proteases may be used in combination.

The Aspergillus fungus-derived protease is preferably Aspergillus oryzae-derived protease.

The Aspergillus fungus-derived protease has both endo-type protease and exo-type protease activities at pH 8 and 10° C., has a total protease activity of 5,000 U/g or more, and has an exo-type protease activity that accounts for the total protease activity. A protease having an exo-type protease activity (exo-type protease activity ratio) of 1.0% or more is particularly preferred.
A more preferable Aspergillus fungus-derived protease has a total protease activity of 7,000 U/g or more, 8000 U/g or more, or 9000 U/g or more, and an exo-type protease activity ratio of 1.2% or more and 1.5%. or more, or 1.6% or more.

Measurement of the total protease activity can be performed by a conventionally known method, for example, according to the 1st report of "Method for Measuring Protease Activity" in "Voluntary Standards for Existing Additives (4th Edition)".
The measurement of exo-type protease activity (aminopeptidase activity) can also be performed by a conventionally known method, for example, it can be performed according to "Aminopeptidase activity measurement method" of "Voluntary Standards for Existing Additives (4th Edition)". can.

As the above-mentioned protease derived from fungi belonging to the genus Aspergillus, a commercially available product may be used, or an appropriately obtained protease may be used. Commercially available enzymes include, for example, Cochrase P from Mitsubishi Chemical Foods Corporation.

[Alkaline protease derived from bacteria belonging to the genus Bacillus]
The Bacillus bacterium-derived alkaline protease contained in the enzyme composition according to the present invention may be derived from any Bacillus bacterium, such as Bacillus amyloliquefaciens and Bacillus cereus. ), Bacillus clausii, Bacillus intermedius, Bacillus lentus, Bacillus licheniformis, Bacillus stearothermophilus, Bacillus stearothermophilus Alkaline proteases from Bacillus subtilis, and Bacillus thermoproteolyticus.
As the alkaline protease derived from bacteria belonging to the genus Bacillus, one type of protease may be used, or two or more types of proteases may be used in combination.

The alkaline protease derived from bacteria belonging to the genus Bacillus is preferably alkaline protease derived from Bacillus subtilis.

As the aforementioned alkaline protease derived from a bacterium belonging to the genus Bacillus, a commercially available product may be used, or an appropriately obtained product may be used. Commercially available enzymes include, for example:
Protin SD-AY10 (derived from Bacillus licheniformis; Amano Enzyme Co., Ltd.)
Delbolase (derived from Bacillus licheniformis; DSM Japan Co., Ltd.)
Esperase (derived from Bacillus sp.; Novozymes Japan K.K.)
Savinase (derived from Bacillus sp.; Novozymes Japan K.K.)
Evalase (derived from Bacillus sp.; Novozymes Japan K.K.)
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.)

Known techniques can be used to produce proteases derived from fungi belonging to the genus Aspergillus and alkaline proteases derived from bacteria belonging to the genus Bacillus. Specifically, it can be produced by culturing a protease-producing microorganism and recovering the protease from the culture. A protease-producing microorganism may be one that originally produces a protease or one that has been modified to produce a protease. Protease-producing microorganisms can be obtained, for example, by introducing a gene encoding the protease into the microorganism so that it can be expressed. Introduction of a gene can be achieved, for example, by introducing a vector carrying the same gene into a microorganism, or by introducing the gene onto the chromosome of the microorganism. Amino acid sequences of various proteases derived from bacteria belonging to the genus Bacillus or fungi belonging to the genus Aspergillus and nucleotide sequences of genes encoding them can be obtained from public databases such as NCBI (http://www.ncbi.nlm.nih.gov/). can be obtained. Culture conditions for the microorganism are not particularly limited as long as the microorganism can grow and protease can be produced. Microorganisms can be cultured, for example, under normal conditions for culturing microorganisms such as bacteria and fungi. The protease may be used as a purified product, or as a culture of a protease-producing microorganism, a culture supernatant separated from the culture, a bacterial cell isolated from the culture, a processed product of the bacterial cell, etc. may be used.
As the Aspergillus fungus-derived protease and the Bacillus bacterium-derived alkaline protease, homologues of the known proteases exemplified above may be used. Homologs are not particularly limited as long as they are found in fungi of the genus Aspergillus and bacteria of the genus Bacillus and have the desired protease activity.
As the Aspergillus fungus-derived protease and the Bacillus bacterium-derived alkaline protease, known proteases such as those exemplified above or artificial modifications of their homologues may be used. Artificial variants are not particularly limited as long as they have the desired protease activity.

[Other enzymes]
The enzyme composition according to the present invention may contain various proteases in addition to the Bacillus-derived alkaline protease and the Aspergillus-derived protease.

Additional proteases include, for example, entopeptidases, papain, bromelain, ficin, actinidin, subtilisin, thermolysin, trypsin, cathebsin, chymotrypsin, rennet, and the like. Examples of exopeptidase include leucine aminopeptidase, aminopeptidase M, carboxypeptidase A and Y, and the like.

The enzyme composition according to the present invention may contain various enzymes such as amylase, lipase, phospholipase, esterase, glutaminase, transglutaminase, oxidase and nuclease in addition to protease.
In addition, the enzyme composition according to the present invention may contain additives that are commonly added to foods.

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

[Wildlife meat]
Wildlife meat is not particularly limited, and may be, for example, livestock meat such as beef, pork, horse, sheep, wild boar, and deer, and poultry meat such as chicken, turkey, duck, and goose. INDUSTRIAL APPLICABILITY The present invention can be suitably applied to poultry meat such as parent birds (adult chickens), breeding chickens, parent pigs, dairy cows, and the like, which have a long breeding period and have hard flesh. The meat part is not particularly limited, but since there is a possibility that the effect of improving meat quality can be greatly obtained, hard proteins such as shins, shoulders, necks, tongues, cheeks, thighs, tails, legs, breast, chicken wings, etc. A part containing a large amount of water or a part with a large amount of moisture loss due to cooking is preferable.

[Protease treatment]
By treating poultry meat with an enzyme composition containing an alkaline protease derived from a bacterium belonging to the genus Bacillus and a protease derived from a fungus belonging to the genus Aspergillus, meat quality (particularly "softness") is improved, and treatment with each protease alone is improved. On the other hand, it is possible to obtain a synergistic flavor improvement effect by significantly improving the "umami" while suppressing the "bitterness/unpleasant taste".

The method of treating the 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 eaten. In general, methods such as immersing meat in a solution of the enzyme composition, injecting the solution into the meat, tumbling in the same solution, or sprinkling the powder or solution of the enzyme composition on the meat are adopted. be.
The alkaline protease derived from bacteria belonging to the genus Bacillus and the protease derived from fungus belonging to the genus Aspergillus are preferably applied to meat at the same time.

The treatment with the enzyme composition is preferably carried out by keeping the meat in contact with the enzyme under optimal enzyme conditions for a certain period of time so that the enzyme reaction proceeds sufficiently and the quality of the meat is maintained. .
The temperature is between 0 and 40°C, preferably between 0 and 25°C.
The pH is between 5 and 10, preferably between 7 and 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 processed, and the desired meat quality and flavor.
After enzymatic treatment, it is preferable to immediately heat-treat or freeze to stop the progress of the enzymatic reaction.

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

[Test Example 1: Sensory evaluation 1]
Parent chicken thigh meat was treated with the enzyme composition, and sensory evaluation of meat quality and flavor was performed.

(1) Enzyme As an alkaline protease derived from bacteria belonging to the genus Bacillus, commercially available enzyme AP (Orientase 22BF, HBI Co., Ltd.) derived from Bacillus subtilis was used.
Commercially available enzymes A, B, C, D, F, G, and H were used as proteases derived from fungi of the genus Aspergillus. As the enzyme A, coclase P from Mitsubishi Chemical Foods Co., Ltd., which is an alkaline protease derived from Aspergillus oryzae, was used.
As enzyme E, commercially available papain (purified papain, Mitsubishi Chemical Foods Co., Ltd.) was used.

(2) Activity measurement Total protease activity and aminopeptidase activity (exo-type protease activity) of enzymes A to D and F to H were measured.
The measurement of the total protease activity was carried out according to the first report of "Protease activity measurement method" of "Voluntary Standards for Existing Additives (4th Edition)".
The aminopeptidase activity was measured in accordance with the "Method for measuring aminopeptidase activity" of "Voluntary 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 solution).

Table 1 shows the measurement results. As shown in the table, enzymes A, F, and G had a total protease activity of 5,000 U/g or more, and the exo-type protease activity accounted for 1.0% or more of the total protease activity.

(3) Enzyme treatment The thawed parent chicken thigh meat was immersed in a pH adjuster solution with or without an enzyme added, and sealed in a vacuum pack. The volume of the pH adjuster solution was 30% (w/w) of the parent chicken thigh meat. The pH adjuster solution used was obtained by dissolving the pH adjuster whose composition is shown in Table 2 in water at 3.5% (w/v). The concentrations of enzymes added to the pH adjuster solution were 0.05% (w/v) for enzyme AP and 0.1% (w/v) for enzymes A to H.
The treatment was performed at 4°C using a vacuum tumbling device (4 sets of 25 minutes of rotation (vacuum condition) and 5 minutes of rest (vacuum open)).
Baking was performed for 8 minutes at 250°C (40% steam output) using a steam convection oven.

(4) Sensory evaluation The "softness" and "umami" of the sample treated with the enzyme-added pH adjuster solution were evaluated based on the sample treated with the enzyme-free pH adjuster solution. If better than the standard, it was evaluated as ◯, if it was equivalent, it was evaluated as Δ, and if it was inferior, it was evaluated as ×. Table 3 shows the results.

N.T.：Not Tested.

NT: Not Tested.

Samples treated with Enzyme AP alone had better "softness" than the reference, but equal "umami" to the reference. In addition, the samples treated with Enzyme A alone were comparable to the standard in terms of "softness" and "umami". However, in the sample treated with Enzyme AP in combination with Enzyme A, both "softness" and "umami" were better than the standard.
It was clarified that the combined use of Enzyme AP and Enzyme A can improve the umami taste compared to the use of each enzyme alone.
Similarly, it was confirmed that both the "softness" and "umami" of Enzymes F and G in combination with Enzyme AP were better than the standard.

Concerning Enzymes B, C, D and H, no improvement in "umami" was observed by combined use with Enzyme AP as compared to single treatment with Enzyme AP.
Regarding Enzymes H and E, samples treated with Enzyme AP alone had the same level of umami as the standard, but those treated with Enzyme AP in combination were inferior to the standard, and the flavor was lower than that of the combination with Enzyme AP. There was a tendency to

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

Parent chicken thigh meat that had been subjected to enzyme treatment (enzyme AP alone or enzyme AP and enzyme A in combination) in the same manner as in Test Example 1 was cut into pieces of about 3 cm square and subjected to evaluation.
"Tenderness of meat part", "umami", "easy to bite through skin", "salty taste", "overall meat flavor" and "preferred taste" of samples treated with enzyme-added pH adjuster solutions. (Comprehensive evaluation)” was evaluated based on the sample treated with the pH adjuster solution to which no enzyme was added. Evaluation was performed by a panel of 10 people.
For each evaluation item, 3 is equivalent to the standard, 4 is slightly better than the standard, 5 is better than the standard, 2 is slightly inferior to the standard, and 1 is inferior to the standard.

Table 4 shows the average score of 10 panelists.

Samples treated with Enzyme AP alone tended to improve "softness of meat" and "umami" compared to the standard. Samples treated with a combination of Enzyme AP and Enzyme A showed further improvement in "tenderness of meat" and "umami", and also improved in "preferability (overall evaluation)" compared to the standard. .

[Test Example 3: Taste sensor measurement]
Taste sensor measurement of parent chicken thigh meat treated with the enzyme composition was entrusted to an external testing institution.

Add 5 times the amount of water to parent chicken thigh meat (baked meat) that has been subjected to enzyme treatment (enzyme AP alone, or enzyme AP and enzyme A in combination) in the same manner as in Test Example 1, and grind to remove the solid content. It was used for the measurement except

The measurement results are shown in FIG. 1 and Table 5. In FIG. 1, the horizontal axis indicates the response value of AT0 (bitter/off-taste/early taste), and the vertical axis indicates the response value of AAE (umami/aftertaste). The response value was corrected so that the response value of the sample treated with the pH adjuster solution to which no enzyme was added was 0. A numerical difference of 1 in the response value corresponds to a 20% difference in component concentration between "bitterness/unpleasant taste/first taste" or "umami/aftertaste" and is considered sufficient to give humans different taste sensations.

The same effect was shown by the response value of the sensor for the combined treatment with enzyme AP and enzyme A, which had been confirmed by the sensory evaluation of Test Examples 1 and 2 to improve "umami and aftertaste." The AAE response values for treatment with Enzyme AP alone and Enzyme A alone were 0.65 and 0.08, respectively, and the AAE response value for treatment with Enzyme AP and Enzyme A in combination was 1.9. The AAE response value for treatment with Enzyme AP and Enzyme A combined (1.9) is significantly higher than the sum of AAE response values for treatment with Enzyme AP alone and Enzyme A alone (0.73), and is 1 or more than the sum. Improved.

Furthermore, the combined treatment with Enzyme AP and Enzyme A also exhibited the effect of suppressing the "bitterness/miscellaneous taste" caused by the treatment with Enzyme AP alone. The AT0 response value was 0.98 when treated with enzyme AP alone, whereas the AT0 response value when treated with both enzyme AP and enzyme A was suppressed to 0.23.
It was shown that the combined use of Enzyme AP and Enzyme A not only improved umami and aftertaste, but also suppressed bitterness, undesired taste, and initial taste.

On the other hand, for Enzymes B, C and D, no improvement in "umami" was observed when used in combination with Enzyme AP.
Regarding Enzyme E, "Umami" was the same between single treatment with Enzyme AP and combined treatment with Enzyme AP.

Claims (2)

A method for improving the meat quality and flavor of game meat by treating the game game meat with an enzyme composition comprising a protease, the method comprising:
The enzyme composition contains an alkaline protease derived from a bacterium belonging to the genus Bacillus and a protease derived from a fungus belonging to the genus Aspergillus,
The protease derived from a fungus belonging to the genus Aspergillus has both endo-type protease and exo-type protease activities at pH 8 and 10° C., has a total protease activity of 5,000 U/g or more, and accounts for the total protease activity. A method, wherein the exo-protease activity is 1.0% or more. An enzyme composition for improving the quality and flavor of poultry meat, comprising a protease,
containing an alkaline protease derived from a bacterium belonging to the genus Bacillus and a protease derived from a fungus belonging to the genus Aspergillus,
The protease derived from a fungus belonging to the genus Aspergillus has both endo-type protease and exo-type protease activities at pH 8 and 10° C., has a total protease activity of 5,000 U/g or more, and accounts for the total protease activity. An enzyme composition having an exo-protease activity of 1.0% or more.

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