JP7707565B2 - Method and enzyme composition for improving meat quality and flavor - Google Patents

Description

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

Previously, attempts have been made to improve the quality and flavor of poultry and animal meat by treating the meat with an enzyme composition containing protease (protease treatment) (see Patent Document 1). For example, meat from chickens (parent birds) that have finished laying eggs and are used as minced meat because the meat is tough, can be treated with protease to make it softer, which is expected to expand its uses.

In addition, protease treatment is expected to increase the amount of amino acids contained in meat, improving its flavor (especially its umami taste). However, it is known that depending on the type of protease used, an unpleasant odor or bitter taste may result from decomposition products (see Patent Document 2).

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

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

In order to solve the above problems, the present invention provides the following [1] to [9].
[1] A method for improving the quality and flavor of poultry or game meat by treating the poultry or game meat with an enzyme composition containing a protease, comprising:
The enzyme composition contains an alkaline protease derived from a bacterium of the genus Bacillus and a protease derived from a fungus of the genus Aspergillus,
The method, wherein the protease derived from a fungus of the genus Aspergillus has both endoprotease and exoprotease activity under conditions of pH 8 and 10°C, has a total protease activity of 5,000 U/g or more, and has exoprotease activity accounting for 1.0% or more of the total protease activity.
[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 poultry and animal meat having improved meat quality and flavor, comprising:
treating the poultry or animal meat with an enzyme composition comprising a protease;
The enzyme composition contains an alkaline protease derived from a bacterium of the genus Bacillus and a protease derived from a fungus of the genus Aspergillus,
The protease derived from a fungus of the genus Aspergillus has both endoprotease and exoprotease activity under conditions of pH 8 and 10°C, has a total protease activity of 5,000 U/g or more, and has exoprotease activity accounting for 1.0% or more of the total protease activity.
[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 method according to [4] or [5], wherein the alkaline protease derived from a bacillus bacterium is an alkaline protease derived from Bacillus subtilis.

[7] An enzyme composition for improving the quality and flavor of poultry and animal meat, comprising a protease,
The alkaline protease is derived from a bacillus bacterium and a protease is derived from an Aspergillus fungus,
An enzyme composition, wherein the protease derived from the Aspergillus fungus has both endoprotease and exoprotease activity under conditions of pH 8 and 10°C, the total protease activity is 5,000 U/g or more, and the exoprotease activity accounts for 1.0% or more of the total protease activity.
[8] The enzyme composition according to [7], wherein the protease derived from a 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.

In the present invention, "meat quality" refers to the quality of meat that affects its palatability. "Meat quality" particularly includes "tenderness of the meat," "ease of biting the skin," "juiciness," "water retention," "appearance (appearance)," etc.

"Flavor" refers to the aroma and taste of food and drink. "Flavor" specifically includes "umami" (richness), "salty," "meaty aroma," "bitterness/off-flavors," "sweetness," "sourness," etc.

The present invention provides a protease treatment technology for poultry and animal meat that can soften the meat and improve its flavor.

1 shows the results of taste sensor measurement of chicken thigh meat treated with an enzyme composition (Test Example 3). The horizontal axis shows the response value of AT0 (bitterness/off-flavor, initial taste), and the vertical axis shows the response value of AAE (umami, aftertaste).

The following describes a preferred embodiment of the present invention. Note that the embodiment described below is an example of a typical embodiment of the present invention, and is not intended to narrow the scope of the present invention.

[Protease derived from Aspergillus fungi]
The protease derived from a fungus of the genus Aspergillus contained in the enzyme composition of the present invention may be any protease derived from a fungus of the genus Aspergillus, and examples thereof include proteases derived from Aspergillus fumigatus, Aspergillus melleus, Aspergillus niger, Aspergillus oryzae, and Aspergillus sojae.
The protease derived from a fungus of the genus Aspergillus may be used alone or in combination of two or more kinds.

The protease derived from the Aspergillus fungus is preferably protease derived from Aspergillus oryzae.

It is particularly preferred that the protease derived from a fungus of the genus Aspergillus has both endoprotease and exoprotease activity under conditions of pH 8 and 10°C, has a total protease activity of 5,000 U/g or more, and has exoprotease activity (exoprotease activity ratio) of 1.0% or more in the total protease activity.
More preferably, the protease derived from an Aspergillus fungus 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 exoprotease activity ratio of 1.2% or more, 1.5% or more, or 1.6% or more.

The total protease activity can be measured by a conventionally known method, for example, according to the "Protease Activity Measurement Method" Report 1 of the "Voluntary Standards for Existing Food Additives (4th Edition)."
Exoprotease activity (aminopeptidase activity) can also be measured by a conventionally known method, for example, in accordance with the "Aminopeptidase Activity Measurement Method" in the "Voluntary Standards for Existing Food Additives (4th Edition)."

The above-mentioned protease derived from the fungus of the genus Aspergillus may be a commercially available product, or may be obtained by appropriate production. An example of a commercially available enzyme is Coclase P from Mitsubishi Chemical Foods Corporation.

[Alkaline protease derived from Bacillus bacteria]
The alkaline protease derived from a bacterium of the genus Bacillus contained in the enzyme composition according to the present invention may be any alkaline protease derived from a bacterium of the genus Bacillus, and examples thereof 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 a bacterium of the genus Bacillus may be used either alone or in combination of two or more kinds.

The alkaline protease derived from a bacillus bacterium is preferably an alkaline protease derived from Bacillus subtilis.

The alkaline protease derived from the Bacillus bacterium may be a commercially available product or may be obtained by appropriate production. Examples of commercially available enzymes include the following:
Protin SD-AY10 (derived from Bacillus licheniformis; Amano Enzyme Co., Ltd.)
Delborase (derived from Bacillus licheniformis; DSM Japan, Ltd.)
Esperase (derived from Bacillus sp.; Novozymes Japan, Ltd.)
Savinase (derived from Bacillus sp.; Novozymes Japan, Ltd.)
Evalase (derived from Bacillus sp.; Novozymes Japan, Ltd.)
Alcalase (derived from Bacillus licheniformis; Novozymes Japan, 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; HIV, Inc.)

A known method can be used to produce a protease derived from a fungus of the genus Aspergillus and an alkaline protease derived from a bacterium of the genus Bacillus. Specifically, the protease can be produced by culturing a microorganism that produces a protease and recovering the protease from the culture. The microorganism that produces the protease may be one that inherently produces a protease, or may be one that has been modified to produce a protease. The microorganism that produces the protease can be obtained, for example, by introducing a gene that codes for the protease into the microorganism so that the gene can be expressed. The introduction of the gene can be achieved, for example, by introducing a vector carrying the gene into the microorganism or by introducing the gene into the chromosome of the microorganism. The amino acid sequences of various proteases derived from bacteria of the genus Bacillus or fungi of the genus Aspergillus and the nucleotide sequences of the genes that code for them can be obtained, for example, from public databases such as NCBI (http://www.ncbi.nlm.nih.gov/). The culture conditions for the microorganism are not particularly limited as long as the microorganism can grow and the protease can be produced. The microorganism can be cultured under normal conditions for culturing microorganisms such as bacteria and fungi. The protease may be used as a purified product, or may be used as a culture of a microorganism producing the protease, a culture supernatant separated from the culture, bacterial cells separated from the culture, or a treated product of the bacterial cells.
As the protease derived from a fungus of the genus Aspergillus and the alkaline protease derived from a bacterium of the genus Bacillus, a homologue of the known protease exemplified above may be used. The homologue is not particularly limited as long as it is found in a fungus of the genus Aspergillus or a bacterium of the genus Bacillus and has the desired protease activity.
As the protease derived from a fungus of the genus Aspergillus and the alkaline protease derived from a bacterium of the genus Bacillus, known proteases such as those exemplified above or artificially modified versions of their homologues may be used. The artificially modified versions 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 types of proteases in addition to the alkaline protease derived from a bacterium of the genus Bacillus and the protease derived from a fungus of the genus Aspergillus.

Additional proteases include, for example, entopeptidases such as papain, bromelain, ficin, actinidin, subtilisin, thermolysin, trypsin, cathepsin, chymotrypsin, and rennet. Also, exopeptidases include leucine aminopeptidase, aminopeptidase M, and carboxypeptidase A and Y.

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.
Furthermore, the enzyme composition according to the present invention may contain additives that are usually incorporated into foods.

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

[Bird and animal meat]
The poultry meat is not particularly limited, and may be, for example, meat from livestock such as beef, pork, horse, sheep, boar, deer, etc., or poultry meat such as chicken, turkey, duck, goose, etc. The present invention is suitably applicable to poultry meat from parent birds (adult chicken meat), breeding chickens, parent pigs, dairy cows, etc., which have been raised for a long time and have tough meat quality. The part of the meat is also not particularly limited, but parts that contain a lot of hard protein, such as the shin, shoulder, neck, tongue, cheek, thigh, tail, foot, breast, and wing, and parts that lose a lot of moisture when cooked are preferred, as they may have a significant effect on improving meat quality.

[Protease treatment]
By treating poultry and game meat with an enzyme composition containing an alkaline protease derived from a Bacillus bacterium and a protease derived from an Aspergillus fungus, it is possible to improve the meat quality (particularly "tenderness") and obtain a synergistic flavor improvement effect by significantly improving the "umami" flavor while suppressing the "bitterness/off-flavors" compared to treatment with either protease alone.

The method of treating the meat with the enzyme composition is not particularly limited, so long as the enzyme acts effectively on the meat from immediately after slaughter until the meat is eaten. Generally, methods such as immersing the meat in a solution of the enzyme composition, injecting the meat with the solution, tumbling in the solution, or sprinkling the meat with a powder or solution of the enzyme composition are used.
It is preferable to allow the alkaline protease derived from a bacterium of the genus Bacillus and the protease derived from a fungus of the genus Aspergillus to act on meat at the same time, but the desired effect may be obtained even if they are allowed to act at different times.

The treatment with the enzyme composition is preferably carried out by maintaining the meat in contact with the enzyme for a certain period of time under optimal conditions for 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 5-10, preferably 7-10.
The time is from 0.5 to 48 hours, preferably from 0.5 to 24 hours.
However, conditions such as temperature, pH and time can be appropriately set depending on 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 immediately carry out a heat treatment or freeze the product to stop the progress of the enzyme reaction.

The amount of the enzyme composition used can be appropriately determined depending on the type, shape and size of the meat to be treated, and the desired meat quality and flavor.
The amount of the protease derived from a fungus of the genus Aspergillus used, for example, when meat is immersed in an enzyme composition solution, can be 0.001-1% (w/v), preferably 0.01-0.5% (w/v), and more preferably 0.05-0.1% (w/v).
The amount of alkaline protease derived from a bacterium of the genus Bacillus used, for example, when meat is immersed in the enzyme composition solution, can be 0.001-1% (w/v), preferably 0.005-0.5% (w/v), and more preferably 0.01-0.05% (w/v).

[Test Example 1: Sensory Evaluation 1]
The thigh meat of parent chickens was treated with the enzyme composition, and the meat quality and flavor were subjected to a sensory evaluation.

(1) Enzyme As the alkaline protease derived from a bacillus bacterium, a commercially available enzyme AP (Orientase 22BF, HIV, Inc.) derived from Bacillus subtilis was used.
The proteases derived from Aspergillus fungi used were commercially available enzymes A, B, C, D, F, G, and H. As enzyme A, Coclase P (Mitsubishi Chemical Foods Corporation), an alkaline protease derived from Aspergillus oryzae, was used.
As the enzyme E, commercially available papain (purified papain, Mitsubishi Chemical Foods Corporation) was used.

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

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

(3) Enzyme treatment Thawed parent chicken thigh meat was immersed in a pH adjuster solution with or without the addition of enzymes, and then vacuum packed. The volume of the pH adjuster solution was 30% (w/w) of the parent chicken thigh meat. The pH adjuster solution was prepared by dissolving the pH adjuster, whose composition is shown in Table 2, in water at 3.5% (w/v). The concentration of enzymes added to the pH adjuster solution was 0.05% (w/v) for Enzyme AP and 0.1% (w/v) for Enzymes A to H.
The treatment was carried out at 4°C using a vacuum tumbling apparatus (four sets of 25 minutes of rotation (vacuum condition) followed by 5 minutes of rest (vacuum release)).
Baking was carried out for 8 minutes at 250°C (steam output 40%) using a steam convection oven.

(4) Sensory evaluation The "softness" and "umami" of the samples treated with the enzyme-added pH adjuster solution were evaluated based on the standard of the sample treated with the enzyme-free pH adjuster solution. If it was better than the standard, it was marked with ◯, if it was the same, it was marked with △, and if it was worse, it was marked with ×. The results are shown in Table 3.

NT: Not tested.

In the sample treated with Enzyme AP alone, the "softness" was better than the standard, but the "umami" was the same as the standard. In the sample treated with Enzyme A alone, the "softness" and "umami" were the same as the standard. However, in the sample treated with Enzyme AP in combination with Enzyme A, both the "softness" and "umami" were better than the standard.
It was revealed that the combination of Enzyme AP and Enzyme A resulted in an improvement in umami flavor compared to the use of either enzyme alone.
Similarly, it was confirmed that when Enzymes F and G were used in combination with Enzyme AP, both "softness" and "flavor" were better than the standard.

For Enzymes B, C, D, and H, the combined use with Enzyme AP did not result in any improvement in "umami" flavor compared to treatment with Enzyme AP alone.
For enzymes H and E, the "umami" taste of the sample treated with enzyme AP alone was equivalent to the standard, but when treated in combination with enzyme AP it was inferior to the standard, and there was a tendency for the flavor to be worse than when treated in combination with enzyme AP.

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

The thigh meat of parent chickens that had been subjected to an enzyme treatment (enzyme AP alone or a combination of enzyme AP and enzyme A) in the same manner as in Test Example 1 was cut into pieces of approximately 3 cm square and subjected to evaluation.
The samples treated with the pH adjuster solution containing the enzyme were evaluated for "tenderness of meat,""umami,""ease of biting the skin,""saltiness,""overall meat aroma," and "likability (overall evaluation)," relative to the samples treated with the pH adjuster solution without the enzyme. The evaluation was performed by a panel of 10 people.
For each evaluation item, a score of 3 was assigned to indicate the same as the standard, 4 to indicate slightly better than the standard, 5 to indicate superior to the standard, 2 to indicate slightly worse than the standard, and 1 to indicate inferior to the standard.

The average scores of the 10 panelists are shown in Table 4.

In samples treated with Enzyme AP alone, there was a tendency for the "tenderness of the meat" and "flavor" to be improved compared to the standard. In samples treated with Enzyme AP in combination with Enzyme A, there was a further improvement in the "tenderness of the meat" and "flavor," and the "liking (overall evaluation)" was also improved compared to the standard.

[Test Example 3: Taste Sensor Measurement]
Taste sensor measurements of the chicken thigh meat treated with the enzyme composition were outsourced to an external testing organization.

Chicken thigh meat (grilled meat) that had been subjected to enzyme treatment (enzyme AP alone or a combination of enzyme AP and enzyme A) in the same manner as in Test Example 1 was mixed with five times the amount of water, pulverized, and the solids were removed before being subjected to measurement.

The measurement results are shown in Figure 1 and Table 5. In Figure 1, the horizontal axis shows the response value for AT0 (bitterness/off-flavors/first taste), and the vertical axis shows the response value for AAE (umami/aftertaste). The response values were corrected so that the response value of the sample treated with the pH adjuster solution without added enzyme was 0. A numerical difference in response values of 1 corresponds to a 20% difference in component concentration for "bitterness/off-flavors/first taste" or "umami/aftertaste", which is considered sufficient to give humans a different taste sensation.

The sensor response value of the treatment using enzyme AP in combination with enzyme A, which was confirmed to have an effect of improving "umami taste and aftertaste" in the sensory evaluation of Test Examples 1 and 2, also demonstrated the same effect. 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 in combination with enzyme A was 1.9. The AAE response value for treatment with enzyme AP in combination with enzyme A (1.9) was significantly higher than the sum of the AAE response values for treatment with enzyme AP alone and enzyme A alone (0.73), and was an improvement of more than 1 above that sum.

Furthermore, the combined treatment with Enzyme AP and Enzyme A also showed the effect of suppressing the "bitterness/off-flavor" that occurred when Enzyme AP was used alone. The AT0 response value when treated with Enzyme AP alone was 0.98, whereas the AT0 response value when treated with Enzyme AP and Enzyme A in combination was suppressed to 0.23.
It was shown that the combined use of Enzyme A and Enzyme AP can improve "umami and aftertaste" while suppressing "bitterness/off-flavors and first tastes."

On the other hand, for Enzymes B, C, and D, no improvement in "umami flavor" was observed when used in combination with Enzyme AP.
For Enzyme E, the "umami" taste was equivalent between the treatment with Enzyme AP alone and the treatment with Enzyme AP in combination, but the "bitterness/off-flavor" actually increased in the treatment with Enzyme AP in combination.

Claims (2)

1. A method for improving the quality and flavor of poultry or game meat by treating the poultry or game meat with an enzyme composition containing a protease, comprising:
The enzyme composition comprises an alkaline protease derived from Bacillus subtilis and a protease derived from a fungus of the genus Aspergillus,
The method, wherein the protease derived from a fungus of the genus Aspergillus has both endoprotease and exoprotease activity under conditions of pH 8 and 10°C, has a total protease activity of 5,000 U/g or more, and has exoprotease activity accounting for 1.0% or more of the total protease activity. An enzyme composition for improving meat quality and flavor of poultry and game meat, comprising a protease,
The present invention comprises an alkaline protease derived from Bacillus subtilis and a protease derived from Aspergillus fungus,
An enzyme composition, wherein the protease derived from the Aspergillus fungus has both endoprotease and exoprotease activity under conditions of pH 8 and 10°C, the total protease activity is 5,000 U/g or more, and the exoprotease activity accounts for 1.0% or more of the total protease activity.