WO2019198973A1 - Debaryomyces hansenii smfm201707 strain for tenderizing meat and improving flavor and method for tenderizing meat and improving flavor of meat by using same - Google Patents

Abstract

The present invention relates to a Debaryomyces hansenii smfm201707 strain having the function of tenderizing meats and improving flavors of the meats and a method for tenderizing meats and improving flavors of the meats by using same. Having the function of tenderizing meats and improving flavors of the meats, the novel strain Debaryomyces hansenii smfm201707 according to the present invention can be easily used for meat aging. In addition, the method for tenderizing meats and improving flavors of the meats according to present invention utilizes the novel strain in meat aging, thus being able not only to increase the tenderness of meats, but also to prepare aged meats having excellent tastes and flavor.

Description

Devariomyces Hanseni SMFM201707 strain for meat softening and flavor enhancement and method for meat softening and flavor enhancement using the same

The present invention relates to a strain of Debaryomyces hansenii smfm201707 having a function of meat softening and flavor enhancement, and a method of softening meat and enhancing flavor using the same.

As the dietary lifestyle is westernized, meat consumption has increased greatly, and in recent years, meat has been consumed in the form of roasted foods such as sirloin and pork belly, using thinly sliced meat. The method is also changing. Accordingly, it is known that consumption of fresh meat is much higher than consumption of processed meat, unlike in the West, where consumption of processed meat is generalized in Korea.

Fresh meat refers to meat immediately after slaughtering the meat obtained from freshly slaughtered cattle at room temperature or low temperature, and consumed within a short time. Fresh meat has the advantage that the meat is fresh, but the meat is fresh for a certain period after the slaughter. Post-stiffness of the muscles occur, such as the meat is tough, the taste, flavor, texture, etc. are disadvantageous. In particular, in the case of frozen meat stored for a long time, there is a problem in that the hygiene problems such as food poisoning, as well as deterioration of quality due to breeding of bacteria because of poor freezing during transportation or purchase.

In order to solve the above problems, in recent years, the demand for aging meat matured through the action of microorganisms or enzymes or interactions between components by leaving the meat for a while under certain conditions, the content of free fatty acids This increases the flavor, the inherent odor of the animal can be removed to satisfy the taste of the intake, and has the advantage of easy digestion compared to fresh meat.

Conventionally, ripening meat is manufactured by varying the ripening period or temperature as a method for ripening meat, using a dry or wet method, or a method of treating proteolytic enzymes, but there is a study on a processing method of ripening meat. Inadequately, most of the existing aging method is still in the range of daily use.

Therefore, there is a need for a study on a method of aging meat to be able to provide aging meat having a high quality meat through aging of low quality meat.

The inventors of the present invention confirmed that the yeast-derived new strain, Debaryomyces hansenii smfm201707, has excellent meat softening and flavor enhancement ability, and can produce mature meat having high quality properties by aging meat. .

Therefore, in the present invention, it is intended to provide a description of the devarionomyces hanseni smfm201707 strain for meat softening and flavor enhancement and a method for meat softening and flavor enhancement using the same.

In order to achieve the above technical problem, the present invention provides a strain of Debariomyces hanseni smfm201707 ( Debaryomyces hansenii smfm201707) having a meat softening and flavor enhancement function, and the strain may be deposited with accession number KCTC18601P. .

In addition, the meat may be beef or pork.

In addition, the present invention provides a composition for softening meat and flavor enhancement comprising the strain described above or a culture solution of the strain.

In addition, the present invention, (a) Devarionomyces Hanseni smfm201707 ( Debaryomyces having a meat softening and flavor enhancement function hansenii smfm201707) treating the meat meat with a strain or a composition for enhancing meat flavor and flavor enhancement comprising the culture medium of the strain; And (b) aging the meat treated with the composition; provides a method for improving meat softening and flavor comprising.

In addition, in the step (b), it is possible to mature the meat meat by dry aging method (dry aging), it is possible to mature the meat meat for 1 to 50 days, supplying a wind speed of more than 0 and 20 m / s or less Under these conditions, the meat can be aged.

Mycobacteria strain Debaryomyces according to the invention hansenii smfm201707 has softening and flavor enhancement functions and can be easily used for ripening of meat.

In addition, according to the meat softening and flavor enhancement method according to the present invention, by utilizing the new strain for the ripening of meat can not only increase the year of the meat, but also can produce mature meat with excellent taste and aroma.

Figure 1 is a new strain Debaryomyces in accordance with the present invention deposit of Hansenii smfm201707 (KCTC18601P).

Figure 2 is a new strain Debaryomyces in accordance with the present invention In order to confirm the proteolytic activity of hansenii smfm201707, the strain milk was inoculated with the new strain at concentrations of (a) 0.5%, (b) 1%, (c) 2%, (d) 4%, and (e) 7%. It is the actual image of the result of proteolytic activity analysis by plate culture.

Figure 3 is a new strain Debaryomyces in accordance with the present invention The microstructural changes in meat after 28 days of dry maturation of the group inoculated with hansenii smfm201707 and the group not inoculated (control) were observed by transmission electron microscopy (TEM).

Hereinafter, the present invention will be described in detail.

The present invention is a novel strain that can affect the meat and increase the year of the meat when it is processed and matured in meat, as well as a novel strain that can provide mature meat with excellent palatability by increasing the taste and flavor. Debariomyces Hanseni smfm201707 ( Debaryomyces for meat aging with flavor enhancement function hansenii smfm201707) strain.

In particular, the Debaryomyces hansenii smfm201707 strain analyzed Debaryomyces as a result of analyzing 23s rRNA The hansenii NRRL Y-7426 showed a 95% agreement with the 1337-bp sequence of the 28s rRNA strain.

Accordingly, the new strain Debaryomyces according to the present invention Hansenii smfm201707 strain was deposited with the Korea Biotechnology Research Institute Biological Resource Center in Yuseong-gu, Daejeon, Korea on August 23, 2017, and was given the accession number KCTC18601P on August 30, 2017, and the new strain Debaryomyces. hansenii smfm201707 strain comprises 26S rRNA comprising the nucleotide sequence of SEQ ID NO: 1 (see FIG. 1).

[SEQ ID NO 1]

Debaryomyces hansenii smfm201707 strain can ripen all types of consumable meats in the market such as fresh meat, chilled meat or freezing meat to increase the year of meat, and also to improve the taste and flavor to produce mature meat with excellent taste. can do.

In particular, the strain may be used to soften various meats derived from birds such as chickens, as well as terrestrial animals such as cattle, pigs, horses, sheep, goats, rabbits, and the like, and to enhance flavor (flavor and flavor). And, preferably, can be aged meat, including beef or pork, more preferably can be utilized for aging beef.

On the other hand, the present invention provides a composition for softening the meat and / or flavor enhancement comprising the strain or its culture.

The composition may be formulated for the purpose of softening the meat softening and enhancing the flavor through conventional methods, for example, may be formulated in the form of a solution, powder or granules and the like, without being limited thereto. It can be formulated to have.

In addition, the composition may be matured by coating or applying to the meat to be subjected to aging, for this purpose, immersing the meat in a solution containing the strain or its culture, or the formulation of a solution, powder, granules, etc. The meat can be treated by coating or applying (eg spraying, misting, powder spraying, granulating spraying, etc.).

In addition, the composition may be processed before aging of the meat or after a certain point after aging, the amount of use and the concentration of the strain can be appropriately adjusted according to the site, weight, ripening place, etc. of the meat to be treated.

Mycobacteria strain Debaryomyces according to the invention as described above hansenii smfm201707 has a softening and flavor enhancement function, which can be easily utilized for the aging of various animal-derived meats.

On the other hand, the present invention, (a) a step for treating meat meat softening and flavor enhancement composition comprising a culture solution of Debaryomyces hansenii smfm201707 strain or culture medium having a meat softening and flavor enhancement function to meat; And (b) aging the meat treated with the composition; provides a method for improving meat softening and flavor comprising.

The step (a) is a step of treating the meat meat softening and flavor enhancing composition comprising a culture solution of Debaryomyces hansenii smfm201707 strain or a culture medium having a meat softening and flavor enhancement function to the meat.

The composition may be formulated for meat softening and / or flavor enhancement in a conventional manner, and may be formulated in the form of a solution, powder or granules, and the like, without being limited thereto.

In this step, the composition may be coated or applied to the meat to be matured to mature the meat, for this purpose, immersed in the composition of the solution formulation containing the strain or its culture, or a solution, powder, The composition having a formulation such as granules can be treated to the meat by a method of coating or applying (for example, spraying, misting, powder spraying, granulating spraying, etc.).

In addition, in this step, the concentration of the new strain contained in the composition may be adjusted according to the site, weight, place of ripening, etc. of the meat to be aged.

The step (b) is a step of aging the meat treated with the composition, when aging the meat treated with the strain Debaryomyces hansenii smfm201707 strain, strain Debaryomyces hansenii smfm201707 affects the meat throughout the maturation period, the age of meat In addition to increasing the flavor of meat, it is possible to produce mature meat which can provide improved palatability.

In addition, the aging step (b) may ripen the meat treated with the composition in a variety of aging methods commonly used for the production of mature meat, in consideration of the age and flavor improvement of the meat, dry aging (dry aging) Is preferred.

In particular, in this step, the aging may be carried out for 1 to 50 days, it may be carried out under the condition that the wind speed of more than 0 20 m / s or less is supplied.

Preferably, the aging may be carried out for 5 to 28 days using a dry aging method of the condition that the wind is supplied at a wind speed of 2.5 to 5 m / s after treating the meat with the composition, according to the present invention Due to the proteolytic activity of the strain, it is possible to soften the meat, and to improve the taste and aroma, to produce dried mature meat of high preference.

According to the meat softening and flavor enhancement method according to the present invention as described above, mycobacteria strain Debaryomyces By using hansenii smfm201707 for the ripening of meat, it is possible not only to increase the year of meat but also to produce mature meat with excellent taste and aroma.

Hereinafter, the present invention will be described in more detail with reference to Examples.

The examples presented are merely illustrative of the invention and are not intended to limit the scope of the invention.

< EXAMPLE  1> Debaryomyces hansenii smfm201707  Isolation and Identification of Strains

(One) Aged  Sensory evaluation

Sensitive meat was prepared by ripening the subgrade parts of grade 3 beef cattle for 28 days using wet ripening method and dry ripening method. The dry ripening method was performed by supplying wind at a specific wind speed (0, 2.5, 5 m / s). It was.

The sensory preferences of the sensitive meats were evaluated by using the evaluation method and ranking method for the emotional meats. The sensory evaluation was performed by selecting 8 consumer panels. To this end, each sample of the sensitive meat was cut into a certain size (5.0 × 2.0 × 1.2 cm, length × width × height) and cooked until the core temperature in the sample reached 72 ℃, and then each panel was ingested. After ingestion, the consumer preference was evaluated by using a nine-point consumer test.

In addition, dry matured meats fed with a wind speed of 2.5 m / s were set as a standard control, so that the sensory panels were familiar with the smells and flavors of the dried matured meats, and then the smells of each sensitive meat sample. And the intensity of the flavor was ranked (ranking test) by rank (1 to 4), the results are shown in Table 1 below.

However, the display shown in Table 1 has the following meaning.

¹ Standard errors of mean (n = 32).

² Sensory panelists ranked the odor and flavor intensity of samples after comparing the sample of Dry 2.5 m / s.

^{a- c} Different letters within same row differ significantly (p <0.05).

Abbreviation: wet-aged for 28 days, Wet; dry-aged for 28 days with 0, 2.5, and 5 m / s of wind velocity, Dry 0, Dry 2.5, and Dry 5, respectively.

As a result of sensory evaluation of the sensitized meat samples after wet and dry ripening, as shown in Table 1, it was confirmed that dry ripened meat showed significantly higher palatability results than wet ripened meat.

In addition, as a result of ranking test of the intensity of flavor and flavor by rank (1 to 4), the aroma and flavor of the dry matured meat (experimental group in which yeast was distributed) was the strongest at 5 m / s wind speed. Also, dry ripening matured at 2.5 m / s wind speed showed high acceptability.

(2) next generation sequencing

Two dry aged meats (1 × 1 cm) dried and matured at each wind speed (0, 2.5, 5 m / s) for 28 days were used for next generation sequencing (NGS). DNA of the microorganism isolated from each sample was extracted and QC (Quality Control) was performed. Sequencing was performed to confirm the distribution of microorganisms with the extracted DNA, which utilized Illuina SBS technology. Statistical analysis was performed using Illumina Miseq's software using the analyzed basic data, and microbial distribution between the groups was compared, and the results are shown in Table 2 below.

As a result of NGS analysis, as shown in Table 2, almost no yeast strain of Debaryomyces Genus , a new strain of the present invention, was detected in the dried matured meat at 0 m / s. In yeast strains matured at wind speeds of 2.5 m / s and 5 m / s, yeast strains of Debaryomyces Genus were distributed at 15.9% and 14.7%, respectively.

(3) Isolation and Identification of Yeast Strains

In order to isolate yeast strains from dry ripened meat, PDA medium containing 10% tartaric acid, which is used for inhibiting the growth of various bacteria, was used. After incubation for 7 days, the grown microbial strain was isolated. To purely separate the first isolated microbial strain, a single colony was taken from each cluster with morphological differences in 1 loop with flame sterilized platinum and inoculated into 10 mL of sterile PDB medium until it grew cloudy. Incubated at 20 ° C. for 4 days. Subsequently, the culture solution was taken 1-loop using flame sterilized platinum teeth and streaked onto sterile PDA medium to incubate at 20 ° C. for 7 days to obtain a purely isolated microbial strain.

23s rRNA analysis was performed to identify purely isolated microorganisms, and primers for sequencing were forward primers, NS1 (5-GTA GTC ATA TGC TTG TCT C-3) and reverse primers. Reverse primer) NS24 (5-AAA CCT TGT TAC GAC TTT TA-3) was used.

As a result of analyzing 23s rRNA of the isolated strain, the isolated strain was Debaryomyces , a kind of yeast. hansenii ) strain. This yeast was found to be high at 15.9% and 14.7%, respectively, in the NGS analysis which confirmed the distribution of microorganisms in dry aged meats dried at both wind speeds of 2.5 m / s and 5 m / s.

In addition, the smell formed during the fermentation process, such as makgeolli in the colonies formed by smearing the sample for each maturation condition was confirmed to be a strain belonging to Table 3.

In addition, as a result of 26S rRNA analysis of the isolated yeast strain, it was confirmed that the strain having the nucleotide sequence shown in SEQ ID NO: 1 below, 95% of D. hansenii NRRL Y-7426 28s rRNA strain and 1337 bp The agreement rate is shown.

[SEQ ID NO 1]

Thus, the novel strain according to the invention Debaryomyces Named hansenii smfm201707, it was registered with the Korea Research Institute of Bioscience and Biotechnology on August 23, 2017, and was given the deposit number KCTC18601P on August 30, 2017 (see FIG. 1).

< EXAMPLE  2> Neomycology  Characterization

(One) Neomycology  Protein resolution measurement

In order to evaluate the protease secretion ability of the new strain, the milk medium (skim milk, Becton, Dickinson and Company) in PDA medium (0.5%, 1%, 2%, 4%, 7%) Each prepared medium was prepared, and the new strain was inoculated into the medium, and cultured at 20 ° C. for 7 days to compare the production of a clear zone and the growth rate of mycelia. It was confirmed, and the results are shown in FIG.

As shown in FIG. 2, a transparent ring was formed around the site of inoculation of the new strain culture medium, and the higher the inoculation concentration was, the larger the size of the clear zone was. As a result, the new strain contained the protein contained in skim milk. It has been thought to have proteolytic activity to decompose and exhibit the sensory characteristics peculiar to dry mature meat during the ripening period of meat by the proteolytic activity of the new strain.

(2) Analysis of Growth Characteristics of Strains by Drying Aging Period

Yeast strain ( Debaryomyces) , a new strain, is used to mature meat. hansenii smfm201707) alone was inoculated into a grade 3 beef cattle as spots, and the differences were observed for each maturation period, and inoculated by culturing as shown below.

1) Yeast strain sample preparation

Inoculate a single colony of yeast strain ( Debaryomyces hansenii ) in 10 mL PDB medium with flame sterilized platinum and incubate for 4 days at 20 ° C until the yeast strain grows thickly . It was added to 10 mL of PDB medium and then incubated at 20 ° C. for 4 days. Final concentration of 10 ⁶ CFU / mL The diluted yeast culture medium twice a step-by-step using PBS to the 9 mL and the yeast strain as a host microorganism Synergy the diluent finally (Debaryomyces hansenii smfm201707) was used as the test solution. At this time, the yeast count was determined by diluting the test stock solution step by step using 0.1% PW solution, dispensing in PDA medium, incubating for 7 days at 10 ℃.

2) Confirmation of yeast change by aging period

D. hansenii smfm201707 new strain prepared in the same manner as above was inoculated into beef cattle, and changes in the number of yeast cells of D. hansenii strain during the aging process for 28 days were observed (Unit: Log CFU / g, mean ±, The results are shown in Table 4. In this case, the experimental group treated with PBS in beef cattle was set as a control group to confirm the change in cell number.

However, the display shown in Table 4 has the following meaning.

¹⁾ Below detection limit

^ac The letters within the same row were significantly different (P <0.05).

^xz The letters within the same column were significantly different (P <0.05).

As shown in Table 4, the analysis of yeast cell number change in each beef cattle treated with the new strain (smfm201707) and the control group shows that the cell number does not increase in the beef treated with the control group, whereas In beef cattle treated with strain D. hansenii , the number of cells increased with the maturation period compared to 1.1 Log CFU / g. I could confirm that.

(3) As a new strain  Aged Aged  Meat evaluation

Shear force was heated in a hot bath until the core temperature of the aged meat sample is 72 ℃ and then allowed to cool at room temperature, was used to form a core (1.27 × 4 cm ² , diameter × length). Using a texture analyzer equipped with Warner-Bratzler shear (TA1, Lloyd Instruments Ltd., Fareham, UK) was measured perpendicular to the direction of muscle fibers until the sample was completely cut, the measurement conditions were probe test speed, 200 mm / min; Shear force was measured by setting the trigger load, 0.1 N, and the shear force measurement results are shown in Table 5 below.

However, the display shown in Table 5 has the following meaning.

¹ Standard error of means (n = 15), ² (n = 6).

^{a- d} The letters within the same row were significantly different (P <0.05).

^{x, y} The letters within the same column were significantly different (P <0.05).

As shown in Table 5, in all the inoculated groups inoculated with the new strain (smfm201707), the shear force decreased until 21 days of dry maturation period, tending to tender meat quality. In addition, the shear force value was confirmed to show a tendency to steadily decrease in the inoculation group inoculated with the new strain (smfm201707) of the present invention. On the other hand, the control group showed a tendency to increase the shear force at the time of 21 days of dry aging period, it can be confirmed that the new strain of the present invention has excellent aging activity because the aging efficiency does not decrease even during long-term aging. there was.

(4) As a new strain  Aged Aged  Meat evaluation

Mycobacteria Debaryomyces on Meat After 28 days of dry maturation of the group vaccinated with hansenii smfm201707 and the non-inoculated group (control), microstructural changes in meat were observed by transmission electron microscopy (TEM) images, and the results are shown in FIG. 3.

Specifically, after aging, samples (1 × 4 cm ² ) were taken, fixed with 2% paraformaldehyde and 2% glutaraldehyde in 0.05 M sodium cacodylate buffer (SCB), washed three times with 0.05 M SCB, and then added 0.05% with 1% osmium tetroxide. Fixed again with M SCB at 4 ° C. for 2 hours. The immobilized samples were washed with deionized distilled water and stained with 0.5% uranyl acetate for 30 minutes at 4 ° C. The samples were then dehydrated by increasing the ethanol concentration and then transitioned and infiltration into propylene oxide and spurr's resin. Thereafter, the mixture was polymerized in spurr's resin for 24 hours, sectioned before TEM analysis, and observed at 80 kV and 20,000 magnification.

Measurement results, unlike the control group, Debaryomyces In the group inoculated with hansenii smfm201707 was observed to the z-disk collapse which means the softening of the meat occurs, which Debaryomyces This may be due to the breakdown of muscle protein due to the proteolytic activity of hansenii smfm201707.

(5) As a new strain  Aged Aged  Reducing Sugar Content ( mM Change measurement

Changes in reducing sugar content according to the dry maturation period of the group inoculated with the new strain Debaryomyces hansenii smfm201707 and the group not inoculated (control) were shown in Table 6 below.

To quantify reducing sugar, first add 5 mL of 80% EtOH (50 ° C.) to 1 g of sample, homogenize and centrifuge (2,265 × g for 10 min; Continent 512R, Hanil Co. Ltd., Korea). It filtered with 1 filter paper. Next, 80% EtOH (50 ° C.) was added again, followed by centrifugation (2,265 × g for 10 min; Continent 512R, Hanil Co. Ltd. Korea), followed by filtration and nitrogen concentration. 2 mL of distilled water was added to the concentrated sample, followed by centrifugation (10,000 × g for 10 min; HM-150IV, Hanil Co. Ltd., Korea) to extract sugar components in meat. After adding 2 mL of the DNS reagent to 1 mL of the extracted sample, the mixture was heated and cooled for 15 minutes in a 90 ° C. constant temperature water bath, and then the absorbance was measured at 550 nm using a spectrophotometer to quantify the reducing sugar content. At this time, a standard curve was prepared using glucose (Sigma Aldrich, USA) as a standard material and converted to quantified reducing sugar content.

	Dry ripening period (days)				SEM 1
	0	14	21	28
Control	9.93 c	17.67 a	15.35 b	17.31 a	0.235
leaven	9.93 b	13.51 a	10.09 b	13.58 a	0.528

¹ Standard error of means (n = 15).

^{a- d} The letters within the same row were significantly different (P <0.05).

Measurement results, Debaryomyces on meat In the group vaccinated with hansenii smfm201707 and the group not inoculated (control), the reducing sugar content increased significantly until 14 days after dry ripening, decreased until 21 days, and increased again on 28 days. The yeast inoculation group showed a lower reducing sugar content than the control group, but it was confirmed that the reducing sugar increased through aging compared to before aging (day 0). In other words, Debaryomyces hansenii smfm201707 was found to have a low reducing sugar content because it was distributed in meat and consumed sugar, but had a positive effect on the year and flavor (data not shown).

(6) As a new strain  Aged Aged Free amino acids  content( mM Change measurement

Mycobacteria Debaryomyces on Meat The change of free amino acid content before and after 28 days of dry ripening of the group vaccinated with hansenii smfm201707 and the group not inoculated (control) was measured, and the results are shown in Table 7 below.

To determine the free amino acid content, 20 mL of 0.6 M perchloric acid was first added to 5 g of the sample, followed by homogenization and centrifugation (3,500 g, 20 min; Combi 514R, Hanil Co. Ltd). After centrifugation, Whatman No. After filtering the supernatant with 1 filter paper to adjust the pH to 7.0 using 6 N sodium hydroxide, the sample was lyophilized and reconstitution with 20 mM phosphate buffer to determine the free amino acid content by NMR analysis.

	Dry maturation control	Dry Aging Group
		Control	leaven
Asp	0.15 c	0.25 a	0.22 b
Glu	0.10 b	0.32 a	0.35 a

^{a, b} The letters within the same row were significantly different (P <0.05).

As a result, the main flavor-related substances, aspartic acid and glutamic acid, which have a significant effect on the formation of meat flavor, were found in both the group inoculated with Debaryomyces hansenii smfm201707 and the group not inoculated (control). It was found to increase significantly after dry ripening.

[Accession number]

Depositary: Korea Research Institute of Bioscience and Biotechnology

Accession number: KCTC18601P

Deposit Date: 20170830

The strain according to the present invention can not only increase the year of meat in the ripening process of meat, but also can provide an improved texture by increasing the flavor by removing off-flavor of meat, effectively utilized in the field of meat industry, including mature meat Can be.

Claims (8)

1. Debaryomyces hansenii smfm201707 strain having a meat softening and flavor enhancing function.
2. The method of claim 1,

   The strain is characterized in that deposited with the accession number KCTC18601P.
3. The method of claim 1,

   The meat is a strain, characterized in that beef or pork.
4. A composition for softening meat and flavor enhancement comprising the strain of claim 1 or a culture solution of the strain.
5. (A) Debaryomyces hansenii smfm201707 (Debaryomyces hansenii smfm201707) strain having a function of meat softening and flavor enhancement or processing for meat softening and flavor enhancement composition comprising a culture solution of the strain to meat; And

   (b) aging the meat treated with the composition; meat softening and flavor enhancement method comprising a.
6. The method of claim 5,

   In the step (b), the meat softening and flavor enhancement method characterized in that the aging of the meat using a dry aging method (dry aging).
7. The method of claim 5,

   In the step (b), the meat softening and flavor enhancement method, characterized in that for aging the meat for 1 to 50 days.
8. The method of claim 5,

   In the step (b), the meat softening and flavor enhancement method characterized in that the meat is matured under the conditions that the wind speed of more than 0 to 20 m / s or less is supplied.

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