KR101859417B1 - Novel Pilaira anomala smfm201611 strain for softening meat and improving flavor, and Method of softening meat and improving flavor using the same - Google Patents

Abstract

The present invention relates to a novel Pilaira anomala strain for tenderizing meat and improving flavor thereof, and a method for tenderizing meat and improving flavor thereof using the same. According to the present invention, it is possible to increase tenderness of meat during a meat aging step and also to improve flavor thereof by removing odor from the meat, thereby ensuring enhanced texture.

Description

[0001] The present invention relates to a novel filaranoma smfm201611 strain for meat softening and flavor enhancement, a method for softening meat flavor and improving flavor using the same,

The present invention relates to a novel filamentola anomalus strain for meat softening and flavor enhancement, and a method for softening food and improving flavor using the same.

Consumer preferences for meat are known as meat color, year, flavor and juiciness. As a method for improving meat quality, various methods such as aging period, different temperature method, dry method, wet method, proteolytic enzyme method and the like are used, but most of the conventional aging methods stay in the range of usual use have.

Aging refers to an operation that leaves a lack of flavor or texture under certain conditions for a while to have the desired flavor and texture. During aging, the action of microorganisms or enzymes, or the interaction between components, results in the production of flavor components and tissue changes, which are desirable foods.

Livestock is rigged for a certain period of time immediately after slaughter. Post stiffness refers to a state in which muscles or joints become stiff after a creature has died. Rigid meat is hard and tasteless, and can not be eaten because it has a lot of water coming out.

However, the muscles with the maximum stiffness are self-extinguishing by the proteolytic enzymes and soften the meat. In addition, when self-digestion begins, the pH of muscle is slightly recovered, water retention is increased, and enzymes act to produce flavors such as amino acids and oleic acid, an unsaturated fatty acid. The phenomenon that makes such a delicious meat is called 'aging'.

Two methods of wet aging and dry aging are used for beef aging. The dry aging method is to store the beef in a well-ventilated aged room without being wrapped and then aged.

Dry-aging, or dry-aging, is a method of evaporating and aging moisture from natural meat using conventional meat aging methods prior to development, unlike wet aging (wet aging), which is a general aging method.

That is, the wet aging is carried out by keeping the moisture of the meat in a plastic pack or a sealed plastic container while keeping the moisture of the meat intact. On the other hand, the dry aging is performed for 2 to 4 weeks Of the total.

Generally, dry aging is a method of aging meat within a controlled temperature and humidity for about 10 to 28 days before slaughtering and cutting. The temperature is 1 to 3 degrees Celsius and the humidity is 50 to 85% It is suitable.

Various methods of aging have been proposed in order to improve meat quality of beef meat and improve price competitiveness. For example, Japanese Patent No. 10-0459858 entitled " meat ripening storage method " A second step of sterilizing the meat to be stored after being sterilized by a sterilizing device, and a third step of storing the aged meat at a temperature of -5 ° C ± 2 ° C.

However, according to the conventional method of beating and aging the beef, when the beer is matured at a low temperature for a certain period of time, the beef becomes dry and the skin becomes hard. Thus, microorganisms that have affected the early stages of ripening can no longer be affected, so that even if the ripening period is prolonged, not only the deep portion of the beef can not be matured, but also there is a problem in that it can not provide good sensory characteristics that consumers can satisfy.

Therefore, it is necessary to develop aging techniques that can improve the meat quality of meat and increase the sensual characteristics (marbling, meat color, texture, juiciness, flavor, palatability, etc.) of meat.

Disclosure of the Invention The present invention has been conceived to solve the above-mentioned problems, and it is an object of the present invention to provide a novel strain having excellent ability to soften the food and improve the flavor, and a method of softening the food and improving the flavor using the same.

In order to solve the above problems,

Pilaira , which has meat-softening and flavor enhancing ability anomala ) smfm201611 strain.

According to the present invention, the strain may be one deposited with accession number KCTC18517P.

According to the present invention, the meat can be beef or pork.

The present invention also provides a composition for softening and softening the flavor of meat flavor comprising the strain or a culture thereof as an active ingredient.

The present invention also relates to a method for improving the flavor and flavor of a food, comprising the steps of: (a) treating the composition for improving meat softening and flavor, which comprises the strain of Pilaira anomala smfm201611 or a culture thereof as an active ingredient, And (b) aging the foodstuff to which the composition has been subjected, to thereby provide a meat softening and flavor enhancing method.

According to the present invention, the strain may be one deposited with accession number KCTC18517P.

According to the present invention, the meat can be beef or pork.

According to the present invention, the aging in step (b) may be a dry aging method.

According to the present invention, the aging of step (b) may be carried out for 1-50 days.

According to the present invention, the aging of step (b) may be carried out at a wind speed of 5 m / s or less.

The use of the strain pfilia anomala smfm201611 having the meat softening and flavor enhancing ability according to the present invention not only increases the year of meat culture in the course of fermentation of the meat, but also improves the texture by removing the odor of the meat and increasing the flavor .

Brief Description of the Drawings Fig. 1 is a diagram showing an 18s rRNA base sequence of a strain of Pilara anomala smfm201611 according to the present invention.
FIG. 2 is a diagram showing a flow diagram of a strain of Pilara anomala smfm201611 according to the present invention.
Fig. 3 is an electron microscope image of a strain of Pilara anomala smfm201611 according to the present invention.
Fig. 4 is a graph showing the distribution of fungi and yeast in dry-aged food (wind speed: 0, 2.5, 5 m / s) at the genus level.
FIG. 5 shows the microstructural changes in the meat after 28 days of inoculation and dry sowing of the strain of Pilara anomala smfm201611 according to the present invention.
6 shows the results of analysis of aflatoxin production ability in order to confirm the safety of P. nomala isolated from dried aged meat.

Hereinafter, the present invention will be described in more detail.

Conventional beef dry aging methods are natural aging at low temperature for a certain period of time. When aged for a long time, the surface of beef is dry and the skin becomes hard, so microorganisms that affect meat softening in the early stage of aging can no longer be affected Even if the fermentation period is prolonged, not only the deep portion of the beef can not be matured but also there is a problem in that it can not provide excellent sensory characteristics which can be satisfied by consumers.

Accordingly, in the present invention, a new strain capable of providing improved texture by increasing flavor and eliminating odor of food by not only increasing the year of meat consumption by affecting meat during fermentation during fermentation, Lt; RTI ID = 0.0 > smfm201611 < / RTI >

The Pilara anomala smfm201611 strain was deposited on Nov. 23, 2016 at the Korea Research Institute of Bioscience & Biotechnology Center located in Yuseong-gu, Daejeon, Republic of Korea and received the deposit number KCTC18517P.

Meat of the present invention refers to meat of all consumed forms which are fresh meat, chilled or frozen, and meat such as beef or pork, which can improve the quality sensation as the year increases.

The present invention also provides a composition for softening and softening the flavor of meat flavor comprising the strain or a culture thereof as an active ingredient.

At this time, the composition may be formulated for softening and flavor enhancing of meats by a conventional method, for example, in the form of a solution, a powder or a granule, but is not particularly limited to the formulation.

In addition, the composition can be treated in the foodstuffs through spraying (for example, spraying, misting, powder spraying, granule spraying, etc.) on the foodstuff to be aged, surface coating, etc., Or after a certain point of time after aging. In addition, the amount thereof to be used can be appropriately determined in accordance with the portion, weight, and aging place of the meat to be treated.

In addition, the present invention provides a method for producing a flavor enhancer, comprising the steps of: (a) treating a composition for improving meat softening and flavor, which comprises a strain of Pilaranomala smfm201611 or a culture thereof as an active ingredient, And (b) aging the foodstuff to which the composition has been subjected, to thereby provide a meat softening and flavor enhancing method.

In this case, when the pineapple Anomalla smfm201611 strain is treated in the food, it may affect the meat during the fermentation period, thereby increasing the year of the meat. In addition, it may improve the flavor by increasing the flavor by removing the odor of the meat. As a new strain, the above-mentioned Pilara anomala smfm201611 strain was deposited on Nov. 23, 2016 at the Korea Research Institute of Bioscience & Biotechnology Center located in Yuseong-gu, Daejeon, Korea, and received the deposit number KCTC18517P.

Meat of the present invention refers to meat of all consumed forms which are fresh meat, chilled or frozen, and meat such as beef or pork, which can improve the quality sensation as the year increases.

The composition for improving meat softening and flavor including the above strain or a culture thereof as an active ingredient can be formulated for meat softening and flavor enhancement by a conventional method and can be prepared, for example, in the form of solution, powder or granule, But is not particularly limited to a formulation.

In addition, the composition can be treated in the foodstuffs through spraying (for example, spraying, misting, powder spraying, granule spraying, etc.) on the foodstuff to be aged, surface coating, etc., Or after a certain point of time after aging. In addition, the amount thereof to be used can be appropriately determined in accordance with the portion, weight, and aging place of the meat to be treated.

The aging of the step (b) may be performed by a dry aging method in consideration of the year and flavor improvement of the food, although the aging of the step (b) is not limited to the general method known in the art .

The aging in step (b) is preferably carried out for 1-50 days.

It is preferable that the aging of step (b) is carried out under the wind speed of 5 m / s or less. When considering the year, flavor, and taste of food, It may be more desirable to be performed in a blocked location.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments and the like. It will be apparent to those skilled in the art, however, that these examples are provided for further illustrating the present invention and that the scope of the present invention is not limited thereto.

Example  One. Pilara Anomalla smfm201611  Isolation and Culture of Strain

On 28th day, Mucoraceae (Family) strain, which shows the effect of meat softening and flavor enhancement, was isolated and cultured from dried aged meat products.

To isolate the fungi, i) single spores of Pilaira anomala strain bloomed on the dry aged meat surface were transferred to a potato dextrose agar (PDA; 4 g patato, 20 g dextrose, 15 g agar per 1 L) For culturing, 1.2% agar was cut into 1 × 1 cm, placed on a slide glass, and monoclonal was inoculated on the corners of the agar film. The PDA medium containing the inoculated agar film was cultured at 4-5 ℃ for 4-5 days. ii) In addition, 50 ml of 0.1% peptone water was added to 25 g of dried aged flesh, stirred with pummeler, and plated on PDA medium. The colonies were selected by 4-5 days of culture at 4-5 ° C.

In order to confirm protein degradation ability through the strain, skim milk was added to the PDA medium, and the cells were inoculated and cultured at 4-5 ° C for 4-5 days. The proteolytic ability of the strain was confirmed by the clear zone formed around the bacteria.

Example  2. Pilara Anomalla smfm201611  Identification of the strain

18s rRNA gene sequence analysis was performed for identification of Pilara anomala strains showing the effects of meat softening and flavor enhancement (Fig. 1).

The samples were submitted to Macrogen Co., Ltd. in the state of a plate culture with the new strain isolate obtained in Example 1, and the sequence obtained through whole sequencing of the 18s rRNA region was transferred to NCBI (Nation Center for Biotechnology Information (Bioinformatics Center). The result of homology search was confirmed through Blast search.

In addition, primers for 18s rRNA gene amplification are described below.

Forward primer: NS1 (5'-GTAGTCATATGCTTGTCTC-3 ')

Reverse primer: NS8 (5'-TCCGCAGGTTCACCTACGGA-3 ')

According to the present invention, smfm201611 As a result of 18s rRNA gene sequencing analysis of the strain, it was confirmed that the strain showed 96% or more homology with the Pilariana anomalus strain IUE 573, and the isolated microorganism was found to be a Pilar ananomala strain. In addition, the isolated strain was observed using an electron microscope, and it is shown in FIG.

Thus, the novel strain according to the present invention was named as pyloranoma smfm201611, and was registered with the Life Resource Center of Korea Biotechnology Research Institute on November 23, 2016, and received the deposit number KCTC18517P.

Experimental Example  1. pH and Shear force  Measure

The pH was measured by adding 9 mL of distilled water to 1 g of sample, homogenizing the sample at 1,130 × g for 1 min using a homogenizer (T10 basic, Ika Works, Staufen, Germany) , Mettler-Toledo international Inc., Seitzerland).

The shear force was used to heat the beef till the depth of the meat reached 72 ° C, cool it at room temperature, and then shape it into a rectangular parallelepiped (5.0 × 2.0 × 1.2 cm, length × width × height). Using a texture analyzer (CT3 10K, Brookfield Engineering Laboratories, USA) equipped with a Warner-Bratzler shear, the measurement was performed until the sample was completely cut perpendicular to the direction of the myofiber. The measurement conditions were maximum cell load, 10 kg; probe test speed, 2.0 mm / s; distance, 30 mm; trigger load, 10 g.

Con Wet Dry 0 Dry 2.5 Dry 5 SEM ¹ pH 5.63 ^b 5.45 ^c 5.96 ^a 5.60 ^b 5.63 ^b 0.030 Shear force (N) 81.45 ^a 48.36 ^bc 42.17 ^c 58.86 ^b 53.34 ^bc 3.381

¹ Standard errors of mean (n = 15).

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

Abbreviation: Non-aged and obtained at 2 days postmortem, Con; 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.

Table 1 shows the meat quality of the beef cattle after wet and dry fermentation. PH during wet aging tended to decrease compared to that of the control, and the pH value increased during dry aging (p <0.05). Especially, pH of 0 m / s dry aging group showed the highest value. Shear force showed significant decrease after aging and lowest shear force at 0 m / s (p <0.05).

Experimental Example  2. Sensory Evaluation

Consumer preference and ranking method were used to evaluate sensory differences between samples. Eight consumer panels were selected for the sensory evaluation. The samples were cut into a certain size (5.0 × 2.0 × 1.2 cm, length × width × height) and cooked until the deep portion of the meat reached 72 ℃. . Consumer preference was assessed using the 9 point scaling method, and 2.5 m / s of dry fermented meat was set as the standard control, so that the sensory panels were known about the flavor and flavor of the dry fermented meat, and then the dry fermentation flavor and flavor intensity 1-4.

Wet Dry 0 Dry 2.5 Dry 5 SEM ¹ Consumer test Overall acceptance 5.38 ^c 6.75 ^a 6.25 ^ab 6.00 ^b 0.199 Ranking test ² Odor 4 2 3 One - Flavor (taste + odor) 4 3 2 One -

¹ Standard errors of mean (n = 32).

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

^{a- c} Different letters within the 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.

The sensory evaluation of the wedge after dry and wet aging is shown in Table 2 above. Compared with wet - aged meat, dry - aged meat showed significantly higher preference. In addition, the dry aged meat (2.5 m / s) was set as the standard control so that the sensory test of the dry aged meat flavor was known, and the dry aging strength was measured in 1-4 ranks. As a result, The fermented flavor and flavor were the strongest, but the taste was the lowest. The degree of preference was the highest at 0 m / s, while the dry fermented flavor and flavor were ranked second and third, respectively.

Experimental Example  3. Next Generation Sequencing, NGS )

Two samples (1 × 1 cm) which were aged according to the aging method (0, 2.5, 5 m / s) for 28 days were used for the analysis, and the DNA was extracted and QC (quality control) was carried out. Sequencing was performed to confirm the microbial distribution of the extracted DNA, which was used by Illuina SBS technology. Statistical analysis was performed using software from Illumina Miseq using the analyzed basic data, and the distribution of microorganisms among the groups was compared and shown in FIG. 4 and Table 3 below.

Dry 0 m / s Dry 2.5 m / s Dry 5 m / s Mucoraceae 99.8 81.0 86.0

The fungi were found to be the most abundant (99.6%) at 0 m / s dry mature, and the existing fungi were identified as Mucoraceae family, and Pilara anomala is also classified as Mucoraceae family.

In order to confirm the enzymatic activity, pila lanoma was cultured on PDA medium containing skim milk. As a result of incubation, the protein was cleaved by cleavage of skim milk by forming a transparent ring around the inoculated area. It is suggested that the protein hydrolyzing ability shows the characteristic sensory characteristics of dried and aged meat during aging.

Experimental Example  4. Pilara Anomalla Inoculation group  Change observation

(1) The shear force changes of the group inoculated with Pfilla anomala smfm201611 and the group not inoculated (control group) according to the dry aging period were measured, and the results are shown in Table 4 below. The shear force was measured by heating the core until the depth of the core reached 72 ° C, cooling at room temperature, and shaping the core (1.54 × 4.0 cm ² ). Using a texture analyzer (CT3 10K, Brookfield Engineering Laboratories, USA) equipped with a Warner-Bratzler shear, the measurement was performed until the sample was completely cut perpendicular to the direction of the myofiber. The measurement conditions were maximum cell load, 10 kg; probe test speed, 2.0 mm / s; distance, 30 mm; trigger load, 0.1 N.

Dry aging period (days) SEM ¹ 0 14 21 28 Control group 35.91 ^a 18.62 ^c 16.28 ^cx 24.53 ^by 0.920 mold 35.91 ^a 19.00 ^c 12.59 ^dy 27.60 ^bx 0.815

¹ Standard error of means (n = 15),

^{a- d} The letters were 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 a result of the measurement, the shear force tended to decrease until 21 days of maturation in both the inoculated smfm201611 group and the untreated group (control group), and according to the present invention, the smell of smfm201611 In the inoculated group, after 21 days of dry aging, the shear force was lower than the control group and the year was better.

(2) The microstructural changes in the meat were observed by TEM (Transmission Electron Microscope) after 28 days of dry fermentation of the smfm201611 and non-inoculated groups (control group) 5. After aging, the specimens were cut and fixed at 1 mm, washed with tertiary distilled water, dyed with 0.5% uranyl acetate, and dehydrated. The dehydrated tissues were substituted with propylene oxide, embedded in a spurr's resin mixture, and polymerized in a dry oven at 70 ° C for 24 hours. The embedded tissues were microtuncated and the sections were observed with a transmission electron microscope.

As a result, z-disk collapse in the group of smfm201611 was observed more frequently than in the control group. The collapse of the z-disk indicates the softening of the flesh, and the proteolytic activity is thought to cause the degradation of muscle protein.

(3) Changes in reducing sugar content were measured according to the duration of dry aging of the group inoculated with Pfilla anomala smfm201611 and those in the non-inoculated group (control group), and the results are shown in Table 5 below. The amount of reducing sugar was determined by adding 80% EtOH (50 ° C) to 1 g of sample, extracting it twice by centrifugation, filtering and concentrating it with nitrogen. Distilled water was added to the concentrated sample and centrifuged to extract the sugar content of the muscle. After adding the DNS reagent to the extracted sample, the sample was cooled at 90 ° C and cooled. Then, the absorbance was measured using a spectrophotometer to determine the reducing sugar content. Standard curves were prepared using glucose (Sigma Aldrich, USA) as a reference material.

Dry aging period (days) SEM ¹ 0 14 21 28 Control group 9.93 ^c 17.67 ^ax 15.35 ^bx 17.31 ^ax 0.235 mold 9.93 ^d 14.46 ^months 11.63 ^cy 13.30 ^by 0.221

¹ Standard error of means (n = 16).

^{a- d} The letters were 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 a result, the reducing sugar content was significantly increased up to 14 days of dry fermentation in both the inoculated group and the non - inoculated group (control group), and decreased to 21 days and increased again on 28 days . The reduced sugar content was lower than that of the control group, but it was confirmed that the reducing sugar was increased by aging comparing with before fermentation (0 day difference). It has been found that smell of smell of smell of smfm201611 in poultry is consumed while consuming sugars, so that the reducing sugar content is low but has a positive effect on the year and flavor (data not shown).

Experimental Example  6. Pilara Anomalla  Identification of Stability of Strain

The aflatoxin production ability was analyzed to confirm the safety of the strain pneumoniae smfm201611 isolated from the dry aged meat, and the results are shown in Fig. Aflatoxin was extracted with 70% methanol, purified with an immunoaffinity column and derivatized with trifluoroacetic acid, and analyzed with a liquid chromatograph equipped with a fluorescence detector. Mycotoxins are secondary metabolites produced by fungi, such as aflatoxin, ochratoxin, and fumonisin. Up to now, about 400 species of aflatoxin have been known, including WHO's International Cancer Institute The International Agency for Research on Cancer (IARC) classifies them as group 1 carcinogens, thus highlighting the risk of toxins. Measurement results, do cyano as discrete pillars in accordance with the present invention, _{aflatoxin (B 1, B 2,} G 1, G 2) from smfm201611 strain stability for this has not been detected bar, toxin was confirmed that high.

Korea Biotechnology Research Institute KCTC18517P 20161123

Claims (10)

Pillara anomalas smfm201611 strain having meat softening and flavor enhancing ability. The method according to claim 1,
Wherein said strain is deposited under accession number KCTC18517P. The method according to claim 1,
Wherein the meat is meat or meat. A composition for enhancing meat softening and flavor enhancement comprising the strain of claim 1 or a culture thereof as an active ingredient. (a) treating a foodstuff softening and flavor improving composition comprising a strain of Pyllaanomala smfm201611 or a culture thereof as an active ingredient having meat meat softening and flavor improving ability; And
(b) aging the meat to which the composition has been subjected. 6. The method of claim 5,
Wherein the strain is deposited under accession number KCTC18517P. 6. The method of claim 5,
Wherein the meat meat is beef or pork meat. 6. The method of claim 5,
Wherein the fermentation of step (b) is a dry aging method. 6. The method of claim 5,
Wherein the fermentation of step (b) is performed for 1-50 days. 6. The method of claim 5,
Wherein the fermentation of step (b) is carried out at a wind speed of 5 m / s or less.

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