KR20240100498A - Novel Pediococcus pentosaceus SMFM2016-GK1 strain and method for producing fermented meat using the same - Google Patents

Abstract

The present invention relates to Pediococcus pentosaceus SMFM2016-GK1 strain having a flavor enhancing function; A starter composition for fermentation and a composition for flavor enhancement containing the above strain; and a method for producing fermented meat using the strain.

Description

Pediococcus pentosaceus SMFM2016-GK1 strain and method for producing fermented meat using the same {Novel Pediococcus pentosaceus SMFM2016-GK1 strain and method for producing fermented meat using the same}

The present invention relates to the Pediococcus pentosaceus SMFM2016-GK1 strain and a method for producing fermented meat using the same.

Lactic acid bacteria, also commonly referred to as lactic acid bacteria, refer to bacteria that produce a significant amount of lactic acid as the final metabolite. Lactic acid bacteria are a type of microorganism that humans have been using for a long time. In Korea, they are widely used in kimchi, and in the West, they are widely used in dairy fermentation.

Recently, research on lactic acid bacteria has been actively conducted, and it is a very important industrial strain that is widely used in fermented meat production, health and functional foods, bowel preparations, pharmaceuticals, and probiotic products. Bacteria belonging to this group include Streptococcus sp. These include Pediococcus sp., Leuconostoc sp., Lactobacillus sp., Sporolactobacillus sp., and Bifidobacterium. In addition, kimchi, Korea's unique fermented vegetable food, is mainly composed of lactic acid bacteria such as Lactobacillus, Pediococcus, Leuconostoc, and Bycella.

The above lactic acid bacteria are Gram-positive bacteria that have anaerobic or hemi-anaerobic properties. They crowd and settle on the intestinal digestive tract wall of animals and use them as an energy source by decomposing nutrients and fiber consumed by the animal, and produce lactic acid and antibiotics. It is known to play an important role in maintaining intestinal health by suppressing the growth of harmful intestinal bacteria.

However, since lactic acid bacteria use live bacteria themselves, their growth is inhibited by the acidic environment where the pH drops below 3 when taken orally, as well as digestive enzymes and bile acids secreted from the stomach and small intestine, which not only reduces the number of live bacteria reaching the large intestine but also reduces the intestinal colonization rate. The problem of deterioration is being pointed out.

In addition, biogenic amines, a toxic substance produced during the food fermentation process using lactic acid bacteria, are known to irritate the nerves and blood vessels of humans and animals, causing symptoms similar to food poisoning. Accordingly, the use of biogenic amine reduction starters is becoming important for the production of hygienic and safe fermented foods, and research on reduction through inhibition of biogenic amine production is being actively conducted.

Accordingly, in order to solve the above problems, the present inventors isolated and identified Pediococcus pentosaceus, which has no biogenic amine production ability and has excellent low-temperature growth ability and acid resistance, from kimchi, a fermented vegetable food unique to Korea, and isolated lactic acid bacteria. The present invention was completed by confirming that fermented meat with excellent taste and aroma can be produced by using as a starter strain.

Republic of Korea Patent No. 1986-0000550

The present invention aims to solve the above-described problems and other problems associated therewith.

An exemplary object of the present invention is to provide a Pediococcus pentosaceus SMFM2016-GK1 strain having low temperature growth tolerance.

Another exemplary object of the present invention is to provide a starter composition for fermentation containing the Pediococcus pentosaceus SMFM2016-GK1 strain.

Another exemplary object of the present invention is to provide a composition for flavor enhancement containing the Pediococcus pentosaceus SMFM2016-GK1 strain.

Another exemplary object of the present invention is a method for producing fermented meat comprising adding a starter composition containing a Pediococcus pentosaceus strain to meat; To provide fermented meat manufactured by the above method.

The technical problem to be achieved according to the technical idea of the invention disclosed in this specification is not limited to the problem to solve the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below. There will be.

This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present application may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Additionally, the scope of the present application cannot be considered limited by the specific description described below.

As an aspect for achieving the above object, the present invention provides a Pediococcus pentosaceus SMFM2016-GK1 strain having low temperature growth tolerance.

The term "Pediococcus pentosaceus" of the present invention refers to a lactic acid bacterium of the Pediococcus genus of the Lactobacillaceae family, and the Pediococcus pentosaceus SMFM2016-GK1 strain of the present invention refers to kimchi. This is a strain isolated and identified from .

In the present invention, the Pediococcus pentosaceus SMFM2016-GK1 strain has been deposited under the accession number KACC 81233BP.

The term "low-temperature growth tolerance" in the present invention refers to improved low-temperature growth ability, and the "low-temperature growth ability" refers to growth ability at a temperature of 25°C or lower. In this case, the lower temperature limit related to low-temperature growth ability is 5°C, preferably 10°C, and more preferably 15°C.

In the present invention, the Pediococcus pentosaceus SMFM2016-GK1 strain is a strain that does not have the ability to produce biogenic amines.

The biogenic amine is a nitrogen compound produced by amino acid decarboxylation during the metabolic process of microorganisms, plants, and animals. If consumed in excess, it can cause skin inflammation, allergies, and in severe cases, food poisoning, palpitations, etc. These include, but are not limited to, histamine, tyramine, putrescine, and cadaverine.

In the present invention, the Pediococcus pentosaceus SMFM2016-GK1 strain is a strain resistant to acid.

The acid-resistant strain refers to a strain with improved acid resistance, and “acid resistance” refers to a strain with a high survival rate under conditions of pH 5, preferably pH 3.5, but is not limited thereto.

As another aspect for achieving the above object, the present invention provides a starter composition for fermentation containing Pediococcus pentosaceus SMFM2016-GK1 strain. The Pediococcus pentosaceus SMFM2016-GK1 strain is as described above.

The term "starter" in the present invention refers to a microbial culture medium used when producing a fermented product. Among microorganisms, bacteria, molds, yeast, etc. are generally used as starters, and they can be used alone or in combination. .

The starter of the present invention can be used by mixing Pediococcus pentosaceus SMFM2016-GK1 strain and Debariomyces hansei strain.

The starter of the present invention may be a combination of Pediococcus pentosaceus SMFM2016-GK1 strain and Debariomyces hansei strain in a volume ratio of 1:2 to 2:1, specifically 1:1. It may be combined in a ratio of, but is not limited to this.

The starter of the present invention may further include Penicillium nalgiobens.

The starter can be used to ferment various meats derived from land animals such as cows, pigs, horses, sheep, goats, rabbits, etc., as well as birds such as chickens, and to enhance the flavor (taste and aroma).

The composition may be formulated for fermentation through a conventional method, and specifically, may be formulated in the form of a solution, powder, or granule, but is not limited thereto.

As another aspect for achieving the above object, the present invention provides a composition for flavor enhancement comprising Pediococcus pentosaceus SMFM2016-GK1 strain. The Pediococcus pentosaceus SMFM2016-GK1 strain is as described above.

The composition may be formulated for use in enhancing flavor through conventional methods. Specifically, the composition may be formulated in the form of a solution, powder, or granule, but is not limited thereto.

The composition may include additives commonly used to enhance flavor.

As another aspect for achieving the above object, the present invention provides a method for producing fermented meat comprising adding a starter composition containing a Pediococcus pentosaceus strain to meat. The Pediococcus pentosaceus SMFM2016-GK1 strain and starter are as described above.

In the above method, the starter containing the strain can be coated or applied to the meat subject to fermentation to ferment the meat, and for this purpose, the meat is immersed in a composition of a solution formulation containing the strain or its culture medium, or the meat is immersed in a solution formulation containing the strain or its culture medium. , a composition having a formulation such as powder or granule can be treated on meat by coating or applying (e.g., spraying, misting, powder spraying, granule spraying, etc.).

In addition, in the above method, the concentration of the strain can be appropriately adjusted depending on the area subject to fermentation, weight, fermentation site, etc.

In the method, the fermenting step can produce fermented meat that can provide improved palatability by allowing the strain to increase flavor during the fermentation period.

The fermentation can ferment meat treated with the composition using various fermentation methods commonly used for producing fermented meat.

Another exemplary object of the present invention is to provide fermented meat produced by a fermented meat production method comprising adding a starter composition containing a Pediococcus pentosaceus strain to meat.

In the present invention, fermented meat is not particularly limited in its type, but includes not only general meat before processing such as beef, pork, and chicken, but also processed meat products such as ham, sausages, and bacon, and packaged meat.

When the Pediococcus pentosaceus SMFM2016-GK1 strain according to the present invention was used for meat fermentation, it was confirmed that the flavor in fermented meat was improved. That is, the Pediococcus pentosaceus SMFM2016-GK1 strain, which has been confirmed to have a flavor enhancing function, can be used for fermentation of meat.

However, the effects according to an embodiment of the technology disclosed in this specification are not limited to those mentioned above, and other effects not mentioned may be clearly understood by those skilled in the art from the description below.

In order to more fully understand the drawings cited in this specification, a brief description of each drawing is provided.
Figure 1 is a diagram showing the results of confirming the acid resistance of the isolated strain.
Figure 2 is a diagram showing the results of confirming the low-temperature growth ability (15°C) of the isolated strain.
Figure 3 is a diagram showing the results of confirming the low-temperature growth ability (25°C) of the isolated strain.
Figure 4 is a diagram showing the results of confirming the growth ability of strains within the model-system.
Figure 5 is a diagram showing the results of confirming the amount of lactic acid produced in the Model-system.

Hereinafter, the present invention will be described in more detail through the following examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1: Pediococcus pentosaceus ( Pediococcus pentosaceus ) Isolation and identification of SMFM2016-GK1 strain

To isolate the Pediococcus pentosaceus SMFM2016-GK1 strain of the present invention, 50 mL of buffered peptone water (BPW) was added to 25 g of kimchi and homogenized for 60 seconds. After decimal dilution of the homogenized solution, the diluted solution was inoculated and smeared on MRS agar (de Man, Rogosa and Sharpe agar) and cultured at 37°C for 48 hours. After isolating colonies showing the characteristics of lactic acid bacteria, microorganisms were identified through 16s rRNA sequencing analysis. As a result of the identification, a new strain, Pediococcus pentosaceus SMFM2016-GK1, was selected.

The selected strains were deposited at the Agricultural Genetic Resources Center of the National Academy of Agricultural Sciences on November 1, 2022, and were given the deposit number KACC 81233BP. The Pediococcus pentosaceus SMFM2016-GK1 strain has the 16S rRNA sequence of SEQ ID NO: 1.

Experimental Example 1: Confirmation of acid resistance of isolated strains

The acid resistance of 10 types of lactic acid bacteria, including the Pediococcus pentosaceus SMFM2016-GK1 strain isolated by the method according to Example 1, was compared.

Specifically, a single colony of lactic acid bacteria was inoculated into MRS broth (de Man, Rogosa and Sharpe broth) and cultured at 37°C for 24 hours, then 0.1 mL was inoculated into new MRS broth and cultured at 37°C for 24 hours. The culture was centrifuged at 1,912 The prepared bacterial solution was plated on MRS agar, cultured at 37°C for 48 hours, and colonies were counted. The number of viable cells on MRS agar with 4% (w/v) NaCl was compared to the number of bacteria on MRS agar without NaCl added. The numbers were compared.

As a result, as shown in Figure 1, low-temperature growth performance experiments were conducted on 8 of the 10 candidate strains, excluding two strains (49 and 67) that showed a significantly low survival rate at pH 3.5.

Experimental Example 2: Confirmation of low-temperature growth ability of isolated strains

The low-temperature growth ability of the eight strains selected in Experimental Example 1 was compared.

Specifically, a single colony of lactic acid bacteria was inoculated into MRS broth and cultured at 37°C for 24 hours, then 0.1 mL was inoculated into new MRS broth and cultured at 37°C for 24 hours. The culture was centrifuged at 1,912 The bacterial solution diluted to 3 to 4 Log CFU/mL was inoculated into MRS broth and cultured at 15°C and 25°C for 48 hours. The culture medium was plated on TSA (tryptic soy agar) and cultured at 37°C for 24 hours to count colonies, and the growth rates were compared at each culture temperature.

As a result, as shown in Figures 2 and 3, two strains (Pediococcus pentosacea) showed significantly high growth ability at the fermented sausage ripening temperature of 15°C (Figure 2) and 25°C (Figure 3). Pediococcus pentosaceus SMFM2016-GK1 and 9) were selected.

Experimental Example 3: Confirmation of fermentation characteristics of strains in model-system

To confirm the fermentation characteristics of the two strains selected in Experimental Example 2, a medium with a composition similar to that of fermented meat [Model-system: beef extract (12g/L), glucose (10g/L), NaCl (20g/L) was used. , dipotassium hydrophosphate (2g/L), MgSO ₄ ·7H ₂ O (0.15g/L), nitrite (0.15g/L)] and pH 5.8, the growth of lactic acid bacteria and the amount of lactic acid production were compared.

As a result, as shown in Figures 4 and 5, the two lactic acid bacteria strains ( Pediococcus pentosaceus SMFM2016-GK1 and 9) reached the maximum growth number on the 1st day of culture and gradually decreased after the 3rd day. showed a tendency to do so. Additionally, as the culture period increased, the amount of lactic acid produced also increased, reaching a value of 2.4% on the final 7th day.

Experimental Example 4: Confirmation of biogenic amine production ability of isolated strain

In order to confirm the biogenic amine production ability of the two strains selected in Experimental Example 2, the production amount of four types of biogenic amines (tyramine, histamine, cadaverine, and putrescine) was confirmed through HPLC analysis.

As a result, as shown in Table 1 , the Pediococcus pentosaceus SMFM2016-GK1 strain showed levels below the detection limit (<1ppm) for all four types of biogenic amines, confirming that it does not have the ability to produce biogenic amines. .

item lactic acid bacteria strains Pediococcus pentosaceus
( Pediococcus pentosaceus ) SMFM2016-GK1 9 histamine <1 <1 tyramine <1 10.608±0.957 putrescine <1 <1 Cadaverine <1 <1

Experimental Example 5: Confirmation of flavor enhancement effect of fermented sausage

Experimental Example 5-1: Production of fermented sausage

Fermented sausage was prepared by inoculating Pediococcus pentosaceus SMFM2016-GK1.

Specifically, for the production of fermented sausage, 0.06% of the starter containing the Pediococcus pentosaceus SMFM2016-GK1 strain was added compared to the raw meat to produce fermented sausage and aged for 4 weeks. The control group was prepared using lactic acid bacteria and mold starters (Lactobacillus powder (21 mixed lactic acid bacteria, Biotopia, Chuncheon, Korea) and Starterkulturen Edelschimmel (Almi, Austria)) sold for fermented meat production.

Experimental Example 5-2: Confirmation of changes in flavor characteristics of fermented sausage

Changes in flavor characteristics were confirmed by analyzing the electronic tongue analysis results for fermented sausages in the fourth week of fermentation.

As a result, as shown in Table 2, the fermented sausage inoculated with starter had higher saltiness and umami taste and lower sourness than the control. According to the results of previous research that domestic consumers prefer the salty taste of fermented sausage but not the sour taste, the mixed starter of the present invention was found to be suitable for producing fermented sausage with a taste preferred by domestic consumers (Table 2_AHS: sensor for sourness. PKS: sensor for sweetness, CTS: sensor for saltness, NMS: sensor for umami, CPS: sensor for standard, ANS: sensor for bitterness, SCS: sensor for standard)

Sample sensor AHS P.K.S. CTS NMS CPS ANS SCS starter 5.5 5.2 6.5 7.4 5.4 6.4 6.4 control 6.5 6.8 5.5 4.6 6.6 5.6 5.6

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.

Rural Development Administration National Agricultural Science Microbial Bank (KACC) KACC81233BP 20221101

Sequence list electronic file attached

Claims (9)

Pediococcus pentosaceus SMFM2016-GK1 strain with low-temperature growth tolerance. According to paragraph 1,
The strain was deposited under the accession number KACC 81233BP. According to paragraph 1,
The strain is a strain that does not have the ability to produce biogenic amines. According to paragraph 1,
The strain is resistant to acid. According to paragraph 1,
The strain has a flavor enhancing function. Pediococcus pentosaceus Starter composition for fermentation comprising SMFM2016-GK1 strain. Pediococcus pentosaceus A composition for flavor enhancement comprising the SMFM2016-GK1 strain. A method for producing fermented meat comprising adding a starter composition containing the Pediococcus pentosaceus strain according to claim 6 to meat. Fermented meat manufactured by the manufacturing method of paragraph 8.