KR100837857B1 - Bacillus sp., isolated from meolchi-jeot, having fibrinolytic, immunostimulating activities and jeot-kal products containing thereof - Google Patents

Abstract

A novel strain is provided to show fibrinolytic activity and immuno-stimulating activity, thereby being used in order to prepare salted(pickled) anchovies with excellent taste and functionality or other foods such as kimchi or processed foods with excellent functionality. A novel Bacillus licheniformis FA0407 isolated from salted(pickled) anchovies is deposited on Dec. 20, 2004 as a deposition no. KCTC 10752BP. A method for preparing salted anchovies comprises the steps of: (a) mixing 5-6 parts by weight of raw anchovy, 3-4 parts by weight of salt, 1-2 parts by weight of purified water and the Bacillus licheniformis FA0407; and (b) ripening the mixture at room temperature for at least 6 months.

Description

Bacillus sp., Isolated from Meolchi-jeot, having Fibrinolytic, Immunostimulating Activities and Jeot-kal Products Containing Thereof}

1 shows a manufacturing process diagram of the present invention.

Figure 2a is a photograph showing the protein degradation capacity of the microorganism separated during the fermentation anchovy salt fermentation process is a photograph showing the protein degradation capacity in the medium containing 1% reduced skim milk.

Figure 2b is a photograph showing the measurement of protein degradation ability of microorganisms isolated during fermentation of anchovy salt, it is a photograph showing the protein degradation capacity using the fibrin plate method.

Figure 3 is a photograph showing the application of commercially available anchovy salt of FA0507 strains. The left photo shows the thrombi solubility test of five commercial anchovy salts. The middle photo shows the thrombi solubility test of FA0507 strain, washing buffer and dilution water. The picture on the right is a picture showing the thrombolytic ability test by mixing FA0507 strains with three commercially available anchovy salt.

4 is a graph showing the NO production concentration of macrophages treated with FA0407.

Figure 5 is a graph showing the IL-1a production concentration of macrophages treated with FA0407.

The present invention relates to Bacillus ( Bacillus ) bacteria having high thrombolytic ability and immune activity isolated from anchovy salt and salted salt containing the same. More specifically, the purpose of the present invention is to search for Bacillus strains having high thrombolytic and immune cell activity health functional activities from anchovy salt, and to utilize Bacillus strains with healthy functional activity in anchovy salt products.

Anchovy salted salted anchovy is a salted salted fish that has been aged for more than 3 months. Anchovy (named Engraulis japonica, Anchovy from the Mediterranean) is herring anchovy and saltwater fish. The body is thin, long and flat. Coastal sashimi (沿岸 性 回 游魚), whose stock is plankton. The spawning season lasts for almost a year, but the penis is two times from spring to summer and autumn. Eggs are elongated and transparent. Both adult and larvae produce a lot of fish and are caught with enamel and hatch nets. It is made of dried foods, salted fish, and stewed foods. It is also used as a bait for bonito fishing. It is distributed in Korea, China, and Japan.

When the most mature anchovies are caught in the south and southwest coasts of Korea in May-June, the good anchovies are steamed, dried, dried, and sold on the market. The rest of the anchovies are made with salted fish. In particular, anchovies are mainly immersed in Gyeongsangnam-do, Jeollanam-do, Chungcheongnam-do, and Jeju-do, and many markets are formed there. When soaking anchovy salt, mix raw anchovy and salt in a ratio of 5: 3, put it in a jar and seal it. Anchovy salted kimchi can be added to decoct when you soak it. It can be used to boil or eat seaweed. Raw anchovies are eaten raw, and the raw anchovies are trimmed to remove the thorns.

Salted seafood is a traditional fermented food that has a distinctive taste and texture by adding a large amount of salt to the muscles and intestines of fish and shellfish and then decomposing the protein by the enzyme action of autophagy or microorganisms during aging. Microorganisms involved in fermentation and fermentation of salted fish are salt-resistant bacteria that can grow at high salt concentrations and are known to affect the nutrition and flavor of products. As for the health functional activity of salted microorganisms, there have been published research cases related to salted microorganisms showing thrombolytic activity, but there are not many studies yet. Patents related to the thrombolytic activity and immunological activity of Cheonggukjang microorganism have already been seen. Although the state has been registered by the researchers (Invention Patent 10-0588521), there have been no examples of the application of the health functional active salted microorganisms to the salted fish products.

In the present invention, Bacillus strains having excellent protein resolution and high immune activity among 326 microorganisms isolated during fermentation or fermentation of anchovy are selected, and applied to commercially available anchovy salt to confirm its functionality. It was.

The present invention measures the general bacterial counts from anchovy and purely isolates the microorganisms from the standard agar medium to select strains with excellent protein degradation ability from skim milk medium, high thrombolytic activity, and good immuno-activating FA0407 strain (Fig. 1). Reference).

The present invention represents a new strain Bacillus licheniformis FA0407 KCTC 10752BP (deposited Dec. 20, 2004 to the Korea Biotechnology Research Institute Gene Bank) isolated from anchovy salt.

The new strain Bacillus licheniformis FA0407 (KCTC 10752BP) isolated from anchovy salt of the present invention has excellent protein resolution.

The new strain Bacillus licheniformis FA0407 (KCTC 10752BP) isolated from anchovy salt of the present invention has excellent thrombolytic activity.

The bacterium Bacillus licheniformis FA0407 (KCTC 10752BP) isolated from anchovy salt of the present invention has excellent immune activity.

The present invention includes a method of using Bacillus licheniformis FA0407 (KCTC 10752BP) strain as a medicament within a pharmaceutically acceptable range.

The present invention includes a method of using Bacillus licheniformis FA0407 (KCTC 10752BP) strain as a health functional food within a food acceptable range.

The present invention includes anchovy salt containing Bacillus licheniformis FA0407 (KCTC 10752BP) strain.

The present invention includes a method for preparing anchovy salted fish containing Bacillus licheniformis FA0407 (KCTC 10752BP) strain.

Anchovy salt containing Bacillus licheniformis FA0407 (KCTC 10752BP) strain of the present invention by mixing 5-6 parts by weight of raw anchovy, 3-4 parts by weight of salt, 1-2 parts by weight of purified water and Bacillus licheniformis FA0407 (KCTC 10752BP) strain It can be prepared by aging at room temperature for 6 months or more, preferably 6-12 months.

Hereinafter, the content of the present invention will be described in detail through examples and test examples. However, these are intended to explain the present invention in more detail, and the scope of the present invention is not limited thereto.

<Example 1>

60kg of anchovy salt from anchovy manufacturer (Kyungdong Market) was transferred to a plastic container, and then stored in a well-ventilated, non-lighting, semi-underground room next to the Korea Food Research Institute building on the 0th and 15th days. Samples are taken at monthly intervals and the results of the investigation of the bacterial counts and general chemical composition of anchovy samples are shown in Table 1.

Table 1. Aerobic and Physicochemical Changes of Anchovy Sauce during Fermentation

Plates with 25-50 colonies were selected from the standard agar media with the complete count of bacteria, and 50 microorganisms were isolated from each sample, and the morphological properties of the colonies and cells were examined. staining, Catalase test, Oxidase test) were performed. Changes of microbial flora in five months of fermentation are shown in Table 2. As shown in Table 2, after two months of fermentation of anchovy salt, Gram negative, Catalytic negative, Oxidase positive bacillus and cocci were the main microflora. It can be seen that.

Table 2. Changes in Number of Cells During Fermentation

<Example 2>

Using the isolated microorganisms, proteolytic ability was measured in standard agar medium supplemented with skim milk 1%. Four strains with the highest proteolytic activity were selected and fibrin plate (0.01M phosphate buffered saline (pH 7.25), fibrinogen 0.3 %, thrombin 20 units) was used for thrombolytic dissolution. The results of measuring protein degradation and thrombolytic activity of these strains are as shown in FIGS. 2A and 2B. As can be seen in Figure 2a, Figure 2b it can be seen that the highest protein degradation and thrombolytic activity of strain FA0407.

Figure 2a is a photograph showing the protein degradation capacity of the microorganism separated during the fermentation anchovy salt fermentation process is a photograph showing the protein degradation capacity in the medium containing 1% reduced skim milk.

Figure 2b is a photograph showing the protein degradation capacity of the microorganisms isolated during the fermentation process of anchovy salt is a photograph showing the protein degradation capacity using the fibrin plate method.

FA0407 strains with high thrombolytic activity were applied to commercially available anchovy salt, and the results of the study were shown as shown in FIG. 3. Five kinds of anchovy saltfish (A, B, C, D, E) on the market were purchased and three products (A, B, C) that do not exhibit thrombolytic ability were mixed with FA0507 strain to have a concentration of 10 ⁷ . As a result, it can be seen that all three products show high thrombolytic activity (see FIG. 3).

<Example 3>

The amount of NO production and IL-1a production were measured to determine the immunoactivating ability of FA0507 strains that were found to have thrombolytic ability by application to commercial anchovy salt. As a result of the measurement, as shown in FIGS. 4 and 5, the amount of NO produced is increased in proportion to the concentration (see FIG. 4), and the amount of IL-1a is increased in proportion to the concentration (see FIG. 5). This shows that there is an immunoactivating effect of strain FA0407 (see FIGS. 4 and 5).

As a result of biochemical test and partial 16S rRNA sequencing of strain FA0407 having high protein degradation, thrombolytic and immuno-activity, the strain FA0407 was identified as Bacillus licheniformis . Table 3 shows the results of the identification of FA0407 strain.

Table 3. Identification results of strain FA0407

Cell shape Gram staining Catalae reaction Oxidase reaction Growth at 30 ℃ 37 ℃ rod +--+ + Sugar fermentation: D-arabinose L-arabinose Ribose Galactose Glucose Fructose Mannose Rhamnose Mannitol Sorbitol methyl-a, D-Glucoside N-acethyl-glucosamine Esculin Salicin Cellobiose Maltose Lactose Sucrose Starch Glycogen Xylitol Gentiobiose D-tagatose  -+ + + + + +-+ + +-+ + + + + + + +---+

As described above, Bacillus sp. Investigations on the health functional activity using FA0407 strains not only have thrombolytic activity and immune activity, but also have thrombolytic activity even when added to anchovy salt. It seems that the production of. The FA0407 strain, identified as Bacillus licheniformis , was deposited with the KCTC 10752BP on December 20, 2004 to the Korea Biotechnology Research Institute Gene Bank.

<Application example 1>

Buy fresh anchovies from Chujado and buy 5.5 parts by weight of fresh anchovy, 3.5 parts by weight of salt, 1 part by weight of purified water, and Bacillus licheniformis FA0407 (KCTC 10752BP) strains were mixed and aged at room temperature for at least 4 months to obtain anchovy.

<Application example 2>

6 parts by weight of fresh anchovies, 4 parts by weight of salt, 2 parts by weight of purified water, and Bacillus licheniformis FA0407 (KCTC 10752BP) strains were mixed at room temperature and aged for at least 6 months to obtain anchovy.

<Test Example>; Sensory evaluation

As an application example of the present invention, the anchovy taste, fishy taste, and sourness of anchovy marinated and anchovy chopsticks on the market (commodity) were compared.

Since the strains of the present invention have thrombolytic activity and immunological activity, using these strains can not only produce anchovy salt with excellent taste and functional activity, but also provide food with excellent functionality even when used in other kimchi or processed foods. can do

Claims (8)

Bacillus licheniformis FA0407 KCTC 10752BP isolated from anchovy salted fish (deposited Dec. 20, 2004 to the Genetic Bank of Korea Research Institute of Bioscience and Biotechnology). The new strain according to claim 1, wherein the new strain is excellent in protein resolution. The new strain according to claim 1, wherein the new strain has excellent thrombolytic activity. The new strain according to claim 1, wherein the new strain has excellent immunological activity. Bacillus licheniformis Anchovy salt containing FA0407 (KCTC 10752BP) strain. delete delete 5 to 6 parts by weight of raw anchovy, 3 to 4 parts by weight of salt, 1 to 2 parts by weight of purified water and Bacillus licheniformis FA0407 (KCTC 10752BP) by mixing the strain for the production of anchovy salt, characterized in that aged at room temperature for 6 months or more.

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