KR20160144630A - Bacillus licheniformis strain KJ-10 and uses thereof - Google Patents

Abstract

Provided are a Bacillus licheniformis KJ-10 strain (KACC92035P), and a fermented food including the same. The Bacillus licheniformis KJ-10 strain exhibits excellent blood clot-dissolving and enzymatic activities, and produces the small amount of volatile ammonia and amine when producing fast-fermented bean paste while having biogenic amine degradability. Also, provided are a microbial preparation including the strain or a cultured fluid thereof as an active ingredient, a method for producing the microbial preparation including a step of culturing the strain, and a fermented food including the strain.

Description

Bacillus licheniformis strain KJ-10 strain and its use {Bacillus licheniformis strain KJ-10 and uses thereof}

The present invention relates to a Bacillus licheniformis KJ-10 strain having biosgenic amine-decomposing ability and enzymatic activity and thrombolytic ability, and more particularly to a Bacillus licheniformis KJ-10 strain having a biogenic amine- Bacillus licheniformis KJ-10 strain (KACC 92035P), which is excellent in thrombolytic and cytotoxicological properties and produces less volatile ammonia and amines when producing chungkukjang, a microorganism preparation containing the strain or a culture thereof as an active ingredient, And a fermented food comprising the strain.

Cheonggukjang is one of the traditional fermented foods of Korea, which is one of the traditional fermented foods that have been traditionally used to date with soy sauce, soybean paste, and kochujang, and it plays an important role as a source food of essential amino acids and fatty acids, . In addition to nutritional research on soybean fermented foods such as Chungkukjang, functional studies such as thrombolytic, antioxidant, hypotensive, and atherogenic inhibition have been actively conducted in soybean fermented foods from all over the world.

Chungkookjang has the advantages of short fermentation period compared to other chapters of Korea. It has fermentation process and it is able to ferment the proteins, oligosaccharides, isoflavones, lipids Etc. are decomposed into amino acids, free sugars, isoflavone aglycones, fatty acids and the like which are easily digestible, and secondary products having antioxidative and thrombolytic functions are produced. Cholangiocarcinol has been reported to decrease cholesterol, antioxidant, hypertension, antibacterial and anticancer effects, and it has recently been reconsidered as a functional food by the action of Bacillus.

Soybean products mainly made from soybeans have been produced and used by self-produced before the 1960s (Agricultural, Shrimp and Newspaper Foods, 1995), but the mass-produced products can be easily found on the market due to the development of the food industry. Recently, however, the demand for Chungkookjang has increased as the traditional manufacturing method has become more and more convenient by purchasing goods through the Internet and using courier service rather than commercialized Chungkookjang. The number of products with homogeneous quality is small because the production methods and microorganisms used in the traditional manufacturing method are not the same in each province. In particular, quality varies depending on the type of strains used in fermentation of Cheonggukjang, so it is important to select and use high-performance microorganisms for production of high-quality Cheonggukjang.

Chungkookjang has excellent nutritional and physiological functions, but its consumption is limited due to the content of odor and biogenic amine produced during fermentation. The odor of Chungkookjang is caused by volatile amines produced by the action of microorganisms such as fermenting microorganisms in foods with high protein content, and the biogenic amines are basic nitrogen compounds, which are generally formed by decarboxylation with amino acids as precursors . The formation of biogenic amines in foods affects not only smell but also health. It also affects blood pressure and can cause an allergic reaction to a single person when there is a certain degree of abnormality in the food, causing problems on migraine and bowel. Recently, studies on biogenic amines produced by food microorganisms and studies on reduction of biogenic amines in traditional fields have been actively conducted.

Therefore, the present invention seeks to isolate and isolate strains from traditional fermented chongkukjang to select and characterize microorganisms having excellent functions. 7 strains of B. cereus toxin genes were identified to confirm the safety of strains isolated from Chungkookjang prepared by traditional fermentation method and to confirm secretion of substances capable of inhibiting contaminants, , Biogenamic amine decomposition ability, sensory evaluation and volatile amine test. In order to use as a basic data for setting the quality index of Cheonggukjang in the food industry such as fermentation starter of Chongkukjang, we are going to provide the best selected strains.

Related prior arts include Korean Patent No. 10-1379230 (Bacillus licheniformis SCK B11 strain having antimicrobial activity against harmful microorganisms and biogenic amine-degrading activity of soybean and its use) and Korean Patent Laid-Open No. 10-1300191 Bacillus licheniformis strains having histamine resolving ability and uses thereof).

An object of the present invention is to provide a Bacillus licheniformis KJ-10 strain having excellent thrombolytic and enzymatic activity, capable of decomposing biorenic amines and producing less volatile ammonia and amines when producing chungkukjang, and a microorganism preparation and a fermented food containing the same. .

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

In order to achieve the above object, the present invention provides a strain of Bacillus subtilis JP-5 containing the nucleotide sequence of SEQ ID NO: 1 and deposited at Accession No. KACC 92036P.

The strain is characterized by its thrombolytic ability and enzyme activity.

The strain is characterized in that it has the ability to decompose at least one of histamine and tyramine in the biogenic amine.

The enzyme is characterized by being at least one of amylase, cellulase, protease, and lipase.

The strain is characterized in that it does not contain a toxin gene.

The toxin genes include nheA, nheB, nheC , hemolytic enterotoxin genes hblA , hblC And cytok < RTI ID = 0.0 > Or more.

The present invention also provides a microorganism preparation comprising Bacillus licheniformis strain KJ-10 (KACC92035P) or a culture thereof as an active ingredient.

The present invention also provides a method for producing a microorganism preparation comprising culturing a strain of Bacillus licheniformis KJ-10 (KACC92035P).

The present invention also provides a fermented food comprising a strain of Bacillus licheniformis KJ-10 (KACC92035P).

The fermented food is characterized in that it is at least one selected from the group consisting of meju or Korean miso, miso, seasoned miso, kochujang, seasoned kochujang, chunjang, chungkukjang, mixed soup, Korean soy sauce, brewed soy sauce and mixed soy sauce.

The content of volatile ammonia and amine in the fermented food is lower than the content of volatile ammonia and amine in the fermented food not containing the strain.

In the present invention, Bacillus licheniformis KJ-10 isolated from a traditional soybean has excellent biosynic amine decomposition ability, excellent enzyme activity and thrombolytic ability, and produces less volatile ammonia and amine during the production of chungkukjang. Respectively. Therefore, Bacillus licheniformis KJ-10 of the present invention is a safe strain, so it can be used as a useful strain capable of producing high quality products having low functionality and odor by being used together with seeds in the production of traditional soup .

Brief Description of the Drawings Figure 1 is a diagram showing the results of amylase, cellulase, lipase and protease activity of Bacillus licheniformis strain KJ-10, which is a strain of the present invention.
(A) Amylase, (B) Cellulase, (C) Lipase, (D) Protease
2 is a diagram showing the results of thrombolysis of a strain of Bacillus licheniformis KJ-10, which is a strain of the present invention.
3 is a diagram showing the presence or absence of the B. cereus toxin gene of the strain of the present invention, Bacillus licheniformis KJ-10.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail, and a detailed description of known functions and configurations that may unnecessarily obscure the gist of the present invention will be omitted.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail, and a detailed description of known functions and configurations that may unnecessarily obscure the gist of the present invention will be omitted.

The present invention provides a Bacillus licheniformis KJ-10 strain (KACC 92035P) having a biogenic amine-cleaving ability, an excellent enzyme activity and a thrombolytic ability, and less volatile ammonia and amine formation during fermentation of chongkukjang.

The Bacillus licheniformis KJ-10 strain (KACC 92035P) was isolated from a traditional herb, and has no toxin gene, and has a biogenic amine-degrading ability, a thrombolytic activity, and an enzyme activity. The above-mentioned Bacillus licheniformis strain KJ-10 was deposited at the National Institute of Agricultural Science on Feb. 11, 2015 (Accession No .: KACC 92035P)

In a strain according to an embodiment of the present invention, the biogenic amine may be histamine or tyramine, but is not limited thereto.

The biogenic amine of the present invention is formed from an amino acid by the action of an amino acid decarboxylase of a microorganism. When a biogenic amine is ingested from a food in excess of the decomposition limit of the human body, the biogenic amine may cause rash, local skin inflammation, Vomiting, nausea and diarrhea.

In a strain according to an embodiment of the present invention, the enzyme may be, but not limited to, amylase, cellulase, protease, and lipase.

In the case of the strain according to an embodiment of the present invention, the toxin gene may be a toxin gene produced by Bacillus cereus, which is a harmful microorganism occurring in a soybean, but may be nheA , nheB, nheC , hblA, hblC , cytK, plcRpapR Do not.

The present invention also provides a microorganism preparation comprising the Bacillus licheniformis strain KJ-10 or a culture thereof as an active ingredient.

The microbial formulation according to one embodiment of the present invention may be prepared in the form of a liquid, and may be added to the granule formulation in the form of a powdered powder or may be granulated by formulating it. However, the formulation is not particularly limited.

Also, the present invention provides a method for producing a microorganism preparation comprising culturing the Bacillus licheniformis KJ-10 strain. The Bacillus licheniformis strain KJ-10 of the present invention can be cultured by a conventional method known in the art.

The present invention also provides a fermented food comprising the strain Bacillus licheniformis KJ-10.

The food according to one embodiment of the present invention may be a fermented food, and the fermented food may be meju or soup, but is not limited thereto.

In the food according to an embodiment of the present invention, the soy sauce may be, but not limited to, Korean-style miso, miso, seasoned miso, kochujang, seasoning kochujang, spring roll, chonggukjang, mixed roll, Korean-style soy sauce, brewed soy sauce and mixed soy sauce .

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, and it is to be understood by those skilled in the art that the present invention is not limited thereto It will be obvious.

Example 1 Preparation of Bacillus licheniformis ( Bacillus licheniformis) Isolation and identification of KJ-10 (KACC 92035P) strain, selection

A variety of strains isolated by R2A agar and Raka-Ray agar were streaked on R2A agar (R) agar, which was diluted to 0.85% in saline buffer. And cultured on a plate medium at 28 ° C for 1 to 2 days.

The KJ-10 strain was isolated from the pure isolates by enzymatic activity (amylase, cellulase, protease, lipase), enzymatic resolution, presence of toxic genes, biogenic amine degradation ability and production of volatile ammonia and amine during fermentation of chungkukjang. Were selected.

In order to classify the KJ-10 strain of the present invention, 16S rRNA was amplified using PCR as follows to determine the nucleotide sequence (SEQ ID NO: 1).

In order to classify isolated strains, 16S rRNA was amplified using PCR as follows to determine the base sequence. Genomic DNA was extracted (DNA purification kit, Promega, USA) from Geno Tech Corp. (Genotech, Deajeon, Korea) to amplify the 16S rRNA nucleotide sequence, and a universal PCR primer 27F (5 ' -AGAGTTTGATCCTGGCTCAG-3 '; SEQ ID NO: 2) and 1492R (5'-GGTTACCTTGTTACGACTT-3'; SEQ ID NO: 3). Using the 16S rRNA seqeunce obtained from the above experiment, the homology between the isolates and the registered strains was compared using the Blast Search program of Ezbiocloud (http://www.ezbiocloud.net/eztaxon) provided by Chunlab, Inc. . In order to determine the phylogenetic location of the isolates, we used the Advenced Blast Search program of the National Center for Biotechnology Information (NCBI, http://www.ncbi.nlm.nih.gov/) The nucleotide sequences were analyzed using the MegAlign program (DNA STAR Lagergene, version 8) and the Mega 6.0 program neighbor-joining method.

The homology of the 16S rRNA sequence of the KJ-10 strain was compared and found to be similar to that of Bacillus licheniformis . The strain of the present invention was named Bacillus licheniformis KJ-10 and deposited under the accession number KACC 92035P on Feb. 11, 2015 at the National Institute of Agricultural Science (National Institute of Agricultural Science and Technology, KACC).

≪ Example 2 > Analysis of enzyme activity

In order to investigate the enzyme activity of the strain Bacillus licheniformis KJ-10 of the present invention, the enzymatic activity and thrombolytic activity of amylase, cellulase, protease and lipase were determined in a selection medium .

- amylase activity

R2A (Difco) was added with 0.5% soluble starch, which is an amylase production inducer. Two strains were inoculated into the plate culture medium and incubated at 28 ° C for 24 hours. The culture medium was stained with Gram's iodine solution (1.0 g of iodine crystal, 0.2 g of potassium iodine, and 30 ml of distilled water) Was confirmed through a clear zone.

- cellulase activity

The R2A medium supplemented with 0.4% CMC (carboxymethyl cellulose) was inoculated in a confluent culture method, cultured at 28 ° C for 24 hours, stained with 0.1% congo red solution, washed with 1M NaCl, Respectively.

- protease activity

2% skim milk and 3% agar were sterilized to prepare a culture medium. Then, the strain was inoculated with two strains in a confluent culture method and cultured at 28 ° C for 24 hours to confirm a clear zone.

- lipase activity

To 450 ml of the R2A agar, 50 ml of the mixture [5 ml of glycerol tributyrate, 1 ml of 0.5 M CaCl ₂ , 2 ml of 5 M NaCl, 2.5 g of gum arabic (from acacia tree)] was mixed using a mixer After sterilization using a high pressure steam sterilizer at 121 ° C for 15 minutes, a solid medium was prepared and the bacteria were inoculated in a confluent culture method to confirm a clear zone.

- thrombotic resolution

Fibrin plate method. 0.5% fibrinogen (containing 0.067M sodium phosphaste buffer) was prepared, and the mixture was completely dissolved at 37 ° C for 30 minutes. Thrombin (1%) was added thereto. After 8 hours of inoculation with the strain on the hard medium, the solubility of fibrin in the medium was confirmed.

Brief Description of the Drawings Fig. 1 is a schematic view showing an example of the amylase (Fig. 1 (A)), cellulase (Fig. 1 (B)), lipase of Bacillus licheniformis strain KJ- (Fig. 1 (C)) and protease (Fig. 1 (D)). FIG. 2 is a photograph showing the result of thrombolysis of the strain Bacillus licheniformis KJ-10 of the present invention by the fibrin plate method.

As shown in FIG. 1 and Table 1, the Bacillus licheniformis KJ-10 strain of the present invention showed high activity in both amylase, cellulase, protease and lipase, which are fermentation-related enzymes, Bacillus subtilis KACC16479 and Bacillus subtilis A, respectively.

Enzyme activity of KJ-10
Strain
Blast Enzyme activity Amy. Cell. Pro. Lip. Fibrin. KJ-10 B. licheniformis + + + + + KACC16749 B. subtilis subsp . subtilis + + + + - A B. subtilis subsp . subtilis + + + + (+)

Symbols: -; negative activity, (+); weak activity, +; positive activity, Amy .; amylase, Cell; cellulase, Pro .; protease, Lip .; lipase KACC 16749, A: control strain

≪ Example 2 > Bacillus cereus ( B.cereus ) Check for toxin gene

In order to ensure the safety of the strain Bacillus licheniformis KJ-10 of the present invention, Bacillus cereus , 6 genes for diarrhea toxin and 1 gene for diarrhea toxin expression transcriptional regulation were identified by PCR. Control strains were purchased from KACC Bacillus cereus Standard strain (KACC 11204) was used.

The toxin genes of Bacillus cereus used in this experiment are as shown in Table 2, and include 3 genes ( nheA ; nheB ; nheC ) of non-hemolytic enterotoxin, 2 types of hemolytic enterotoxin genes ( hblA ; hblC ) Cytotoxin K gene (cytK), and diarrhea toxin expression transcriptional regulatory protein, phospholipase C regulatory gene (plcRpapR) of the protein, conditions for PCR, after 2 minutes at 94 ℃ initial denaturation, 94 ℃ 1 bungan denaturation, 56 ℃, 58 ℃, The amplification was carried out at 72 ° C for 5 minutes. The amplification was carried out at 72 ° C for 30 minutes (72 ° C for 1 minute, 72 ° C for 1 minute).

Bacillus cereus toxic gene sequence and binding temperature. Primer pair Sequence Target sequence
(GeneBank accession no.) Size
(bp) Annealing temp. nheAF ATATGCGCAAAATGTAATTGCTCCA
(SEQ ID NO: 4) Non-hemolytic enterotoxin,
nheA (AY835995) 935
56 ℃
nheAR TGCCTTCTTCAACATTTGTTTGAATTT
(SEQ ID NO: 5) nheBF GACCAGCAGGATTCCCAGATGTAAT
(SEQ ID NO: 6) Non-hemolytic enterotonin,
nheB (AY835995) 972
62 ° C
nheBR CCACGCCTTCATGTAATTTTTCTGT
(SEQ ID NO: 7) nheCF ACCAGTAGCGACTTTTGCAAGTGAA
(SEQ ID NO: 8) Non-hemolytic enterotoxin,
nheC (AY835995)
930
62 ° C
nheCR TTTTGAGCTGCATTCTCAATATGCC
(SEQ ID NO: 9) HBlAF ACCAGTAGCGACTTTTGCAAGTGAA
(SEQ ID NO: 10) Hemolytic enterotoxin,
HBla (AY822584)
909
56 ℃
HBlAF TTTTGAGCTGCATTCTCAATATGCC
(SEQ ID NO: 11) hblCF ATCAATACTCTCGCAACACCAACTG
(SEQ ID NO: 12) Hemolytic enterotoxin,
hblC (AY822584)
957
62 ° C
hblCR ATGTGCTCGTTGCTCTGCTGTTAAT
(SEQ ID NO: 13) cytKF CCGCTGTTTTTGCTAGTAGTGCTGT
(SEQ ID NO: 14) Cytotoxin K,
cytK (DQ019311)

901
58 ℃
cytKR ACGTCTTTTACGTTGTTTCCAACCC
(SEQ ID NO: 15) plcRF CRGGYGCRGTATACCCAAGT
(SEQ ID NO: 16) Phospholipase C regulatory protein, ( plcRpapR (58 ) 888
58 ℃
plcRR TGAAATACCCCATGYCATYG
(SEQ ID NO: 17)

Bacillus licheniformis KJ-10 is not only a phylogenetically close to Bacillus cereus but also a strain isolated from chungkukjang, which is produced by a conventional fermentation method, not by inoculating a strain. Therefore, by confirming presence or absence of a Bacillus cereus toxin gene And to secure the safety of the test strains. As shown in FIG. 3, the Bacillus licheniformis KJ-10 strain did not have six diarrheal toxin genes of Bacillus cereus and one gene for regulating diarrhea toxin expression transcription.

≪ Example 3 > Quantitative analysis of resolving ability of biogenic amine

The histamine and tyramine resolutions of the Bacillus licheniformis KJ-10 of the present invention were quantitatively analyzed by HPLC to determine the biodegenerative activity.

The strain was inoculated with nutrient broth containing 1% histamine and tyramine and then cultured at 38 ° C for 20 days. After incubation, centrifugation was carried out at 1300 rpm for 5 minutes. To 200 상 of the supernatant, 2 내부 of internal standard solution 2M Diaminoctane was added.

After adding 300 μl Na ₂ CO ₃ solution (100 mg / ml in water) and 800 μl dansyl chloride solution (7.5 mg / ml in acetone), the mixture was dansilated at 60 ° C for 20 minutes.

After adding 100 μl of proline solution (50 mg / ml in water), the mixture was allowed to stand at 60 ° C for 10 minutes and then cooled at 5 ° C. Finally, 100 μl of toluene was added and shaken, followed by centrifugation for 5 minutes, followed by HPLC analysis using the supernatant.

The analysis conditions of the HPLC for the above experiment are as follows.

Amines were extracted using an Alliance e2695 HPLC system (Waters Co., Milford, MA, USA) and a 474 fluorescence detector (Waters Co., Milford, Mass., USA).

The flow rate was 1 ml / min using a C18 column (Waters, X Bridge C18, 4.6 X 250 mm 3.5 μm), and the concentration gradient of the mobile phase is shown in Table 3 below.

The concentration gradient of the HPLC mobile phase used in the present invention min A (Water)% B (Acetonitrile)% 0 60 40 5 60 40 35 15 85 40 15 85 54 0 100 55 60 40 60 60 40

The resolving power of 1% histamine and tyramine after 20 days of KJ-10 was confirmed. As shown in Table 4, histidine resolving power of 77.5% and tyramine resolving power of 23.8% were shown.

Biogenic amine resolution of KJ-10
Strain
Blast Histamine Tyramine before after 20days before after 20days KJ-10 B. licheniformis 54402959 12235980 68760490 52370219

< Example  4> volatile Amine  And ammonia measurement

To measure volatile ammonia and amines of fermented Chungkukjang using Bacillus licheniformis KJ-10, fermented Chungkookjang fermented with Bacillus licheniformis KJ-10 was first prepared. Bacillus licheniformis KACC16749 and Bacillus licheniformis A were used as control strains. 1.5 times water was added to the washed soybeans and the mixture was heated at 121 ° C for 20 minutes. 10 g of boiled soybeans maintained at a temperature of 40 ° C. were mixed with 250 μl of each strain culture (1 × 10 9 CFU / ml) and cultured at 40 ° C. for 72 hours to prepare chungkukjang using each strain. The content of ammonia and amine in 50 ml of gas inside the silicon container was measured using a detector tube type gas detector (GASTEC, GV-100S, Japan) after shaking for 1 minute and waiting for 5 minutes. At this time, an ammonia detection tube (Ammonia; NH3, No.3L, Japan) and an amine detection tube (Amines (Methylamine); R? NH2 No.180, Japan) were used.

As a result of the measurement, as shown in Table 5, 21 ppm ammonia and 13 ppm amine were found in 50 ml of gaseous Bacillus licheniformis KJ-10 in the gas, which was lower than that of the strain isolated from commercially available Cheonggukjang.

Strain
Blast ppm / 50 ml Ammonia Amines KJ-10 B. licheniformis 21 13 KACC16749 B. subtilis subsp . subtilis 6.5 13 A B. subtilis subsp . subtilis > 35 > 120

Symbols:>; excess

Based on the above results, the present invention provides a Bacillus licheniformis KJ-10 strain having excellent thrombolytic and enzymatic activity, having a biogenic amine-decomposing ability and less volatile ammonia and amine production during the production of chungkukjang, and a fermented food . &Lt; / RTI &gt;

Korea Agricultural Microbiology Resource Center KACC92035P 20150311

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Bacillus licheniformis strain KJ-10 and uses thereof <130> P2015-0041 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1451 <212> RNA <213> Bacillus licheniformis KJ-10 <400> 1 gcagtcgagc ggaccgacgg gtagcttgct cccttaggtc agcggcggac gggtgagtaa 60 cacgtgggta acctgcctgt aagactggga taactccggg aaaccggggc taataccgga 120 tgcttgattg aaccgcatgg ttcaatcata aaaggtggct tttagctacc acttacagat 180 ggacccgcgg cgcattagct agttggtgag gtaacggctc accaaggcga cgatgcgtag 240 ccgacctgag agggtgatcg gccacactgg gactgagaca cggcccagac tcctacggga 300 ggcagcagta gggaatcttc cgcaatggac gaaagtctga cggagcaacg ccgcgtgagt 360 gatgaaggtt ttcggatcgt aaaactctgt tgttagggaa gaacaagtac cgttcgaata 420 gggcggtacc ttgacggtac ctaaccagaa agccacggct aactacgtgc cagcagccgc 480 ggtaatacgt aggtggcaag cgttgtccgg aattattggg cgtaaagcgc gcgcaggcgg 540 tttcttaagt ctgatgtgaa agcccccggc tcaaccgggg agggtcattg gaaactgggg 600 aacttgagtg cagaagagga gagtggaatt ccacgtgtag cggtgaaatg cgtagagatg 660 tggaggaaca ccagtggcga aggcgactct ctggtctgta actgacgctg aggcgcgaaa 720 gcgtggggag cgaacaggat tagataccct ggtagtccac gccgtaaacg atgagtgcta 780 agtgttagag ggtttccgcc ctttagtgct gcagcaaacg cattaagcac tccgcctggg 840 gagtacggtc gcaagactga aactcaaagg aattgacggg ggcccgcaca agcggtggag 900 catgtggttt aattcgaagc aacgcgaaga accttaccag gtcttgacat cctctgacaa 960 ccctagagat agggcttccc cttcgggggc agagtgacag gtggtgcatg gttgtcgtca 1020 gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg atcttagttg 1080 ccagcattca gttgggcact ctaaggtgac tgccggtgac aaaccggagg aaggtgggga 1140 tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac acgtgctaca atgggcagaa 1200 caaagggcag cgaagccgcg aggctaagcc aatcccacaa atctgttctc agttcggatc 1260 gcagtctgca actcgactgc gtgaagctgg aatcgctagt aatcgcggat cagcatgccg 1320 cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccacgaga gtttgtaaca 1380 cccgaagtcg gtgaggtaac cttttggagc cagccgccga aggtgggaca gatgattggg 1440 gtgaagtcgt a 1451

Claims (11)

Bacillus licheniformis KJ-10 strain containing the nucleotide sequence of SEQ ID NO: 1 and deposited with Accession No. KACC92035P.
The method according to claim 1,
Bacillus licheniformis KJ-10 strain (KACC92035P) with thrombolytic and enzymatic activity.
The method according to claim 1,
Bacillus licheniformis strain KJ-10 (KACC92035P), which has the ability to degrade either histamine or tyramine among biogenic amines.
3. The method of claim 2,
The enzyme is Bacillus licheniformis KJ-10 (KACC92035P), which is one or more of amylase, cellulase, protease, and lipase.
The method according to claim 1,
Bacillus licheniformis KJ-10 strain (KACC92035P) which does not contain toxin gene.
6. The method of claim 5,
The toxin gene is the gene of the enterotoxin Non-hemolytic nheA, nheB, nheC, Hemolytic enterotoxin gene of hblA, hblC and Cytotoxin K gene cytK Bacillus licheniformis KJ-10 strain (KACC92035P).
A microbial preparation comprising the Bacillus licheniformis strain KJ-10 (KACC92035P) or a culture thereof as an active ingredient according to any one of claims 1 to 6.
A method for producing a microbial agent comprising culturing a Bacillus licheniformis KJ-10 strain (KACC92035P) according to any one of claims 1 to 6.
A fermented food product comprising the Bacillus licheniformis KJ-10 strain (KACC92035P) according to any one of claims 1 to 6.
10. The method of claim 9,
Wherein the fermented food is at least one selected from the group consisting of meju or Korean miso, miso, seasoned miso, kochujang, seasoned kochujang, chunjang, chungkukjang, mixed jang, Korean traditional soy sauce, brewed soy sauce and mixed soy sauce.
10. The method of claim 9,
Wherein the content of volatile ammonia and amine in the fermented food is lower than the content of volatile ammonia and amine in the fermented food not containing the strain.

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