KR20150090416A - Bacillus licheniformis SCDB 34 strain having activity inhibiting growth of film-forming yeast isolated from traditionally fermented soybean product and uses therof - Google Patents

Abstract

The present invention relates to: a Bacillus licheniformis SCDB 34 strain (donation number KCCM11493P) having an activity in inhibiting growth of film-forming yeast; a composition for fermenting soybean paste containing a concentrate of a cultured material of the strain or a cultured liquid of the strain as an active ingredient; a method for producing fermented soybean paste, which comprises a step of inoculating and fermenting the strain in steamed soybeans; and fermented soybean paste produced through the method. A problem related to film formation which occurs when low salt fermented soybean paste is produced is relieved, so that the present invention can be industrially used.

Description

Bacillus licheniformis SCDB 34 strain having an ability to inhibit proliferation of cultured yeast isolated from a traditional pasture and its use {Bacillus licheniformis SCDB 34 strain having an activity inhibiting growth of yeast strain from traditionally fermented soybean product and uses therof}

The present invention relates to a strain of Bacillus licheniformis SCDB 34 having an ability to inhibit the growth of shoot-forming yeast isolated from a traditional pasture, and more particularly to a strain of Bacillus licheniformis SCDB 34 having a film-forming yeast growth- Bacillus A method for fermenting soybean paste comprising the step of fermenting soybean strain with fermented soybeans, comprising the step of fermenting the fermented soybean strain with a strain of Saccharomyces cerevisiae strain SCDB 34 (accession number KCCM11493P), a culture of the strain or a concentrate of the culture medium of the strain as an active ingredient, And a doenjang prepared by the above-mentioned method.

It is an important task to secure the safety of the soup by the corrosion which is essential for the Korean table. Due to the food culture habits developed by high salt fermented foods such as kimchi, soy sauce, and fermented seafood, Koreans consume the highest amount of salt in the world (12.2g per day) (sodium content: 4.8g) Organization) recommended amount (5 g / day). Since overdose of salt is associated with cardiovascular disease, stomach cancer, kidney stones, etc., efforts are being made nationwide to reduce its intake. According to the 2011 National Health and Nutrition Examination Survey, the main source of sodium intake is 17.7% from salt, 22.1% from kimchi (cabbage kimchi, bamboo kimchi and kakdugi), 20.6% from soy sauce (soybean paste, Of the total daily intake of salt is reported to eat more than 40%. As an improvement measure, the Korea Food and Drug Administration recommends that soy sauce be reduced by 0.4 g per 100 g of soy sauce. However, when fermenting protein and fat-rich soybeans with low salt, many kinds of noxious bacteria that inhibited proliferation due to salt grow rapidly, and it is easy to form acid film together with decay. In addition to lowering the merchantability of the fermented product, the acid film changes the community characteristics of the fermenting microorganisms due to the interruption of the oxygen supply, and causes self-digestion of the acid film to cause the growth of several spoilage bacteria. In order to inhibit the growth of spoilage bacteria, alcohol or natural spices having antimicrobial activity can be added, but there is a disadvantage that the inherent flavor of the soup is disappeared. As a solution to this problem, it would be most appropriate to select antagonistic fermenting bacteria against the rotifer that grows in low salt and then use it in the fermentation process. For this purpose, it is necessary to identify the acid-forming bacteria that are formed during the production of low-salt flour. However, until now, there has been no report except that the acid-forming yeast of the genus Zygosaccharomyces has been identified in commonly used high salt fermented soy sauce. Some strains are the genus Bacillus that the main role in fermenting together in the traditional sauces and in Aspergillus (Aspergillus) (Bacillus) that many bacterial community of the advantages involved in fermentation, especially the Bacillus genus are the antibiotics or surfactants By secretion Have been reported to have antibacterial and antifungal activity. The present invention relates to a method for isolating acid film-forming fungi that have proliferated during the production of low-salt doenjang, separating beneficial Bacillus species having antagonistic ability against these fungi, and producing hygienic low-salt products I aimed to try.

Korean Patent No. 0368183 discloses a strain of Bacillus licheniformis and one of its uses, Korean Patent Publication No. 2009-0120081 discloses a strain of Bacillus licheniformis useful for the production of chungkukjang, And Chungkukjang powder. However, the present invention is not limited to the Bacillus licheniformis SCDB 34 strain having the ability to inhibit the growth of clustering yeast isolated from the conventional bacterium of the present invention.

The present invention relates to a method for isolating Bacillus licheniformis SCDB 34 having an ability to inhibit the growth of shoot-forming yeast from a conventional herbaceous plant. The present invention relates to the isolation of Bacillus licheniformis SCDB 34, It is an object of the present invention to provide an effective strain capable of producing a low salt content by confirming that it has a resistance to the Kiakiidiae swallow and has a ribonucine synthetic gene showing antimicrobial properties.

In order to solve the above problems, the present invention provides a method for inhibiting proliferation of a film-forming yeast, comprising the steps of culturing Bacillus subtilis licheniformis ) SCDB 34 strain (KCCM11493P).

The present invention also provides a composition for fermenting soybean sprouts containing the strain, the culture of the strain, or a concentrate of the culture solution of the strain as an active ingredient.

In addition, the present invention provides a method for producing doenjang comprising the step of fermenting the strain by inoculating the expanded soybean.

In addition, the present invention provides a doenjang prepared by the above-described method.

In the present invention, it was confirmed that the strain Bacillus licheniformis SCDB 34 isolated from the traditional soy sauce has a ribonin synthase gene which not only effectively inhibits the growth of yeast forming an acid film in the production of low salt miso, but also has antimicrobial properties Respectively. By using the strain of the present invention, it is possible to industrially be used by solving the problem of the formation of the acid film occurring in the production of low salt miso.

1 is a scanning electron microscope (SEM) image of a membrane-forming yeast strain SCS FY10.
FIG. 2 is a diagram showing the single base difference in the ITS region nucleotide sequence of the film-forming yeast strains. FIG. The ITS region sequence (EF568018) of the P. kudriavzevii standard strain was used for the alignment of the ITS region.
Fig. 3 shows the relationship between yeast strains which are closely related to the film-forming yeast strains isolated from soybean paste prepared by soaking with 8% low salt water, based on the ITS nucleotide sequence (484 bp) It is a schematic diagram drawn by the Maximum Likelihood method.
Figure 4 is a flow diagram drawn by the Maximum Likelihood method based on the ITS nucleotide sequence to indicate the location of a strain isolated from a conventional species in relation to the genus Bacillus .

In order to accomplish the object of the present invention, the present invention provides a method for inhibiting proliferation of a film-forming yeast, which comprises culturing Bacillus licheniformis licheniformis ) SCDB 34 strain.

The Bacillus licheniformis SCDB 34 strain having a film-forming yeast proliferation inhibiting ability according to the present invention was isolated from a traditional pulp. Of the strains isolated from the traditional strains, pestresin, which is a biogenic amine, has high proteolytic activity and suppresses Bacillus cereus, which is a harmful microorganism, and has the highest activity as a fermentation strain, and named as SCDB 34 strain .

The strain Bacillus licheniformis SCDB 34 of the present invention was deposited at the Korean Microorganism Conservation Center on Dec. 3, 2013 (Accession No .: KCCM11493P).

The present invention also provides a composition for fermenting soybean sprouts containing the strain, the culture of the strain, or a concentrate of the culture solution of the strain as an active ingredient.

The present invention also provides a method of manufacturing a soybean paste comprising fermenting the strain with soybeans added thereto and soybean paste produced by the method.

The method for producing the doenjang using the strain of the present invention can be produced by a method known in the art.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Materials and methods

1. Preparation of meju and doenjang separation

For the preparation of doenjang, 20kg of meju bean (Sohn Changsan) was immersed in water, immersed in water at 121 ℃ for 30 min and cooled. Aspergillus oryzae ( Aspergillus oryzae) isolated from traditional red pepper paste oryzae SKM7 spores (3-7 x 10 ⁷ cfu / ml) and Bacillus sp. licheniformis) SCK B11 culture broth ^{(2 ~ 10 × 10 8 cfu} / ㎖) each 0.1% (v / w) and 0.05% (v / w) sikyeotgo was inoculated at a concentration at the same time the fermentation at 30 ℃ for 48 hours, this 60 ℃ And dried for 24 hours. Aspergillus oryzae SKM7 was cultured on a PDA medium at 28 ° C for 14 days, and spores were collected using 10 ml of 0.01% Triton X-100, followed by filtration. Bacillus licheniformis SCK B11 was inoculated after culturing in NB medium for 18 hours at 37 ° C. After adding 6 ℓ of 8% (w / v) and 14% (w / v) of saline to each grain of 2kg unit, they were immersed in fermented water for about 45 days, To remove miso.

The isolated soybean paste was aged at room temperature for 6 weeks and then subjected to high performance liquid chromatography (HPLC) to analyze organic acids.

2. Isolation and identification of acid-forming microorganisms

To separate the acid-forming microorganisms, the acid membrane was separated from each sample, diluted with distilled water, inoculated into 3M Petrifilm for yeast and mold isolation, and cultured at 27 ° C for 48 hours. The formed colonies were inoculated into PDA, cultured, separated again in new PDA, and confirmed by electron microscope.

Genomic DNA was extracted using a genomic DNA prep kit (Solgent Co., Ltd.) for isolation of the microorganisms, and the universal primer ITS1F (5'-TCCGTAGGTGAACCTGCGG-3 ' ; SEQ ID NO: 2) and ITS4R (5'-TCCTCCGCTTATTGATATGC-3 '; SEQ ID NO: 3). Amplification was performed at 95 ° C for 2 minutes, followed by 95 ° C for 20 seconds, 50 ° C for 40 seconds, 1 minute reaction was repeated 30 times and reaction was terminated at 72 ° C for 5 minutes. The reaction products were confirmed by electrophoresis on a 1.5% agarose gel, purified using a PCR purification kit (Qiagen, Germany), and analyzed for nucleotide sequences with Macrogen (Seoul, Korea).

ITS gene sequences of the standard strains were obtained from StrainInfo's Advanced Search and GenBank database, and CLUSTAL W was used for mutual comparison with the eight microbial ITS nucleotide sequences of the present invention. The phylogenetic analysis was performed using the Maximum Likelihood method based on the Tamura-Nei model after aligning the ITS gene sequences of the strains and calibrating to minimize gaps by visual observation and manual manipulation Respectively. The bootstrap analysis was performed 1,000 times to calculate the statistical reliability for each branch in each of the calculated tree branches, and the MEGA program was used for the system analysis and the bootstrap analysis.

3. Membrane-forming yeast ( firm - forming yeast ) With anti-proliferative activity Bacillus  Selection of strain

In order to select strains suitable for low salt fermentation, Bacillus sp. Bacillus strains with excellent fermentation ability were firstly selected and tested for antimutagenic effect against acid - forming yeast. First, to isolate strains of Bacillus sp. Having excellent fermentation ability, isolates of Bacillus cereus belonging to the Bacillus cereus group were selected by using chromogenic Bacillus cereus selective medium (Oxoid, UK) The presence or absence of 10 Bacillus cereus toxin synthetase genes that produce diarrhea, vomiting and respiratory toxin, the presence of biogenic amines in pH indicator containing media, the ability to degrade biogenic amines by HPLC, (2012, Korean J. Microbiol., 48, 220-224) based on the amount of glutamate produced, protease and amylase production ability.

The strains selected in the above procedure were examined for inhibiting the growth of shoot-forming yeast. The selected strains were cultured in PDB at 28 占 폚 for 18 hours, 200 占 퐇 were taken evenly on the surface of the PDA culture dish, and then put on a 6 mm diameter paper disk (Toyo Roshi, Japan). The selected strains were cultured in NB medium at 37 ° C. for 48 hours, and then centrifuged and 20 μl of the supernatant was dispensed on a paper disk. After further culturing in a 30 ° C. thermostat for 24 hours, a transparent pore diameter Were selected as the best isolates.

4. Selected Bacillus  Identification of the strain

For biochemical identification of Bacillus subtilis strains with inhibitory ability against acid forming yeast, they were inoculated on NA medium and cultured at 37 ° C for 18 hours. The colonies were diluted in NB medium and injected into a BCL ID card (bioMerieux Vitek Inc., Hazelwood, USA) consisting of 46 dry media and biochemical reagents. After 14 hours, the culture or reaction results were analyzed using VITEK 2 Were compared with biochemical databases of standard strains stored in Compact ™ software (bioMerieux Vitek).

For the molecular genetic identification of the selected strains, 16S rRNA gene (5'-AGAGTTTGATCCTGGCTCAG-3 '; SEQ ID NO: 4) and 1,492R And then 1,443 bp containing a variable base region important for identification was decoded using a Big Die Terminator v3.1 cycle sequencing kit. Sequence analysis was performed using the Maximum Likelihood method based on the Tamura-Nei model after aligning the 16S rRNA gene sequences of the strains and correcting them to minimize gaps by visual observation and manual manipulation. . The bootstrap analysis was performed 1,000 times to calculate the statistical reliability for each branch in each of the calculated tree branches, and the MEGA program was used for the system analysis and the bootstrap analysis.

5. Investigation of the presence of selected antimicrobial surfactant

In order to determine whether the starting bacteria contained the lichenysin or surfactin gene, a representative antibacterial surfactant of Bacillus subtilis or Bacillus subtilis , Bacillus licheniformis, synthase gene, the selection lchAA, lchAB, lchAC and Bacillus subtilis standing pectin synthase gene srfAA, srfAB, srfAC was performed by the PCR. Using the primers shown in Table 1, initial denaturation at 94 ° C for 2 minutes, denaturation at 94 ° C for 1 minute, binding at 56 ° C for 1 minute, and 72 ° C for 1 minute were repeated 30 times and final amplification at 72 ° C for 5 minutes The presence of the antibacterial surfactant was investigated.

List of primers used to confirm the presence of antimicrobial surfactant order
number primer Base sequence Target gene
(GeneBank accession no) Amplification product
Size (bp) 6 lchAF ACGGCCGATCAGGAGCTTTC Lichenysinsynthetase, lchAA
(AJ005061) 557 7 lchAR TCTCAGCGCCTTCGATCTGC 8 lchBF TTTGACCCGGAGCTCGTTGA Lichenysinsynthetase, lchAB
(AJ005061) 706 9 lchBR CTGAGGGCGGAAAGCAGGAT 10 lchCF CATGTATACGGGCCGACGGA Lichenysinsynthetase, lchAC
(AJ005061) 1173 11 lchCR CTGAAGGCCGGAGATGGCTT 12 srfAF CAGCGGCAGCGGATTAAATG Surfactinsynthetase AA, srfAA
(NC000964) 1025 13 SRFAR GGCCTTCAAAATCGCCTGCT 14 srfBF CGGTGTGTCATGGCGGATTT Surfactinsynthetase AB, srfAB
(NC000964) 696 15 srfBR TCGAAAGCGGACGGTTCAAA 16 srfCF TTCACTGTCGGAGGCGGAAA Surfactinsynthetase AC, srfAC
(NC000964) 933 17 srfCR ACCGGCAGATAGGCTGCTCC

Example  1. Salinity of Doenjang according to salt concentration

Salinity of soybean paste prepared by adding 8% (w / v) and 14% (w / v) of mannitol in water was measured for each grain Meju prepared by inoculation with Aspergillus oryzae and Bacillus licheniformis. As a result of measurement with a salinity meter (Takemura, Japan), 8 ± 0.4% (w / w) of salt was added to soy sauce prepared by soaking in 8% low salt water and 14 ± 0.5% (w / w) of salt.

Example  2. Characterization of Doenjang according to salt concentration

In the case of soybean paste prepared by soaking in 14% high salt water, the soybean paste prepared by soaking in 8% low salt water showed no distinctive flavor and flavor All had a white film on the surface and smelled of stale gods.

As a result of analysis of organic acids by HPLC (Agilent, USA), the content of succinic acid, lactic acid and acetic acid was 66 ± 4 mg / kg in soybean paste prepared by immersion in 14% 221 ± 7 mg and 1238 ± 8 mg, respectively. In the soybean paste prepared by soaking with 8% low salt water, 86 ± 5 mg, 680 ± 9 mg and 2864 ± 13 mg were found in soybean paste. , Lactic acid was about 3 times, and acetic acid was about 2.3 times.

Example  3. Isolation and identification of microorganisms

Eight kinds of yeast were isolated from the acid film of doenjang prepared by soaking with 8% low salt water. The morphology and size of the yeast were confirmed by electron microscope, and they were elliptical and had a size of about 1.7-2.5 탆 3.1-5.2 탆 (Fig. 1). As a result of CLUSTAL W, the common ITS region (484 bp) of the 8 strains was completely identical except for one sequence difference, as compared with the 476-723 bp nucleotide sequence obtained by amplifying the ITS region for identification of the isolated yeast (FIG. 2), and when they were compared with each other by the BLASTN search, they all belonged to the yeast of the genus Pichia . In order to identify the strains, we analyzed the phylogenetic relationships of strains using the ITS region sequences of the strains found in the Strain Info database and the MEGA program database. As a result, all eight yeasts were found to be Pichia Kidriabs Pichia kudriavzevii ) (Fig. 3). In particular, the discovery of the genus Pichia as the main membrane forming yeast of low salt doenjang has the significance that the present invention is the first.

Example  4. Antioxidant yeast ( firm - forming yeast ) With anti-proliferative activity Bacillus  Selection of strain

In order to select the fermenting strains which inhibit the growth of the yeast that form the acid film in the low salt fermented soybeans while maintaining the unique flavor of the traditional fermented soybean, 11 kinds of Bacillus , (Table 2). SCDB 34, the most important strain of these strains, was selected to investigate the inhibitory effect on growth inhibition of yeasts.

Table 3 shows the inhibitory power of acid-forming yeast by SCDB 34. SCDB 34 showed antagonistic ability against all membrane yeasts. Since the cultured strain was centrifuged and only the supernatant from which the cells had been removed was dispensed onto the paper disk, the substance inhibiting the proliferation of acid membrane forming yeast was presumed to be the extracellular release material of SCDB 34 strain.

Separate Bacillus (Bacillus) properties of sp Strain Deactivation of putrescine (%) Thiamine decomposition (%) Bacillus cereus inhibition Glutamate content
(mg / g) Bacillus cereus toxin gene Protease activity ^a Amylase activity ^b SCKB 17 64 ± 2 99 ± 1 ++ 0.96 + 0.12 none +++ +++ SCKB 18 56 ± 1 95 ± 1 ++ 1.28 ± 0.09 none ++++ +++ SCDB 19 68 ± 2 92 ± 3 ++ 0.96 + - 0.05 none ++++ +++ SCCB 32 88 ± 2 94 ± 2 +++ 4.47 ± 0.08 none +++ +++ SCKB 33 78 ± 3 92 ± 2 ++ 3.84 0.10 none +++ +++ SCDB 34 83 ± 1 91 ± 3 +++ 3.83 ± 0.08 none ++++ +++ SCDB 35 83 ± 3 88 ± 1 ++ 3.21 ± 0.13 none ++++ +++ SCDB 36 84 ± 2 91 ± 2 ++ 2.24 ± 0.15 none ++++ +++ SCCB 37 98 ± 1 90 ± 1 +++ 3.20 ± 0.08 none +++ +++ SCSB 38 90 ± 3 87 ± 1 ++ 3.23 ± 0.07 none ++ +++ SCSB 39 94 ± 2 91 ± 2 ++ 4.79 ± 0.12 none ++++ +++

^a halo radius. ++++; 10 mm or more, +++; 7-9 mm; ++, 5-6mm

^b Halo radius. +++; 7-9mm

Inhibition of growth of acid membrane-forming yeast by Bacillus strains isolated from doenjang Acid film formation
leaven Relative inhibition range by Bacillus strains ^a SCDB 34 SCS FY6 1.6 ± 0.05 SCS FY7 1.2 ± 0.01 SCS FY8 1.2 ± 0.01 SCS FY9 1.3 ± 0.03 SCS FY10 1.6 + 0.04 SCS FY11 1.4 ± 0.02 SCS FY12 1.4 ± 0.01 SCS FY13 1.5 ± 0.01

^a halo radius / paper disk radius

Example  5. Selected Bacillus  Identification of the strain

The selected Bacillus strain was identified using BCL ID card and identified as Bacillus licheniformis with a high probability of 95% (Table 4). The analysis of the base sequence of the 16S rRNA gene (SEQ ID NO: 1) It was identified as Bacillus licheniformis (Fig. 4).

Test reaction Test reaction β-Xylosidase - D-Mannitol + L-Lysine arylamidase - D-Mannose + L-Aspartate arylamidase - D-Melezitose - Leucinearylamidase + N-Acetyl-D-glucosamine (-) Phenylalanine arylamidase + Palatinose + L-Prolinearylamidase - L-Rhammose - β-Galactosidase + β-Gluse + L-Pyrrollydonylarylamidase + β-Mannosidase + α-Galactosidase - Phosphorylcholine esterase - Alanine arylamidase - Pyruvate (-) Tyrosine arylamidase + α-Glucosidae + β-N-Acetyl-glucosaminidase - D-Tagatose + Ala-Phe-Pro-arylamidase - D-Trehalose + Cyclodextrin + InuLin - D-Galactose - D-Glucose + Glycogen + D-Ribose + Myoinositol + Putrescine assimilation - Methyl-a-D-glucopyranose + Growth in 6.5% NaCl + Ellman - Kanamycin resistance - Methyl-D-xylosidase - Oleandomycin resistance - α-Mannosidase - Esculin hydrolase + Maltotriose + Tetrazolium red + Glycine arylamidase + Polymyxin B resistance +

Example  6. Investigation of the presence of selected antimicrobial surfactant

As a result, it was confirmed that the selected strain had a typical antimicrobial surfactant, richenicin or succinic acid gene. As a result, it possessed a richenicin synthetic gene (Table 5).

Presence of Surfactant Synthetic Gene of Selected Bacillus Strain Strain Surfactant synthetic gene ^a lchAA lchAB lchAC srfA srfB srfC Bacillus Lee Kenny Formis SCDB 34 + + + - - -

^a +; Exists; it does not exist.

Korea Microorganism Conservation Center (overseas) KCCM11493P 20131203

<110> Sunchang Research Center for Fermentation Microbes (SRCM) <120> Bacillus licheniformis SCDB 34 strain having activity inhibiting          growth of film-forming yeast isolated from traditionally          fermented soybean product and uses therof <130> PN13438 <160> 17 <170> Kopatentin 2.0 <210> 1 <211> 1454 <212> DNA <213> Bacillus licheniformis <400> 1 ctggcggcgt gcctaataca tgcaagtcga gcggaccgac gggagcttgc tcccttaggt 60 cagcggcgga cgggtgagta acacgtgggt aacctgcctg taagactggg ataactccgg 120 gaaaccgggg ctaataccgg atgcttgatt gaaccgcatg gttcaatcat aaaaggtggc 180 ttttagctac cacttacaga tggacccgcg gcgcattagc tagttggtga ggtaacggct 240 caccaaggcg acgatgcgta gccgacctga gagggtgatc ggccacactg ggactgagac 300 acggcccaga ctcctacggg aggcagcagt agggaatctt ccgcaatgga cgaaagtctg 360 acggagcaac gccgcgtgag tgatgaaggt tttcggatcg taaaactctg ttgttaggga 420 agaacaagta ccgttcgaat agggcggtac cttgacggta cctaaccaga aagccacggc 480 taactacgtg ccagcagccg cggtaatacg taggtggcaa gcgttgtccg gaattattgg 540 gcgtaaagcg cgcgcaggcg gtttcttaag tctgatgtga aagcccccgg ctcaaccggg 600 gagggtcatt ggaaactggg gaacttgagt gcagaagagg agagtggaat tccacgtgta 660 gcggtgaaat gcgtagagat gtggaggaac accagtggcg aaggcgactc tctggtctgt 720 aactgacgct gaggcgcgaa agcgtgggga gcgaacagga ttagataccc tggtagtcca 780 cgccgtaaac gatgagtgct aagtgttaga gggtttccgc cctttagtgc tgcagcaaac 840 gcattaagca ctccgcctgg ggagtacggt cgcaagactg aaactcaaag gaattgacgg 900 gggcccgcac aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacca 960 ggtcttgaca tcctctgaca accctagaga tagggcttcc ccttcggggg cagagtgaca 1020 ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 1080 cgcaaccctt gatcttagtt gccagcattc agttgggcac tctaaggtga ctgccggtga 1140 caaaccggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca 1200 cacgtgctac aatgggcaga acaaagggca gcgaagccgc gaggctaagc caatcccaca 1260 aatctgttct cagttcggat cgcagtctgc aactcgactg cgtgaagctg gaatcgctag 1320 taatcgcgga tcagcatgcc gcggtggaat acgttcccgg gccttgtaca caccgcccgt 1380 cacaccacga agagtttgta acacccgaag tcgggtgagg taaccttttt ggagccagcc 1440 gccgaaggtg ggac 1454 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 tccgtaggtg aacctgcgg 19 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 tcctccgctt attgatatgc 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 agagtttgat cctggctcag 20 <210> 5 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 ggttaccttg ttacgactt 19 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 6 acggccgatc aggagctttc 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 7 tctcagcgcc ttcgatctgc 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 8 tttgacccgg agctcgttga 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 9 ctgagggcgg aaagcaggat 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 10 catgtatacg ggccgacgga 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 11 ctgaaggccg gagatggctt 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 12 cagcggcagc ggattaaatg 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 13 ggccttcaaa atcgcctgct 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 14 cggtgtgtca tggcggattt 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 15 tcgaaagcgg acggttcaaa 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 16 ttcactgtcg gaggcggaaa 20 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 17 accggcagat aggctgctcc 20

Claims (4)

Lodges forming yeast (film-forming yeast) has a proliferation inhibitory effect, Bacillus Lee Kenny Po Ms (Bacillus licheniformis ) SCDB 34 strain (KCCM11493P). The Bacillus licheniformis SCDB 34 strain according to claim 1, wherein the film-forming yeast is Pichia kudriavzevii . A composition for fermenting doenjang containing the strain of claim 1, a culture of the strain, or a concentrated liquid of the culture medium of the strain as an active ingredient. A soybean paste prepared by inoculating and fermenting the strain of claim 1 in soy sauce.

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