• Journal of Life Science
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• v.19 no.7
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• pp.994-1002
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• 2009

A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.282-291
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• 1999

Lactic acid bacteria were isolated from Kimchi and screened for bacteriocin. A total of 99 strains showed antimicrobial activity when grown on solid media, yet only 10 showed antimicrobial activity in liquid media. Strain H-559, identified as Lactococcus lactis subsp. lactis, exhibited the strongest inhibitory activity and was active against pathogenic bacteria including Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus as well as other lactic acid bacteria. The antimicrobial substance produced by L. lactis subsp. lactis H-559 was confirmed to be a bacteriocin by the treatment of $\alpha$-chymotrypsin, and protease type Ⅸ and ⅩIV. The bacteriocin activity remained stable between pH 2.0 and pH 11.0 and during heating for 10 min at $100^{\circ}C$. The bacteriocin production started in the exponential phase and stopped in the stationary phase. L. lactis subsp. lactis H-559 showed the highest bacteriocin activity at a culture temperature of $25^{\circ}C$, and an inverse relationship between the bacteriocin productivity and mean growth rate at different culture temperatures was observed. The mean growth rate and bacteriocin productivity of L. lactis subsp. lactis H-559 increased as the initial pH of the media increased.

• Journal of Food Hygiene and Safety
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• v.22 no.3
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• pp.151-158
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• 2007

Among 68 strains of lactic acid bacteria (LAB) isolated from Dongchimi, a strain K11 was selected due to its bactericidal activity against Escherichia coli O157 The strain K11 was identified as Lactobacillus plantarum, based on physiological and biochemical characteristics. In the late exponential phase, La. plantarum K11 showed maximum bacteriocin activity (12,800 BU/mL) and maintained until the early stationary phase. The bacteriocin activity was completely inactivated by all the proteolytic enzymes such as pepsin, protease, proteinase K, papain, chymotrypsin, and trypsin, but the activity was not affected by catalase, a-amylase, lysozyme, and lipase, suggesting proteinaceous nature of the bacteriocin. Additionally, this activity was not affected in the pH range from 3.0 to 9.0 and under storage conditions like 30 days at -20,4, or $25^{\circ}C$. Although the bacteriocin activity was absolutely lost after 15 min treatment at 121, it was relatively stable at $70^{\circ}C$ for 60 min or $100^{\circ}C$ for 30 min. The activity was disappeared by treatment with acetone, benzene, ethanol, or methanol, but it was not affected by treatment with chloroform or hexane. The antibacterial activity of the bacteriocin was good against some LAB including Lactobacillus spp., Enterococcus spp., and Streptococcus spp., but not against food-borne pathogens such as Bacillus spp., Listeria spp., and Staphylococcus spp. as well as yeasts and molds. Especially, some intestinal bacteria such as Enterobacter aerogenes and E. coli were significantly affected by the bacteriocin of La, plantarum K11. Furthermore, the addition of 640 BU/mL resulted in the complete clearance of E. coli O157 after 10 hr.

• Korean Journal of Microbiology
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• v.55 no.3
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• pp.306-308
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• 2019

A number of Bacillus strains are known to have antimicrobial activity useful in various fields. In order to prevent Propionibacterium acnes, which is one of the factors of acne, we selected Bacillus halotolerans F41-3 which have high antimicrobial activities against P. acnes. We conducted complete genome sequencing of B. halotolerans F41-3 and analyzed genomic characteristics. This genome size is 4,144,458 bp with a G + C content of 43.76%, 4,145 total genes and 3,686 protein coding genes. Among the genes, we found that gene clulster of subtilosin, a kind of bacteriocin, synthesis and gene cluster of nickel transportation. Both of them may influence inhibition of P. acnes.

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.481-485
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• 2010

Lactobacillus plantarum strain KK3 with tannase activity was isolated from Gochunipkimchi (red pepper leaf kimchi) and showed a high antagonistic activity against five kinds of food pathogens. Strain KK3 secreted antibacterial compound into culture medium and 24-h culture in MRS broth at $30^{\circ}C$ was enough for the antibacterial compound production. The crude antibacterial compound prepared from culture supernatant inhibited the growth of some Gram-negative bacteria and Bacillus cereus but not Listeria monocytogenes. The antibacterial activity was sensitive to proteinase K treatment, confirming its proteinaceous nature (bacteriocin). The crude bacteriocin was active in the pH range 3.5-8.5 and extremely stable after 15 min of heat treatment at $121^{\circ}C$. The strain KK3 produced equally active bacteriocin in Chinese cabbage juice as it produced in MRS broth.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1493-1501
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• 2015

A Bacillus species, EMD4, with strong antibacterial activity was isolated from ganjang (soy sauce) and identified as B. subtilis. B. subtilis EMD4 strongly inhibited the growth of B. cereus ATCC14579 and B. thuringiensis ATCC33679. The antibacterial activity was stable at pH 3-9 but inactive at pH 10 and above. The activity was fully retained after 15 min at 80℃ but reduced by 50% after 15 min at 90℃. The activity was completely destroyed by proteinase K and protease treatment, indicating its proteinaceous nature. The bacteriocin (BacEMD4) was partially purified from culture supernatant by ammonium sulfate precipitation, and Q-Sepharose and Sephadex G-50 column chromatographies. The specific activity was increased from 769.2 AU/mg protein to 8,347.8 AU/mg protein and the final yield was 12.6%. The size of BacEMD4 was determined to be 3.5 kDa by Tricine SDS-PAGE. The N-terminal amino acid sequence was similar with that of Subtilosin A. Nucleotide sequencing of the cloned gene confirmed that BacEMD4 was Subtilosin A. BacEMD4 showed bactericidal activity against B. cereus ATCC14579.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.471-481
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• 2016

The primary objective of this study was to investigate the effects of the bacteriocin-producing heterofermentative lactic acid bacteria (LAB) and acid-resistant yeast isolated from Mukeunji, a Korean ripened kimchi on shelf-life extension and quality improvement of sourdough. According to gene sequence analysis the heterofermentative LAB that showed the antimicrobial activity against bread-spoilage Bacillus strains were identified as Leuconostoc mesenteroides LAS112, Lactobacillus brevis LAS129, and L. mesenteroides subsp. dextranicum LAB137. In addition, the yeasts that were able to grow at acidic pH were identified as Pichia membranifaciens YS05, Pichia fermentans YS19, and Pichia anomala YS26. During sourdough fermentation the levels of acetic acid and bacteriocin produced by L. brevis LAS129 strain were higher than those of L. mesenteroides LAS112 and L. mesenteroides subsp. dextranicum LAS 137 strains, whereas LAS112 strain produced the highest levels of lactic acid. The maximum bacteriocin activity (640 AU/g) against Bacillus subtilis ATCC 35421 was obtained in sourdough fermented by mixed culture of L. brevis LAS129 and P. membranifaciens YS05 or P. anomala YS26. After 24 h of fermentation at $30^{\circ}C$, the viable cell counts of LAS129 ($10^9CFU/g$) in sourdough were higher than those of the YS05 or YS26 ($10^7CFU/g$). Meanwhile, the viable cells of bread-spoilage strain in sourdough fermented with these strains were significantly (P < 0.05) lower than the control group.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.131-137
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• 2021

This study aimed at isolating lactic acid bacteria with anti-MRSA (methicillin-resistant Staphylococcus aureus) bacteriocin activity from fermented shrimp. We selected three strains, named Weissella sp. S1, S2, and S3, using analysis based on 16S rRNA gene sequences. All strains showed appropriate growth in an MRS medium containing 5% (w/v) NaCl and showed antibacterial activities against Bacillus cereus, Escherichia coli, Staphylococcus aureus, and MRSA. The strains exhibited similar growth rates at 0-5% NaCl, with approximate reduction in growth rate observed at 9% NaCl. Weissella sp. S1, S2, and S3 exhibited maximum growth rates at pH 7, 9, and 8, respectively. The crude bacteriocin was prepared from Weissella sp. S3 and subjected to characterization. The remaining activities after 30 min of exposure at each temperature were 100%, beyond 75%, and 49% at 4℃ and 37℃, 50℃ and 70℃, and 100℃, respectively. The remaining activities after 24 h of exposure at each pH were 100%, 75%, and 49% at pH 3 and 5, 7 and 9, and 10, respectively. Use of 50% (v/v) ethanol or isopropanol treatment did not diminish the antibacterial activity of the bacteriocin, while the 50% (v/v) hexane treatment reduced the activity by 51%. The molecular weight of the bacteriocin was nearly 6 kDa that was quantified using tricine-SDS-PAGE. Our findings suggest that Weissella sp. S3 may be considered a probiotic and useful source of antimicrobial substances in the development of bio-preservatives for food or in MRSA treatment.

• Journal of Food Hygiene and Safety
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• v.15 no.3
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• pp.271-276
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• 2000

A amylase producing bacteria were isolated from tofu residue and identified as Bacillus sp. according to the morphological and biochemical properties, which were named Bacillus sp. GM7330 and Bacillus sp. GM7312. The cultural condition for the production of amylase was showed on 5% tofu residue added 3% glucose and 0.15% yeast extract. And incubated during 72 hrs at 30。C, Bacillus sp. GM7330 and Bacillus sp. GM7312 were producing amylase of 488 units and 341 units.

• Microbiology and Biotechnology Letters
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• v.38 no.1
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• pp.84-92
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• 2010

Antibacterial compound from Bacillus subtilis MJP1 was purified using C18 Sep-Pak cartridge, ion exchange chromatography, and gel filtration chromatography. The purified antibacterial compound showed antibacterial activity against Listeria monocytogenes, Bacillus subtilis, Staphylococcus aureus subsp. aureus, and Enterococcus faecalis. The purified antibacterial compound was found to be stable at $100^{\circ}C$ for 5 min and in the pH range of 3.0~9.0, but it was unstable at pH 10.0. It was inactivated by proteinase K and pronase E, and heat treatment at $121^{\circ}C$ for 15 min, but it was stable with lipase and $\alpha$-amylase treatment, which indicated its proteineous nature. Ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compound and confirmed the existence of two peptides (3356.54 Da, 3400.5244 Da).