• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.259-265
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• 1994

A bacteriocin-producing lactic acid bacteria was isolated from contaminated milk products, which was identified by using the API50 CH kit as Lactococcus lactis subsp. lactis with reliability of 98%. Fatty and analysia of the cell membrane showed that this strain contained same fatty acids profiles as type strain, Lactococcus lactis subsp. lactis ATCC 19435. The bacteriocin of Lactococcus sp. HY 449 showed relatively wide range of inhibition spectrum against gram positive and some gram negative bacteria such as Escherichia coli and maintained the inhibitory activity between pH2.0 and pH9.0 The thermostability of this bacteriocin was higher in acidic solution than in distilled water and was stable at 60$\circ $C for 1 hour.

• Journal of Life Science
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• v.27 no.2
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• pp.202-210
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• 2017

As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

• Journal of Food Hygiene and Safety
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• v.20 no.4
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• pp.211-216
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• 2005

The bacteriocin produced by E. faecium MJ-14 was precipitated with $50\%$ saturated ammonium sulfate in MRS broth and then the precipitated protein was dissolved in 20 mM sodium phosphate buffer (pH 6.0). The crude bacteriocin was purified by CM-sepharose CL 6B and Sephacry S-100 column chormatograhy. In this case, the purification fold of the bacteriocin was 114, therefore, its activity was 127,293 BU/mg of specific activity. Result from SDS-PAGE of the purified bacteriocin, it was obtained two protein bands of 4.3 kDa and 5.8 kDa having antilisterial activity.

• Journal of Microbiology and Biotechnology
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• v.32 no.11
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• pp.1462-1470
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• 2022

Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0-70℃ and pH 2-10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.663-667
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• 2007

Pediococcus acidilactici produces bacteriocin, which kills Listeria monocytogenes. The bactericidal mode of action of the bacteriocin against L. monocytogenes V7 was investigated by transmission electron microscopy. The bacteriocin was purified partially from the cell-free extract using Micro-Cel and cation-exchange chromatography, and the specific activity was increased 1,791 fold. The bacteriocin (6,400 AU/ml) was inoculated with L. monocytogenes V7 and incubated for 0.5h, 1h, 3h, and 6h. The bacteriocin was found to destroy most of the cell wall and released most of the inclusions in the cells after 6 h of incubation. These results suggest that the bactericidal effect of the bacteriocin was due to bacterial lysis.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.183-189
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• 1992

Cultural conditions of Lactococcus sp. 1112-1, a bacteriocin producing strain, were studied for enhancing its production with regard to environmental and nutritional factors. Optimal compositions of culture medium for bacteriocin production were glucose 20 g/l as carbon source, casein acid hydrolyzate 15 g/l as nitrogen source, and sodium acetate 3 g/l, ammonium citrate 2 g/l as morganic salt with other basal components. The optimal pH of medium and fermentation temperature were 6.2 and $35^{\circ}C$, respectively. This strain required exclusively riboflavin and pantothenic acid for growth and bacteriocin production. In a 1l batch culture, stationary phase emerged after 8.5 hours of fermentation when 1.81 g/l of biomass was accumulated. The maximum antimicrobial activity was 3,894 IU/ml after 12 hours.

• Food Science of Animal Resources
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• v.31 no.2
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• pp.176-182
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• 2011

The bacteriocin-producing lactic acid bacteria Enterococcus faecium DB1 was isolated from Korean traditional gajami sikhae. Culture conditions were optimized by response surface methodology (RSM) to maximize bacteriocin DB1 production. E. faecium DB1 displayed the highest bacteriocin activity when grown in modified MRS medium containing sucrose, rather than glucose, as a carbon source. The effects of temperature, initial pH, and sucrose concentration were tested to determine the optimum conditions for maximum bacteriocin production by E. faecium DB1. A central composite design was used to control the three variables in the experiment. RSM revealed that the optimum values for bacteriocin production were 27.66 g/L sucrose, temperature of $34.37^{\circ}C$, and an initial pH of 6.54. A 2.08-fold increase in bacteriocin production was obtained with sucrose-containing MRS medium compared to production in standard MRS medium.

• Food Science of Animal Resources
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• v.39 no.6
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• pp.1015-1020
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• 2019

Staphylococcus aureus is a representative pathogenic bacterium carefully controlled in the dairy industry because it causes bovine mastitis and thus, can enter the dairy chain. Furthermore, the emergence of multi-drug resistant S. aureus is a big problem. We previously isolated a Lactococcus lactis strain producing a bacteriocin that exhibited strong antimicrobial activity against S. aureus. In this study, we investigated the synergistic inhibition of S. aureus by the bacteriocin and a bacteriophage (SAP84) which is specific to the organism. The bacteriocin (12.5-100 AU/mL) inhibited the growth of S. aureus KCTC 3881 in a dose-dependent manner, as did the bacteriophage SAP84 (0.001-1 MOI; multiplicity of infection). Co-treatment with the bacteriocin (100 AU/mL) and the bacteriophage (0.1 MOI) significantly inhibited the growth of S. aureus compared to each treatment alone (bacteriocin or bacteriophage), indicating the two components showed synergistic inhibition of S. aureus. Therefore, the bacteriocin and bacteriophage combination can be used as a good strategy for controlling pathogenic bacteria.

• The Plant Pathology Journal
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• v.17 no.5
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• pp.249-256
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• 2001

Xanthomonas axonopodis pv. glycines 8ra produces a bacteriocin called glycinecinA, which specifically inhibits the growth of bacteria belonging to Xanthomonas species. GlycinecinA was produced by culturing Escherichia coli DH5 containing biosynthetic genes for glycinecinA, and was tested for its control effect against X. vesicatoria on red pepper and X. oryzae pv. oryzae on rice. The bacteriocin activity was much higher in the cell extract than in the supernatant. It reached a maximum level at the stationary phase, ws maintained up to 2 months at room temperature and approximately 10 months at $4^{\circ}$. The optimum concentration of glycinecinA for the control in the greenhouse and in the field was 12,800 AU/ml. In this study, the activity of glycinecinA on rice and red pepper leaves continued for 7-8 days, during which the pathogen populations remained at low levels. Bacterial leaf spot of red pepper and bacterial leaf blight of rice were significantly reduced by the bacteriocin treatments. The control efficacy was as high as, or even higher than, the chemical treatment of copper hydroxide. These results suggest that the bacteriocin is a potential control agent for bacterial diseases.