• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.53-60
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• 2000

Strain BH5 was isolated from naturally fermented Kimchi and identified as a bacteriocin producer, which has bactericidal activity against Micrococcus flavus ATCC 10240. Strain BH5 was identified tentatively as Lactococcus lactis by the API test and some characteristics. Lactococcus lactis BH5 showed a broad spectrum of activity against most of the non-pathogenic and pathogenic microorganisms tested by the modified deferred method. The activity of lacticin BH5, named tentatively as the bacteriocin produced by Lactococcus lactis BH5, was detected at the mid-log growth phase, reached its maximum during the early stationary phase, and decreased after the late stationary phase. Lacticin BH5 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms as tested by the spot-on-lawn method. Its antimicrobial activity on sensitive indicator cells was completely disappeared by protease XIV or ${\alpha}$-chymotrypsin. The inhibitory activities of lacticin BH5 were detected during treatments up to 100$^{\circ}C$ for 30 min. Lacticin BH5 was very stable over a pH range of 2.0 to 9.0 and was stable with all the organic solvents examined. The cell concentration and bacteriocin production in strain BH5 were maximum when grown at 30$^{\circ}C$ in a modified MRS medium supplemented with 0.5% tryptone, 1.0% yeast extract, and 0.5% beef extract as nitrogen sources. It demonstrated a typical bactericidal mode of inhibition against Micrococcus flavus ATCC 10240. Lacticin BH5 was purified through ammonium sulfate precipitation, ethanol precipitation, and CM-Sepharose column chromatography. The apparent molecular mass of lacticin BH5 was estimated to be in the region of 3.7 kDa, by the direct detection of bactericidal activity after SDS-PAGE. Mutant strain NO141 which was isolated by nitrosoguanidine mutagenesis produced about 4 fold more bacteriocin than the wild type.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1092-1100
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• 2012

Ten Bacillus strains with antimicrobial activities were isolated from Cheonggukjang produced at different parts in Korea. They all inhibited Listeria monocytogenes ATCC 19111 and nine inhibited Bacillus cereus ATCC 14579. Four isolates (W42, H27, SKE 12, and K21) showing strong inhibiting activities were identified as B. subtilis. B. subtilis W42 was the most inhibiting strain. The antimicrobial activity of culture supernatant from B. subtilis W42 was destroyed completely by proteinase K treatment, indicating that a bacteriocin was the responsible agent. The bacteriocin, Bac W42, was most stable at pH 7 and stable between pH 3-6 and 8-9. Bac W42 was stable up to $80^{\circ}C$. BHI (brain heart infusion) and TSB (tryptic soy broth) were the best media for the activity (320 AU/ml) followed by LB (160 AU/ml). Bac W42 was partially purified by column chromatographies. The specific activity was increased from 1,151.2 AU/ml to 9,043.5 AU/ml and the final yield was 26.3%. Bac W42 was 5.4 kDa in size as determined by SDS-PAGE. Bac W42 showed bactericidal activity against L. monocytogenes ATCC 19111.

• Journal of Life Science
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• v.25 no.2
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• pp.180-188
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• 2015

The bacterial strain isolated from Kimchi showed antibacterial activity against Micrococcus luteus IAM 1056. The selected strain was identified as Lactococcus lactis by 16S rRNA nucleotide sequence analysis and named as Lactococcus sp. KD 28. The treatment of culture supernatant with proteinase K removed antibacterial activity, indicating its proteinaceous nature, a bacteriocin. This bacteriocin was sensitive to hydrolytic enzymes such as ${\alpha}$-chymotrypsion, trypsin, proteinase K, lipase, ${\alpha}$-amylase and subtilisin A. The bacteriocin was highly thermostable and resistant to heating at $80^{\circ}C$ for up to an hour but 50 % of the total activity was remained at $100^{\circ}C$ for 30 min. The pH range from 2.0 to 8.0 had no effect on bacteriocin activity and it was not affected by solvents such as acetonitrile, isopropanol, methanol, chloroform and acetone up to 50% concentration. The bacteriocin showed antibacterial activity against M. luteus IAM 1056, Lactobacillus delbrueckii subsp. lactis KCTC 1058, Enterococcus faecium KCTC 3095, Bacillus cereus KCTC 1013, B. subtilis KCTC 1023, Listeria ivanovii subsp. ivanovii KCTC 3444, Staphylococcus aureus subsp. aureus KCTC 1916, B. megaterium KCTC 1098 and B. sphaericus KCTC 1184. The bacteriocin was purified through ammonium sulfate concentration, SP-Sepharose chromatography and RP-HPLC. The molecular weight was estimated to be about 3.4 kDa by tricine-SDS-PAGE analysis.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.554-557
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• 2000

Strain JV3 was isolated from commercial Swiss cheese products and identified as a bacteriocin producer, which has bactericidal activity against Leuconostoc mesenteroides KCCM 11324. Strain JW3 was identified tentatively as Lactococcus lactis by the API test. The activity of lacticin JW3, named tentatively as the bacteriocin produced by Lactococcus lactis JW3, was detected during the mid-log growth phase, and reached a maximum during the early stationary phase, and decreased after the late stationary phase. Its antimicrobial activity on sensitive indicator cells was completely disappeared by protease IV. The inhibitory activities of lacticin JW3 were detected during treatments of up to $121\'^{circ}C$ for 15 min. Lacticin JW3 was very stable over a pH range of 2.0 to 9.0 The apparent molecular mass of lacticin JW3 was estimated to be in the region of 3-3.5kDa, which was determined by the direct detection of bactericidal activity after SDS-PAGE.

• Animal cells and systems
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• v.12 no.1
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• pp.41-46
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• 2008

A bacterium strain called Bacillus coagulans was isolated from an industrial wastewater drainage and selected for its antimicrobial activities against bacteria and fungi. Characterization studies strongly suggested that this strain is Bacillus coagulans. Antimicrobial activity was found against gram-positive, gram-negative bacteria and yeast strain. Maximal activity was observed after 24 h when incubated at $30^{\circ}C$ and pH 8. The activity was found to be stable at $75^{\circ}C$ for 30 min and at pH range of 2-12. Analysis of the antimicrobial compound by SDS-PAGE suggested a molecular mass of approximately 7.5 KDa. The substance was characterized as a bacteriocin, because of its proteinaceous nature and low molecular weight. Our bacteriocin could potentially be used as a food preservative, because of its thermostable property and broad antimicrobial spectrum.

• Microbiology and Biotechnology Letters
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• v.22 no.6
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• pp.700-706
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• 1994

Lactic acid bacteria in Kimchi fermentation were tested for inhibitory activity against Gram positive bacteria and Gram negative bacteria. The Lactobacillus brevis (KCCM 35464) was found to produce a antimicrobial substance. It showed relatively wide range of inhibition spectrum against gram positive and gram negative bacteria and maintained the inhibitory activity between pH 4.0 and pH 9.0. The antimicrobial substance was obtained in the stationary growth phase and was purified by gel chromatography. The inhibitory effect of the antimicrobial substance on sensitive bacterial strains was determined by filter paper test. The activity of antimicrobial substa- nce was stable at 75$\circ$C. On the basis of its electrophoretic pattern is SDS-PAGE, antimicrobial substance appeared as a single band of 59 KDalton.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.3
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• pp.441-445
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• 1993

Ten bacteriocin-producing Enterococcus faecium strains with urease activity ($1.10-6.2nkat.mL^{-1}$) were isolated from the rumen of 2-8 weeks old calves. All strains were resistant aginst disodium arsenate at a minimal inhibition concentration - MIC $5g.L^{-1}$ and mercury chloride ($MIC=10-20mg.L^{-1}$). Eight strains were resistant against silver nitrate ($MIC=40-50mg.L^{-1}$) and three against antibiotics used. The resistance against six antibiotics was found in A23 strain. Values of adherence index ranged from 5.02 to 20.4 enterococci adhered per one epithelial cell of rumen wall. All isolates produced bacteriocins which inhibited the growth at least of one of five indicator organisms. The EF1 strain with a good affinity to the epithelial cell ($15.2{\pm}1.2$) produced bacteriocin substance with antimicrobial activity against grampositive and gramnegative indicator bacteria.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.740-744
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• 2003

The antimicrobial substance produced by Lactobacillus bulgaricus was inactivated by pretense, which confirmed it as a bacteriocin and referred to 'bulgaricin HJ'. The bulgaricin HJ showed the inhibitory activity against mastitis pathogens, gram-positive and gram-negative bacteria. The optimal conditions for the production of bulgaricin HJ were at the temperature of $30^{\circ}C$ and 10 h after cultivation of L. bulgaricus. Staph. and Strep. agalactiae, common bovine mastitis pathogens, were treated with bulgaricin HJ by the agar well diffusion method and showed antimirobial activities to the bovine mastitis pathogens. The activity of the bulgaricin HJ was maintained at pH 6-7 and $100^{\circ}C$ for 60 min against the mastitis pathogens. The bulgaricin HJ was determined as class IV bacteriocin by various enzyme treatments. Colony forming units analysis with indicator strains by the treatments of bulgaricin HJ indicates that the mode of bacteriocin action was bactericidal rather than bacteriostatic.

• Preventive Nutrition and Food Science
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• v.15 no.1
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• pp.74-77
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• 2010

Antibiotic-resistant Staphylococcus aureus is beginning to pose a social issue. Thus, there is an urgent need for the development of effective anti-staphylococcal agents to eradicate antibiotic-resistant S. aureus in food systems and to treat the pathogen in clinical areas. To address this need, lactic acid bacteria (LAB) from kimchi were screened for the production of anti-staphylococcal bacteriocin. From this screening, a bacteriocin generated by the MK3 strain, which was identified by 16S rRNA gene sequence analysis as Enterococcus faecium, demonstrated antimicrobial activity against an S. aureus strain, and was designated enterocin MK3. Enterocin MK3 also demonstrated activity against other gram-positive bacteria, including several LAB and Listeria monocytogenes, but not gram-negative Escherichia coli. The molecular mass of enterocin MK3 was estimated as approximately 6.5 kDa on an SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) gel.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1393-1400
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• 2009

A bacteriocin-producing strain PI80 was isolated from the gut of Penaeus indicus (Indian white shrimp) and identified as Streptococcus phocae PI80. The bacteriocin was purified from a culture supernatant to homogeneity as confirmed by Tricine SDS-PAGE. Reverse-phase HPLC analysis revealed a single active fraction eluted at 12.94 min, and MALDI-TOF mass spectrometry analysis showed the molecular mass to be 9.244 kDa. This molecular mass does not correspond to previously described streptococcal bacteriocins. The purified bacteriocin was named phocaecin PI80 from its producer strain, as this is the first report of bacteriocin production by Streptococcus phocae. The bacteriocin exhibited a broad spectrum of activity and inhibited important pathogens: Listeria monocytogenes, Vibrio parahaemolyticus, and V. fischeri. The antibacterial substance was also sensitive to proteolytic enzymes: trypsin, protease, pepsin, and chymotrypsin, yet insensitive to catalase, peroxidase, and diastase, confirming that the inhibition was due to a proteinaceous molecule (i.e., the bacteriocin), and not due to hydrogen peroxide or diacetyl. Phocaecin PI80 moderately tolerated heat treatment (up to $70^{\circ}C$ for 10 min) and resisted certain solvents (acetone, ethanol, and butanol). A massive leakage of $K^+$ ions from E. coli $DH5\alpha$, L. monocytogenes, and V. parahaemolyticus was induced by phocaecin PI80, as measured by Inductively Coupled Plasma Optical Emission Spectrometry (ICPOES). Therefore, the results of this study show that phocaecin PI80 may be a useful tool for inhibiting L. monocytogenes in seafood products that do not usually undergo adequate heat treatment, whereas the cells of Streptococcus phocae PI80 could be used to control vibriosis in shrimp farming.