• Title/Summary/Keyword: bacteriocin

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Antilisterial Effect of Bacteriocin SH01, Obtained from Enterococcus faecium SH01, in Ground Beef

  • Kim, Min-Ju;Jung, Miran;Kim, Wang June
    • Food Science of Animal Resources
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    • v.35 no.2
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    • pp.211-215
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    • 2015
  • From the previous study, Enterococcus faecium SH01 was isolated from mukeunji, an over-ripened kimchi, and it produced bacteriocin SH01. Bacteriocin SH01 showed an inhibitory effect against Listeria monocytogenes ATCC 19111, a bacterial strain causing human listeriosis. Crude bacteriocin SH01 was purified by ammonium sulfate precipitation and its inhibitory activity at two concentrations (500 and 1,000 AU/g) against Listeria monocytogenes ATCC 19111 was investigated in ground beef at increasing temperatures (5, 10, 15, and 20℃) for 8 d. The number of Listeria monocytogenes ATCC 19111 significantly decreased (p<0.05) as the concentration of bacteriocin increased from 500 to 1,000 AU/g. Intrinsic crude protease activities in ground beef were examined and increased as the temperature increased. Experiments varying both the concentrations of added bacteriocin SH01 and temperature demonstrated a maximum inhibition (2.33 log reduction of bacteria) in samples containing 1,000 AU/g of bacteriocin SH01 incubated at 20℃. When the crude bacteriocin SH01 solution (1,280 AU/mL) was incubated with crude protease solutions at different temperatures, its activity decreased by only half (640 AU/mL), as assessed in an agar well diffusion assay. The finding that the antilisterial activity of bacteriocin SH01 increased with temperature can be explained by the fact that higher temperatures increase bacterial membrane fluidity, thereby promoting the cellular penetration of bacteriocin SH01 into L. monocytogenes. Bacteriocin SH01 may be an excellent candidate as a biopreservative for controlling L. monocytogenes growth in ground beef.

Characteristics and Partial Purification of a Bacteriocin Produced by Pediococcus damnosus JNU 534 (Pediococcus damnosus JNU 534가 생산하는 박테리오신의 특성 및 정제)

  • Lee, Jae-Won;Han, Su-Min;Yun, Bo-Hyun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.952-959
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    • 2011
  • A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

Isolation of Lactococci Inhibiting Listeria monocytogenes from Kimchi Habitat and Its Identification by 16S rDNA Analysis (김치 서식처에서 Listeria monocytogenes를 억제하는 lactococci의 분리와 16S rDNA분석에 의한 동정)

  • 박은주;한홍의;민봉희
    • The Korean Journal of Ecology
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    • v.22 no.1
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    • pp.45-50
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    • 1999
  • A bacteriocin-producing strain was isolated from kimchi at the early stage of kimchi fermentation. It was identified as Lactococcus lactis by morphological, cultural and physiological characteristics and partial sequence of 16S rDNA. The bacteriocin from isolate had antimicrobial activity against gram positive pathogenic bacteria, such as Listeria monocytogenes. Staphylococcus aureus and several strains of lactic acid bacteria but not to gram negative bacteria, Yersinia enterocolitica. The bacteriocin was sensitive to protease, protease ⅩⅣ, a-chymotrypsin and pepsin but not to lipase, trypsin and lysozyme. The bacteriocin activity was stable at pH 2-11 and temperature of 100 for 10 min. Thus, Listeria monocytogenes could be inhibited by Lactococcus lactis at early stage of fermentation.

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Characterization of Bacteriocin Production by Lactococcus lactis LAB3113 Isolated from Kimchi

  • Shin, Jong-Yeun;Cheol Ahn
    • Preventive Nutrition and Food Science
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    • v.2 no.2
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    • pp.101-108
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    • 1997
  • A lactic acid bacterium LAB3113, isolated from traditionally fermented Kimchi was found to produce bacteriocin whose activity was very specific toward lactobacilli and not effective against any other bacteria. Lactobacilli affected by the inhibitory substance included Lactobacillus delbrueckii-lactis, L. johnsonii, L. gsseri, and L. curvatus. Based upon biochemical and physiological characteristics, LAB3113 was classified as Lactococcus lactis, and its bacteriocin was named as lactococcin K3113. Lactococcus lactis. LAB3113 produced bacteriocin at th early stage of growth and the concentration of the bacteriocin did not decrease even after alt stationalry phase. Optimal temperature of bacteriocin production was $25^{\circ}C$ at the initial pH 7.0. Partially purified lactococcin K3113 was completely inactivated by protease, but not affected by lipase, lysozyme and RNase. The bacteriocin was very heat-stable even after autoclaving for 20 min. It was also stable in pH changes, an was not affected by th presence of solvents. lacotococcin K3113 appeared to act in bactericidal mode against L. delbrueckii-lactis ATCC4797. Molecular weight of lactococcin K3113 was calibrated as 10,500 dal by SDS-PAGE an activity staining. Lactococcus lactis LAB3113 had four residential plasmids of 3.7kb, 11.2kb, 15.5kb, and 48kh in molecular sizes. Plasmid profile analysis of mutant strain revealed that 15.5 kb plasmid was re-sponsible for the production of lactococcin K3113 and its immunity to the bacteriocin.

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Antimicrobial Effects of a Bacteriocin Mixture from Lactic Acid Bacteria against Foodborne Pathogens (복합 박테리오신의 항균활성 및 축산식품 저장성 증진 효과)

  • 한경식;오세종;문용일
    • Food Science of Animal Resources
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    • v.22 no.2
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    • pp.164-171
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    • 2002
  • The purpose of this study was to evaluate inhibitory activity of a bacteriocin mixture from lactic acid bacteria(LAB) against foodborne pathogens. Each bacteriocin solutions were prepared by growing nine strains of bacteriocin producers in MRS broth for 18~24 h followed by centrifugation(8000$\times$g, 20 min, 4$^{\circ}C$). Bacteriocins were purified from ammonium sulfate precipitation and were resuspended in 50 mM phosphate buffer(pH 7.0). Nine bacteriocins were mixed together and then allowed to freeze at -2$0^{\circ}C$. The mixture of nine bacteriocins showed enhanced inhibitory activity compared to each of bacteriocins and inhibited the Gram negative pathogens including Escherichia coli 0157:H7, Klebsiella pneumoniae, Pseudomonas chlororaphis and Shigella sonnei. The mixture of bacteriocin solutions was significantly lower than controls when a freeze-dried bacteriocin mixture was added to frank sausage, Mozzarella cheese and pork loin. With addition of bacteriocin mixture, total mesophilic bacteria in pork loin were constant over storage period, whereas total mesophilic bacteria in Mozzarella cheese and frank sausang slightly increased. Total viable cells of control group increased during storage without bacteriocin treatment. Volatile base nitrogen content of pork loin during storage also increased significantly without bacteriocin treatment. The bacteriocin mixture was capable of inhibiting pathogenic and spoilage microorganisms and extending the shelf-life of cheese and meat products during storage.

Characterization of the Bacteriocin from Enterococcus faecium CJNU 2008 (Enterococcus faecium CJNU 2008 균주 생산 박테리오신의 특성 규명)

  • Seo, Souk-Jin;Yang, Jung-Mo;Moon, Gi-Seong
    • Journal of Food Hygiene and Safety
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    • v.33 no.6
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    • pp.516-520
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    • 2018
  • Bacteriocin is a proteinaceous compound produced by microorganisms showing antimicrobial activities. In this study, the physicochemical properties of the bacteriocin produced by Enterococcus faecium CJNU 2008 strain were characterized. Partially purified bacteriocin showed stabilities against heat treatments at $100^{\circ}C$ for 30 min and $121^{\circ}C$ for 15 min and against solvents treatments such as methanol, ethanol, acetone, acetonitrile and chloroform. The bacteriocin also exhibited stabilities against lipase and ${\alpha}-amylase$ treatments but the stability was abolished at protease treatment, indicating that the antimicrobial agent from E. faecium CJNU 2008 was a proteinaceous bacteriocin. The bacteriocin also showed bactericidal mode of action against Listeria monocytogenes. The molecular mass of the bacteriocin was estimated to be under 6.5 kDa by a tricine-SDS-PAGE analysis. The bacteriocin was purified by HPLC. Further studies toward biochemical analysis of the bacteriocin are needed in near future.

Isolation of Leuconostoc mesenteroides subsp. mesenteroides DU-0608 with Antibacterial Activity from Kimchi and Characterization of Its Bacteriocin

  • Cha, Dong-Soo;Ha, Duk-Mo
    • Journal of Microbiology and Biotechnology
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    • v.6 no.4
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    • pp.270-277
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    • 1996
  • A bacteriocin-producing strain, DU-0608, was isolated from Kimchi and identified as Leuconostoc mesenteroides subsp. mesenteroides. The bacteriocin from isolate was inhibitory against Listeria monocytogenes, Micrococcus luteus and several strains of lactic acid bacteria. The bacteriocin was inactivated by pepsin, trypsin, $\alpha$-chymotrypsin, protease, $\alpha$-amylase and lipase, but not by catalase or by heating at $100^{\circ}C$ for 60 min. The molecular weight of the bacteriocin was estimated approximately 6 kDa. The inhibitory effect was bactericidal and rapid. Following treatment with isolate bacteriocin, cells of indicator strain (Lactobacillus sake JCM 1157) were damaged at the end regions of the cell wall, whereas the cells treated with nisin were damaged at many places around the cell wall.

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Antibacterial Activity and Characteristics of Bacteriocin Produced by Lactobacillus plantarum LMG 7945 (Lactobacillus plantarum LMG 7945가 생산하는 bacteriocin의 항균력과 그 특성)

  • 김상현;이명숙;장동석
    • Journal of Food Hygiene and Safety
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    • v.10 no.2
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    • pp.65-71
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    • 1995
  • Bacteriocins from lactic acid bacteria have attracted much attention in recent years because of their useful worth in increasing safety and extending shelf life of foods. These substances show an inhibitory effect against some food spoilage bacteria and food-borne pathogens. The inhibitory effect fo the bacteriocin produces by lactic acid bacteria against Listeria monocytogenes(L. monocytogenes) was examined in this study. The culture supernatants of 5 kinds of bacteria among the 10 kinds of testes lactic acid bacteria had the inhibitory activity against Listeria sp., various Gram positive and Gram negative bacteria. Bacteriocin produced by Lactobacillus plantarum(Lact. plantarum) LMG 7945 was the most active toward L. monocytogenes. Bacteriocin production of the Lact. plantarum LMG 7945 cultured on MRS broth was increased late logarithmic phase over early stationary phase. This bacteriocin was stable at heat treatment and acidic pH relatively; The activity was retained after heating at 121$^{\circ}C$ for 15min and was active in the pH range of 2~4 but was lost above pH 5.

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Isolation and Characterization of Lacticin 10790, a New Bacteriocin Produced by Lactococcus lactis subsp. cremoris KFCC 10790

  • Joo, Nam-Eok;Kim, Il-Han;Yoo, Jin-Young;Lee, Yong-Eok
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.539-543
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    • 2000
  • A new bacteriocin, named lacticin 10790, was purified from Lactococcus lactis subsp. cremoris KFCC 10790 by sequential adsorption, immobilized metal-affinity, cation-exchange, and $C_{18}$ reverse-phase chromatographies. The molecular mass of the bacteriocin was estimated to be between 3,000 and 3,500 Da. Lacticin 10790 showed a broad antimicrobial spectrum against many gram-positive bacteria. The bacteriocin was stable to heat and in the pH range between 2 and 6. Lacticin 10790 was destroyed by digestion with proteases and exhibited a bactericidal mode of action. An amino acid composition analysis of purified lacticin 10790 revealed a high concentration of hydrophobic amino acids. The N terminus of the bacteriocin was found to be blocked, upon analysis by Edman degradation. The results suggest that lacticin 10790 is a class I bacteriocin.

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Optimizing Conditions for the Growth and Bacteriocin Production of Lactococcus sp.HY 449 Usin Response Surface Methodology (반응표면 분석에 의한 Lactococcus sp. HY 449의 성장 및 Bacteriocin 생산의 최적화)

  • Kim, Sang-Kyo;Oh, Se-Jong;Lee, Sang-Jun;Baek, Young-Jin;Park, Yun-Hee
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.522-530
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    • 1994
  • Bacteriocin-producing lactic acid bacteria, Lactococcus sp. HY 449, was isolated from dairy products. Using response surface methodology, the various concentrations of medium compo- nents (tryptone, glucose, yeast extract, tween 80, and initial pH) were tested to find the optimum conditions for maximum bacteriocin production and growth of Lactococcus sp. HY 449. Central composite design was used to control the concentrations of medium components in the experiment. Bacteriocin production and cell growth of Lactococcus sp. HY 449 were most affected by glucose and yeast extract. Estimated optimum growth conditions of Lactococcus sp. HY 449 were as follows; tryptone 1.08%, glucose 1.129%, yeast extract 0.674%, tween 80 0.11%, and initial pH 7.19. Also estimated optimum conditions for bacteriocin production were tryptone 0.937%, glucose 1.108%, yeast extract 0.163%, tween 80 0.09%, and initial pH 6.98.

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