• Title/Summary/Keyword: Lactococcus sp

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Purification and Partial Amino Acid Sequence of a Bacteriocin Produced by Lactococcus, sp. HY449 (Lactococcus sp. HY449가 생산한 Bacterisocin의 정제)

  • 오세종;이상준;김경태;김상교;박연희;백영진
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.155-161
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    • 2001
  • A bactriocin produced by Lactrococcus sp. HY449 was purified by sequential purfication steps such as n-propanol-acetone precipitation ion -exchange chromatography using CM-Sequential CL6B. gel filtration chromatography using Sephacry HR100 and reverse-phase chromatography using pro RPC HR 5/10. Reverse-phase chromatography the final step of the purfication yielded a single symmetrical peak of bacteriocin activity The purification resulted in final yield of 3.25% and 413.35 fold increase of the specific activity of bacteriocin. The active fraction from reverse-phase chromatography was used for N-terminal amino acid analysis . The purified bacteriocin contained isoleucine, leucine, methionine, and glycine at but N-terminal end no aromatic amino acids. Calculation of the number of amino acid residues in the bacteriocin revealed that it is consisted of 32 residues assuming the molecular weight of bacteriocin to be about 3.6kDa. Edman degrandation elucidated amino acid residues of the first four of the N-terminus to be $NH_2$-Ile-Leu-Pro-GIn.

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An outbreak of Lactococcus garvieae Infection in Cage-cultured Red Lip Mullet Chelon haematocheilus with Green Liver Syndrome

  • Han, Hyun-Ja;Lee, Nam Sil;Kim, Myoung Sug;Jung, Sung Hee
    • Fisheries and Aquatic Sciences
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    • v.18 no.3
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    • pp.333-339
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    • 2015
  • Red lip mullet Chelon haematocheilus (body weight = $468{\pm}91g$) which became sick during an outbreak of disease at mariculture facilities at Cheonsu Bay, Korea, during July-August 2013, were examined to identify the cause of the disease. Diseased mullets displayed green liver syndrome, and Lactococcus garvieae were isolated from their internal organs. Argulus sp., Trichodina sp., and/or Vibrio spp. were also discovered in some infected fish. Histopathological examination revealed that fatty liver syndrome with hepatocyte degeneration, reflected in heterokaryons, inflammatory lesions, and melanomacrophage centers ($MMC_S$), had caused fibrosis around the kidney, spleen, and blood vessels. After the outbreak, visceral fat and green liver syndrome in the mullets were consistently observed throughout the year in the same mariculture facilities, indicating that the cultured mullets suffered a chronic metabolic disorder. Although Vibrio spp. were also isolated from some individuals, L. garvieae, which is known to be a causative agent of red lip mullet mortality, was isolated from all diseased individuals. This is the first report of L. garvieae infection in cultured red lip mullet.

Bacteriocin Production by Streptococcus sp. J-C1 Isolated from Kimchi (김치에서 분리한 Streptococcus sp. J-C1의 bacteriocin 생산)

  • Jo, Young-Bae;Cho, Young-Im;Baik, Hyung-Suk;Jun, Hong-Ki
    • Journal of Life Science
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    • v.6 no.4
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    • pp.270-277
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    • 1996
  • Streptococcus sp J-C1 producing bacteriocin was isolated from Kimchi. The optimum conditions for bacteriocin production by Streptococcus sp. J-C1 were evaluated. For the maximum yield of bacteriocin production by Streptococcus sp. J-C1, the cell should be harvested at the late stationary phase and the temperature, pH and NaCl concentration should be 25$\circ$C, pH 8 and without the addition of NaCl, respectively. Sucrose should be used as a carbon source and organic nitrogen such as peptone should be used as a nitorgen source for the best yield. The production of bacteriocin is related to the cell growth of Streptococcus sp. J-C1. The bacteriocin from Streptococcus sp. J-C1 was active for gram positive microorganisms such as Lactobacillus sp., Leuconoctoc sp., Lactococcus sp., Streptococcus mutans, Staphylococcus aureus amd Bacillus subtilis and also active for gram negetive bacteria such as Acetobacter aceti. Antibacterial activity of the bacteriocin was completely disappeared by protease treatment.

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Development of a Food-Grade Integration Vector for Heterologous Gene Expression and Protein Secretion in Lactococcus lactis

  • Jeong, Do-Won;Lee, Jong-Hoon;Kim, Kyoung-Heon;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1799-1808
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    • 2006
  • A food-grade integration vector based on site-specific recombination was constructed. The 5.7-kb vector, pIMA20, contained an integrase gene and a phage attachment site originating from bacteriophage A2, with the ${\alpha}$-galactosidase gene from Lactobacillus plantarum KCTC 3104 as a selection marker. pIMA20 was also equipped with a controllable promoter of nisA ($P_{nisA}$) and a signal peptide-encoding sequence of usp45 ($SP_{usp45}$) for the production and secretion of foreign proteins. pIMA20 and its derivatives mediated site-specific integration into the attB-like site on the Lactococcus lactis NZ9800 chromosome. The vector-integrated recombinant lactococci were easily detected by the appearance of blue colonies on a medium containing $X-{\alpha}-gal$ and also by their ability to grow on a medium containing melibiose as the sole carbon source. Recombinant lactococci maintained these traits in the absence of selection pressure during 100 generations. The ${\alpha}-amylase$ gene from Bacillus licheniformis, lacking a signal peptide-encoding. sequence, was inserted downstream of $P_{nisA}\;and\;SP_{usp45}$ in pIMA20, and the plasmid was integrated into the L. lactis chromosome. ${\alpha}-Amylase$ was successfully produced and secreted by the recombinant L. lactis, controlled by the addition and concentration of nisin.

Antimicrobial activity of hydrolysis compound of cactus stem (Opuntia ficus-indica var. saboten Makino) against fish pathogenic bacteria (손바닥선인장 줄기 가수분해 분획물의 어류 병원성 세균에 대한 항균활성)

  • Jung, Sung-Hee;Lee, Nam-Sil;Jee, Bo-Young;Park, Mi-Seon
    • Journal of fish pathology
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    • v.21 no.1
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    • pp.29-34
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    • 2008
  • In this study, we examined in vitro antimicrobial activity of the hydrolysis compound (MBT-01108) of cactus stem (Opuntia ficus-indica var. saboten Makino) against 18 strains of fish pathogenic bacteria, Edwardsiella tarda, Listonella anguillarum, Vibrio sp., Lactococcus garvieae, L. raffinose, Streptococcus parauberis and S. iniae. Antibiotic susceptibility of the same bacteria was also assayed to commercial discs containing various antibacterials. According to these results, MBT-01108 showed the excellent antimicrobial activity against the tested pathogenic bacteria.

Isolation and Properties of Bacteriocin-producing Microorganisms (Bacteriocin 생산균주의 분리 및 성질)

  • 유진영;이이선;남영중;정건섭
    • Microbiology and Biotechnology Letters
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    • v.19 no.1
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    • pp.8-13
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    • 1991
  • Bacteriocin-producing microorganisms were screened from raw milk and tested their antimicrobial activities against Lactobacillus plantarum ATCC 8014 as target organism, Antimicrobial substances isolated showed broad antimicrobial spectra against Gram positives and negatives. Strain 1112-1 was selected as a test organism due to its highest antimicrobial activity among the isolates. Antimicrobial substance produced by 1112-1 completely suppressed the growth of Lactobacillus plantarum at 230 IUIml and showed 11% growth inhibition of E. coli at 500 IUIrnl level. The antimicrobial substance was found to be proteinaceous material which was inactivated by carboxypeptidase, elastase, alpha amylase, amyloglucosidase, pronase, protease IV, alpha chymotrypsin, ficin, cellulase, phosphatase and lipase. The molecular weight was estimated by SDS-PAGE as 5,900. The isolate 1112-1 was identified as one of the related strains of Lactococcus sp. The strain was different from Lactococcus lactis in the following characteristics: late positive in maltose and sucrose fermentation; positive in mannitol and salicin fermentation; negative in lactose fermentation.

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김치에서 분리한 Lactococcus sp. JC-3 bacteriocin의 특성

  • Kim, Yeong-Hwa;Kim, Mi-Ryeong;Park, Geun-Yeong;Jeon, Hong-Gi;Kim, Seong-Gu
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.623-625
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    • 2000
  • Bacteriocin-producing lactic acid bacteria was isolated from Kimchi using MRS as selective media and Lactobacillus delbruekii subsp. delbruekii as an indicator strain. Strain JC-3 was tentatively identified as Lactococcus latis subsp. lactis through the API test and the bacteriocin produced by JC-3 showed the inhibitory activity against Grampositive pathogens and other lactic acid bacteria. The antimicrobial substance was inactivated by Protamax, Aroase AP-10, Neutrase, R-AMANO and was confirmed to be heating at $100^{\circ}C$. However, it was lost at high pH values showed the highest bacteriocin activity at a culture temperature of $30^{\circ}C$. The bacteriocin was partially purified by ammonium sulfate precipitation, Sep-pak $C_{18}$ cartridge. The apparent molecular mass of the bacteriocin was about 8 Kda, which was determined through the direct detection of bactericidal activity using SDS -PAGE.

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Inhibitory Effects of Candidate Probiotic Bacteria on the Growth of Fish Pathogenic Bacteria, Streptococcus sp. (Candidate Probiotic Bacteria의 어류병원성 Streptococcus sp. 성장에 대한 억제 효과)

  • Lee, Minyeong;Kim, Eunheui
    • Journal of fish pathology
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    • v.27 no.2
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    • pp.107-114
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    • 2014
  • For the treatments and protection of bacterial fish disease, many requirements are needed for aquatic probiotics so that they are effective in aquaculture animals but are also harmless to humans. In the present study, among 17 candidate probiotic bacteria (CPB) obtained from the edible part of the shellfish, Bacillus sp. CPB-St (CPB-St) were selected and in vitro evaluated for the possibility as a probiotic strain for the control of fish streptococcosis which frequently occurs in the olive flounder farms. CPB-St showed inhibitory effects on the growth of various fish pathogenic bacteria, Streptococcus sp., S. parauberis, S. iniae, Lactococcus garvieae and L. piscium by the double layer test ranging about 18~26 mm of clear zone. Inhibitory activity of CPB-St to Streptococcus sp. was observed 6 hours after and the growth of Streptococcus sp. was decreased to 8~55 folds in the co-culture of CPB-St with Streptococcus sp.. The safety of CPB-St to fish and survival of CPB-St in the intestine were assessed in the olive flounder, Paralichthys olivaceus. Fish mortality was not observed in artificial infection with CPB-St for 2 weeks. CPB-St was entirely excreted from the stomach and intestine 24 hours after oral injection. This results indicate that CPB-St has potential applications as a probiotic for the control of fish streptococcosis in aquaculture.