• Title/Summary/Keyword: Vibrio sp.

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Development of a thermo-stabel ${\beta}-agarase$ from marine organism

  • Lee, Sang-Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.31-32
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    • 2005
  • Neoagaro-oligosaccharides are produced only by enzymatic degradation of agarose by ${\beta}-agarase.^{1)}$ Neoagaro-oligosaccharides inhibit the growth of bacteria, slow the rate of degradation of starch, are used as low-calorie additives to improve food quality, and have macrophage-stimulating activity. Furthermore, neoagarobiose is a rare reagent that has both moisturizing effect on skin and whitening effect on melanoma $cells.^{2)}$ An agar-degrading marine bacterium was isolated from the sea water at the northeast coast in Cheju island, Korea. The strain was gram negative, aerobic, and motile rod. The 16S rRNA of the strain had the closest match of 98% homology, with that from Agarivorans albus. On the basis of several phenotypic characters and a phylogenetic analysis, this strain was designated Agarivorans sp. JA-1. In solid agar plate, Agarivorans sp. JA-1 produced a diffusible agarase that caused agar softening around the colonies. Agarivorans sp. JA-1 was cultured for 36 hr in marine broth 2216 (Difco, USA) and the supernatant that containing an extracellular ${\beta}-agarase$ was prepared by centrifugation of culture media. The enzyme exhibited relatively strong activity at $40^{\circ}C$ and was stable up to $60^{\circ}C$. Using PCR primers derived from the ${\beta}-agarase$ gene of Vibrio sp., the gene encoding ${\beta}-agarase$ from Agarivorans sp. JA-1 was cloned and sequenced. The structural gene consists of 2931 bp encoding 976 amino acids with a predicted molecular weight of 107,360 Da. The deduced amino acid sequence showed 99% and 34% homology to $agaA^{2)}$ and $agaB^{2)}$ genes for ${\beta}-agarase$ from Vibrio sp., respectively. The expression plasmid for ${\beta}-agarase$ gene of Agarivorans sp. JA-1 is being constructed and the recombinant enzyme will be biochemically characterized.

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Isolation of Plasmid DNA in Pediococci from Kimchi (김치의 Pediococci에 존재하는 Plasmid DNA 분리)

  • Park, Yun-Hee;Ryu, Uk-Sang;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.31 no.1
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    • pp.33-37
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    • 1988
  • Three species of Pediococci, Pediococcus pentosaceus, Pediococcus acidilactici and Pediococcus halophilus were isolated from Kimchi. P. pentosaceus and P. acidilactici showed inhibitory activity against Streptococcus faecalis, Pseudomonas sp., P20 and Vibrio parahaemolyticus. However, the growth of all test organisms was not inhibited by P. halophilus. Ten strains contained one to seven plasmids, ranging in size from 1 to 60 megadaltons.

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Purification and Characterization of a New κ-Carrageenase from the Marine Bacterium Vibrio sp. NJ-2

  • Zhu, Benwei;Ning, Limin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.255-262
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    • 2016
  • The carrageenan-degrading marine bacterium Vibrio sp. strain NJ-2 was isolated from rotten red algae, and κ-carrageenase with high activity was purified from the culture supernatant. The purified enzyme with molecular mass of 33 kDa showed the maximal activity of 937 U/mg at 40℃ and pH 8.0. It maintained 80% of total activity below 40℃ and between pH 6.0 and 10.0. The kinetics experiment showed the Km and Vmax values were 2.54 g/ml and 138.89 mmol/min/mg, respectively. The thin layer chromatography and ESI-MS analysis of hydrolysates indicated that the enzyme can endolytically depolymerize the κ-carrageenan into oligosaccharides with degrees of depolymerization of 2-8. Owing to its high activity, it could be a valuable tool to produce κ-carrageenan oligosaccharides with various biological activities.

Production of Tetrodotoxin Using Novel Marine Microorganism Isolated from Intestine of Pufferfish (복어 장내에서 분리한 신규 해양 미생물을 이용한 Tetrodotoxin 생산)

  • 윤성준;송성광;이명자;정동윤;김희숙;김동수;이은열
    • Journal of Life Science
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    • v.10 no.2
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    • pp.196-201
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    • 2000
  • The production of tetrodotoxin (TTX) using Vibrio sp. YE-101, a novel marine microorganism isolated from the intestine of pufferfish, was investigated. Culture condition was optimized for the enhanced production of TTX using response surface methodology. The experimental sets of environmental conditions including pH, temperature and NaCl concentration were designed using central composite experimental design. The optimal conditions of pH, temperature and NaCl concentration were determined to be 8.1, 29.2℃, and 2.6% (w/v) respectively. The relative growth extent could be enhanced up to 80%, and final mouse unit (MU) value of TTX was also enhanced up to 87% by response surface optimization.

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Isolation and characterization of the outer membrane vesicle (OMV) protein from Vibrio anguillarum O1 (Vibrio anguillarum O1이 생산하는 Outer Membrane Vesicle (OMV)의 분리 및 OMV 내의 단백질 특성)

  • Hong, Gyeong-Eun;Kim, Dong-Gyun;Min, Mun-Kyeong;Kong, In-Soo
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.2
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    • pp.123-125
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    • 2007
  • Vibrio anguillarum is a gram-negative bacterium that causes vibriosis in approximately 80 different fish species. V. anguillarum produces several exotoxins are correlated with the pathogenesis of vibriosis. This study is focused on the composition of the outer membrane vesicle. Most of gram-negative bacteria produce outer membrane vesicle (OMV) during cell growth. OMV was formed from the outer membrane surface of cell and than released to extracellular environment. OMV consists of outer membrane lipids, outer membrane protein (OMP), LPS, and soluble periplasmic components. Also, they contain toxins, adhesions, and immunomodulatory. Many gram-negative bacteria were studied out forming OMV. In Vibrio sp., formation of OMV by electron microscopy has been reported from V. cholerae and V. parahaemolyticus. In present study, we isolated OMV from V. anguillarum and OMV protein was separated by SDS-PAGE. Magor band was sliced and analyzed by MALDI-TOF. The major protein band of 38kDa was identified as OmpU by MALDI-TOF MS analysis.

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Antibacterial Property of Ecklonia cava Extract against Marine Bacterial Pathogens (해양 유해세균에 대한 감태 추출물의 항균특성)

  • Kim, Ji-Hoon;Kim, Se-Bong;Hwang, Hye-Jin;Kim, Young-Mog;Lee, Myung-Suk
    • Journal of Food Hygiene and Safety
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    • v.31 no.5
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    • pp.380-385
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    • 2016
  • The object of this study was to evaluate antibacterial activity of edible seaweed extracts against marine bacterial pathogens including Edwardsiella tarda, Streptococcus iniae, Streptococcus parauberis, Vibrio anguillarum, Vibiro harveyi and Vibrio scophthalm, which are associated with human or fish infectious disease. Ecklonia cava methanolic extract showed a strong and broad spectrum antibacterial activity against marine bacterial pathogens used in this study. Among solvent-soluble fractions of the E. cava extract, the ethyl acetate (EtOAc) soluble fraction showed the strongest antibacterial activity against marine bacterial pathogens tested in this study with MIC in the range of $128-256{\mu}g/mL$. Furthermore, HPLC analysis revealed that the soluble fraction contains abundant dieckol, a phlorotannin compound, compared to other solvent soluble fractions, suggesting that phlorotannins including dieckol would be a key antibacterial agent against marine bacterial pathogens.

NADP+-Dependent Dehydrogenase SCO3486 and Cycloisomerase SCO3480: Key Enzymes for 3,6-Anhydro-ʟ-Galactose Catabolism in Streptomyces coelicolor A3(2)

  • Tsevelkhorloo, Maral;Kim, Sang Hoon;Kang, Dae-Kyung;Lee, Chang-Ro;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.756-763
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    • 2021
  • Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.

Disinfection of Culture Water Supply by Ozonization I. Susceptibility of Some Fish-Pathogenic Bacteria Isolated From Cultured Marine Fish (오존처리법에 의한 양어용수 살균에 대하여 I. 해산어류 병원세균의 오존 감수성)

  • Oh, Myung-Joo;Kim, Heung-Yoon;Cho, Hyun-Soh
    • Journal of fish pathology
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    • v.12 no.1
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    • pp.42-48
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    • 1999
  • The disinfectant effects of total residual oxidants (TROs) produced by ozonization of natural sea water were investigated against fish pathogenic bacteria isolated from flounder and red seabream. The concentration of 0.1 mg TROs/liter was stable for 20 min in filtered natural seawater, and those of 0.3 and 0.5 mg TROs/liter were also stable for more 1 hr. Disinfectant effects of TRO against Edwardsiella tarda, Vibrio sp., Streptococcus sp. and Staphylococcus sp. were observed with a concentration of 0.1 mg/liter for 180 sec, and the treatment killed more than 99.9% of bacterial cells. With TROs of 0.3 to 0.5 mg/ liter, the viable cells of the bacteria were reduced by more than 99.99% in 60 sec.

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