• Title/Summary/Keyword: Halophilic

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Enzymatic and Energetic Properties of an Aerobic Respiratory Chain­Linked NADH Oxidase System in Marine Bacterium Vibrio natriegens

  • Kang, Ji-Won;Kim, Young-Jae
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1080-1086
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    • 2005
  • Membranes prepared from Vibrio natriegens oxidized both NADH and deamino-NADH as substrates. The maximum activity of the membrane-bound NADH oxidase was obtained at about pH 8.5 in the presence of 0.2 M NaCl, whereas that of the NADH:ubiquinone oxidoreductase was obtained at about pH 7.5 in the presence of 0.2 M NaCl. Electron transfer from NADH or deamino-NADH to ubiquinone-l or oxygen generated a considerable membrane potential (${\Delta}{\psi}$), which occurred even in the presence of $20{\mu}M$ carbonylcyanide m-chlorophenylhydrazone (CCCP). However, the ${\Delta}{\psi}$ was completely collapsed by the combined addition of $10{\mu}M$ CCCP and $20{\mu}M$ monensin. On the other hand, the activity of the NADH oxidase and the ${\Delta}{\psi}$ generated by the NADH oxidase system were inhibited by about $90\%$ with $10{\mu}M$ HQNO, whereas the activity of the NADH:ubiquinone oxidoreductase and the ${\Delta}{\psi}$ generated at the NADH:ubiquinone oxidoreductase segment were inhibited by about $60\%$. Interestingly, the activity of the NADH:ubiquinone oxidoreductase and the ${\Delta}{\psi}$ generated at the NADH:ubiquinone oxidoreductase segment were resistant to the respiratory chain inhibitors such as rotenone, capsaicin, and $AgNO_3$, and the activity of the NADH oxidase and the ${\Delta}{\psi}$ generated by the NADH oxidase system were very sensitive only to $AgNO_3$. It was concluded, therefore, that V. natriegens cells possess a $AgNO_3$-resistant respiratory $Na^+$ pump that is different from the $AgNO_3$-sensitive respiratory $Na^+$ pump of a marine bacterium, Vibrio alginolyticus.

Effect of NaCl on Halomonas subglaciescola DH-1 Incapable of Growing at Non-Salinity (Halomonas subglaciescola DH-1의 생장에 미치는 염화나트륨의 영향)

  • Na, Byung-Kwan;Yoo, Young-Sun;Park, Doo-Hyun
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.298-303
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    • 2007
  • A halophilic bacterium, H. subglaciescola DH-1, grew at 2.0 M salinity, but did not grow at 0.8 M salinity when cultivated at higher temperature ($40^{\circ}C$) than optimum ($30^{\circ}C$). When the cell extract of strain DH-1 was heated at $50^{\circ}C$ for 60 min in the absence of NaCl, isocitrate dehydrogenase and malate dehydrogenase lost their activities, but when it was heated in the presence of 2.0 M NaCl, the activity was maintained. Meanwhile, the cell extract of E. coli did not catalyze the reduction of $NAD^+$ to NADH coupled with the oxidation of isocitrate and malate at higher salinities than 1.0 M. The pH range for DH-1 was 7 to 10, and that for E. coli was 5 to 9. DH-1 was not grown in conditions with sodium salts other than NaCl.

Studies on Mycoflora and Mycotoxins of Cowpea Cultivars (동부 콩열매에 있는 진균류와 균독소에 관한 연구)

  • Zohri, A.A.;Khayria, M.;Gawad, Abd. El.
    • The Korean Journal of Mycology
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    • v.20 no.3
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    • pp.252-258
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    • 1992
  • Thirty three species and two species varieties belonging to 14 genera of fungi were collected from 20 cowpea cultivars on glucose Czapek's agar (11 genera and 25 species+1 var.) and glucose-Czapek's agar supplemented with 10% NaCl (7 genera and 18 species+2 var.) at $28{\pm}2^{\circ}C$. The total count of fungi were 6716 colonies/g in all cowpea cultivars. On glucose-Czapek's agar and identified; Aspergillus flavus, A. niger, A. sydowii, A. flavus var. columnaris, A. terreus, Penicillium chrysogenum, Emericella nidutans and Rhizopus stolonifer. The total count of halotolerant or halophilic fungi was 3515 colonies/g on 10% NaCl-glueose-Czapek's agar and identified; the most common species were: A. flavus, A. sydowii, A. tamarii A. flavipes, A. niger, A. flavus var. columnaris, A. ochraceus, A. oryzae and P. chrvsogenusm. Thin layer chrormatographic analysis of chloroform extracts of the different seed samples revealed that four cultivars were naturally contaminated with aflatoxins $B_1,\;B_2,\;G_1$ and $G_2$, $(45-112\;{\mu}g/kg)$.

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Microbiological Characteristics of Alcoholfermenting Wild Yeast, Aureobasidium pullulans P-1 and Its Makgeolli Fermentation Characteristics (알코올 발효성 야생 효모, Aureobasidium pullulans P-1의 균학적 특성과 막걸리 발효 특성)

  • Hong, Yong-Cheol;Han, Sang-Min;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.46 no.3
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    • pp.307-314
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    • 2018
  • The goal of this study was to investigate the microbiological characteristics of the ethanol-producing wild yeast, Aureobasidium pullulans P-1, isolated from flowers growing near the Yedang reservoir, Chungnam province, Korea, and in addition, to optimize its fermentation ability for the production of Makgeolli. A. pullulans P-1 was oval-shaped, and formed ascospores and pseudomycelium. The P-1 strain was a halophilic and sugar tolerant yeast which grew in 15% NaCl and 50% glucose-containing yeast extract-peptone-dextrose media. The P-1 strain was also resistant to 20% ethanol. Changes of the physicochemical properties during Makgeolli fermentation by A. pullulans P-1 were investigated. A maximum of 8.45% ethanol was obtained when a mixture of cooked rice, 150% water, and 35% ipguk per cooked rice was fermented by 5% A. pullulans P-1 culture broth at $25^{\circ}C$ for 10 days. Antihypertensive angiotensin I-converting enzyme inhibitory activity in the Makgeolli ferment produced by A. pullulans P-1 reached a high of 71.1% after 10 days.

Production of Carotenoid from Halophilic Erythrobacter sp. and characterization of Physiological Properties (해양미생물 Erythrobacter 속으로부터 Carotenoid의 생산 및 그 생리활성)

  • 김종덕;강동수;김민용;노승배;최명락;송상호;백승한;서효진;김대현
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.143-151
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    • 2001
  • A marine bacterium producing carotenoid was isolated from the Yosu coastal area of South Korea, which was recorded as MCK-1. It was identified as Erythrobacter sp. Optimium conditions of marine carotenoid fermentation from Erythrobacter sp. were pH 6.0, a temperature of $25^{\circ}C$, 16 mM mannitol as a carbon source, 0.5% tryptone as a nitrogen source, 0.1 mM $Fe^{+2}$ ion as a mineral source and 1$\mu$M of cyanocobalamine as a growth factor in a jar-fermentor. Erythrobacter sp. was produced 351.27 mg/100mL of the marine carotenoid in these optimum conditions. This marine carotenoid was composed of 4 different conpounds, like as notoxanthin (61.4%), can thaxanthin (24.6%), fucoxanthin (8.2%), and zeaxanthin (5.8%). Physiological properties including antibacterial activity, cytotoxic effect, antioxidative effect and free radical scavenging activity were characterized with crude carotenoid. Carotenoid exhibited no antibacterial activity against E. coli and lactobacillus bulgaricus, but showed cytotoxic effect against cancer cells such as HepG2 (Hepatocellular carcinoma, human, ATCC HB-8065) and HeLa (Cervical carcinoma, human, ATCC CCL-2) cells. The impediment ratios for HepG2 and HeLa cell were 37.14% and 33.78%, respectively. This carotenoid expressed a strong antioxidative effect (77%) against CCL-13 5 $\mu\textrm{g}$/mL and 50 $\mu\textrm{g}$/mL crude carotenoid, respectively.

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Bacteriocin ("Vulnificin") Typing of Vibrio vulnificus

  • Ha, Tai-You;Lee, Jeong-Ho;Lee, Hern-Ku;Whang, Hee-Sung;Park, Jae-Seung
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.225-237
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    • 2000
  • Vibrio vulnificus, a halophilic vibrio is an estuarine gram-negative bacteria that is associated with severe and frequently fatal wound infections and life-threatening septicemia. Bacteriocins are defined as antibacterial substance produced by various species of bacteria which are usually active against closely related organisms. Bacteriocins have found widespread application in epidemiological studies as specific markers of bacteria. It was proposed by Ha et al. (1990. J. Korean. Soc. Microbiol. 25: 586.) to give the bacteriocins produced by V. vulnificus the name "vulnificins". In the present study, a total of 72 strains of V. vulnificus isolated from patients and oysters were subjected to screen potential producers and indicators of vulnificin, applying ultraviolet induction method. Sensitivity of several strains of Serratia marcesans, Pseudomonas aeruginosa, Shigella flexneri, Salmonella typhi and Yersinia enterocolitica to vulnificins were also examined out. All the tested strains of V. vulnificus produced vulnificins active against indicator strains with various different inhibitory patterns. The spectrum of vulnificin activity and sensitive spectrum of indicator strains were considerably broad. Interestingly, almost all strains of S. marcescens, P. aeruginosa, Salmonella sp., Shigella sp. and Y. enterocolitica tested were sensitive to 1-7 vulnificin(s). Taken together, the present study demonstrated that all of the isolates of V. vulnificus produced vulnificins and that 8 good vulnificin producers and 10 good indicators were detected. These strains can be employed efficiently for establishing vulnificin typing scheme of V. vulnificus and for the detection of bacteriocinogeny and sensitivity in V. vulnificus. Biological role of vulnificin remains to be further elucidated.

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A New Salt-Tolerant Thermostable Cellulase from a Marine Bacillus sp. Strain

  • dos Santos, Yago Queiroz;de Veras, Bruno Oliveira;de Franca, Anderson Felipe Jacome;Gorlach-Lira, Krystyna;Velasques, Jannaina;Migliolo, Ludovico;dos Santos, Elizeu Antunes
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1078-1085
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    • 2018
  • A salt-tolerant cellulase secreted by a marine Bacillus sp. SR22 strain with wide resistance to temperature and pH was purified and characterized. Its approximate mass was 37 kDa. The endoglucanase, named as Bc22Cel, was purified by ammonium sulfate precipitation, gel filtration chromatography, and extraction from the gel after non-reducing sodium dodecyl sufate-polyacrylamide gel electrophoresis. The optimal pH value and temperature of Bc22Cel were 6.5 and $60^{\circ}C$, respectively. The purified Bc22Cel showed a considerable halophilic property, being able to maintain more than 70% of residual activity even when pre-incubated with 1.5 M NaCl for 1 h. Kinetic analysis of the purified enzyme showed the $K_m$ and $V_{max}$ to be 0.704 mg/ml and $29.85{\mu}mol{\cdot}ml^{-1}{\cdot}min^{-1}$, respectively. Taken together, the present data indicate Bc22Cel as a potential and useful candidate for industrial applications, such as the bioconversion of sugarcane bagasse to its derivatives.

Characteristics of Urease from Vibrio parahaemolyticus Possessing tah and the Genes Isolated in Korea

  • Kim, Young-Hee;Kim, Jong-Sook
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.279-285
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    • 2001
  • Vibrio parahaemolyticus is a halophilic bacterium associated with seafood gastroenteritis. An unusual strain of Kanagawa-positive urease producing Vibrio parahaemolyticus O1:K1 was isolated from the environment and identified . A polymerase chain reaction assay revealed that this strain harbored both the tdh and the genes. The urease from this strain was studied. Maximum urease production was induced in LB medium containing 0.2% urea, 0.5% glucose, 2% NaCl and pH 5.5 with 6h of culti-vation at 37$\^{C}$ under aeration. Purification of urease was achieved by the process of whole cell lysate, 65% ammonium sulphate precipitation, DEAE-cellulose ion exchange column chromatography, Sepharose CL-6B gel filtration and oxirane activated Sepharose 6B-urea affinity chromatography with 203 fold purification and 2.2% yield. Analysis of the purified enzyme by SDS-PAGE demonstrated the presence of the subunits with a molecular weight of 85kDa, 59kDa, 41kDa and the molecular weight for the native enzyme by nondenaturing PAGE and gel filtration chromatography was 255kDa. The purified urease was stable at pH 7.5 and the opeimal pH in HEPES buffer was 8.0 The enzyme was stable at 60$\^{C}$ for 2 h with a residual activity of 32% . The addition of 10$\mu$M if NiCl$_2$maintained stability for 30 min. The Km value of the purified enzyme was 35.6 mM in urea substrate. The TD$\_$50/(median toxic dose) of the purified urease was 2.5$\mu\textrm{g}$/ml on human leukemia cells.

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Bacteriological Studies on the Distribution of Pathogenic Enterobacteria in the Natural Environments in Seoul(1978) (서울 시내 자연환경내에 있어서의 병원성 장내세균 분포에 관한 연구)

  • Lee, Chong-Hoon;Ko, Kwang-Kjune;Lim, Byung-Uk;Moon, Ki-Sung
    • The Journal of the Korean Society for Microbiology
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    • v.14 no.1
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    • pp.1-9
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    • 1979
  • A bacteriological survey was carried out to get hold of the distribution of pathogenic enteric bacteria in Korea. The total number of 2,013 specimens were obtained from various sources; 1,407 specimens from marine products circulated in the markets, sewage, and 606 rectal swabs from persons. All the specimens were collected in Seoul, Chumunjin(Kangwondo), and Gwangcheon(Chungcheongnam-do) during 1978. The isolation and identification of enteric pathogens from the specimens were performed by means of bacteriological studies. 1. The isolation rates of the pathogenic enterobacteria among the total 2,013 specimens are as follows: Salmonella species 0.05%(1 strain), Shigella species 0.50%(10 strains), and Vibrio parahaemolyticus 0.35%(7 strains). 2. One salmonella strain was isolated from marine products circulated in the market in Seoul. Its serotype was identified as group $C_1$. 3. Ten shigella strains were isolated from various sources: 0.45% from natural environments and 0.05% from rectal swabs. The distribution of shigella serotype was identified as Sh. boydii 90%(9 strains), Sh. sonnei 10%(1 strain). 4. Seven strains of V. parahaemolyticus were isolated from natural environments. In addition, 40 strains of halophilic vibrios nontypable with anti-K antisera were also isolated. Of the 7 strains, the 2 strains were agglutinated with type K-32, each 1 strain of the others with K-17, K-19, K-36, K-39, K-56.

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Isolation of Vibrio vulnificus Serotype Strains for Vaccine Preparation (Vibrio vulnificus 백신제조원의 혈청형균주 분리)

  • Ju, Jin-Wo
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.4
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    • pp.393-402
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    • 1987
  • The halophilic bacterium Vibrio vulnificus, previously called lactose-positive(L+) Vibrio and Beneckea vulnifica, causes acute, fulminating wound infections and septicemia in humans. Septicemia is very serious infection with a fatality rate of about 50%. Most patients with primary septicemia due to V. vulnificus have preexisting liver disease. V. vulnificus also cause severe wound infections usually after trauma and exposure to marine animals or the marine environment. The mortality rate is not nearly as high as in primary septicemia caused by this organism. In most cases human disease results from ingestion of contaminated seafood or from infection of a wound, frequently of seawater or crab origin. The author made an attempt to isolation of the V vulnificus from seawater, seamud, fish, shellfish, and algae on the southern sea of Korea from January to September in 1987, using for the purpose of vaccine preparation. The author investigated for bacteriological identification, hemolysis and determination of serotypes of isolated V. vulnificus strains. Eighty-five strains(5.9%) out of 1450 specimens collected of V. vulnificus were isolated. The distribution of the 85 isolates were as follows: 21 strains from seawater, 11 strains from seamud, 28 strains from fish, 19 strains from shellfish, and 6 strains from algae, respectively. All 85 isolates were positive reaction on human blood agar. The distribution of serotypes of V. vulnificus isolates were O1 to O8: 13 strains of O1, 6 strains of O2, 11 strains of O3, 9 strains of O4, 10 strains of O5, 7 strains of O6, 15 strains of O7, and 10 strains of O8, respectively. Eighty-one strains showed agglutination with O antisera, but 4 strains failed to show agglutination. In this study, the author suspected that serotypes of V. vulnificus isolates distributed also in the seaside of Korea as well as in most seaside of the world, and new serotypes were in existence in the seaside of Korea except reported up to now.

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