• Title/Summary/Keyword: Klebsiella oxytoca

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Characterization of the Plasmid-Encoded Arsenic Salts Resistance Determinant from Klebsiella oxytoca D12

  • Rhie, Ho-Gun;Lee, Sung-Jae;Lee, Ho-Sa
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.593-598
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    • 2004
  • The arsenical resistance (ars) operon was cloned from a 67-kilobase pair (kb) plasmid, which was previously shown to be responsible for arsenic salts resistance in K. oxytoca D12. When plasmid pAE48, carrying the ars operon, was transformed into E. coli, transformed cells displayed enhanced survival in the presence of 4 mM arsenite, 50 mM arsenate, or 0.4 mM antimonite. The nucleotide sequence of the 5.6-kb fragment encoding arsenical resistance revealed five open reading frames (ORFs), which were predicted to encode polypeptides of 12.8 (arsR), 13.4 (arsD), 62.6 (arsA), 45 (arsB), and 16.7 (arse) kilodaltons (kDa). Each ORF was preceded by a ribosome binding site. A putative promoter-like sequence was identified upstream of arsR, and a possible termination site was found downstream of arsC. When the deduced amino acid sequences of the K. oxytoca Dl2 Ars proteins were compared with the amino acid sequences of the E. coli R773 Ars proteins, a significant amino acid similarity was observed (87.9% for ArsR, 89.2% for ArsD, 83.2% for ArsA, 92.6% for ArsB, and 91.3% for ArsC), suggesting an evolutionary relationship of the ars genes of E. coli plasmid R773 and K. oxytoca Dl2.

Genetic Characteristics of Arsenic Compounds-Resistant Bacteria Isolated from Stream Water (하천에서 분리한 비소 내성세균의 유전적 특성)

  • 정미경;이호자
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.63-68
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    • 1991
  • Several arsenic compound-resistant bacteria were isolated from Jung-Rang stream. The isolates, D-3, D-12, and D-14 were characterized phenotypically and genetically, and identified as Serratia liquefaciens, Klebsiella oxytoca, and Klebsiella pneumoniae, respectively. A plasmid of 67kb was found in Klebsiella oxytoca D-12 and designated as pMH12. Transfer of this plasmid from D-12 to E. coli HB101 was occurred, and the resulting transconjugant strains expressed the same level of heavy metal resistance as the donor strain. The physical presence of this plasmid in transconjugant was detected with agarose gel electrophoresis. Arsenite-sensitive derivatives of isolate D-12 were obtained with Mitomycin C treatment which cured pMH12. Antibiotics and heavy metal resistances were also examined to be used as a proper marker for the isolates in gene cloning. Isolate D-12 has resistance to several heavy metal ions such as $Cd^{2+}$ , $Zn^{2+}$ and $Hg^{ 2+}$ Also, all the other arsenite resistant isolates showed resistance to several heavy metal ions and antibiotics.

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Isolation and Characterization of Benzene-degrading Bacteria. (Benzene 분해 세균의 분리와 특성연구)

  • 김정현;유재근;이형환
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.379-383
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    • 1988
  • To evaluate the treatability of activated sludge induced by benzene with microorganisms, isolation and characterization of benzene-degrading microorganisms were carried out. Six bacterial isolates from the activated sludge were identified ; Pseudomonas fluorescens, Enterobacter agglomerans, Enterobacter cloacae, Klebsiella oxytoca, Citrobacter freundii and Klebsiella pneumoniae. P. fluorescens degraded 55% of benzene contained in the medium as a sole carbon source, E. cloacae 24%, E. agglomerans 41%, and K. oxytoca 32%. Optimal temperature, pH and benzene concentration for growth of P. fluorescens appeared to be 31$^{\circ}C$, pH 7.0, and 300mg benzene per liter. When the P. fluorescens was dominant in the activated sludge induced by benzene, the indicator protozoa was Aspidisca sp. When concentration of benzene was about 387mg per liter, the growths of Aspidisca sp. and Litonotus sp. were high. Protozoa, Litonotus sp. and Vorticella sp. did not grow over 1600mg of benzene per liter.

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Characterization of a Chromosomal Nickel Resistance Determinant from Klebsiella oxytoca CCUG 15788

  • Park, Jae-Sun;Lee, Sung-Jae;Rhie, Ho-Gun;Lee, Ho-Sa
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1040-1043
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    • 2008
  • Klebsiella oxytoca CCUG 15788 is resistant to $Ni^{2+}$ at a concentration of 10 mM and grows in an inducible manner when exposed to lower concentrations of $Ni^{2+}$. The complete genomic sequence of a 4.2-kb HindIII-digested fragment of this strain was determined from genomic DNA. It was shown to contain four nickel resistance genes (nirA, nirB, nirC, and nirD) encoding transporter and transmembrane proteins for nickel resistance. When the plasmid pKOHI4, encoding nirABCD, was transformed into Escherichia coli JM109, the cells were able to grow in Tris-buffered mineral medium containing 3 mM nickel. TnphoA'-1 insertion mutants in the four nickel genes nirA, nirB, nirC, and nirD showed nickel sensitivity. The nir genes were heterogeneously expressed in E. coli, suggesting functional roles of these genes in nickel resistance.

Characterization of a Novel Lipopolysaccharide Biosurfactant from Klebsiella oxitoca

  • Kim, Pil;Kim, Jung-Hoe
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.494-499
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    • 2005
  • The chemical, physical, and emulsifying properties of BSF-1, which is an extracellular lipopolysaccharide biosurfactant produced by Klebsiella oxytoca strain BSF-1, were studied. BSF-1 was found to be composed mainly of carbohydrate and fatty acids. The average molecular weight was $1,700{\sim}2,000 kDa$. The polysaccharide fraction contained L-rhamnose, D-galactose, D-glucose, and D-glucuronic acid at a molar ratio of 3:1: 1:1. The fatty acid content was 1.1 % (w/w) and consisted mainly of palmitic acid (C16:0), 3-hydroxylauric acid (3-OH-C12:0), and lauric acid (C12:0). In terms of thermal properties, BSF-1 was revealed to have inter- and intra-molecular hydrogen bonds. The hydrodynamic volume (intrinsic viscosity) of BSF-1 was 22.8dL/g. BSF-1 could be maintained as a stable emulsion for 48 h through a low-level reduction in surface tension. The optimal emulsification temperature was $30^{\circ}C$. Emulsification by BSF-1 was efficient at both acidic and neutral pH values.

Isolation, Purification and Characterization of the β-Xylosidase from Klebsiella sp. Sc. (Klebsiella sp. Sc가 생산하는 β-xylosidase의 분리, 정제 및 특성)

  • Lee, Yong-Seok;Park, In-Hye;Ahn, Soon-Cheol;Choi, Yong-Lark
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1801-1806
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    • 2010
  • A $\beta$-xylosidase encoding gene from Klebsiella sp. Sc was cloned in Escherichia coli. The $\beta$-xylosidase gene consisted of an open reading frame of 1,680 nucleotides and encodes 559 amino acids with a deduced molecular weight of 63 kDa. The deduced amino acid sequence of the $\beta$-xylosidase from Klebsiella sp. Sc exhibits 90% identities and 95% positives compared to those from Klebsiella oxytoca (KOX), Lactobacillus lactis (LAC, 82%, 90%), Bacillus longum (BLON, 69%, 81%) and Escherichia coli (ECOLI, 47%, 63%). The $\beta$-xylosidase was purified by GST-fusion purification system. The pH and temperature optima of the enzyme were 6.6 and $55^{\circ}C$, respectively. The $\beta$-xylosidase hydrolyzes xylobiose to xylose.