• Title/Summary/Keyword: Klebsiella oxytoca D12

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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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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.

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.

Trans-acting regulation and Arsenite sensing properties of ars operon

  • Lee, Soo-Chan;Lee, Sung-Jae;Lee, Ho-Sa
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2001.05a
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    • pp.141-141
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    • 2001
  • The arsenic resistance operon from pMH12 in Klebsiella oxytoca contains two regulatory genes. The first open reading frame for arsR extend up to 348 bp and has a translational product corresponding to a protein of 116 amino acid residue polypeptide with a molecular mass of 13 kDa. And the second ORF for arsD extend up to 360 bp and express a protein of 120 amino anid residue polypeptide with 13kDa. (omitted)

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Effect of ArsA, Arsenite-Specific ATPase, on Inhibition of Cell Division in Escherichia coli

  • Lee, Sung-Jae;Lee, Soo-Chan;Choi, Seung-Ho;Chung, Mi-Kyung;Rhie, Ho-Gun;Lee, Ho-Sa
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.825-830
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    • 2001
  • Escherichia coli, which harbored the ars operon from a plasmid pMH12 of Klebsiella oxytoca D12, showed filamentation due to the expression of ars genes in the presence of arsenite. The continued DNA replication in the absence of cell division was revealed, since nucleoids abound with DAPI appeared to be arranged in chains. In contrast to overexpression of arsA, its frame-shift mutant and knock-out mutant lost filamentation in the presence of arsenite, which suggested that ars-induced division block was dependent on expression of arsA. ArsA-induced division inhibition was not a consequence of an inhibition of DNA replication, and the inability of arsenite to induce an SOS response indicated that arsA-mediated division inhibition was dependent on the expression of the gene product encoded by the minB operon. ArsA is a peripheral membrane protein with an ATP-binding domain, which is homologous to MinD that requires ATP-dependent efflux. These results suggested that ArsA could possibly recruit MinC to the membrane and modulate cytoplasmic FtsZ to block assembly at the middle of the cell.

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