• Title/Summary/Keyword: Pseudomonas rhodesiae KK1

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Identification and Molecular Characterization of Superoxide Dismutase Genes in Pseudomonas rhodesiae KK1 Capable of Polycyclic Aromatic Hydrocarbon Degradation (PAH를 분해할 수 있는 Pseudomonas rhodesiae KK1의 SOD 유전자의 동정 및 분자학적 특성 분석)

  • Lee, Dong-Heon;Oh, Kye-Heon;Kim, Seung Il;Kahng, Hyung-Yeel
    • Journal of Life Science
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    • v.26 no.1
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    • pp.75-82
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    • 2016
  • Pseudomonas rhodesiae KK1 has been reported to degrade polycyclic aromatic hydrocarbons (PAHs), such as anthracene, naphthalene, and phenanthrene, which are considered major environmental contaminants. Interestingly, antioxidant genes, including superoxide dismutase, are known to be expressed at different levels in response to environmental contaminants. This study was performed to identify the superoxide dismutase gene in strain KK1, which may be indirectly involved with degradation of PAHs, as well as to investigate the expression pattern of the superoxide dismutase gene in cells grown on different PAHs. Two types of superoxide dismutase genes responsible for the antioxidant defense mechanism, Mn-superoxide dismutase (sodA) and Fe-superoxide dismutase (sodB), were identified in P. rhodesiae KK1. The sodA gene in strain KK1 shared 95% similarity, based on 141 amino acids, with the Mn-sod of P. fluorescens Pf-5. The sodB strain, based on 135 amino acids, shared 99% similarity with the Fe-sod of P. fluorescens Pf-5. Southern hybridization using the sod gene fragment as a probe showed that at least two copies of superoxide dismutase genes exist in strain KK1. RT-PCR analysis revealed that the sodA and sodB genes were more strongly expressed in response to naphthalene and phenanthrene than to anthracene. Interestingly, sodA and sodB activities were revealed to be maintained in cells grown on all of the tested substrates, including glucose.

Molecular Characteristics of Pseudomonas rhodesiae Strain KK1 in Response to Phenanthrene

  • Kahng, Hyung-Yeel;Nam, Kyoung-Phile
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.729-734
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    • 2002
  • Radiorespirometric analysis revealed that Pseudomonas sp. strain KKI isolated from a soil contaminated with petroleum hydrocarbons was able to catabolize polycyclic aromatic hydrocarbons such as phenanthrene and naphthalene. The rate and extent of phenanthrene mineralization was markedly enhanced when the cells were pregrown on either naphthalene or phenanthrene, compared to the cells grown on universal carbon sources (i.e., TSA medium). Deduced amino acid sequence of the Rieske-type iron-sulfur center of a putative phenanthrene dioxygenase (PhnAl) obtained from the strain KKI shared significant homology with DxnAl (dioxin dioxygenase) from Spingomonas sp. RW1, BphA1b (biphenyl dioxygenase) from Spingomonas aromaticivorans F199, and PhnAc (phenanthrene diokygenase) from Burkholderia sp. RP007 or Alcaligenes faecalis AFK2. Northern hybridization using the dioxygenase gene fragment cloned from KKI showed that the expression of the putative phn dioxygenase gene reached the highest level in cells grown in the minimal medium containing phenanthrene and $KNO_3$, and the expression of the phn gene was repressed in cells grown with glucose. In addition to the metabolic change, phospholipid ester-linked fatty acids (PLFA) analysis revealed that the total cellular fatty acid composition of KKI was significantly changed in response to phenanthrene. Fatty acids such as 14:0, 16:0 3OH, 17:0 cyclo, 18:1$\omega$7c, 19:0 cyclo increased in phenanthrene-exposed cells, while fatty acids such as 10:0 3OH, 12:0, 12:0 2OH, 12:0 3OH, 16:1$\omega$7c, 15:0 iso 2OH, 16:0, 18:1$\omega$6c, 18:0 decreased.