Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Characterization of a Gene Cluster for Cyclohexanone Oxidation in Rhodococcus sp. TK6

  • Choi Jun-Ho (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Kim Tae-Kang (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Kim Young-Mog (Institute of Agricultural Science & Technology, Kyungpook National University) ;
  • Kim Won-Chan (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Park Kunbawui (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Rhee In-Koo (Department of Agricultural Chemistry, Kyungpook National University)
  • Published : 2006.04.01
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Abstract

A gene cluster for cyclohexanone oxidation was cloned from Rhodococcus sp. TK6, which is capable of growth on cyclohexanone as the sole carbon source. The 9,185-bp DNA sequence analysis revealed seven potential open reading frames (ORFs), designated as ssd-chnR-chnD-chnC-chnB-chnE-partial pcd. The chnBCDE genes encode enzymes for the four-step conversion of cyclohexanone to adipic acid, catalyzed by cyclohexanone monooxygenase (ChnB), $\varepsilon-caprolactone$ hydrolase (ChnC), 6-hydroxyhexanoate dehydrogenase (ChnD), and 6-oxohexanoate dehydrogenase (ChnE). Furthermore, the presence of a regulatory element in the downstream region of the chnD gene supports the notion that chnR is a putative regulatory gene. Among them, the activity of ChnB was confirmed and characterized, following their expression and purification in Escherichia coli harboring the modified chnB gene (chnB gene with 6 successive codons for His at the 3' terminus).

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References

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