Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Characterization of Cyclohexanol Dehydrogenase Gene from Rhodococcus sp. TK6

  • CHOI JUN-HO (Division of Applied Biology and Chemistry, Kyungpook National University) ;
  • KIM TAE-KANG (Division of Applied Biology and Chemistry, Kyungpook National University) ;
  • KIM YOUNG-MOG (Institute of Agricultural Science & Technology, Kyungpook National University) ;
  • KIM WON-CHAN (Division of Applied Biology and Chemistry, Kyungpook National University) ;
  • JOO GIL-JAE (Institute of Agricultural Science & Technology, Kyungpook National University) ;
  • LEE KYEONG-YEOLL (Division of Applied Biology and Chemistry, Kyungpook National University) ;
  • RHEE IN-KOO (Division of Applied Biology and Chemistry, Kyungpook National University)
  • Published : 2005.12.01
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Abstract

The cyclohexanol dehydrogenase (ChnA), produced by Rhodococcus sp. TK6, which is capable of growth on cyclohexanol as the sole carbon source, has been previously purified and characterized. However, the current study cloned the complete gene (chnA) for ChnA and its flanking regions using a combination of a polymerase chain reaction (PCR) based on the N-terminal amino acid sequence of the purified ChnA and plaque hybridization from a phage library of Rhodococcus sp. TK6. A sequence analysis of the 5,965-bp DNA fragment revealed five potential open reading frames (ORFs) designated as partial pte (phosphotriesterase), acs (acyl-CoA synthetase), scd (short chain dehydrogenase), stp (sugar transporter), and chnA (cyclohexanol dehydrogenase), respectively. The deduced amino acid sequence of the chnA gene exhibited a similarity of up to $53\%$ with members of the short-chain dehydrogenase/reductase (SDR) family. The chnA gene was expressed using the pET21 a(+) system in Escherichia coli. The activity of the expressed ChnA was then confirmed (13.6 U/mg of protein) and its properties investigated.

Keywords

References

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