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Journal of Bacteriology, December 2003, p. 7120-7128, Vol. 185, No. 24
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.24.7120-7128.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Propane Monooxygenase and NAD+-Dependent Secondary Alcohol Dehydrogenase in Propane Metabolism by Gordonia sp. Strain TY-5

Tetsuya Kotani, Tazuko Yamamoto, Hiroya Yurimoto, Yasuyoshi Sakai, and Nobuo Kato*

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa Sakyo-ku, Kyoto 606-8502, Japan

Received 7 July 2003/ Accepted 18 September 2003

A new isolate, Gordonia sp. strain TY-5, is capable of growth on propane and n-alkanes with C13 to C22 carbon chains as the sole source of carbon. In whole-cell reactions, significant propane oxidation to 2-propanol was detected. A gene cluster designated prmABCD, which encodes the components of a putative dinuclear-iron-containing multicomponent monooxygenase, including the large and small subunits of the hydroxylase, an NADH-dependent acceptor oxidoreductase, and a coupling protein, was cloned and sequenced. A mutant with prmB disrupted (prmB::Kanr) lost the ability to grow on propane, and Northern blot analysis revealed that polycistronic transcription of the prm genes was induced during its growth on propane. These results indicate that the prmABCD gene products play an essential role in propane oxidation by the bacterium. Downstream of the prm genes, an open reading frame (adh1) encoding an NAD+-dependent secondary alcohol dehydrogenase was identified, and the protein was purified and characterized. The Northern blot analysis results and growth properties of a disrupted mutant (adh1::Kanr) indicate that Adh1 plays a major role in propane metabolism. Two additional NAD+-dependent secondary alcohol dehydrogenases (Adh2 and Adh3) were also found to be involved in 2-propanol oxidation. On the basis of these results, we conclude that Gordonia sp. strain TY-5 oxidizes propane by monooxygenase-mediated subterminal oxidation via 2-propanol.

* Corresponding author. Mailing address: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan. Phone: 81 75 753 6385. Fax: 81 75 753 6385. E-mail: nkato{at}kais.kyoto-u.ac.jp.

Journal of Bacteriology, December 2003, p. 7120-7128, Vol. 185, No. 24
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.24.7120-7128.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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