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Applied and Environmental Microbiology, November 1999, p. 5158-5162, Vol. 65, No. 11
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification of a Transcriptional Activator (ChnR) and a 6-Oxohexanoate Dehydrogenase (ChnE) in the Cyclohexanol Catabolic Pathway in Acinetobacter sp. Strain NCIMB 9871 and Localization of the Genes That Encode Themdagger

Hiroaki Iwaki,1 Yoshie Hasegawa,1,* Masahiro Teraoka,1 Tai Tokuyama,1 Hélène Bergeron,2 and Peter C. K. Lau2

Department of Biotechnology, Faculty of Engineering & High Technology Research Center, Kansai University, Suita, Osaka 564-8680, Japan,1 and Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec H4P 2R2, Canada2

Received 5 March 1999/Accepted 8 August 1999

We identified chnR, a gene encoding an AraC-XylS type of transcriptional activator that regulates the expression of chnB, the structural gene for cyclohexanone monooxygenase (CHMO) in Acinetobacter sp. strain NCIMB 9871. The gene sequence of chnE, which encodes an NADP+-linked 6-oxohexanoate dehydrogenase, the enzyme catalyzing the fifth step of cyclohexanol degradation, was also determined. The gene arrangement is chnB-chnE-chnR. The predicted molecular masses of the three polypeptides were verified by radiolabeling by using the T7 expression system. Inducible expression of cloned chnB in Escherichia coli depended upon the presence of chnR. A transcriptional chnB::lacZ fusion experiment revealed that cyclohexanone induces chnB expression in E. coli, in which a 22-fold increase in activity was observed.

* Corresponding author. Mailing address: Department of Biotechnology, Faculty of Engineering & High Technology Research Center, Kansai University, Suita, Osaka 564-8680, Japan. Phone: (06) 6368-0909. Fax: (06) 6388-8609. E-mail: yoshie{at}ipcku.kansai-u.ac.jp.

dagger This publication is issued as NRCC number 43271.

Applied and Environmental Microbiology, November 1999, p. 5158-5162, Vol. 65, No. 11
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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