This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An erratum has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Iwaki, H.
Right arrow Articles by Lau, P. C. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Iwaki, H.
Right arrow Articles by Lau, P. C. K.
Agricola
Right arrow Articles by Iwaki, H.
Right arrow Articles by Lau, P. C. K.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2002, p. 5671-5684, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5671-5684.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Cloning and Characterization of a Gene Cluster Involved in Cyclopentanol Metabolism in Comamonas sp. Strain NCIMB 9872 and Biotransformations Effected by Escherichia coli-Expressed Cyclopentanone 1,2-Monooxygenase{dagger}

Hiroaki Iwaki,1 Yoshie Hasegawa,2 Shaozhao Wang,3 Margaret M. Kayser,3 and Peter C. K. Lau1*

Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec H4P 2R2,1 Department of Physical Sciences, University of New Brunswick, Saint John, New Brunswick E2L 4L5, Canada,3 Department of Biotechnology, Faculty of Engineering and High Technology Research Center, Kansai University, Suita, Osaka 564-8680, Japan2

Received 22 May 2002/ Accepted 23 August 2002

Cyclopentanone 1,2-monooxygenase, a flavoprotein produced by Pseudomonas sp. strain NCIMB 9872 upon induction by cyclopentanol or cyclopentanone (M. Griffin and P. W. Trudgill, Biochem. J. 129:595-603, 1972), has been utilized as a biocatalyst in Baeyer-Villiger oxidations. To further explore this biocatalytic potential and to discover new genes, we have cloned and sequenced a 16-kb chromosomal locus of strain 9872 that is herein reclassified as belonging to the genus Comamonas. Sequence analysis revealed a cluster of genes and six potential open reading frames designated and grouped in at least four possible transcriptional units as (orf11-orf10-orf9)-(cpnE-cpnD-orf6-cpnC)-(cpnR-cpnB-cpnA)-(orf3-orf4 [partial 3' end]). The cpnABCDE genes encode enzymes for the five-step conversion of cyclopentanol to glutaric acid catalyzed by cyclopentanol dehydrogenase, cyclopentanone 1,2-monooxygenase, a ring-opening 5-valerolactone hydrolase, 5-hydroxyvalerate dehydrogenase, and 5-oxovalerate dehydrogenase, respectively. Inactivation of cpnB by using a lacZ-Kmr cassette resulted in a strain that was not capable of growth on cyclopentanol or cyclopentanone as a sole carbon and energy source. The presence of {sigma}54-dependent regulatory elements in front of the divergently transcribed cpnB and cpnC genes supports the notion that cpnR is a regulatory gene of the NtrC type. Knowledge of the nucleotide sequence of the cpn genes was used to construct isopropyl-ß-thio-D-galactoside-inducible clones of Escherichia coli cells that overproduce the five enzymes of the cpn pathway. The substrate specificities of CpnA and CpnB were studied in particular to evaluate the potential of these enzymes and establish the latter recombinant strain as a bioreagent for Baeyer-Villiger oxidations. Although frequently nonenantioselective, cyclopentanone 1,2-monooxygenase was found to exhibit a broader substrate range than the related cyclohexanone 1,2-monooxygenase from Acinetobacter sp. strain NCIMB 9871. However, in a few cases opposite enantioselectivity was observed between the two biocatalysts.

* Corresponding author. Mailing address: Biotechnology Research Institute, National Research Council Canada, 6100 Royalmount Ave., Montreal, Quebec H4P 2R2, Canada. Phone: (514) 496-6325. Fax: (514) 496-6265. E-mail: peter.lau{at}nrc.ca.

{dagger} This publication is issued as NRCC number 44669.

Applied and Environmental Microbiology, November 2002, p. 5671-5684, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5671-5684.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Rehdorf, J., Zimmer, C. L., Bornscheuer, U. T. (2009). Cloning, Expression, Characterization, and Biocatalytic Investigation of the 4-Hydroxyacetophenone Monooxygenase from Pseudomonas putida JD1. Appl. Environ. Microbiol. 75: 3106-3114 [Abstract] [Full Text]  
  • Kweon, O., Kim, S.-J., Jones, R. C., Freeman, J. P., Adjei, M. D., Edmondson, R. D., Cerniglia, C. E. (2007). A Polyomic Approach To Elucidate the Fluoranthene-Degradative Pathway in Mycobacterium vanbaalenii PYR-1. J. Bacteriol. 189: 4635-4647 [Abstract] [Full Text]  
  • Onaca, C., Kieninger, M., Engesser, K.-H., Altenbuchner, J. (2007). Degradation of Alkyl Methyl Ketones by Pseudomonas veronii MEK700. J. Bacteriol. 189: 3759-3767 [Abstract] [Full Text]  
  • Kotani, T., Yurimoto, H., Kato, N., Sakai, Y. (2007). Novel Acetone Metabolism in a Propane-Utilizing Bacterium, Gordonia sp. Strain TY-5. J. Bacteriol. 189: 886-893 [Abstract] [Full Text]  
  • Iwaki, H., Wang, S., Grosse, S., Bergeron, H., Nagahashi, A., Lertvorachon, J., Yang, J., Konishi, Y., Hasegawa, Y., Lau, P. C. K. (2006). Pseudomonad Cyclopentadecanone Monooxygenase Displaying an Uncommon Spectrum of Baeyer-Villiger Oxidations of Cyclic Ketones. Appl. Environ. Microbiol. 72: 2707-2720 [Abstract] [Full Text]  
  • Malito, E., Alfieri, A., Fraaije, M. W., Mattevi, A. (2004). Crystal structure of a Baeyer-Villiger monooxygenase. Proc. Natl. Acad. Sci. USA 101: 13157-13162 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»