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Applied and Environmental Microbiology, January 2001, p. 307-316, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.307-316.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Chloromethane Utilization Gene Cluster from Hyphomicrobium chloromethanicum Strain CM2T and Development of Functional Gene Probes To Detect Halomethane-Degrading Bacteria

Craig McAnulla,1 Claire A. Woodall,1 Ian R. McDonald,1,* Alex Studer,2 Stephane Vuilleumier,2 Thomas Leisinger,2 and J. Colin Murrell1

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, England,1 and Mikrobiologisches Institute, ETH Zurich, CH-8092 Zurich, Switzerland2

Received 3 July 2000/Accepted 4 October 2000

Hyphomicrobium chloromethanicum CM2T, an aerobic methylotrophic member of the alpha  subclass of the class proteobacteria, can grow with chloromethane as the sole carbon and energy source. H. chloromethanicum possesses an inducible enzyme system for utilization of chloromethane, in which two polypeptides (67-kDa CmuA and 35-kDa CmuB) are expressed. Previously, four genes, cmuA, cmuB, cmuC, and purU, were shown to be essential for growth of Methylobacterium chloromethanicum on chloromethane. The cmuA and cmuB genes were used as probes to identify homologs in H. chloromethanicum. A cmu gene cluster (9.5 kb) in H. chloromethanicum contained 10 open reading frames: folD (partial), pduX, orf153, orf207, orf225, cmuB, cmuC, cmuA, fmdB, and paaE (partial). CmuA from H. chloromethanicum (67 kDa) showed high identity to CmuA from M. chloromethanicum and contains an N-terminal methyltransferase domain and a C-terminal corrinoid-binding domain. CmuB from H. chloromethanicum is related to a family of methyl transfer proteins and to the CmuB methyltransferase from M. chloromethanicum. CmuC from H. chloromethanicum shows identity to CmuC from M. chloromethanicum and is a putative methyltransferase. folD codes for a methylene-tetrahydrofolate cyclohydrolase, which may be involved in the C1 transfer pathway for carbon assimilation and CO2 production, and paaE codes for a putative redox active protein. Molecular analyses and some preliminary biochemical data indicated that the chloromethane utilization pathway in H. chloromethanicum is similar to the corrinoid-dependent methyl transfer system in M. chloromethanicum. PCR primers were developed for successful amplification of cmuA genes from newly isolated chloromethane utilizers and enrichment cultures.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, England. Phone: 44 24 765 28362. Fax: 44 24 765 23568. E-mail: IMcDonald{at}bio.warwick.ac.uk.

Applied and Environmental Microbiology, January 2001, p. 307-316, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.307-316.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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  • Borodina, E., McDonald, I. R., Murrell, J. C. (2004). Chloromethane-Dependent Expression of the cmu Gene Cluster of Hyphomicrobium chloromethanicum. Appl. Environ. Microbiol. 70: 4177-4186 [Abstract] [Full Text]  
  • Studer, A., McAnulla, C., Buchele, R., Leisinger, T., Vuilleumier, S. (2002). Chloromethane-Induced Genes Define a Third C1 Utilization Pathway in Methylobacterium chloromethanicum CM4. J. Bacteriol. 184: 3476-3484 [Abstract] [Full Text]  
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