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Applied and Environmental Microbiology, October 2002, p. 4764-4771, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4764-4771.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Cloning, Sequencing, and Characterization of the Hexahydro-1,3,5-Trinitro-1,3,5-Triazine Degradation Gene Cluster from Rhodococcus rhodochrous

Helena M. B. Seth-Smith,1 Susan J. Rosser,1 Amrik Basran,1 Emma R. Travis,1 Eric R. Dabbs,2 Steve Nicklin,3 and Neil C. Bruce1*

Institute of Biotechnology, University of Cambridge, Cambridge CB2 1QT,1 Defence Science and Technology Laboratory, Fort Halstead, Sevenoaks, Kent TN14 7BP, United Kingdom,3 University of the Witwatersrand, Johannesburg, South Africa2

Received 16 April 2002/ Accepted 3 July 2002

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is a high explosive which presents an environmental hazard as a major land and groundwater contaminant. Rhodococcus rhodochrous strain 11Y was isolated from explosive contaminated land and is capable of degrading RDX when provided as the sole source of nitrogen for growth. Products of RDX degradation in resting-cell incubations were analyzed and found to include nitrite, formaldehyde, and formate. No ammonium was excreted into the medium, and no dead-end metabolites were observed. The gene responsible for the degradation of RDX in strain 11Y is a constitutively expressed cytochrome P450-like gene, xplA, which is found in a gene cluster with an adrenodoxin reductase homologue, xplB. The cytochrome P450 also has a flavodoxin domain at the N terminus. This study is the first to present a gene which has been identified as being responsible for RDX biodegradation. The mechanism of action of XplA on RDX is thought to involve initial denitration followed by spontaneous ring cleavage and mineralization.

* Corresponding author. Present address: Centre for Novel Agricultural Products, Department of Biology, University of York, P.O. Box 373, York YO10 5YW, United Kingdom. Phone: 44 (0) 1904 432618. Fax: 44 (0) 1904 432928. E-mail: ncb5{at}york.ac.uk.

Applied and Environmental Microbiology, October 2002, p. 4764-4771, Vol. 68, No. 10
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.10.4764-4771.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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