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Applied and Environmental Microbiology, August 2004, p. 4880-4888, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4880-4888.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Molecular Identification of the Catabolic Vinyl Chloride Reductase from Dehalococcoides sp. Strain VS and Its Environmental Distribution

Jochen A. Müller,1,{dagger} Bettina M. Rosner,1,{ddagger} Gregory von Abendroth,1 Galit Meshulam-Simon,1 Perry L. McCarty,1 and Alfred M. Spormann1,2*

Department of Civil and Environmental Engineering,1 Departments of Biological Sciences and of Geological and Environmental Sciences, Stanford University, Stanford, California 943052

Received 13 February 2004/ Accepted 4 May 2004

Reductive dehalogenation of vinyl chloride (VC) to ethene is the key step in complete anaerobic degradation of chlorinated ethenes. VC-reductive dehalogenase was partially purified from a highly enriched culture of the VC-respiring Dehalococcoides sp. strain VS. The enzyme reduced VC and all dichloroethene (DCE) isomers, but not tetrachloroethene (PCE) or trichloroethene (TCE), at high rates. By using reversed genetics, the corresponding gene (vcrA) was isolated and characterized. Based on the predicted amino acid sequence, VC reductase is a novel member of the family of corrinoid/iron-sulfur cluster containing reductive dehalogenases. The vcrA gene was found to be cotranscribed with vcrB, encoding a small hydrophobic protein presumably acting as membrane anchor for VC reductase, and vcrC, encoding a protein with similarity to transcriptional regulators of the NosR/NirI family. The vcrAB genes were subsequently found to be present and expressed in other cultures containing VC-respiring Dehalococcoides organisms and could be detected in water samples from a field site contaminated with chlorinated ethenes. Therefore, the vcrA gene identified here may be a useful molecular target for evaluating, predicting, and monitoring in situ reductive VC dehalogenation.

* Corresponding author. Mailing address: Department of Civil and Environmental Engineering, Clark Center East Wing, E250A, Stanford University, Stanford, CA 94305-5429. Phone: (650) 723-3668. Fax: (650) 725-3164. E-mail: Spormann{at}stanford.edu.

{dagger} Present address: Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, MD 21202.

{ddagger} Present address: Dow Chemical Company, San Diego, CA 92121.

Applied and Environmental Microbiology, August 2004, p. 4880-4888, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4880-4888.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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