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Applied and Environmental Microbiology, December 2000, p. 5141-5147, Vol. 66, No. 12
0099-2240/00/$04.00+0

Trichloroethene Reductive Dehalogenase from Dehalococcoides ethenogenes: Sequence of tceA and Substrate Range Characterization

Jon K. Magnuson,1,* Margaret F. Romine,1 David R. Burris,2 and Mark T. Kingsley1

Battelle/Pacific Northwest National Laboratory, Richland, Washington 99352,1 and Air Force Research Laboratory, Tyndall Air Force Base, Florida 324032

Received 25 April 2000/Accepted 15 September 2000

The anaerobic bacterium Dehalococcoides ethenogenes is the only known organism that can completely dechlorinate tetrachloroethene or trichloroethene (TCE) to ethene via dehalorespiration. One of two corrinoid-containing enzymes responsible for this pathway, TCE reductive dehalogenase (TCE-RDase) catalyzes the dechlorination of TCE to ethene. TCE-RDase dehalogenated 1,2-dichloroethane and 1,2-dibromoethane to ethene at rates of 7.5 and 30 µmol/min/mg, respectively, similar to the rates for TCE, cis-dichloroethene (DCE), and 1,1-DCE. A variety of other haloalkanes and haloalkenes containing three to five carbon atoms were dehalogenated at lower rates. The gene encoding TCE-RDase, tceA, was cloned and sequenced via an inverse PCR approach. Sequence comparisons of tceA to proteins in the public databases revealed weak sequence similarity confined to the C-terminal region, which contains the eight-iron ferredoxin cluster binding motif, (CXXCXXCXXXCP)2. Direct N-terminal sequencing of the mature enzyme indicated that the first 42 amino acids constitute a signal sequence containing the twin-arginine motif, RRXFXK, associated with the Sec-independent membrane translocation system. This information coupled with membrane localization studies indicated that TCE-RDase is located on the exterior of the cytoplasmic membrane. Like the case for the two other RDases that have been cloned and sequenced, a small open reading frame, tceB, is proposed to be involved with membrane association of TCE-RDase and is predicted to be cotranscribed with tceA.

* Corresponding author. Mailing address: Battelle/PNNL, MSIN: K2-21, 902 Battelle Blvd., P.O. Box 999, Richland, WA 99352. Phone: (509) 372-4119. Fax: (509) 375-2009. E-mail: jon.magnuson{at}pnl.gov.

Applied and Environmental Microbiology, December 2000, p. 5141-5147, Vol. 66, No. 12
0099-2240/00/$04.00+0


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