Trichloroethene reductive dehalogenase from Dehalococcoides ethenogenes: sequence of tceA and substrate range characterization.
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 micromol/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.
Magnuson
J K
JK
Battelle/Pacific Northwest National Laboratory, Richland, Washington 99352, USA. jon.magnuson@pnl.gov
Romine
M F
MF
Burris
D R
DR
Kingsley
M T
MT
D000595:Amino Acid Sequence; D001421:Bacteria, Anaerobic / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D001673:Biodegradation, Environmental; D017931:DNA Primers / Q000235:genetics; D005798:Genes, Bacterial; D006846:Hydrocarbons, Halogenated / Q000378:metabolism; D006867:Hydrolases / Q000235:genetics* / Q000378:metabolism*; D008566:Membranes / Q000201:enzymology; D008969:Molecular Sequence Data; D010088:Oxidoreductases; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D017931:DNA Primers; D006846:Hydrocarbons, Halogenated; D010088:Oxidoreductases; C112990:tetrachloroethene dehalogenase; D006867:Hydrolases