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Applied and Environmental Microbiology, May 2009, p. 2684-2693, Vol. 75, No. 9
0099-2240/09/$08.00+0     doi:10.1128/AEM.02037-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene{triangledown}

Ariel Grostern1 and Elizabeth A. Edwards2*

Department of Botany,1 Department of Chemical Engineering, University of Toronto, Toronto, Ontario, Canada2

Received 2 September 2008/ Accepted 25 February 2009

Dehalobacter and "Dehalococcoides" spp. were previously shown to be involved in the biotransformation of 1,1,2-trichloroethane (1,1,2-TCA) and 1,2-dichloroethane (1,2-DCA) to ethene in a mixed anaerobic enrichment culture. Here we report the further enrichment and characterization of a Dehalobacter sp. from this mixed culture in coculture with an Acetobacterium sp. Through a series of serial transfers and dilutions with acetate, H2, and 1,2-DCA, a stable coculture of Acetobacterium and Dehalobacter spp. was obtained, where Dehalobacter grew during dechlorination. The isolated Acetobacterium strain did not dechlorinate 1,2-DCA. Quantitative PCR with specific primers showed that Dehalobacter cells did not grow in the absence of a chlorinated electron acceptor and that the growth yield with 1,2-DCA was 6.9 (±0.7) x 107 16S rRNA gene copies/µmol 1,2-DCA degraded. PCR with degenerate primers targeting reductive dehalogenase genes detected three distinct Dehalobacter/Desulfitobacterium-type sequences in the mixed-parent culture, but only one of these was present in the 1,2-DCA-H2 coculture. Reverse transcriptase PCR revealed the transcription of this dehalogenase gene specifically during the dechlorination of 1,2-DCA. The 1,2-DCA-H2 coculture could dechlorinate 1,2-DCA but not 1,1,2-TCA, nor could it dechlorinate chlorinated ethenes. As a collective, the genus Dehalobacter has been show to dechlorinate many diverse compounds, but individual species seem to each have a narrow substrate range.

* Corresponding author. Mailing address: University of Toronto, 200 College St., Toronto, ON M5S 3E5, Canada. Phone: (416) 946-3506. Fax: (416) 978-8605. E-mail: elizabeth.edwards{at}utoronto.ca

{triangledown} Published ahead of print on 6 March 2009.

Applied and Environmental Microbiology, May 2009, p. 2684-2693, Vol. 75, No. 9
0099-2240/09/$08.00+0     doi:10.1128/AEM.02037-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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