Characterization of a Dehalobacter coculture that dechlorinates 1,2-dichloroethane to ethene and identification of the putative reductive dehalogenase gene

Appl Environ Microbiol. 2009 May;75(9):2684-93. doi: 10.1128/AEM.02037-08. Epub 2009 Mar 6.

Abstract

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, H(2), 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 10(7) 16S rRNA gene copies/mumol 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-H(2) coculture. Reverse transcriptase PCR revealed the transcription of this dehalogenase gene specifically during the dechlorination of 1,2-DCA. The 1,2-DCA-H(2) 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acetobacterium / growth & development
  • Acetobacterium / metabolism*
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Coculture Techniques
  • Colony Count, Microbial / methods
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Ethylene Dichlorides / metabolism*
  • Ethylenes / metabolism*
  • Hydrolases / genetics*
  • Hydrolases / metabolism
  • Molecular Sequence Data
  • Peptococcaceae / growth & development
  • Peptococcaceae / metabolism*
  • Sequence Analysis, DNA
  • Trichloroethanes / metabolism

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Ethylene Dichlorides
  • Ethylenes
  • Trichloroethanes
  • 1,1,2-trichloroethane
  • ethylene dichloride
  • ethylene
  • Hydrolases

Associated data

  • GENBANK/FJ010189
  • GENBANK/FJ010190
  • GENBANK/FJ010191
  • GENBANK/FJ157998