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Reductive dechlorination of tetrachloroethene by a stepwise catalysis of different organohalide respiring bacteria and reductive dehalogenases

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Abstract

The enrichment culture SL2 dechlorinating tetrachloroethene (PCE) to ethene with strong trichloroethene (TCE) accumulation prior to cis-1,2-dichloroethene (cis-DCE) formation was analyzed for the presence of organohalide respiring bacteria and reductive dehalogenase genes (rdhA). Sulfurospirillum-affiliated bacteria were identified to be involved in PCE dechlorination to cis-DCE whereas “Dehalococcoides”-affiliated bacteria mainly dechlorinated cis-DCE to ethene. Two rdhA genes highly similar to tetrachloroethene reductive dehalogenase genes (pceA) of S. multivorans and S. halorespirans were present as well as an rdhA gene very similar to the trichloroethene reductive dehalogenase gene (tceA) of “Dehalococcoides ethenogenes” strain 195. A single strand conformation polymorphism (SSCP) method was developed allowing the simultaneous detection of the three rdhA genes and the estimation of their abundance. SSCP analysis of different SL2 cultures showed that one pceA gene was expressed during PCE dechlorination whereas the second was expressed during TCE dechlorination. The tceA gene was involved in cis-DCE dechlorination to ethene. Analysis of the internal transcribed spacer region between the 16S and 23S rRNA genes revealed two distinct sequences originating from Sulfurospirillum suggesting that two Sulfurospirillum populations were present in SL2. Whether each Sulfurospirillum population was catalyzing a different dechlorination step could however not be elucidated.

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Acknowledgments

M.-P. Charnay was financed during her visit to LBE by the Ministry of agriculture of France (AgroParisTech). Part of this work was supported by the Swiss National Science Foundation grants 3152-55413 and 3100-066957, by a grant of the Swiss Federal Office for Education and Science (contract 99.0362) in the framework of the EU project MAROC (EVK1-1999-00023), and by the project BIOTOOL (GOCE-003998) of the European Commission under the Sixth Framework Programme. We are grateful to Magdalena Sanchez for the excellent technical support and to Janneke Krooneman from Bioclear BV, Groningen, The Netherlands, for supplying the sample used for enrichment inoculation.

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Author notes

  1. Marie-Paule Charnay

    Present address: Université de Lausanne, Ecole de Biologie, Quartier UNIL-Sorge, Bâtiment Biophore, 1015, Lausanne, Switzerland

  2. Christophe Regeard

    Present address: CNRS UMR 8621, Institut de Génétique et Microbiologie, Bat. 409, Université Paris Sud, 91405, Orsay, France

  3. Katia Rouzeau-Szynalski

    Present address: Product Technology Center-Nestlé, Konolfingen, Switzerland

  4. Pierre Rossi

    Present address: EPFL ENAC IIE CEL, Station 2, 1015, Lausanne, Switzerland

Authors and Affiliations

  1. Ecole Polytechnique Fédérale de Lausanne (EPFL), ENAC IIE—Laboratory for Environmental Biotechnology, Station 6, 1015, Lausanne, Switzerland

    Julien Maillard, Marie-Paule Charnay, Christophe Regeard, Emmanuelle Rohrbach-Brandt, Katia Rouzeau-Szynalski, Pierre Rossi & Christof Holliger

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Correspondence to Christof Holliger.

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Maillard, J., Charnay, MP., Regeard, C. et al. Reductive dechlorination of tetrachloroethene by a stepwise catalysis of different organohalide respiring bacteria and reductive dehalogenases. Biodegradation 22, 949–960 (2011). https://doi.org/10.1007/s10532-011-9454-4

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