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Applied and Environmental Microbiology, February 2004, p. 1169-1175, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1169-1175.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Detection and Characterization of a Dehalogenating Microorganism by Terminal Restriction Fragment Length Polymorphism Fingerprinting of 16S rRNA in a Sulfidogenic, 2-Bromophenol-Utilizing Enrichment

Donna E. Fennell ,^1,, Sung-Keun Rhee,^2,, Young-Beom Ahn,¹ Max M. Häggblom,¹ and Lee J. Kerkhof^2*

Department of Biochemistry and Microbiology and Biotechnology Center for Agriculture and the Environment,¹ Institute for Marine and Coastal Sciences, Rutgers, The State University of New Jersey, New Brunswick, New Jersey 08901²

Received 7 July 2003/ Accepted 23 October 2003

Terminal restriction fragment length polymorphism analysis of reverse-transcribed 16S rRNA during periods of community flux was used as a tool to delineate the roles of the members of a 2-bromophenol-degrading, sulfate-reducing consortium. Starved, washed cultures were amended with 2-bromophenol plus sulfate, 2-bromophenol plus hydrogen, phenol plus sulfate, or phenol with no electron acceptor and were monitored for substrate use. In the presence of sulfate, 2-bromophenol and phenol were completely degraded. In the absence of sulfate, 2-bromophenol was dehalogenated and phenol accumulated. Direct terminal restriction fragment length polymorphism fingerprinting of the 16S rRNA in the various subcultures indicated that phylotype 2BP-48 (a Desulfovibrio-like sequence) was responsible for the dehalogenation of 2-bromophenol. A stable coculture was established which contained predominantly 2BP-48 and a second Desulfovibrio-like bacterium (designated BP212 based on terminal restriction fragment length polymorphism fingerprinting) that was capable of dehalogenating 2-bromophenol to phenol. Strain 2BP-48 in the coculture could couple reductive dehalogenation to growth with 2-bromophenol, 2,6-dibromophenol, or 2-iodophenol and lactate or formate as the electron donor. In addition to halophenols, strain 2BP-48 appears to use sulfate, sulfite, and thiosulfate as electron acceptors and is capable of simultaneous sulfidogenesis and reductive dehalogenation in the presence of sulfate.

Present address: Department of Environmental Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901.

D. E. Fennell and S.-K. Rhee contributed equally to this work.

Present address: BioDiversity Information Laboratory, Biological Resources Center, Korea Research Institute of Bioscience & Biotechnology, Yuseong-gu, Daejon 305-333, South Korea.

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