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Applied and Environmental Microbiology, May 2003, p. 2964-2974, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2964-2974.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of Two Tetrachloroethene-Reducing, Acetate-Oxidizing Anaerobic Bacteria and Their Description as Desulfuromonas michiganensis sp. nov.

Youlboong Sung,1 Kirsti M. Ritalahti,1,2 Robert A. Sanford,3 John W. Urbance,2 Shannon J. Flynn,2 James M. Tiedje,2 and Frank E. Löffler1,4*

School of Civil and Environmental Engineering,1 School of Biology, Georgia Institute of Technology, Atlanta, Georgia 30332-0512,4 Center for Microbial Ecology, Michigan State University, East Lansing, Michigan 48824-1325,2 Department of Civil and Environmental Engineering, University of Illinois, Urbana, Illinois 61801-23523

Received 30 September 2002/ Accepted 30 January 2003

Two tetrachlorethene (PCE)-dechlorinating populations, designated strains BB1 and BRS1, were isolated from pristine river sediment and chloroethene-contaminated aquifer material, respectively. PCE-to-cis-1,2-dichloroethene-dechlorinating activity could be transferred in defined basal salts medium with acetate as the electron donor and PCE as the electron acceptor. Taxonomic analysis based on 16S rRNA gene sequencing placed both isolates within the Desulfuromonas cluster in the {delta} subdivision of the Proteobacteria. PCE was dechlorinated at rates of at least 139 nmol min-1 mg of protein-1 at pH values between 7.0 and 7.5 and temperatures between 25 and 30°C. Dechlorination also occurred at 10°C. The electron donors that supported dechlorination included acetate, lactate, pyruvate, succinate, malate, and fumarate but not hydrogen, formate, ethanol, propionate, or sulfide. Growth occurred with malate or fumarate alone, whereas oxidation of the other electron donors depended strictly on the presence of fumarate, malate, ferric iron, sulfur, PCE, or TCE as an electron acceptor. Nitrate, sulfate, sulfite, thiosulfate, and other chlorinated compounds were not used as electron acceptors. Sulfite had a strong inhibitory effect on growth and dechlorination. Alternate electron acceptors (e.g., fumarate or ferric iron) did not inhibit PCE dechlorination and were consumed concomitantly. The putative fumarate, PCE, and ferric iron reductases were induced by their respective substrates and were not constitutively present. Sulfide was required for growth. Both strains tolerated high concentrations of PCE, and dechlorination occurred in the presence of free-phase PCE (dense non-aqueous-phase liquids). Repeated growth with acetate and fumarate as substrates yielded a BB1 variant that had lost the ability to dechlorinate PCE. Due to the 16S rRNA gene sequence differences with the closest relatives and the unique phenotypic characteristics, we propose that the new isolates are members of a new species, Desulfuromonas michiganensis, within the Desulfuromonas cluster of the Geobacteraceae.

* Corresponding author. Mailing address: School of Civil and Environmental Engineering, 311 Ferst Dr., Room 3228, Georgia Institute of Technology, Atlanta, GA 30332-0512. Phone: (404) 894-0279. Fax: (404) 894-8266. E-mail: frank.loeffler{at}ce.gatech.edu.

Applied and Environmental Microbiology, May 2003, p. 2964-2974, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2964-2974.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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