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Applied and Environmental Microbiology, December 2002, p. 6013-6020, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6013-6020.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Isolation and Characterization of Metal-Reducing Thermoanaerobacter Strains from Deep Subsurface Environments of the Piceance Basin, Colorado

Yul Roh, Shi V. Liu,{dagger} Guangshan Li, Heshu Huang, Tommy J. Phelps, and Jizhong Zhou*

Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831-6038

Received 21 December 2001/ Accepted 29 August 2002

Five bacterial strains were isolated from anaerobic enrichment cultures that had originated from inoculations with samples collected from the deep subsurface environments of the millions-of-years-old, geologically and hydrologically isolated Piceance Basin in Colorado. Small-subunit rRNA gene-based analyses indicated that all of these bacteria were closely related to Thermoanaerobacter ethanolicus, with similarities of 99.4 to 99.5%. Three isolates (X513, X514, and X561) from the five bacterial strains were used to examine physiological characteristics. These thermophilic bacteria were able to use acetate, glucose, hydrogen, lactate, pyruvate, succinate, and xylose as electron donors while reducing Fe(III), cobalt(III), chromium(VI), manganese(IV), and uranium(VI) at 60°C. One of the isolates (X514) was also able to utilize hydrogen as an electron donor for Fe(III) reduction. These bacteria exhibited diverse mineral precipitation capabilities, including the formation of magnetite (Fe3O4), siderite (FeCO3), rhodochrosite (MnCO3), and uraninite (UO2). The gas composition of the incubation headspace and the ionic composition of the incubation medium exerted profound influences on the types of minerals formed. The susceptibility of the thermophilic Fe(III)-reducing cultures to metabolic inhibitors specific for ferric reductase, hydrogenase, and electron transport indicated that iron reduction by these bacteria is an enzymatic process.

* Corresponding author. Mailing address: Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6038. Phone: (865) 576-7544. Fax: (865) 576-8646. E-mail: zhouj{at}ornl.gov. {dagger} Present address: National Exposure Research Laboratory, Environmental Protection Agency, Research Triangle Park, NC 27711.

{dagger} Present address: National Exposure Research Laboratory, Environmental Protection Agency, Research Triangle Park, NC 27711.

Applied and Environmental Microbiology, December 2002, p. 6013-6020, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6013-6020.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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