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Applied and Environmental Microbiology, November 2007, p. 7067-7074, Vol. 73, No. 21
0099-2240/07/$08.00+0 doi:10.1128/AEM.01161-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Michele Breary, and
Timothy R. McDermott*
Thermal Biology Institute and Department of Land Resources and Environmental Sciences, Montana State University, Bozeman, Montana 59717
Received 23 May 2007/ Accepted 30 August 2007
Previous studies in an acid-sulfate-chloride spring in Yellowstone National Park found that microbial arsenite [As(III)] oxidation is absent in regions of the spring outflow channel where H2S exceeds
5 µM and served as a backdrop for continued efforts in the present study. Ex situ assays with microbial mat samples demonstrated immediate As(III) oxidation activity when H2S was absent or at low concentrations, suggesting the presence of As(III) oxidase enzymes that could be reactivated if H2S is removed. Cultivation experiments initiated with mat samples taken from along the H2S gradient in the outflow channel resulted in the isolation of an As(III)-oxidizing chemolithotroph from the low-H2S region of the gradient. The isolate was phylogenetically related to Acidicaldus and was characterized in vitro for spring-relevant properties, which were then compared to its distribution pattern in the spring as determined by denaturing gradient gel electrophoresis and quantitative PCR. While neither temperature nor oxygen requirements appeared to be related to the occurrence of this organism within the outflow channel, H2S concentration appeared to be an important constraint. This was verified by in vitro pure-culture modeling and kinetic experiments, which suggested that H2S inhibition of As(III) oxidation is uncompetitive in nature. In summary, the studies reported herein illustrate that H2S is a potent inhibitor of As(III) oxidation and will influence the niche opportunities and population distribution of As(III) chemolithotrophs.
Published ahead of print on 7 September 2007.
Present address: Department of Chemistry and Biochemistry, California Polytechnic State University, San Luis Obispo, CA 93407.
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