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Applied and Environmental Microbiology, September 2004, p. 5595-5602, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5595-5602.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Alkaline Anaerobic Respiration: Isolation and Characterization of a Novel Alkaliphilic and Metal-Reducing Bacterium

Qi Ye,^1, Yul Roh,² Susan L. Carroll,² Benjamin Blair,³ Jizhong Zhou,² Chuanlun L. Zhang,^1, and Matthew W. Fields^2,4*

Department of Geology, University of Missouri, Columbia, Missouri,¹ Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee,² Department of Biology, Jacksonville State University, Jacksonville, Alabama,³ Department of Microbiology, Miami University, Oxford, Ohio⁴

Received 11 December 2003/ Accepted 16 May 2004

Iron-reducing enrichments were obtained from leachate ponds at the U.S. Borax Company in Boron, Calif. Based on partial small-subunit (SSU) rRNA gene sequences (approximately 500 nucleotides), six isolates shared 98.9% nucleotide identity. As a representative, the isolate QYMF was selected for further analysis. QYMF could be grown with Fe(III)-citrate, Fe(III)-EDTA, Co(III)-EDTA, or Cr(VI) as electron acceptors, and yeast extract and lactate could serve as electron donors. Growth during iron reduction occurred over the pH range of 7.5 to 11.0 (optimum, pH 9.5), a sodium chloride range of 0 to 80 g/liter (optimum, 20 g/liter), and a temperature range of 4 to 45°C (optimum, approximately 35°C), and iron precipitates were formed. QYMF was a strict anaerobe that could be grown in the presence of borax, and the cells were straight rods that produced endospores. Sodium chloride and yeast extract stimulated growth. Phylogenetic analysis of the SSU rRNA gene indicated that the bacterium was a low-G+C gram-positive microorganism and had 96 and 92% nucleotide identity with Alkaliphilus transvaalensis and Alkaliphilus crotonatoxidans, respectively. The major phospholipid fatty acids were 14:1, 16:17c, and 16:0, which were different from those of other alkaliphiles but similar to those of reported iron-reducing bacteria. The results demonstrated that the isolate might represent a novel metal-reducing alkaliphilic species. The name Alkaliphilus metalliredigens sp. nov. is proposed. The isolation and activity of metal-reducing bacteria from borax-contaminated leachate ponds suggest that bioremediation of metal-contaminated alkaline environments may be feasible and have implications for alkaline anaerobic respiration.

Present address: Department of Marine Sciences, University of Georgia, Athens, GA 30602.

Present address: Savannah River Ecology Laboratory, University of Georgia, Aiken, SC 29802.

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