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Applied and Environmental Microbiology, January 2003, p. 107-112, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.107-112.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Isolation and Characterization of a Sulfur-Oxidizing Chemolithotroph Growing on Crude Oil under Anaerobic Conditions

Yumiko Kodama and Kazuya Watanabe*

Marine Biotechnology Institute, Kamaishi Laboratories, Heita, Kamaishi City, Iwate 026-0001, Japan

Received 23 May 2002/ Accepted 30 September 2002

Molecular approaches have shown that a group of bacteria (called cluster 1 bacteria) affiliated with the {varepsilon} subclass of the class Proteobacteria constituted major populations in underground crude-oil storage cavities. In order to unveil their physiology and ecological niche, this study isolated bacterial strains (exemplified by strain YK-1) affiliated with the cluster 1 bacteria from an oil storage cavity at Kuji in Iwate, Japan. 16S rRNA gene sequence analysis indicated that its closest relative was Thiomicrospira denitrificans (90% identity). Growth experiments under anaerobic conditions showed that strain YK-1 was a sulfur-oxidizing obligate chemolithotroph utilizing sulfide, elemental sulfur, thiosulfate, and hydrogen as electron donors and nitrate as an electron acceptor. Oxygen also supported its growth only under microaerobic conditions. Strain YK-1 could not grow on nitrite, and nitrite was the final product of nitrate reduction. Neither sugars, organic acids (including acetate), nor hydrocarbons could serve as carbon and energy sources. A typical stoichiometry of its energy metabolism followed an equation: S2- + 4NO3- -> SO42- + 4NO2- ({Delta}G0 = -534 kJ mol-1). In a difference from other anaerobic sulfur-oxidizing bacteria, this bacterium was sensitive to NaCl; growth in medium containing more than 1% NaCl was negligible. When YK-1 was grown anaerobically in a sulfur-depleted inorganic medium overlaid with crude oil, sulfate was produced, corresponding to its growth. On the contrary, YK-1 could not utilize crude oil as a carbon source. These results suggest that the cluster 1 bacteria yielded energy for growth in oil storage cavities by oxidizing petroleum sulfur compounds. Based on its physiology, ecological interactions with other members of the groundwater community are discussed.

* Corresponding author. Mailing address: Marine Biotechnology Institute, 3-75-1 Heita, Kamaishi, Iwate 026-0001, Japan. Phone: 81-193-26-5781. Fax: 81-193-26-6592. E-mail: kazuya.watanabe{at}mbio.jp.

Applied and Environmental Microbiology, January 2003, p. 107-112, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.107-112.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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