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Methanogenic Bacteria from the Bondyuzhskoe Oil Field: General Characterization and Analysis of Stable-Carbon Isotopic Fractionation

Belyaev, S. S. and Wolkin, R. and Kenealy, W. R. and DeNiro, M. J. and Epstein, S. and Zeikus, J. G. (1983) Methanogenic Bacteria from the Bondyuzhskoe Oil Field: General Characterization and Analysis of Stable-Carbon Isotopic Fractionation. Applied and Environmental Microbiology, 45 (2). pp. 691-697. ISSN 0099-2240.

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Selective enrichment culture techniques were employed to obtain mixed cultures of methanogenic rods and sarcina from surface flooding waters and deep subsurface (∼1650 m) oil-bearing sedimentary rocks and formation waters sampled from an old oil field in the U.S.S.R. previously reported to display active biological methanogenesis. The methanogens were selectively isolated as colonies on agar petri dishes that were incubated in a novel container. The general cellular and growth features of three Methanobacterium isolates were determined. These strains grew optimally at 37 to 45°C in anaerobic pressure tube cultures with a doubling time of 16 to 18 h on H2-CO2 and proliferated as autotrophs. Acetate addition significantly enhanced the final cell yield. Growth of these strains was completely inhibited by either 0.6 g of sodium sulfide per liter or 31.0 of sodium chloride per liter, but growth was not inhibited by either 0.3 g of sodium sulfide per liter or 1.0 g of sodium sulfate per liter. One novel isolate, Methanobacterium sp. strain ivanov, was grown on H2-CO2, and the stable-carbon isotopic fractionations that occurred during synthesis of methane, cell carbon, and lipids were determined. The results of this study were used to examine the anomalous relationship between the isotopic and chemical compositions of natural gas occurring in the deep subsurface environment of the oil field.

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Additional Information:Copyright © 1983, American Society for Microbiology Received 20 June 1982/Accepted 21 October 1982 This research was supported by the College of Agricultural and Life Sciences, University of Wisconsin, Madison, and National Science Foundation EAR 79-26239. We gratefully acknowledge the research fellowship awarded to S.S.B. from the cooperative agreement between the U.S.S.R. and U.S.A. National Academies of Sciences. R.W. and W.R.K. were supported by predoctoral traineeships from the National Institutes of Health and by Cellular and Molecular Biology training grant 53 GM07215. We thank A. N. Fuex of Shell Development Corp., Houston, Tex. for helpful discussions.
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Deposited On:13 Mar 2006
Last Modified:02 Oct 2019 22:50

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