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Applied and Environmental Microbiology, December 1999, p. 5532-5540, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Initial Reactions in Anaerobic Alkane Degradation by a Sulfate Reducer, Strain AK-01

Chi Ming So and L. Y. Young^*

Biotechnology Center for Agriculture and the Environment and Department of Environmental Sciences, Cook College, Rutgers, The State University of New Jersey, New Brunswick, New Jersey 08901-8520

Received 28 June 1999/Accepted 28 September 1999

An alkane-degrading, sulfate-reducing bacterial strain, AK-01, isolated from a petroleum-contaminated sediment was studied to elucidate its mechanism of alkane metabolism. Total cellular fatty acids of AK-01 were predominantly C even when it was grown on C-even alkanes and were predominantly C odd when grown on C-odd alkanes, suggesting that the bacterium anaerobically oxidizes alkanes to fatty acids. Among these fatty acids, some 2-, 4-, and 6-methylated fatty acids were specifically found only when AK-01 was grown on alkanes, and their chain lengths always correlated with those of the alkanes. When [1,2-¹³C₂]hexadecane or perdeuterated pentadecane was used as the growth substrate, ¹³C-labeled 2-Me-16:0, 4-Me-18:0, and 6-Me-20:0 fatty acids or deuterated 2-Me-15:0, 4-Me-17:0, and 6-Me-19:0 fatty acids were recovered, respectively, confirming that these monomethylated fatty acids were alkane derived. Examination of the ¹³C-labeled 2-, 4-, and 6-methylated fatty acids by mass spectrometry showed that each of them contained two ¹³C atoms, located at the methyl group and the adjacent carbon, thus indicating that the methyl group was the original terminal carbon of the [1,2-¹³C₂]hexadecane. For perdeuterated pentadecane, the presence of three deuterium atoms, on the methyl group and its adjacent carbon, in each of the deuterated 2-, 4-, and 6-methylated fatty acids further supported the hypothesis that the methyl group was the terminal carbon of the alkane. Thus, exogenous carbon appears to be initially added to an alkane subterminally at the C-2 position such that the original terminal carbon of the alkane becomes a methyl group on the subsequently formed fatty acid. The carbon addition reaction, however, does not appear to be a direct carboxylation of inorganic bicarbonate. A pathway for anaerobic metabolism of alkanes by strain AK-01 is proposed.

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^* Corresponding author. Mailing address: Biotechnology Center for Agriculture and the Environment, Foran Hall, Cook College, Rutgers, The State University of New Jersey, 59 Dudley Rd., New Brunswick, NJ 08901-8520. Phone: (732) 932-8165, ext. 312. Fax: (732) 932-0312. E-mail: lyoung{at}aesop.rutgers.edu.

Present address: Department of Biology, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong SAR, People's Republic of China.

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Applied and Environmental Microbiology, December 1999, p. 5532-5540, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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