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Applied and Environmental Microbiology, March 2009, p. 1339-1344, Vol. 75, No. 5
0099-2240/09/$08.00+0     doi:10.1128/AEM.02491-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Anaerobic Biodegradation of n-Hexadecane by a Nitrate-Reducing Consortium{triangledown} ,{dagger}

Amy V. Callaghan,1,{ddagger} Meghan Tierney,1 Craig D. Phelps,1,2 and L. Y. Young1,2*

Biotechnology Center for Agriculture and the Environment, Rutgers University, School of Environmental and Biological Sciences, New Brunswick, New Jersey 08901-8521,1 Department of Environmental Sciences, Rutgers University, School of Environmental and Biological Sciences, New Brunswick, New Jersey 08901-85212

Received 30 October 2008/ Accepted 22 December 2008

Nitrate-reducing enrichments, amended with n-hexadecane, were established with petroleum-contaminated sediment from Onondaga Lake. Cultures were serially diluted to yield a sediment-free consortium. Clone libraries and denaturing gradient gel electrophoresis analysis of 16S rRNA gene community PCR products indicated the presence of uncultured alpha- and betaproteobacteria similar to those detected in contaminated, denitrifying environments. Cultures were incubated with H34-hexadecane, fully deuterated hexadecane (d34-hexadecane), or H34-hexadecane and NaH13CO3. Gas chromatography-mass spectrometry analysis of silylated metabolites resulted in the identification of [H29]pentadecanoic acid, [H25]tridecanoic acid, [1-13C]pentadecanoic acid, [3-13C]heptadecanoic acid, [3-13C]10-methylheptadecanoic acid, and d27-pentadecanoic, d25-, and d24-tridecanoic acids. The identification of these metabolites suggests a carbon addition at the C-3 position of hexadecane, with subsequent β-oxidation and transformation reactions (chain elongation and C-10 methylation) that predominantly produce fatty acids with odd numbers of carbons. Mineralization of [1-14C]hexadecane was demonstrated based on the recovery of 14CO2 in active cultures.

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

{triangledown} Published ahead of print on 29 December 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} Present address: Department of Botany and Microbiology, University of Oklahoma, Norman, OK 73019.

Applied and Environmental Microbiology, March 2009, p. 1339-1344, Vol. 75, No. 5
0099-2240/09/$08.00+0     doi:10.1128/AEM.02491-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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