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Applied and Environmental Microbiology, January 2003, p. 191-198, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.191-198.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Carbon and Hydrogen Isotopic Fractionation during Anaerobic Biodegradation of Benzene

Silvia A. Mancini,1 Ania C. Ulrich,2 Georges Lacrampe-Couloume,1 Brent Sleep,3 Elizabeth A. Edwards,2 and Barbara Sherwood Lollar1*

Stable Isotope Laboratory, Department of Geology,1 Department of Chemical Engineering and Applied Chemistry,2 Department of Civil Engineering, University of Toronto, Toronto, Canada3

Received 5 June 2002/ Accepted 2 September 2002

Compound-specific isotope analysis has the potential to distinguish physical from biological attenuation processes in the subsurface. In this study, carbon and hydrogen isotopic fractionation effects during biodegradation of benzene under anaerobic conditions with different terminal-electron-accepting processes are reported for the first time. Different enrichment factors ({varepsilon}) for carbon (range of -1.9 to -3.6{per thousand}) and hydrogen (range of -29 to -79{per thousand}) fractionation were observed during biodegradation of benzene under nitrate-reducing, sulfate-reducing, and methanogenic conditions. These differences are not related to differences in initial biomass or in rates of biodegradation. Carbon isotopic enrichment factors for anaerobic benzene biodegradation in this study are comparable to those previously published for aerobic benzene biodegradation. In contrast, hydrogen enrichment factors determined for anaerobic benzene biodegradation are significantly larger than those previously published for benzene biodegradation under aerobic conditions. A fundamental difference in the previously proposed initial step of aerobic versus proposed anaerobic biodegradation pathways may account for these differences in hydrogen isotopic fractionation. Potentially, C-H bond breakage in the initial step of the anaerobic benzene biodegradation pathway may account for the large fractionation observed compared to that in aerobic benzene biodegradation. Despite some differences in reported enrichment factors between cultures with different terminal-electron-accepting processes, carbon and hydrogen isotope analysis has the potential to provide direct evidence of anaerobic biodegradation of benzene in the field.

* Corresponding author. Mailing address: Stable Isotope Laboratory, Department of Geology, University of Toronto, 22 Russell St., Toronto, Ontario, Canada M5S 3B1. Phone: (416) 978-0770. Fax: (416) 978-3938. E-mail: bsl{at}quartz.geology.utoronto.ca.

Applied and Environmental Microbiology, January 2003, p. 191-198, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.191-198.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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