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Applied and Environmental Microbiology, May 2004, p. 2935-2940, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2935-2940.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Stable Isotope Fractionation Caused by Glycyl Radical Enzymes during Bacterial Degradation of Aromatic Compounds

Barbara Morasch,1,2 Hans H. Richnow,3 Andrea Vieth,3 Bernhard Schink,1 and Rainer U. Meckenstock2,4*

Fakultät für Biologie, Universität Konstanz, D-78457 Konstanz,1 Zentrum für Angewandte Geowissenschaften, Eberhard-Karls-Universität Tübingen, D-72076 Tübingen,2 UFZ-Umweltforschungszentrum Leipzig-Halle GmbH, D-04318 Leipzig,3 Institut für Grundwasserökologie, GSF-Forschungszentrum für Umwelt und Gesundheit, D-85764 Neuherberg, Germany4

Received 4 August 2003/ Accepted 22 January 2004

Stable isotope fractionation was studied during the degradation of m-xylene, o-xylene, m-cresol, and p-cresol with two pure cultures of sulfate-reducing bacteria. Degradation of all four compounds is initiated by a fumarate addition reaction by a glycyl radical enzyme, analogous to the well-studied benzylsuccinate synthase reaction in toluene degradation. The extent of stable carbon isotope fractionation caused by these radical-type reactions was between enrichment factors ({varepsilon}) of –1.5 and –3.9, which is in the same order of magnitude as data provided before for anaerobic toluene degradation. Based on our results, an analysis of isotope fractionation should be applicable for the evaluation of in situ bioremediation of all contaminants degraded by glycyl radical enzyme mechanisms that are smaller than 14 carbon atoms. In order to compare carbon isotope fractionations upon the degradation of various substrates whose numbers of carbon atoms differ, intrinsic {varepsilon} ({varepsilon}intrinsic) were calculated. A comparison of {varepsilon}intrinsic at the single carbon atoms of the molecule where the benzylsuccinate synthase reaction took place with compound-specific {varepsilon} elucidated that both varied on average to the same extent. Despite variations during the degradation of different substrates, the range of {varepsilon} found for glycyl radical reactions was reasonably narrow to propose that rough estimates of biodegradation in situ might be given by using an average {varepsilon} if no fractionation factor is available for single compounds.

* Corresponding author. Mailing address: Institut für Grundwasserökologie, GSF-Gesellschaft für Gesundheit und Umwelt, Ingolstädter Landstrasse 1, D-85764 Neuherberg, Germany. Phone: 49-(0)89-31872560. Fax: 49-(0)89-3187-3361. E-mail: rainer.meckenstock{at}gsf.de.

Applied and Environmental Microbiology, May 2004, p. 2935-2940, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2935-2940.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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