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Applied and Environmental Microbiology, September 2009, p. 5621-5630, Vol. 75, No. 17
0099-2240/09/$08.00+0 doi:10.1128/AEM.02948-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Department of Geosciences, Guyot Hall, Princeton University, Princeton, New Jersey 08544,1 Molecular Biology Department, Moffett Lab, Princeton University, Princeton, New Jersey,2 Department of Geology, Indiana University, Bloomington, Indiana 47405-1405,3 Department of Earth Sciences, University of Western Ontario, London, Ontario, Canada N6A 5B7,4 Center for Biomarker Analysis, University of Tennessee, Knoxville, Tennessee,5 School of Geology and Geophysics, University of Oklahoma, Norman, Oklahoma6
Received 27 December 2008/ Accepted 19 June 2009
Values of
34S (
, where
34SHS and
indicate the differences in the isotopic compositions of the HS– and SO42– in the eluent, respectively) for many modern marine sediments are in the range of –55 to –75
, much greater than the –2 to –46
34S (kinetic isotope enrichment) values commonly observed for microbial sulfate reduction in laboratory batch culture and chemostat experiments. It has been proposed that at extremely low sulfate reduction rates under hypersulfidic conditions with a nonlimited supply of sulfate, isotopic enrichment in laboratory culture experiments should increase to the levels recorded in nature. We examined the effect of extremely low sulfate reduction rates and electron donor limitation on S isotope fractionation by culturing a thermophilic, sulfate-reducing bacterium, Desulfotomaculum putei, in a biomass-recycling culture vessel, or "retentostat." The cell-specific rate of sulfate reduction and the specific growth rate decreased progressively from the exponential phase to the maintenance phase, yielding average maintenance coefficients of 10–16 to 10–18 mol of SO4 cell–1 h–1 toward the end of the experiments. Overall S mass and isotopic balance were conserved during the experiment. The differences in the
34S values of the sulfate and sulfide eluting from the retentostat were significantly larger, attaining a maximum
34S of –20.9
, than the –9.7
observed during the batch culture experiment, but differences did not attain the values observed in marine sediments.
Published ahead of print on 26 June 2009.
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