The purpose of this thesis was to experimentally investigate the influence of increasing sulfide concentrations on the 34S isotope enrichment in sulfate during dissimilatory sulfate reduction (DSR). Two independent batch culture experiments with different maximum sulfide concentrations of up to 20 mM in the first and up to 40 mM in the second experiment were conducted using the marine sulfate reducer Desulfobacter latus. A comparison of the results from both experiments revealed a distinct offset towards more positive δ34S(SO42-) values in the 'high-sulfide' experiment, compared to the 'low-sulfide' experiment. While a Rayleigh type fractionation model was able to match the slopes - i.e., enrichment factors - of both experiments, it failed to reproduce the proper y-axis intercept in the 'high-sulfide' experiment. I therefore propose a new fractionation model that allows for a backward flow of ambient H2S into the bacterial cell and a subsequent enzymatically mediated oxidation of H2S to sulfate. The new backward flow increases with elevated H2S concentrations and is described as a first order rate constant. Unlike a Rayleigh type fractionation model, my model explains the slope and y-intercept of both experiments with a single parameter set. The new model with H2S-reflux further suggests that it can be used to determine growth kinetic parameters like the half-saturation constant through δ34S measurements. These findings support the hypothesis of microbially mediated, bi-directional S-fluxes between oxidized and reduced sulfur species. Because the S-transport during DSR appears to be bi-directional, great care must be taken when evaluating culture experiments with a Rayleigh type fractionation model, owing to the fact that an evident S-backward flow violates the prerequisites for applying the Rayleigh model. A variable S-backward flow results in variable enrichment factors which increased from -11 (no H2S) to ≈-17 ‰ (40 mM of H2S) in my experiments. I show for the first time the significance of a bi-directional H2S transport across the cell membrane during DSR and its consequences for the 34S-isotope fractionation in sulfate.

The purpose of this thesis was to experimentally investigate the influence of increasing sulfide concentrations on the 34S isotope enrichment in sulfate during dissimilatory sulfate reduction (DSR). Two independent batch culture experiments with different maximum sulfide concentrations of up to 20 mM in the first and up to 40 mM in the second experiment were conducted using the marine sulfate reducer Desulfobacter latus. A comparison of the results from both experiments revealed a distinct offset towards more positive δ34S(SO42-) values in the 'high-sulfide' experiment, compared to the 'low-sulfide' experiment. While a Rayleigh type fractionation model was able to match the slopes - i.e., enrichment factors - of both experiments, it failed to reproduce the proper y-axis intercept in the 'high-sulfide' experiment. I therefore propose a new fractionation model that allows for a backward flow of ambient H2S into the bacterial cell and a subsequent enzymatically mediated oxidation of H2S to sulfate. The new backward flow increases with elevated H2S concentrations and is described as a first order rate constant. Unlike a Rayleigh type fractionation model, my model explains the slope and y-intercept of both experiments with a single parameter set. The new model with H2S-reflux further suggests that it can be used to determine growth kinetic parameters like the half-saturation constant through δ34S measurements. These findings support the hypothesis of microbially mediated, bi-directional S-fluxes between oxidized and reduced sulfur species. Because the S-transport during DSR appears to be bi-directional, great care must be taken when evaluating culture experiments with a Rayleigh type fractionation model, owing to the fact that an evident S-backward flow violates the prerequisites for applying the Rayleigh model. A variable S-backward flow results in variable enrichment factors which increased from -11 (no H2S) to ≈-17 ‰ (40 mM of H2S) in my experiments. I show for the first time the significance of a bi-directional H2S transport across the cell membrane during DSR and its consequences for the 34S-isotope fractionation in sulfate.