Physiology of Multiple Sulfur Isotope Fractionation during Microbial Sulfate Reduction

Microbial sulfate reduction (MSR) utilizes sulfate as an electron acceptor and produces sulfide that is depleted in heavy isotopes of sulfur relative to starting sulfate. The fractionation of S-isotopes is commonly used to trace the biogeochemical cycling of sulfur in nature, but a mechanistic understanding of factors that control the range of isotope fractionation is still lacking. This thesis investigates links between the physiology of sulfate reducing bacteria in pure cultures and multiple sulfur isotope ("S, "S, 34S, and 36S) fractionation during MSR in batch and continuous culture experiments. Experiments address the influence of nutrient and electron donor conditions, including organic carbon, nitrogen, and iron, in cultures of a newly isolated marine sulfate reducing bacterium (DMSS-1). An actively growing culture of DMSS-1 produced sulfide depleted in 3"S by 6 to 66%o, depending on the availability and chemistry of organic electron donors. The magnitude of isotope effect correlated well with the cell specific sulfate reduction rate (csSRR), and the largest isotope effects occurred when cultures grew slowly on glucose, a recalcitrant organic substrate. These findings bridge the long-standing discrepancy between the upper limit for S-isotope effect in laboratory cultures and the corresponding observations in nature and indicate that the large (>46 %o) fractionation of S-isotopes does not unambiguously record the oxidative sulfurrecycling. When the availability of iron was limited, the increase in S-isotope fractionation was accompanied by a decrease in the cytochrome c content as well as csSRR. In contrast, growth in nitrogenlimited cultures increased both csSRR and S-isotope fractionation. The influence of individual enzymes and electron carriers involved in sulfate respiration on the fractionation of S-isotopes was also investigated in cultures of mutant strains of Desulfovibrio vulgaris Hildenborough. The mutant lacking Type I tetraheme cytochrome c3 fractionated 34S/S ratio 50% greater relative to the wild type. The increasing S-isotope fractionation accompanied the evolution of H2 in the headspace and the decreasing csSRR. These results further demonstrate that the flow of electrons to terminal reductases imparts the primary control on the magnitude of the fractionation of S-isotopes, suggested by culture experiments using DMSS-1.