Effect of electron donors on the fractionation of sulfur isotopes by a marine Desulfovibrio sp

► S-isotope effects in pure culture of a sulfate reducing bacterium depend on electron donor. ► Limited availability of electron donor increases the isotope effect. ► Large isotope effects during growth in refractory organic compounds. ► S-isotope effects increase with declining csSRR. ► Expanded range of multiple S-isotope effects attributable to microbial sulfate reduction. Sulfur isotope effects produced by microbial dissimilatory sulfate reduction are used to reconstruct the coupled cycling of carbon and sulfur through geologic time, to constrain the evolution of sulfur-based metabolisms, and to track the oxygenation of Earth’s surface. In this study, we investigate how the coupling of carbon and sulfur metabolisms in batch and continuous cultures of a recently isolated marine sulfate reducing bacterium DMSS-1, a Desulfovibrio sp ., influences the fractionation of sulfur isotopes. DMSS-1 grown in batch culture on seven different electron donors (ethanol, glycerol, fructose, glucose, lactate, malate and pyruvate) fractionates 34S/ 32S ratio from 6‰ to 44‰, demonstrating that the fractionations by an actively growing culture of a single incomplete oxidizing sulfate reducing microbe can span almost the entire range of previously reported values in defined cultures. The magnitude of isotope effect correlates well with cell specific sulfate reduction rates (from 0.7 to 26.1 fmol/cell/day). DMSS-1 grown on lactate in continuous culture produces a larger isotope effect (21–37‰) than the lactate-grown batch culture (6‰), indicating that the isotope effect also depends on the supply rate of the electron donor and microbial growth rate. The largest isotope effect in continuous culture is accompanied by measurable changes in cell length and cellular yield that suggest starvation. The use of multiple sulfur isotopes in the model of metabolic fluxes of sulfur shows that the loss of sulfate from the cell and the intracellular reoxidation of reduced sulfur species contribute to the increase in isotope effects in a correlated manner. Isotope fractionations produced during sulfate reduction in the pure culture of DMSS-1 expand the previously reported range of triple sulfur isotope effects ( 32S, 33S, and 34S) by marine sulfate reducing bacteria, implying that microbial sulfur disproportionation may have a smaller 33S isotopic fingerprint than previously thought.