The role of intermediate sulfur species (ISS) for isotopic fractionation processes during abiotic and chemolithoautotrophic sulfide oxidation in a natural environment

DFG PL 302/10-1

Sulfur isotope fractionation (34S/32S) has been used since the late 1940s to study the chemical and biological sulfur cycle. While large isotope fractionations during bacterial sulfate reduction were used successfully to interpret, e.g., accumulation of sulfate in ancient oceans or the evolution of early life, much less is known about fractionation during sulfide oxidation. The fractionation between the two end-members sulfide and sulfate is commonly much smaller and inconsistencies exist whether substrate or product are enriched. These inconsistencies are explained by a lack of knowledge on oxidation pathways and rates as well as intermediate sulfur species, such as elemental sulfur, polysulfides, thiosulfate, sulfite, or metalloid-sulfide complexes (e.g. thioarsenates), potentially acting as 34S sinks.

In the proposed project, we will develop a method for sulfur species-selective isotope analysis based on separation by preparative chromatography. Separation of Sn2- and S0 will be achieved after derivatization with methyl triflate on a C18 column, separation of the other sulfur species in an alkaline eluent on an AS16 column. Sulfur in the collected fractions will be extracted directly with activated copper chips (Sn2-, S0), or precipitated as ZnS (S2-) or BaSO4 and analyzed by routine methods as SO2. Results of this species-selective approach will be compared to those from previous techniques of end-member pool determinations and sequential precipitations.

The method will be applied to sulfide oxidation profiles at neutral to alkaline hot springs at Yellowstone National Park, USA, where we detected intermediate sulfur species as important species. Determining 34S/32S only in sulfide and sulfate, our previous study has shown different fractionation patterns for two hot spring drainages with sulfide oxidation profiles that seemed similar from a geochemical perspective. The reasons for the different isotopic trends are unclear. In the present project, we will differentiate species-selective abiotic versus biotic fractionation using on-site incubation experiments with the chemolithotrophic sulfur-oxidizing bacteria Thermocrinis ruber as model organism. For selected samples, we will test whether 33S and 36S further elucidate species-selective sulfide oxidation patterns. We expect that lower source sulfide concentrations increase elemental sulfur disproportionation, thus increase redox cycling and isotope fractionation. We also expect that the larger the concentration of intermediate sulfur species, including thioarsenates, the larger the isotope fractionation. Following fractionation in species-selective pools, we will be able to clarify previously reported inconsistencies of 34S enrichment in substrate or product, elucidate sulfide oxidation pathways and rates, and reveal details about sulfur metabolism. Our new methodology and field-based data will be a basis for more consistent studies on sulfide oxidation in the future.