Indiana University-Purdue University Indianapolis (IUPUI) / The reaction between elemental sulfur and sulfide is a lynchpin in the biotic and abiotic cycling of sulfur. This dissertation is focused on the reactivity of elemental sulfur nanoparticles (S8weimarn, S8raffo) among other forms of elemental sulfur (S8aq, S8aq-surfactant, α-S8), and how the variation of their surface area, character and coatings reflect on the analytical, physical-chemical and geochemical processes involving sulfur cycling. A comprehensive electrochemical investigation utilizing mercury-surface electrodes showed that elemental sulfur compounds are represented by three main voltammetric signals, corresponding to potentials at -1.2V, -0.8V, and -0.6V in the absence of organics at circumneutral pH. Dissolved S8aq-surfactant signals can be found from -0.3V up to -1.0V, depending on the surfactant in the system. Variations in current response resulted from differences in electron transfer efficiency among the forms of S8, due to their molecular structural variability. Based on this observation a new reaction pathway between S8 and Hg-surface electrodes is proposed, involving an amalgam-forming intermediate step. The kinetics of the nucleophilic dissolution of S8nano by sulfide, forming polysulfides, were investigated under varying surface area, surface character and presence or absence of surfactant coatings on S8nano. Hydrophobic S8weimarn and hydrophilic S8raffo show kinetic rate laws of 𝑟𝑆8𝑤𝑒𝑖𝑚𝑎𝑟𝑛 = 10−11.33 (𝑒 −700.65 𝑅𝑇 ) (Molar(S8)/second/dm-1) and𝑟𝑆8𝑟𝑎𝑓𝑓𝑜 = 10−4.11 𝑖−0.35 (𝑒 −615.77 𝑅𝑇 ) (Molar(S8)/second), respectively. The presence of surfactant molecules can influence the reaction pathways by dissolving S8nano and releasing S8aqsurfactant, evolving the rate-limiting step as a function of the degree of the solubilization of S8nano. The reaction rate of S8biological can be compared with those of S8raffo and S8weimarn in circumneutral pH values and T=50oC, making the forms of S8nano successful abiotic analogue models of microbially produced S8biological. Field observations and geochemical kinetic modeling in the geothermal features of Yellowstone indicate that the nucleophilic dissolution reaction appears to be a key abiotic pathway for the cycling of sulfur species and the enhancement of elemental sulfur bioavailability. Furthermore, in situ and ex situ voltammetry in the same geothermal waters disclosed chaotic variability in chemical gradients of sulfide (observed over small temporal and spatial scales) which can be considered as an ecological stressor capable of influencing single cell physiology and microbial community adaptation.

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We present research investigating how, with in depth knowledge of the community, microbial communities may be harnessed for bioremediation of hazardous water contaminants. We focused on the bacterial reduction of perchlorate, a common water contaminant. For this we studied the structure and capabilities of a novel sulfur-utilizing, perchlorate-reducing bacterial (SUPeRB) consortium. Initially, we characterized the minimal consortium that retained functional capabilities, using 16S rRNA and functional gene analysis. A diverse functional consortium dominated by Beta-Proteobacteria of the family Rhodocyclaceae and sulfur-oxidizing Epsilon-Proteobacteria was found. We also examined the optimal growth conditions under which perchlorate degradation occurred and uncovered the upper limits of this function. Bacterial isolates were screened for function and the presence of functional genes. We expanded to bioreactor studies at bench- and pilot-scale, and first used a perchlorate-reducing, bench-scale bioreactor to probe the stability of the microbial ecosystem. During stable reactor function, a core consortium of Beta- and Epsilon-Proteobacteria reduced perchlorate and the co-contaminant nitrate. A disturbance of the vi consortium led to a failure in function and to higher system diversity. This suggests that the SUPeRB consortium was not metabolically flexible and high population diversity was necessary for a return to stable function. In a pilot-scale bioreactor we determined that the SUPeRB consortium could stably degrade low levels of perchlorate to below the EPA maximum recommended limit. Field conditions, such as temperature extremes and intermittent perchlorate feed, did not negatively impact overall function. When all reactor consortia were compared we observed that the volume of the reactor and the initial inoculum were not as important to stable reactor function as the acclimatization of the consortium to the system and maintenance of favorable conditions within the reactor. In summary we found that the SUPeRB consortium successfully degraded perchlorate in multiple systems. The study of this novel consortium expands our knowledge of the metabolic capabilities of perchlorate-reducing bacteria and suggests potential evolutionary pathways for perchlorate-reduction by microorganisms. The SUPeRB consortium may be used to establish bioremediation systems for perchlorate and other environmental contaminants.

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The cytosolic iron sulfur cluster assembly (CIA) pathway is responsible for the maturation of >40 cytosolic and nuclear iron sulfur (FeS) proteins critical for fundamental processes such as DNA replication, transcription, and translation. The final stages of the pathway require the CIA targeting complex, which is composed of Cia1, Cia2, and Met18. This large multiprotein complex is proposed to recognize apo-enzyme substrates and insert their FeS clusters. However, it is unclear how these substrates are identified and how the CIA targeting complex mediates cofactor insertion. In this thesis, I mapped the protein-protein interaction sites critical for formation of the CIA targeting complex and discovered the first peptide motif that is both necessary and sufficient for recognition of a subset of FeS proteins by the CIA system. Cia1’s seventh beta-propeller blade was found to bind to Cia2, while Cia2’s fifth conserved region mainly interacts with Cia1, via an in vitro affinity co-purification assay. A quantitative MicroScale Thermophoresis assay supported these findings, in addition this approach affirmed that Cia2’s N-terminal intrinsically disordered domain and hyperreactive cysteine are dispensable for CIA targeting complex assembly. In collaboration with the Drennan Lab at MIT, Met18 was discovered to form a hexamer via cryo-EM. Met18 is proposed to arrange into a hexamer before its CIA-related function. Hexamer formation and Cia2 binding depend on Met18’s C-terminus, whereas Leu1 recognition relies on Met18’s N-terminus. A C-terminal W motif was demonstrated as both necessary and sufficient for identification of a subset of FeS proteins by the CIA targeting complex. A bioinformatics analysis revealed roughly 20% of CIA client proteins, including substrates, factors, and adaptors, terminate in a conserved [LTQ]-[DE]-[W]-COO- motif. CIA recognition depends on the C-terminal aromatic side chain and the carboxy terminus. This tripeptide motif is also sufficient for identification by the CIA system when attached to SUMO. Moreover, a series of competition experiments showed that the CIA targeting complex contains distinct, non-overlapping binding sites for client proteins where Cia1 serves as the docking site for the C-terminal W motif. Altogether, the first recognition motif is defined for one in five of CIA client proteins. / 2024-11-03T00:00:00Z

<p> S^32/S^34 ratios have been determined between some coexisting sulfides from the Heath Steele B-1 orebody and five other stratiform deposits in the Bathurst-Newcastle area, New Brunswick, and in crushed ore samples and pyrites from three across deposit profiles in the B-1 orebody. Zn, Cu, Pb and Ag abundances have been established across the orebody. Fractionations of S^32/S^34 ratios between coexisting sulfides have been determined in heating experiments at temperatures of 350 to 500°C and isotope effects measured in aqueous precipitation and exchange experiments at 25°C. S^32/S^34 fractionations between given coexisting sulfides from all the sulfide deposits are found to show only small variations. Isotope exchange is generally rapid in dry sulfide systems. Metal sulfides in aqueous solutions are slightly enriched in S^32 relative to the H2S with which they precipitate or communicate. It is concluded that the sulfide deposits have been regionally metamorphosed. The origins that have been proposed for the deposits so far are discussed and a model is suggested to explain the sulfur isotope and base metal abundances in the B-1 orebody.</p> / Thesis / Doctor of Philosophy (PhD)