A deep-sea sulfate reducing bacterium directs the formation of zero-valent sulfur via sulfide oxidation

Zero-valent sulfur (ZVS) is a critical intermediate in the biogeochemical sulfur cycle. Up to date, sulfur oxidizing bacteria have been demonstrated to dominate the formation of ZVS. In contrast, formation of ZVS mediated by sulfate reducing bacteria (SRB) has been rarely reported. Here, we report for the first time that a typical sulfate reducing bacterium Desulfovibrio marinus CS1 directs the formation of ZVS via sulfide oxidation. In combination with proteomic analysis and protein activity assays, thiosulfate reductase (PhsA) and sulfide: quinone oxidoreductase (SQR) were demonstrated to play key roles in driving ZVS formation. In this process, PhsA catalyzed thiosulfate to form sulfide, which was then oxidized by SQR to form ZVS. Consistently, the expressions of PhsA and SQR were significantly up-regulated in strain CS1 when cultured in the deep-sea cold seep, strongly indicating strain CS1 might form ZVS in its real inhabiting niches. Notably, homologs of phsA and sqr widely distributed in the metagenomes of deep-sea SRB. Given the high abundance of SRB in cold seeps, it is reasonable to propose that SRB might greatly contribute to the formation of ZVS in the deep-sea environments. Our findings add a new aspect to the current understanding of the source of ZVS.

Engineering bacteria for biogenic synthesis of chalcogenide nanomaterials

SummaryMicrobes naturally build nanoscale structures, including structures assembled from inorganic materials. Here we combine the natural capabilities of microbes with engineered genetic control circuits to demonstrate the ability to control biological synthesis of chalcogenide nanomaterials in a heterologous host. We transferred reductase genes from both Shewanella sp. ANA-3 and Salmonella enterica serovar Typhimurium into an heterologous host (Escherichia coli) and examined the mechanisms that regulate the properties of biogenic nanomaterials. Expression of arsenic reductase genes and thiosulfate reductase genes in E. coli resulted in the synthesis of arsenic sulfide nanomaterials. In addition to processing the starting materials via redox enzymes, cellular components also nucleated the formation of arsenic sulfide nanomaterials. The shape of the nanomaterial was influenced by the bacterial culture, with the synthetic E. coli strain producing nanospheres and conditioned media or cultures of wild type Shewanella sp. producing nanofibers. The diameter of these nanofibers also depended on the biological context of synthesis. These results demonstrate the potential for biogenic synthesis of nanomaterials with controlled properties by combining the natural capabilities of wild microbes with the tools from synthetic biology.

Engineering Hydrogen Sulfide Production and Cadmium Removal by Expression of the Thiosulfate Reductase Gene (phsABC) from Salmonella enterica Serovar Typhimurium in Escherichia coli

ABSTRACT The thiosulfate reductase gene (phsABC) fromSalmonella enterica serovar Typhimurium was expressed inEscherichia coli to overproduce hydrogen sulfide from thiosulfate for heavy metal removal (or precipitation). A 5.1-kb DNA fragment containing phsABC was inserted into the pMB1-based, high-copy, isopropyl-β-d-thiogalactopyranoside-inducible expression vector pTrc99A and the RK2-based, medium-copy,m-toluate-inducible expression vector pJB866, resulting in plasmids pSB74 and pSB77. A 3.7-kb DNA fragment, excluding putative promoter and regulatory regions, was inserted into the same vectors, making plasmids pSB103 and pSB107. E. coli DH5α strains harboring the phsABC constructs showed higher thiosulfate reductase activity and produced significantly more sulfide than the control strains under both aerobic and anaerobic conditions. Among the four phsABC constructs, E. coli DH5α (pSB74) produced thiosulfate reductase at the highest level and removed the most cadmium from solution under anaerobic conditions: 98% of all concentrations up to 150 μM and 91% of 200 μM. In contrast, a negative control did not produce any measurable sulfide and removed very little cadmium from solution. Energy-dispersive X-ray spectroscopy revealed that the metal removed from solution precipitated as a complex of cadmium and sulfur, most likely cadmium sulfide.