Biomolecular and Isotopic Signatures Related to Cr(VI) Reduction by a Sulfate-Reducing Bacterium Isolated from the Hanford 100H Aquifer

Chromium contamination of groundwater is widespread within the Dept. of Energy (DOE) complex. At DOE's Hanford 100H area, we have conducted Cr bioremediation (in situ reductive immobilization) studies involving injection of a lactate-containing polymer, and have observed sequential use of the dissolved electron acceptors present in groundwater (namely, oxygen, nitrate, and sulfate). Sulfate-reducing bacteria are of particular interest for chromate reduction because they can reduce Cr(VI) enzymatically (e.g., using cytochrome c₃ or thioredoxin reductase) and abiotically with hydrogen sulfide, the end product of their respiration. In this poster, we use studies of a sulfate-reducing bacterium isolated from the Hanford 100H aquifer, Desulfovibrio vulgaris strain RCH1, to explore (a) isotopic signatures that might allow us to distinguish between enzymatic and sulfide-mediated Cr(VI) reduction and (b) biomolecular signatures (gene or transcript copy number of diagnostic genes) that might be used as proxies of in situ metabolic rates. In order to differentiate between the mechanisms of Cr reduction by sulfate reducers, we analyzed the isotopic fractionation during Cr(VI) reduction by strain RCH1. Cell suspension studies of strain RCH1 demonstrated that Cr(VI) reduction could occur in the presence of lactate (electron donor) alone or with both lactate and sulfate. Cr(VI) reduction in the presence of lactate and sulfate was 25-30% more rapid than enzymatic Cr reduction when only lactate was added, suggesting that biogenic hydrogen sulfide increases the specific rate of Cr(VI) reduction beyond purely enzymatic activity. Cr isotopic measurements showed different fractionation behavior for the lactate-only and lactate+sulfate systems, with fractionation (epsilon) values of 2.3 and 1.66 per mil, respectively. In order to determine whether gene or transcript copy number for diagnostic sulfate and chromate reduction genes could serve as proxies to estimate in situ metabolic rates, chemostat studies were conducted with strain RCH1. Genes assayed by qPCR and RT-qPCR included aprB (APS reductase, beta subunit), dsrA (dissimilatory sulfite reductase, alpha subunit), cyc3 (cytochrome c₃), and trxB (thioredoxin reductase). Strong linear relationships were observed between sulfate reduction rates and the gene and transcript copy numbers of all of the targeted genes. These results suggest that use of gene and transcript copy numbers in groundwater samples may be a useful approach for estimating in situ metabolic rates of sulfate-reducing bacteria during Cr bioremediation.