Singh, R; Dong, H; Liu, D; Zhao, L; Marts, AR; Farquhar, E; Tierney, DL; Almquist, CB; Briggs, BR
Despite the significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70°C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr(6+) concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr(6+) concentrations with the 1, 5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr(6+) solutions by this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr(6+) concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr(6+) to cells at this concentration range. At these higher Cr(6+) concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr(6+) bioreduction rates decreased with increased initial concentrations of Cr(6+) from 13.3 to1.9 µM h(-1). X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr(6+) to insoluble Cr(3+) precipitates, which was confirmed as amorphous chromium hydroxide by X-ray diffraction and selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr(3+). Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr(6+) exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of this study demonstrate the ability of M. thermautotrophicus cells to reduce toxic Cr(6+) to less toxic Cr(3+) and its potential application in metal bioremediation, especially at high temperature subsurface radioactive waste disposal sites, where the temperature may reach ∼70°C.