Evaluation of in vitro Reduction of Hexavalent Chromium by Cell-Free Extract of Arthrobacter sp. SUK 1201

Abstract

Aims: This study aims at to evaluate the hexavalent chromium [Cr(VI)] reduction potential of crude cell-free extracts of chromium resistant and reducing bacterium Arthrobacter sp. SUK 1201 and determination of optimum conditions for Cr(VI) reduction for possible bioremediation of Cr pollutants. Place and Duration of Study: Chromium reduction studies with Arthrobacter sp. SUK 1201, was undertaken in the Microbiology Laboratory, Department of Botany, University of Calcutta, Kolkata during 2010-2012. Methodology: Cell-free extract was prepared from freshly grown cell mass of Arthrobacter sp. SUK 1201 following the standard procedure. Cell mass suspended in Tris-HCl was sonicated (120 KHz for 30 min), centrifuged (12,000×g at 4ºC for 10 min) and the supernatant (S12) was used as the cell- free extract (CFE). Chromate reductase activity of the CFE was assayed colorimetrically using 1, 5-diphenylcarbazide as the complexing reagent. Results: Chromate reductase activity of CFE of Arthrobacter sp. SUK 1201 was constitutive in nature and reduced Cr(VI) with decreasing efficiency as the concentration of Cr(VI) was increased. Its Km and Vmax were 263.45 μM Cr(VI) and 17.5 U mg-1 protein respectively. Reduction of Cr(VI) was optimal at pH 7 and 32ºC but was extremely thermolabile. NADH was the most suitable electron donor, and the chromate reduction was enhanced by Cu(II) and Fe(III), but inhibited by Hg(II). Among the different inhibitors tested, 2, 4-dinitrophenol (DNP) restored nearly 96.4% reductase activity, while carbonyl cyanide-m-chloro phenyl hydrazone (CCCP) was most inhibitory to the process. Conclusion: It has been established that the Cr(VI) reduction potential of the cell-free extract of Arthrobacter sp. SUK 1201 is promising and could be exploited in the bioremediation of toxic hexavalent chromium.