In the present study, hexavalent chromium (Cr(VI)) reduction potential of chromium reductase associated with the cell-free extracts (CFE) of Arthrobacter rhombi-RE species was evaluated. Arthrobacter rhombi-RE, an efficient Cr(VI) reducing bacterium, was enriched and isolated from a chromium-contaminated site. Chromium reductase activity of Arthrobacter rhombi-RE strain was associated with the cell-free extract and the contribution of extracellular enzymes to Cr(VI) reduction was negligible. NADH enhanced the chromium reductase activity. The enzyme activity was optimal at a pH of 5.5 and a temperature of 30 degrees C. Among the ten electron donors screened, sodium pyruvate was the most effective one followed by NADH and propionic acid. Michaelis-Menten constant, K-m, and maximum reaction rate, V-max, obtained from the Lineweaver-Burk plot were 48 mu M and 4.09 nM/mg protein/min, respectively, in presence of NADH as electron donor and 170.5 mu M and 4.29 nM/mg protein/min, respectively, in presence of sodium pyruvate as electron donor. Ca2+ enhanced the enzyme activity while Hg2+, Cd2+, Ba2+, and Zn2+ inhibited the enzyme activity. Among the various immobilization matrices screened, calcium alginate beads seemed to be the most effective one. Though immobilized enzyme system was able to reduce Cr(VI), the performance was not very encouraging in continuous mode of operation.