A soluble Cr(VI) reductase was purified from the Cr(VI) reducing strain Escherichia coli ATCC33456 by ammonium sulfate fractionation, and chromatographies on Q-Sepharose FF, Cibacron blue 3GA dye affinity, Mono-Q 5/5, and Superdex 200 HR 10/30 columns. The estimated molecular mass of the purified enzyme was 27 kDa on SDS-polyacrylamide gel electrophoresis and 54 kDa on gel filtration, thus indicating a dimeric structure. The isoelectric point of the enzyme was pH 4.85. The optimum reaction pH and storage pH were both 7.0, the optimum reaction temperature was $37^{circ}C$, and the storage temperature was $4^{circ}C$. NADH and NADPH both served as electron donors for the reductase, with $V_{max}$ of 68.3 ${mu}M$ Cr(VI)/min/mg protein and Km of 7.6 $mu$M using HADH, and Vmax of 42.3 ${mu}M$ Cr(VI)/min/mg protein and Km of 14.6 $muM$ using NADPH. When 1 mM EDTA was added, the Cr(VI) reducing activity increased 4-fold.