[Purification and characterization of 2-carbonyl reductase from marine bacteria Bacillus sp]

ANADPH-dependent 2-Oxoaldehyde reductase was isolated and purified from a marine bacteria Bacillus sp. The purification procedure involved ammonium sulfate fractionation and Q Sepharose FF, Hydroxyapatite, Sephadex G-100 column chromatographies. The specific activity of the purified enzyme was increased by 141.1 folds over crude extract and the recovery yield was 11.4%. 2-Oxoaldehyde compounds were found to be speciall good substrates. The optimum pH of the enzyme activity was 6.2-6.6. The Km coefficient for 3-deoxyglucosone was 2.5 mmol/L. The molecular weight of the enzyme was estimated to be 33 kD The enzyme activity is stable below 30 degrees C and pH 5.0-8.0. EDTA, beta-mercaptoethanol and dithiothreitol enhanced the enzyme activity. On the other hand, the enzyme activity was partially lost by idoacetic acid or N-ethylmaleimide.