Purification and Characterization of 3- Deoxyglucosone Metabolizing Enzyme From Bacillus sp-2.

A NADPH dependent 3 DG metabolizing enzyme was isolated and purified to electrophoretic homogeneity from Bacillus sp 2 by combined consecutive treatment consisting of ammonium sulfate fractionation, Q Sepharose FF, Sephadex G 100(Ⅰ), Hydroxyapatite and Sephadex G 100(Ⅱ) column chromatographies. The specific activity of purified 3 DG metabolizing enzyme was 63 75 U/mg . The molecular weight of the enzyme was about 32 ku. 2 Oxoaldehyde compounds were found to be specifically good substrate for this reductase. The optimum pH of the enzyme activity was 6 2. The enzyme was stable in the pH range from 5 to 8 and in the temperature range from 25℃ to 30℃. The K m for 3 DG was 2 3 mmol/L. Suitable amount of EDTA、β mercaptoethanol and dithiothreitol enhanced the enzyme activity, but the activity of the enzyme was partially lost by adding iodoacetic acid or N ethylmaleimide.