Purification and biochemical characterization of a novel ene- reductase from Kazachstania exigua HSC6 for dihydro-β-ionone from β-ionone

Purpose We purified and characterized a novel ene-reductase (KaDBR1) from Kazachstania exigua HSC6 for the synthesis of dihydro- β -ionone from β -ionone. Methods KaDBR1 was purified to homogeneity by ammonium sulfate precipitation and phenyl-Sepharose Fast Flow and Q-Sepharose chromatography. The purified enzyme was characterized by measuring the amount of dihydro- β -ionone from β -ionone with LC–MS analysis method. Results The molecular mass of KaDBR1 was estimated to be 45 kDa by SDS-PAGE. The purified KaDBR1 enzyme had optimal activity at 60 °C and pH 6.0. The addition of 5 mM Mg 2+ , Ca 2+ , Al 3+ , Na + , and dithiothreitol increased the activity of KaDBR1 by 25%, 18%, 34%, 20%, and 23%, respectively. KaDBR1 favored NADH over NADPH as a cofactor, and its catalytic efficiency ( kcat/Km ) toward β-ionone using NADH was 8.1-fold greater than when using NADPH. Conclusion Owing to its unique properties, KaDBR1 is a potential candidate for the enzymatic biotransformation of β -ionone to dihydro- β -ionone in biotechnology applications.