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-beta-ionone from beta-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-beta-ionone from beta-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 ?degrees C and pH 6.0. The addition of 5 mM Mg2+, Ca2+, Al3+, 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 beta-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 beta-ionone to dihydro-beta-ionone in biotechnology applications.

Automated summary

A novel ene-reductase (KaDBR1) was purified and characterized from Kazachstania exigua HSC6 for the synthesis of dihydro-beta-ionone from beta-ionone. The purified enzyme had optimal activity at 60 degrees C and pH 6.0, and it favored NADH over NADPH as a cofactor. The catalytic efficiency of KaDBR1 using NADH was 8.1-fold greater than when using NADPH. Owing to its unique properties, KaDBR1 is a potential candidate for the enzymatic biotransformation of beta-ionone to dihydro-beta-ionone in biotechnology applications.