Coenzyme Binding Site Analysis of an Isopropanol Dehydrogenase with Wide Substrate Spectrum and Excellent Organic Solvent Tolerance

• Published in: Applied biochemistry and biotechnology Vol. 190; no. 1; pp. 18 - 29
• Main Authors: Jiang, Wei, Fang, Bai-Shan
• Format: Journal Article
• Language: English
• Published: New York Springer US 2020; Springer Nature B.V
• Subjects: Acetone; Acetonitrile; Acetophenone; Adaptability; Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - metabolism; Alcohols; Amines; Amino acids; Binding Sites; Biocatalysts; Biochemistry; Biotechnology; Butanol; Catalysis; Chemistry; Chemistry and Materials Science; Cloning, Molecular; Coenzymes; Coenzymes - metabolism; Dehydrogenase; Dehydrogenases; Docking; Enzymes; Ethanol; Ethylene glycol; Glycerol; Industrial production; Isobutanol; Isopropanol; Kinetics; Molecular Docking Simulation; NAD; NAD - metabolism; NADP; NADP - metabolism; Organic Chemicals - metabolism; Organic solvents; Regeneration; Solvents; Solvents - metabolism; Substrate Specificity; Substrates; Coenzyme regeneration; Butanol; Docking; Organic solvent-tolerant; NAD(P); dependent dehydrogenase; NAD(P)+-dependent dehydrogenase; n-Butanol
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-019-03091-1

NAD(P)H-dependent enzymes are ideal biocatalysts for the industrial production of chiral compounds, such as chiral alcohols, chiral amino acids, and chiral amines; however, efficient strategies for the regeneration of coenzyme are expected as costly of the coenzymes. Herein, a solvent-tolerant isopropanol dehydrogenase (IDH) showing lower similarity (37%) with other proteins was obtained and characterized. The enzyme exhibits high catalysis ability of its substrates methanol, ethanol, ethylene glycol, glycerol, isopropanol,
n
-butanol, isobutanol, and acetone. And it has good adaptability in organic solvents (isopropanol, acetonitrile, acetone, and acetophenone). Interaction force and the corresponding amino acid residues between IDH and NAD
+
or NADP
+
were parsed by docking. The wide substrate spectrum, excellent organic solvent tolerance, and good biocatalytic activity make the excavated enzyme a promising biocatalyst for the production of chiral compounds industrially and the construction of coenzyme regeneration systems in aqueous organic phase or organic phase.