Characterization of alcohol dehydrogenase from Kangiella koreensis and its application to production of all-trans-retinol

• Published in: Biotechnology letters 2015-04, Vol.37 (4), p.849-856
• Main Authors: Hong, Seung-Hye, Ngo, Ho-Phuong-Thuy, Kang, Lin-Woo, Oh, Deok-Kun
• Format: Article
• Language: English
• Subjects: Alcanivoraceae - enzymology; Alcanivoraceae - genetics; Alcanivoraceae - isolation & purification; Alcohol dehydrogenase; Alcohol Dehydrogenase - chemistry; Alcohol Dehydrogenase - genetics; Alcohol Dehydrogenase - metabolism; Amino Acid Sequence; Applied Microbiology; Bacteria; Biochemistry; Biomedical and Life Sciences; Biotechnology; Coenzymes - metabolism; Conversion; Dehydrogenase; Dermatology; Enzyme Stability; Enzymes; Hydrogen-Ion Concentration; hydroquinone; Hydroquinones - metabolism; Kinetics; Life Sciences; methanol; Methanol - metabolism; Methyl alcohol; Microbiology; Molecular Sequence Data; Molecular Weight; NAD (coenzyme); NAD - metabolism; NADH; Optimization; Original Research Paper; Phylogeny; Protein Multimerization; Recombinant; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Sequence Homology, Amino Acid; Skin diseases; Temperature; Vitamin A - metabolism
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-014-1740-x

A recombinant alcohol dehydrogenase (ADH) from Kangiella koreensis was purified as a 40 kDa dimer with a specific activity of 21.3 nmol min⁻¹ mg⁻¹, a Kₘof 1.8 μM, and a kcₐₜof 1.7 min⁻¹for all-trans-retinal using NADH as cofactor. The enzyme showed activity for all-trans-retinol using NAD ⁺ as a cofactor. The reaction conditions for all-trans-retinol production were optimal at pH 6.5 and 60 °C, 2 g enzyme l⁻¹, and 2,200 mg all-trans-retinal l⁻¹in the presence of 5 % (v/v) methanol, 1 % (w/v) hydroquinone, and 10 mM NADH. Under optimized conditions, the ADH produced 600 mg all-trans-retinol l⁻¹after 3 h, with a conversion yield of 27.3 % (w/w) and a productivity of 200 mg l⁻¹ h⁻¹. This is the first report of the characterization of a bacterial ADH for all-trans-retinal and the biotechnological production of all-trans-retinol using ADH.