Purification and characterization of a (R)-1-phenyl-1,3-propanediol-producing enzyme from Trichosporon fermentans AJ-5152 and enzymatic (R)-1-phenyl-1,3-propanediol production

• Published in: Bioscience, biotechnology, and biochemistry Vol. 73; no. 7; pp. 1640 - 1646
• Main Authors: Kira, I.(Ajinomoto Co. Inc., Kawasaki, Kanagawa (Japan). AminoScience Labs.), Onishi, N
• Format: Journal Article
• Language: English
• Published: England Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.07.2009; Oxford University Press
• Subjects: (R)-1-phenyl-1,3-propanediol; 3-hydroxy-1-phenylpropane-1-one; Biocatalysis - drug effects; BIOSINTESIS; BIOSYNTHESE; BIOSYNTHESIS; CHEMICOPHYSICAL PROPERTIES; chiral alcohol; Coenzymes - metabolism; DRUGS; ENZIMAS; ENZYME; Enzyme Stability; ENZYMES; Glucose 1-Dehydrogenase - metabolism; Hydrogen-Ion Concentration; Indicators and Reagents - pharmacology; Ketones - metabolism; MEDICAMENT; MEDICAMENTOS; Molecular Weight; NADP - metabolism; Oxidoreductases - chemistry; Oxidoreductases - isolation & purification; Oxidoreductases - metabolism; Propanols - metabolism; PROPIEDADES FISICOQUIMICAS; PROPRIETE PHYSICOCHIMIQUE; Propylene Glycols - metabolism; PROTEIN SYNTHESIS; PURIFICACION; PURIFICATION; SINTESIS DE PROTEINAS; stereospecific oxidoreductase; Substrate Specificity; SYNTHESE PROTEIQUE; Temperature; TRICHOSPORON; Trichosporon - enzymology; Trichosporon - metabolism; Trichosporon fermentans
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.90159

An (R)-1-phenyl-1,3-propanediol-producing enzyme was purified from Trichosporon fermentans AJ-5152. It was NADPH-dependent and converted 3-hydroxy-1-phenylpropane-1-one (HPPO) to (R)-1-phenyl-1,3-propanediol [(R)-PPD] with anti-Prelog's specificity. It showed maximum activity at pH 7.0 and 40 °C. Its K m and V max values toward HPPO were 20.1 mM and 3.4 μmol min −1 mg protein −1 respectively. The relative molecular weight of the enzyme was estimated to be 68,000 on gel filtration and 32,000 on SDS-polyacrylamide gel electrophoresis. An (R)-PPD-producing reaction using the (R)-PPD-producing enzyme and an NADPH recycling system was carried out by successive feeding of HPPO. A total (R)-PPD yield of 8.9 g/l was produced in 16 h. The molar yield was 76%, and the optical purity of the (R)-PPD produced was over 99% e.e.