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.
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 degrees C. Its K(m) and V(max) values toward HPPO were 20.1 mM and 3.4 mumol 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.
Kira
Ikuo
I
AminoScience Laboratories, Ajinomoto Co, Inc, Kawasaki, Japan. ikuo_kira@ajinomoto.com
Onishi
Norimasa
N
D055162:Biocatalysis / Q000187:drug effects; D003067:Coenzymes / Q000378:metabolism; D004795:Enzyme Stability; D042843:Glucose 1-Dehydrogenase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000494:pharmacology; D007659:Ketones / Q000378:metabolism; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism; D010088:Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D020005:Propanols / Q000378:metabolism; D011409:Propylene Glycols / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D014250:Trichosporon / Q000201:enzymology* / Q000378:metabolism
C541751:1-phenyl-1,3-propanediol; C541752:3-hydroxy-1-phenylpropane-1-one; D003067:Coenzymes; D007202:Indicators and Reagents; D007659:Ketones; D020005:Propanols; D011409:Propylene Glycols; D009249:NADP; D010088:Oxidoreductases; D042843:Glucose 1-Dehydrogenase