Engineering of phenylacetaldehyde reductase for efficient substrate conversion in concentrated 2-propanol.
Phenylacetaldehyde reductase (PAR) is suitable for the conversion of various aryl ketones and 2-alkanones to corresponding chiral alcohols. 2-Propanol acts as a substrate solvent and hydrogen donor of coupled cofactor regeneration during the conversion of substrates catalyzed by PAR. To improve the conversion efficiency in high concentrations of substrate and 2-propanol, selection of a PAR mutant library and the subsequent rearrangement of mutations were attempted. With only a single selection round and following the manual combination of advantageous mutations, PAR was successfully adapted for the conversion of high concentrations of substrate with concentrated 2-propanol. This method will be widely applicable for the engineering of enzymes potentially valuable for industry.
Makino
Yoshihide
Y
Biotechnology Research Center, Toyama Prefectural University, Kurokawa 5180, Kosugi, Toyama 939-0398, Japan. makino@pu-toyama.ac.jp.
Inoue
Kousuke
K
Dairi
Tohru
T
Itoh
Nobuya
N
D000433:1-Propanol / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D003352:Corynebacterium / Q000201:enzymology / Q000235:genetics; D015723:Gene Library; D005818:Genetic Engineering / Q000379:methods*; D007218:Industrial Microbiology / Q000379:methods; D008969:Molecular Sequence Data; D009154:Mutation; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity
D000433:1-Propanol; D000429:Alcohol Oxidoreductases; C110191:phenylacetaldehyde reductase