title: Efficient synthesis of optically pure alcohols by asymmetric hydrogen-transfer biocatalysis: application of engineered enzymes in a 2-propanol-water medium.
abstract: We describe an efficient method for producing both enantiomers of chiral alcohols by asymmetric hydrogen-transfer bioreduction of ketones in a 2-propanol (IPA)-water medium with E. coli biocatalysts expressing phenylacetaldehyde reductase (PAR: wild-type and mutant enzymes) from Rhodococcus sp. ST-10 and alcohol dehydrogenase from Leifsonia sp. S749 (LSADH). We also describe the detailed properties of mutant PARs, Sar268, and HAR1, which were engineered to have high activity and productivity in media composed of polar organic solvent and water, and the construction of three-dimensional structure of PAR by homology modeling. The K(m) and V(max) values for some substrates and the substrate specificity of mutant PARs were quite different from those of wild-type PAR. The results well explained the increased productivity of engineered PARs in IPA-water medium.
authors: Department of Biotechnology, Faculty of Engineering, Biotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan. nbito@pu-toyama.ac.jp
authors: Nobuya
authors: N
authors: Itoh
authors: Kentaro
authors: K
authors: Isotani
authors: Masatoshi
authors: M
authors: Nakamura
authors: Kousuke
authors: K
authors: Inoue
authors: Yasuhiro
authors: Y
authors: Isogai
authors: Yoshihide
authors: Y
authors: Makino
mesh_terms: D019840:2-Propanol / Q000032:analysis; D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000438:Alcohols / Q000737:chemistry / Q000378:metabolism*; D055162:Biocatalysis; D001709:Biotechnology / Q000379:methods; D003470:Culture Media / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D006859:Hydrogen / Q000378:metabolism; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity; D014867:Water / Q000032:analysis
chemical_list: D000438:Alcohols; D003470:Culture Media; D011994:Recombinant Proteins; D014867:Water; D006859:Hydrogen; D000429:Alcohol Oxidoreductases; C110191:phenylacetaldehyde reductase; D000426:Alcohol Dehydrogenase; D019840:2-Propanol