Purification and characterization of a novel alcohol dehydrogenase from Leifsonia sp. strain S749: a promising biocatalyst for an asymmetric hydrogen transfer bioreduction.
To find microorganisms that could reduce phenyl trifluoromethyl ketone (PTK) to (S)-1-phenyltrifluoroethanol [(S)-PTE], styrene-assimilating bacteria (ca. 900 strains) isolated from soil samples were screened. We found that Leifsonia sp. strain S749 was the most suitable strain for the conversion of PTK to (S)-PTE in the presence of 2-propanol as a hydrogen donor. The enzyme corresponding to the reaction was purified homogeneity, characterized and designated Leifsonia alcohol dehydrogenase (LSADH). The purified enzyme had a molecular weight of 110,000 and was composed of four identical subunits (molecular weight, 26,000). LSADH required NADH as a cofactor, showed little activity with NADPH, and reduced a wide variety of aldehydes and ketones. LSADH catalyzed the enantioselective reduction of some ketones with high enantiomeric excesses (e.e.): PTK to (S)-PTE (>99% e.e.), acetophenone to (R)-1-phenylethanol (99% e.e.), and 2-heptanone to (R)-2-heptanol (>99% e.e.) in the presence of 2-propanol without an additional NADH regeneration system. Therefore, it would be a useful biocatalyst.
Biotechnology Research Center, Toyama Prefectural University, Kosugi, Toyama 939-0398, Japan.
Kousuke
K
Inoue
Yoshihide
Y
Makino
Nobuya
N
Itoh
D019840:2-Propanol / Q000378:metabolism; D000192:Actinomycetales / Q000201:enzymology* / Q000254:growth & development / Q000302:isolation & purification; D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D003470:Culture Media; D006859:Hydrogen / Q000378:metabolism*; D007659:Ketones / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D012988:Soil Microbiology; D020058:Styrene / Q000378:metabolism; D013379:Substrate Specificity
D003470:Culture Media; D007659:Ketones; D009243:NAD; D020058:Styrene; C448317:phenyl trifluoromethyl ketone; D006859:Hydrogen; D000426:Alcohol Dehydrogenase; D019840:2-Propanol