biostudies-literatureZhu YHNational Research Foundation of Korea225-233https://www.ebi.ac.uk/biostudies/studies/S-EPMC6458232biostudies-literatureUnknown59(2)A novel alcohol dehydrogenase from <i>Bartonella apis</i> (BaADH) was heterologous expressed in <i>Escherichia coli</i>. Its biochemical properties were investigated and used to catalyze the synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE), which is a chiral intermediate of the cholesterol-lowering drug atorvastatin. The purified recombinant BaADH displayed 182.4 U/mg of the specific activity using ethyl 4-chloroacetoacetate as substrate under the conditions of 50 °C in pH 7.0 Tris-HCl buffer. It was stable in storage buffers of pH 7 to 9 and retains up to 96.7% of the initial activity after 24 h. The <i>K</i> _m and <i>V</i> _max values of BaADH were 0.11 mM and 190.4 μmol min^-1 mg^-1, respectively. Synthesis of (S)-CHBE catalyzed by BaADH was performed with a cofactor regeneration system using a glucose dehydrogenase, and a conversion of 94.9% can be achieved after 1 h reaction. Homology modeling and substrate docking revealed that a typical catalytic triad is in contact with local water molecules to form a catalytic system. The results indicated this ADH could contribute to the further enzymatic synthesis of (S)-CHBE.Indian journal of microbiologyCloning, Expression and Characterization of a Highly Active Alcohol Dehydrogenase for Production of Ethyl (S)-4-Chloro-3-Hydroxybutyrate.PMC6458232NRF-2018H1D3A2001746Liu CYCai SZhang YWLee JKKim IWZhu YHKalia VCGuo LBfalseCloning, Expression and Characterization of a Highly Active Alcohol Dehydrogenase for Production of Ethyl (S)-4-Chloro-3-Hydroxybutyrate.A novel alcohol dehydrogenase from <i>Bartonella apis</i> (BaADH) was heterologous expressed in <i>Escherichia coli</i>. Its biochemical properties were investigated and used to catalyze the synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE), which is a chiral intermediate of the cholesterol-lowering drug atorvastatin. The purified recombinant BaADH displayed 182.4 U/mg of the specific activity using ethyl 4-chloroacetoacetate as substrate under the conditions of 50 °C in pH 7.0 Tris-HCl buffer. It was stable in storage buffers of pH 7 to 9 and retains up to 96.7% of the initial activity after 24 h. The <i>K</i> _m and <i>V</i> _max values of BaADH were 0.11 mM and 190.4 μmol min^-1 mg^-1, respectively. Synthesis of (S)-CHBE catalyzed by BaADH was performed with a cofactor regeneration system using a glucose dehydrogenase, and a conversion of 94.9% can be achieved after 1 h reaction. Homology modeling and substrate docking revealed that a typical catalytic triad is in contact with local water molecules to form a catalytic system. The results indicated this ADH could contribute to the further enzymatic synthesis of (S)-CHBE.2019-01-01T00:00:00Z2019 Jun2024-02-15T09:42:22.395Z2020-06-04T07:10:35ZS-EPMC64582323103143810.1007/s12088-019-00795-0