title: Cloning, expression, and characterization of an anti-Prelog stereospecific carbonyl reductase from Gluconobacter oxydans DSM2343.
abstract: A new anti-Prelog stereospecific carbonyl reductase (GoKR) from Gluconobacter oxydans DSM2343 was cloned and identified in Escherichia coli. This GoKR formed a homo-tetramer with a subunit size of approximately 27.0kDa. GoKR exhibited full activity with NADPH but not with NADH as a cofactor. The optimal pH and temperature were 9.0 and 30°C, respectively. GoKR reduced various ketones, including aliphatic and aromatic ketones, α- and β-keto esters. Aromatic ketones were reduced to (R)-enantiomers, whereas keto esters were reduced to (S)-hydroxy esters with different enantioselectivities. The data indicate that GoKR does not obey Prelog's rule and exhibits anti-Prelog enantiopreference. Enzyme-substrate-cofactor docking analysis showed that hydride transfer occurred at the si faces of carbonyl group for ethyl 4-chloro-3-oxobutanoate (COBE), which was then selectively reduced to the chiral (S)-alcohol. Excellent enantioselectivities were obtained for reducing COBE and ethyl 2-oxo-4-phenylbutyrate into the corresponding (S)-type products. These products are important for synthesizing HMG-CoA reductase (statins) and angiotensin-converting enzyme inhibitors, respectively.
authors: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China; Center for Biomedicine and Health, Division of Basical Medicine, Hangzhou Normal University, Hangzhou 310012, China.
authors: Rong
authors: R
authors: Chen
authors: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
authors: Xu
authors: X
authors: Liu
authors: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
authors: Jiale
authors: J
authors: Wang
authors: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China. Electronic address: jplin@ecust.edu.cn.
authors: Jinping
authors: J
authors: Lin
authors: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China. Electronic address: dzhwei@ecust.edu.cn.
authors: Dongzhi
authors: D
authors: Wei
mesh_terms: D000090:Acetoacetates / Q000737:chemistry / Q000378:metabolism; D000449:Aldehyde Reductase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000074408:Aldo-Keto Reductases; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001709:Biotechnology; D020134:Catalytic Domain; D003001:Cloning, Molecular; D015870:Gene Expression; D005798:Genes, Bacterial; D020600:Gluconobacter oxydans / Q000201:enzymology* / Q000235:genetics*; D007659:Ketones / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010654:Phenylbutyrates / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D000090:Acetoacetates; D001426:Bacterial Proteins; D007659:Ketones; D010654:Phenylbutyrates; C572921:ethyl 2-oxo-4-phenylbutyrate; C111982:ethyl 4-chloro-3-oxobutanoate; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase
keywords: Anti-Prelog rule; Asymmetric reduction; Carbonyl reductase; Molecular modeling