Abstract
A novel NADPH-dependent reductase (CaCR) from Candida albicans was cloned for the first time. It catalyzed asymmetric reduction to produce ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE). It contained an open reading frame of 843 bp encoding 281 amino acids. When co-expressed with a glucose dehydrogenase in Escherichia coli, recombinant CaCR exhibited an activity of 5.7 U/mg with ethyl 4-chloro-3-oxobutanoate (COBE) as substrate. In the biocatalysis of COBE to (S)-CHBE, 1320 mM (S)-CHBE was obtained without extra NADP+/NADPH in a water/butyl acetate system, and the optical purity of the (S)-isomer was higher than 99% enantiomeric excess.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acetates / chemistry
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Acetoacetates / chemistry
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Acetoacetates / metabolism
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Amino Acid Sequence
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Biocatalysis
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Butyrates / chemical synthesis
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Butyrates / metabolism*
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Candida albicans / enzymology*
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Candida albicans / genetics
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Cloning, Molecular
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Escherichia coli / enzymology
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Escherichia coli / genetics
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Escherichia coli Proteins / chemistry
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism*
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Fungal Proteins / chemistry
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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Gene Expression
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Glucose Dehydrogenases / chemistry
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Glucose Dehydrogenases / genetics
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Glucose Dehydrogenases / metabolism*
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Molecular Sequence Data
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NADP / metabolism
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Open Reading Frames
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Oxidoreductases / chemistry
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Oxidoreductases / genetics
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Oxidoreductases / metabolism*
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Plasmids / chemistry
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Plasmids / genetics
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Stereoisomerism
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Water / chemistry
Substances
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Acetates
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Acetoacetates
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Butyrates
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Escherichia coli Proteins
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Fungal Proteins
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ethyl 4-chloro-3-hydroxybutanoate
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Water
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butyl acetate
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NADP
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ethyl 4-chloro-3-oxobutanoate
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Oxidoreductases
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Glucose Dehydrogenases