Abstract
A gene encoding a stereo-specific secondary alcohol dehydrogenase (CpSADH) that catalyzed the oxidation of (S)-1,3-BDO to 4-hydroxy-2-butanone was cloned from Candida parapsilosis. This CpSADH-gene consisted of 1,009 nucleotides coding for a protein with M(r) 35,964. A recombinant Escherichia coli JM109 strain harboring the expression plasmid, pKK-CPA1, produced (R)-1,3-BDO (93.5% ee., 94.7% yield) from the racemate without any additive to regenerate NAD+ from NADH.
MeSH terms
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Alcohol Dehydrogenase / analysis
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Alcohol Dehydrogenase / biosynthesis
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Alcohol Dehydrogenase / genetics*
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Amino Acid Sequence
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Base Sequence
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Candida / genetics*
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Candida / metabolism
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Chromatography, High Pressure Liquid
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Cloning, Molecular
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DNA Primers
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DNA, Fungal / biosynthesis
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Escherichia coli / genetics*
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Escherichia coli / metabolism
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Gene Expression Regulation, Enzymologic / genetics*
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Molecular Sequence Data
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Plasmids
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / genetics
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Reverse Transcriptase Polymerase Chain Reaction
Substances
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DNA Primers
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DNA, Fungal
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Recombinant Proteins
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Alcohol Dehydrogenase