Cloning and expression in Escherichia coli of a gene coding for a secondary alcohol dehydrogenase from Candida parapsilosis.
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.
Yamamoto
H
H
Tsukuba Research Center, Daicel Chemical Industries, Ltd., Ibaraki, Japan. h-yamamoto@daicel.co.jp
Kawada
N
N
Matsuyama
A
A
Kobayashi
Y
Y
D000426:Alcohol Dehydrogenase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002175:Candida / Q000235:genetics* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D017931:DNA Primers; D004271:DNA, Fungal / Q000096:biosynthesis; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D004271:DNA, Fungal; D011994:Recombinant Proteins; D000426:Alcohol Dehydrogenase