Cloning and expression in Escherichia coli of a gene coding for a secondary alcohol dehydrogenase from Candida parapsilosis

Biosci Biotechnol Biochem. 1999 Jun;63(6):1051-5. doi: 10.1271/bbb.63.1051.

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

  • Alcohol Dehydrogenase / analysis
  • Alcohol Dehydrogenase / biosynthesis
  • Alcohol Dehydrogenase / genetics*
  • Amino Acid Sequence
  • Base Sequence
  • Candida / genetics*
  • Candida / metabolism
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • DNA Primers
  • DNA, Fungal / biosynthesis
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Gene Expression Regulation, Enzymologic / genetics*
  • Molecular Sequence Data
  • Plasmids
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA Primers
  • DNA, Fungal
  • Recombinant Proteins
  • Alcohol Dehydrogenase