Overexpression and characterization of a novel (S)-specific extended short-chain dehydrogenase/reductase from Candida parapsilosis

Bioresour Technol. 2012 Nov:123:690-4. doi: 10.1016/j.biortech.2012.07.060. Epub 2012 Jul 25.

Abstract

The gene encoding a putative protein from Candida parapsilosis CDC317 (CPE) was cloned and overexpressed in Escherichia coli. The protein was amenable to overexpression in E. coli and constituted 35% of the total cell protein content. The optimal activity was determined at pH 5.5 and 40°C with the substrate 4-chloro-3-oxobutanoate ethyl ester (COBE). The optical purity of the product was over 99% enantiomeric excess for the (S)-isomer, and the molar conversion yield of the product was 91.1%. The apparent k(m) value for COBE was 0.19±0.01mM, which is an order of magnitude lower than that of other enzymes in the literature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Biocatalysis
  • Candida / enzymology*
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Fatty Acid Synthases / chemistry
  • Fatty Acid Synthases / isolation & purification
  • Fatty Acid Synthases / metabolism*
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • NADH, NADPH Oxidoreductases / chemistry
  • NADH, NADPH Oxidoreductases / isolation & purification
  • NADH, NADPH Oxidoreductases / metabolism*
  • Oxidoreductases / isolation & purification
  • Oxidoreductases / metabolism*
  • Sequence Alignment
  • Sequence Analysis, Protein
  • Sequence Homology, Amino Acid
  • Substrate Specificity
  • Temperature

Substances

  • Oxidoreductases
  • short chain trans-2-enoyl-CoA reductase
  • NADH, NADPH Oxidoreductases
  • Fatty Acid Synthases