Efficient production of recombinant aldehyde reductase and its application for asymmetric reduction of ethyl 4-chloro-3-oxobutanoate to ethyl (R)-4-chloro-3-hydroxybutanoate

Prep Biochem Biotechnol. 2005;35(3):203-15. doi: 10.1081/PB-200065622.

Abstract

An NADPH-dependent aldehyde reductase (ALR, EC1.1.1.2) gene is cloned from Sporobolomyces salmonicolor ZJUB 105, and inserted into plasmid pQE30 to construct the expression plasmid (pQE30-ALR). A variety of E. coli strains were employed as hosts to obtain transformants with pQE30-ALR, respectively. Among these different types of transformants, the highest enzyme activity of ALR can be produced with E. coli M15 (pQE30-ALR). The bioactivity of ALR could be further improved significantly by the optimization of induction conditions. The results showed that the enzyme activity of ALR reached 6.48 U/mg protein, which is fifteen times higher than that of S. salmonicolor ZJUB 105. This recombinant strain was applied to the asymmetric reduction of ethyl 4-chloro-3-oxobutanoate (COBE) to ethyl (R)-4-chloro-3- hydroxybutanoate (CHBE). The results showed that the yield and optical purity of (R)-CHBE reached 98.5% and 99% e.e. (enantiomeric excess), respectively.

MeSH terms

  • Acetoacetates / metabolism*
  • Aldehyde Reductase / biosynthesis
  • Aldehyde Reductase / genetics
  • Aldehyde Reductase / metabolism*
  • Basidiomycota / enzymology*
  • Basidiomycota / genetics
  • Butyrates / metabolism*
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Fungal Proteins / biosynthesis
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Oxidation-Reduction
  • Plasmids / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Transformation, Bacterial

Substances

  • Acetoacetates
  • Butyrates
  • Fungal Proteins
  • Recombinant Proteins
  • ethyl 4-chloro-3-hydroxybutanoate
  • ethyl 4-chloro-3-oxobutanoate
  • Aldehyde Reductase