An enone reductase from Nicotiana tabacum: cDNA cloning, expression in Escherichia coli, and reduction of enones with the recombinant proteins

Bioorg Chem. 2008 Feb;36(1):23-8. doi: 10.1016/j.bioorg.2007.08.005. Epub 2007 Oct 22.

Abstract

In the course of the purification of enone reductase participating to the reduction of pulegone, two reductases (NtRed-1 and NtRed-2) were isolated from cultured cells of Nicotiana tabacum. The partial amino acid sequences of the reductases revealed that NtRed-1 was allyl-alcohol dehydrogenase (Accession No. BAA89423) and NtRed-2 was malate dehydrogenase (Accession No. CAC12826). cDNA cloning and expression of these reductases in Escherichia coli were performed. Reduction with recombinant proteins was examined with cyclic alpha,beta-unsaturated ketones, such as pulegone, carvone and verbenone, as substrates. It was found that the recombinant NtRed-1 catalyses the hydrogenation of the exocyclic C-C double bond of pulegone.

MeSH terms

  • Amino Acid Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics*
  • DNA, Complementary / isolation & purification
  • Escherichia coli / genetics*
  • Gene Expression Regulation, Enzymologic / genetics*
  • Ketones / chemistry
  • Ketones / metabolism*
  • Molecular Conformation
  • Molecular Sequence Data
  • Oxidation-Reduction
  • Oxidoreductases / chemistry
  • Oxidoreductases / genetics*
  • Oxidoreductases / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Tobacco / enzymology*

Substances

  • DNA, Complementary
  • Ketones
  • Recombinant Proteins
  • Oxidoreductases