title: An enone reductase from Nicotiana tabacum: cDNA cloning, expression in Escherichia coli, and reduction of enones with the recombinant proteins.
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.
authors.affiliation: Natural Science Center for Basic Research and Development, Radioisotope Center, Hiroshima University, 1-4-2 Kagamiyama, Higashi-Hiroshima 739-8526, Japan.
authors.forename: Akihito
authors.initials: A
authors.lastname: Matsushima
authors.forename: Yuya
authors.initials: Y
authors.lastname: Sato
authors.forename: Miki
authors.initials: M
authors.lastname: Otsuka
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Watanabe
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Hirata
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D007659:Ketones / Q000737:chemistry / Q000378:metabolism*; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014026:Nicotiana / Q000201:enzymology*
chemical_list: D018076:DNA, Complementary; D007659:Ketones; D011994:Recombinant Proteins; D010088:Oxidoreductases