title: Purification and characterization of a novel enoyl coenzyme A reductase from Streptomyces collinus.
abstract: A novel NADPH-dependent enoyl reductase, catalyzing the conversion of 1-cyclohexenylcarbonyl coenzyme A (1-cyclohexenylcarbonyl-CoA) to cyclohexylcarbonyl-CoA, was purified to homogeneity from Streptomyces collinus. This enzyme, a dimer with subunits of identical M(r) (36,000), exhibits a Km of 1.5 +/- 0.3 microM for NADPH and 25 +/- 3 microM for 1-cyclohexenylcarbonyl-CoA. It has a pH optimum of 7.5, is most active at 30 degrees C, and is inhibited by both divalent cations and thiol reagents. Two internal peptide sequences were obtained. Ansatrienin A (an antibiotic produced by S. collinus) contains a cyclohexanecarboxylic acid moiety, and it is suggested that the 1-cyclohexenylcarbonyl-CoA reductase described herein catalyzes the final reductive step in the conversion of shikimic acid into this moiety.
authors.affiliation: Department of Biomedicinal Chemistry, School of Pharmacy, University of Maryland, Baltimore 21201.
authors.forename: K A
authors.initials: KA
authors.lastname: Reynolds
authors.forename: P
authors.initials: P
authors.lastname: Wang
authors.forename: K M
authors.initials: KM
authors.lastname: Fox
authors.forename: M K
authors.initials: MK
authors.lastname: Speedie
authors.forename: Y
authors.initials: Y
authors.lastname: Lam
authors.forename: H G
authors.initials: HG
authors.lastname: Floss
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D003509:Cyclohexanecarboxylic Acids / Q000378:metabolism; D004586:Electrophoresis; D044943:Fatty Acid Desaturases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009249:NADP / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D003509:Cyclohexanecarboxylic Acids; D009249:NADP; C074886:1-cyclohexenecarboxylic acid; D044943:Fatty Acid Desaturases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C074885:1-cyclohexenylcarbonyl-CoA reductase, Streptomyces collinus