title: Comparison of acyl-CoA synthetic activities and enantioselectivity toward 2-arylpropanoic acids in firefly luciferases.
abstract: Measurement of thioesterification activities for dodecanoic acid (C12) and ketoprofen was done using five firefly luciferases, from Pyrocoelia miyako (PmL), Photinus pyralis (PpL), Luciola cruciata (LcL), Hotaria parvura (HpL), and Luciola mingrelica (LmL). Among these, PmL, PpL, and LcL showed the expected thioesterification activities toward both substrates. All the enzymes exhibited (R)-enantioselectivity toward ketoprofen, which had same tendency as firefly luciferase from Luciola lateralis (LUC-H). HpL and LmL, however, did not accept ketoprofen, although they had thioesterification activity toward C12. These results indicate that the substrate acceptance of luciferases for the thioesterification reaction varies dramatically relying on the origin of firefly. Hence we focused primarily on PmL and investigated the effect of pH on enzymatic activity. In addition, by determining the kinetic parameters at various pH values, we verified that the k(cat) parameter contributed to the preferential enantioselectivity of this enzyme.
authors: Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo. kato@eng.u-hyogo.ac.jp
authors: Dai-ichiro
authors: D
authors: Kato
authors: Keisuke
authors: K
authors: Yokoyama
authors: Yoshihiro
authors: Y
authors: Hiraishi
authors: Masahiro
authors: M
authors: Takeo
authors: Seiji
authors: S
authors: Negoro
mesh_terms: D000214:Acyl Coenzyme A / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli; D004951:Esterification; D048888:Fireflies / Q000201:enzymology* / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007660:Ketoprofen / Q000378:metabolism*; D007700:Kinetics; D007850:Lauric Acids / Q000378:metabolism*; D049409:Luciferases, Firefly / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D013237:Stereoisomerism; D013379:Substrate Specificity; D013440:Sulfides / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000214:Acyl Coenzyme A; D007527:Isoenzymes; D007850:Lauric Acids; D011994:Recombinant Proteins; D013440:Sulfides; C030358:lauric acid; D007660:Ketoprofen; D049409:Luciferases, Firefly