title: Characterization of Phosphoenolpyruvate Carboxylase from Oceanimonas smirnovii in Escherichia coli.
abstract: In this study, phosphoenolpyruvate carboxylase (PEPC) derived from Oceanimonas smirnovii (OS) was expressed as a soluble protein in Escherichia coli BL21(DE3). We isolated OS-PEPC (a recombinant PEPC protein) by his-tag purification. The purified protein showed a single band upon analysis with SDS-PAGE, and it had an apparent molecular mass of 98 kDa. Pufied OS-PEPC showed a specific activity value of 21.8 ± 0.495 U/mg protein. Especially, OS-PEPC showed the enzymatic activity between 40 and 50 °C. It maintained enzymatic activity in basic pH conditions (pH value, 9-10). We also measured OS-PEPC PEP and HCO3 (-) saturation kinetics and confirmed the effect of divalent cation on OS-PEPC activity.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, 121-742, South Korea.
authors.forename: Soohyun
authors.initials: S
authors.lastname: Park
authors.forename: Wangjun
authors.initials: W
authors.lastname: Lee
authors.forename: Hyeonsoo
authors.initials: H
authors.lastname: Kim
authors.forename: Seung Pil
authors.initials: SP
authors.lastname: Pack
authors.forename: Jinwon
authors.initials: J
authors.lastname: Lee
mesh_terms: D001639:Bicarbonates / Q000378:metabolism; D002413:Cations, Divalent / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D004926:Escherichia coli / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010728:Phosphoenolpyruvate / Q000378:metabolism; D010730:Phosphoenolpyruvate Carboxylase / Q000302:isolation & purification / Q000378:metabolism*; D020560:Proteobacteria / Q000201:enzymology*; D013696:Temperature
chemical_list: D001639:Bicarbonates; D002413:Cations, Divalent; D010728:Phosphoenolpyruvate; D010730:Phosphoenolpyruvate Carboxylase