Characterization of Phosphoenolpyruvate Carboxylase from Oceanimonas smirnovii in Escherichia coli.

Park, Soohyun; Lee, Wangjun; Kim, Hyeonsoo; Pack, Seung; Lee, Jinwon

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 HCO saturation kinetics and confirmed the effect of divalent cation on OS-PEPC activity.

PHOSPHOENOLPYRUVIC carboxylase; ESCHERICHIA coli; BICARBONATE ions; MARINE bacteria; CARBON sequestration

Applied Biochemistry & Biotechnology, 2015, Vol 177, Issue 1, p217

0273-2289

Academic Journal

10.1007/s12010-015-1739-3