High-level expression of maize C4-type phosphoenolpyruvate carboxylase in Escherichia coli and its rapid purification.
Maize C4-type phosphoenolpyruvate carboxylase (PEPC) was expressed in E. coli with the pET32 system. The expressed fusion PEPC was active and its amount comprised more than 10% of total soluble protein. The specific activity increased by about 45-fold, compared with our previous system [S. Yanagisawa and K. Izui, Agric. Biol. Chem., 54, 241-243 (1990)]. The fusion PEPC was rapidly purified with His bind metal chelation resin, showing a single band on SDS-PAGE. Moreover, the tag domain fused at the N-terminus did not have any effect on catalytic and regulatory properties of PEPC.
Dong
L Y
LY
Laboratory of Applied Botany, Faculty of Agriculture, Kyoto University, Japan.
Hata
S
S
Izui
K
K
D002384:Catalysis; D002614:Chelating Agents / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D010730:Phosphoenolpyruvate Carboxylase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003313:Zea mays / Q000201:enzymology*
D002614:Chelating Agents; D010940:Plant Proteins; D011994:Recombinant Proteins; D010730:Phosphoenolpyruvate Carboxylase