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In order to express archaea chaperonin A from hyperthermophilic Aeropyrum pernix K1 in E. coli, the gene (cpnA, 1,871 bp ORF) was amplified by PCR and subcloned into the pET3d vector. When the resulting plasmid, pET-3d-cpnA (6.1 Kb), was transformed into E. coli Rosetta(DE3), the transformant cell successfully expressed cpnA, irrespective of IPTG concentration. In this work, the target protein for coexpression with cpnA was pro-carboxypeptiase B (pro-CPB) from porcine pancreas. The effects of coexpression of archaea chaperonin on the productivity of soluble pro-CPB in E. coli were examined. Based on the IPTG concentration for pro-CPB expression and the culture time for the maximum expression of cpnA, the production of soluble pro-CPB was sharply increased at 6 hr after induction. The soluble pro-CPB was activated
with trypsin and its activity was tested with the synthetic substrate hippuryl-L-Arg. The activity of carboxypeptidase B in the soluble fraction of E. coli Rosetta(DE3) coexpressing cpnA and pro-CPB reached about 2.68 unit/ml.