원문정보
초록
영어
The chaperonin α-subunit gene (ApCpnA, 1,871 bp ORF) from the hyperthermophilic archaeon Aeropyrum pernix K1 was amplified by PCR and subcloned into vector pRSFDuet-1. The constructed pRSFDuet-ApCpnA (5.4 kb) was transformed into E. coli BL21 Rosetta (DE3). The E. coli transformant cell successfully expressed ApCpnA as 60.7 kDa protein in both soluble and insoluble fractions of cell lysate.
The purified ApCpnA showed ATPase activity and its activity was dependent on temperature. When the alginate lyase gene (aly, 1.19 kb) from Pseudoalteromomas elyakovii was coexpressed with ApCpnA, alginate lyase was produced as a soluble and active form, speculating that ApCpnA facilitates the correct folding of alginate lyase. These results suggest that ApCpnA has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.