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영어
This study investigated that Aeropyrum pernix K1 chaperonins (ApCpnA and ApCpnB) can efficiently prevent the thermal aggregation and inactivation of foreign model protein, citrate synthase (CS, 48.9 kDa, dimer) from porcine heart. ApCpnA and ApCpnB, chaperonin genes were amplified by PCR from the A. pernix K1 genomic DNA, and subcloned into pET3d and pET21a vectors, respectively. Each of the constructed pET3d-ApCpnA (6.1 kb) and pET21a- ApCpnB (6.9 kb) was transformed into E. coli Rosetta and BL21 Codonplus
(DE3), and the transformant cells successfully expressed ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa). Each of the recombinant ApCpnA and ApCpnB was purified by heat treatment and anion-exchange chromatography. ApCpnA and ApCpnB prevented the thermal aggregation and inactivation of foreign model protein, CS at 43℃. Especially, the addition of ATP or CTP with ApCpnA and ApCpnB more extensively prevented thermal aggregation and inactivation of CS. Based on these results, we propose that ApCpnA and ApCpnB from A.
pernix K1 can be utilized to enhance the durability and cost effectiveness of high-temperature biocatalysts.