Operation conditions of enzyme refolding by chaperonin and recycle system using ultrafiltration

摘要

To clarify the efficient refolding conditions of enzymes using chaperonin GroE from Escherichia coli, the effects of various factors on the chaperonin-mediated refolding of enzymes from 4M guanidine hydrochloride (GdnHCl) were investigated. Three enzymes, Bacillus subtilis α-amylase, bovine deoxyribonuclease I (Dnase I) and yeast enolase were used as the model systems. In all enzymes, the maximum recovery of activities (90-15/100 with respect to the enzyme activities before denaturation) was attained in the presence of 2mM ATP and 2-5-fold molar excess of GroE 21-mer or GroEL 14-mer over enzyme molecules.α