Refolding of denatured carbonic anhydrase assisted by amide and refolding kinetics
The effect of various amides on the refolding of denat ur ed carbonic anhydrase B (CAB) was investigated at low guanidine hydrochloride (G dmCl) concentration.The competitive model of first-order folding and third-ord er aggregation was found to well describe the dynamic refolding process in the p resence of the additives. The results indicated that the denatured CAB could be properly refolded in the presence of the amides, and the effect of the amides on assisting CAB refolding was found kinetically similar to that of GdmCl. The ref olding rate constant k_N decreased with increasing additive concentration, while there was an optimum additive concentration where a minimum aggregation ra te constant k_A was obtained. At this additive concentration, the value of the ratio of k_N to k_A was maximum, leading to the maximal refolding yield of CAB.