Achievement of high recovered activity and efficient purification and recovery by application of immobilized inhibitor to protease refolding

In order to suppress autolysis which decreases recovered activity), synthes ized inhibitor is applied to protease refolding. In the refolding of Strept omyces griseus trypsin (SGT) from its denatured and reduced state, recovere d activity) reaches only 15% by the ordinary dilution method, while it exce eds 60% in the presence of the gel on which p-aminobenzamidine, an inhibito r to trypsin, is immobilized covalently. Furthermore, by use of the immobil ized inhibitor, purification and recovery of renatured SGT are efficiently performed. In addition, aggregation, the other disadvantageous reaction to renaturation, can also be diminished with coexistence of 1 M urea in this s uspension system. Reusability of the immobilized inhibitor will make it pos sible to develop this refolding into the continuous process.