Reactivation of heterodimer and individual subunits of penicillin acylase from E. coli after inactivation in urea.

Individual subunits of penicillin acylase from E. coli were isolated by gel-filtration under denaturing conditions (8 M urea). Recovery of the catalytic activity of the penicillin acylase heterodimer was studied after removal of urea. In the case of the heterodimer, 40-60% of the initial activity was recovered, whereas the activity of individual subunits was not recovered. Combination of native enzyme subunits with subunits isolated from the enzyme pre-inactivated with the irreversible inhibitor phenylmethylsulfonyl fluoride resulted in heterodimers which were active only in the case of involvement of the beta-subunit of the active enzyme.