Synergy of Ion Exchange and Covalent Reaction: Immobilization of Penicillin G Acylase on Heterofunctional Amino-Vinyl Sulfone Agarose

Agarose-vinyl sulfone (VS) beads have proven to be a good support immobilize several enzymes. However, some enzymes are hardly immobilized on it. This is the case of penicillin G acylase (PGA) from Escherichia coli, which very slowly this (less than 10% in 24 h). enzyme also not significantly adsorbed aminated MANAE-agarose beads, an anionic exchanger. In study, were modified with divinyl (DVS) produce MANAE-vinyl agarose beads. When PGA was support, fully less 1.5 h. cannot released by incubation at high ionic strength, suggesting that rapidly covalent fashion. Considering amount reactive VS groups only marginally increased, results indicated cooperative effect between anion exchange amine probably as first step process, and attachment previously molecules. The reaction molecules proceeds much more efficiently free enzyme, due proximity enzyme. Finally, steps immobilization, incubation, blocking different agents studied determine effects final activity/stability. stability new catalyst improved respect VS-agarose prepared low strength.