STRATEGIES FOR ENZYME STABILIZATION BY INTRAMOLECULAR CROSS-LINKING WITH BIFUNCTIONAL REAGENTS

A original strategy to obtain intramolecular crosslinking is discussed . This strategy consisted of three consecutive steps to direct the rea ction to the formation of intramolecular crosslinks: (a) enzyme are pa rtially modified with the bifunctional reagent in a very controlled fa shion; (b) the excess of reagent is removed; and (c) the modified enzy me is incubated long-term to allow a crosslinking reaction without the competition of additional single-point modifications. In this way, we were able to obtain interesting stabilizations of immobilized derivat ives of penicillin G acylase using glutaraldehyde as a crosslinking re agent under very controlled conditions.