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Strategies for enzyme stabilization by intramolecular crosslinking with bifunctional reagents

Abstract:

A original strategy to obtain intramolecular crosslinking is discussed. This strategy consisted of three consecutive steps to direct the reaction to the formation of intramolecular crosslinks: (a) enzyme are partially modified with the bifunctional reagent in a very controlled fashion; (b) the excess of reagent is removed; and (c) the modified enzyme 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 derivatives of penicillin G acylase using glutaraldehyde as a crosslinking reagent under very controlled conditions. © 1995.