Study on the role of tyrosine side-chains at the active centre of emulsin beta-D-glucosidase.

The role of exposed tyrosine side-chains in enzyme-catalyzed reaction by sweet almond emulsin beta-D-glucosidase (beta-D-glucoside glucohydrolase, EC 3.2.1.21) has been studied using N-acetylimidazole as the specific reagent. The changes in activity, binding affinity and kinetic parameters (Km, V) as a result of acetylation of the phenolic hydroxyl groups have been determined. The acetylation increased the Km values of both beta-glucosidase and beta-galactosidase activities, whereas V remained unchanged. Similarly, the binding affinity for immobilized phenyl beta-D-glucopyranoside decreased appreciably. After the removal of the acetyl groups the enzyme regained 96% of the original activity. It is concluded that the tyrosine moieties, located in the active centre of the enzyme, have both glucoside and galactoside binding functions.