Application of Cycloamylose Ligand Affine Chromatography for the Analysis of Amylolytic Enzymes

Abstract

Abstract The investigation of functional groups participating in enzyme‐catalyzed reactions plays a significant role in the study of the molecular mechanism of these reactions. In order to decide whether a given functional group participates in the formation of the enzyme‐substrate complex it should be known whether there are changes in the binding between enzyme and substrate in the course of modification process. This can be done by the affine chromatographic technique. After explaining the preparation of epoxy‐activated agarose the role of the extend of the spacer and cross‐linking density of agarose gel in binding α‐amylase, β‐amylase and glucoamylase is described. Further subjects of the paper are thermodynamic study of the linkage and the effect of functional group modification on the linkage.