Purification of pancreatic α-amylase by affinity chromatography: A report submitted in partial fulfilment of the requirement for the degree of B.Sc (Hons) in the University of Canterbury

Abstract

The present work was undertaken to examine a model system for a method applicable to the industrial purification of pancreatic α-amylase using the technique of affinity chromatography. This method is based on the specificity of the enzyme for the substrate and the resultant reversible binding. A column of bound substrate will then specifically absorb the enzyme in preference to the other components of a solution. Insoluble starch is used as a substrate for α-amylase in this way, the α- amylase will absorb specifically to the ligand (substrate), but the other pancreatic enzyme should not exhibit appreciable affinity for the ligand and should pass through the column unretarded. Since a-amylase is the major of pancreatin, the purification can be best determined by following the removal of the contaminant enzymes. Trypsin will be used as a representative enzyme for these contaminants and the object of this work will be to show the removal of trypsin activity from the amylase fraction. In this work, a comparative study has also been done on the purification of α-amylase by precipitation of amylase-glycogen complex and the affinity chromatography.