Studies on the Purification of Aspergillus niger Endopolygalacturonase by Agarose Gel Chromatography
Endopolygalacturonase (endo PG) (poly (1,4-D-galacturonide) glycanohydrolase, E.C. 22.214.171.124), a pectolytic enzyme hydrolyzing randomly glycosidic linkages of pectic substances, can be purified by the usual methods. Nevertheless some original purification procedures have been developed using affinity chromatography on cross-linked pectic acid (1) or chromatography on agarose gel (2). On agarose it was observed that purified endo PG was delayed. In view of the resemblance between the anhydrogalactose residues of agarose and the anhydrogalacturonic ones of endo PG substrates, these workers suggested that biospecific interactions between agarose and endo PG were occurring. We used this suggestion to develop a method to purify endo PG from commercial Pectinase produced by Aspergillus niger and obtained from the Rapidase Company.