Purification and characterization of two endopolygalacturonases from Sclerotinia sclerotiorum.

The endopolygalacturonase (EC 3.2.1.15) enzyme produced in vitro by Sclerotinia sclerotiorum were found to consist of numerous isoforms covering a broad pI range. Two of the isoforms, labelled PG2 and PG3, were purified using gel-filtration chromatography, isoelectric focusing and anion-exchange chromatography. The pIs of PG2 and PG3 were, respectively, 4.8 and 4.9. Their molecular weights were similar. Both enzymes hydrolysed 0.9% of the bonds in reaching a 50% reduction in viscosity. However, their enzymic parameters were different. Their amino acid compositions differed only in the aspartic acid-asparagine content. The N-terminal sequences differed at the fourth amino acid only. PG2 and PG3 exhibited a high level of glycosylation compared to a similar enzyme isolated from Aspergillus niger. Antibody raised against PG3 was shown to crossreact with PG2, but not with the enzyme purified from Aspergillus niger.