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Endo-polygalacturonase (endo-PG) of Aspergillus niger was purified by (NH4)2 SO4 precipitation, ion exchange chromatography on CM-Sephadex C-50, preparative isoelectric focusing and Sephadex G-75 gel filtration. The endo-PG I and II were purified approximately 260- and 280-fold, and they were found to have Km values of 3.33 and 1.67mg/ml, respectively, from the LINEWEAVER-BURK plot. The molecular weights of endo-PG I and II were 46000 and 35000, and had an isoelectric point of pH 5.6 and 6.5, respectively. Endo-PG I was most stable at a pH range between 4.0 and 5.5. Endo-PG II was found to be stable at pH 2.5 to 4.0. However, the enzymes had optimum pH at 4.5, and their optimum temperature for the action was 55°C. They were stable at a temperature up to 45°C. Therefore, the two forms of endo-PG might differ slightly in the physical and chemical structure of their molecules in the mechanism of hydrolysis of pectic acid.

Purification and some properties of exo-polygalacturonase from Aspergillus niger cultured in the medium containing satsuma mandarin peel.