Physico-chemical characteristics of immobilized polygalacturonase from <i>Aspergillus niger</> (SA6)

Abstract

Polygalacturonase (PG) was isolated from Aspergillus niger (A. niger) (SA6), partially purified, characterized and immobilized by entrapment using calcium alginate. The polygalacturonase showed two bands on sodium dodecyl sulfate polyacryamide gel electrophoresis (SDS-PAGE) suggesting an “endo and exo” polygalacturonase with apparent molecular weights of 35 and 40 KDa, respectively. The enzyme was purified 9 fold with a yield of 0.18% and specific activity of 246 μmole/min/mg. The apparent K_M and V_max of the immobilized polygalacturonase were11.1 mg/ml and 1.65 μmole/min/mg, respectively. The optimum pH and optimum temperature of the immobilized polygalacturonase were 4.5 and 40°C, respectively. Immobilized polygalacturonase exhibited more stability to changes in pH than the temperature. The activity of the immobilized polygalacturonase reduced to 34.56 and 14.81% of the initial activity in the second and third catalytic cycles, respectively. The half life of the enzyme and the
activity lost per minute on thermal storage were 10 min and 0.0213 μMole of D-galacturonic acid.

Keywords: Polygalacturonase, Aspergillus niger, pectinases, enzymes