Original Articles: 2013 Vol: 5 Issue: 1
Optimization and characterization of polygalacturonase enzyme produced by gamma irradiated Penicillium citrinum
Abstract
The activity of polygalacturonase(PGase) produced by Penicillium citrinum was studied in solid state fermentation (SSF) using sugar beet pulp as substrate. A full factorial design was used to study the effect of 5 variables (yeast extract , pH , inoculum size, incubation period, and incubation temperature) on polygalacturonase production. Maximum polygalacturonase (PGase) production ( 132 U/g dry fermented substrate(U/gdfs) was obtained in 8 days at 30°C and pH 5.5 with yeast extract as the best nitrogen source (1.2%) inoculated with 1 ml of spore suspension (1.8×105 spores/ml). Using the optimized conditions of factorial design, maximum PGase production has been obtained by using 0.7 kGy of gamma radiation with an activity 152.2U/gdfs as compared to the parent strain (unirradiated). The enzyme was partially purified using 75% ammonium sulphate precipitation, dialysis, and gel filteration chromatography on sephadex G-100. The optimum pH and temperature of the enzyme were found to be 6.0 and 40°C respectively. The enzyme was found to be stable in the pH range 4-8 and showed high stability at temperature range 20-60°C. The effect of metal ions on enzyme activity was investigated. It was found that Mg+2, and Zn+2 stimulated PGase activity.