Pectinases find wide application in different industrial applications, e.g., textile, food, tea, paper and pulp, vegetable oil extractions, saccharification of agricultural remains, and in fermented drinks. Three fungal strains were screened for pectinase production. Results indicated that all strains are capable of producing a pectinase enzyme. Penicillium chrysogenum MF318506 showed the highest pectinase activity (0.214 U/ml) on the recommended medium at 30 ◦C at 150 rpm, on 6 days. Different agricultural wastes (orange peel, banana peel, potato peel, pomegranate peel, wheat bran, and rice bran) were screened as a carbon source. The maximum pectinase activity of 0.48U/ml was obtained from Orange peel and was selected for further studies. Screening of important medium components for pectinase production by the Plackett-Burman design (PBD) showed that the maximum pectinase activity of 1.057 U/ml was achieved with 3 % orange peel, 2.0 g/l (NH4)2SO4, 6.0 g/l peptone, 6.0 g/l KH2PO4, 6.0 g/l K2HPO4, 0.1 g/l MgSO4.7H2O, 0.5 g/l KCl and 0.1 g/l FeSO4. Finally, the Box-Behken design (BBD) which uses the response surface methodology (RSM) was applied to further optimization of the selected factors using the PBD. The maximum pectinase activity of 1.292U/ml was observed at 2.5% orange peel and 4.5 g/l peptone. The Statistical optimization, enhanced the pectinase production by 6.04 folds using Orange peel waste as a substrate. The pectinase produced by Penicillium chrysogenum MF318506 has potential applications in different industrial fields.