Optimization of Hydrolysis Condition of Pumpkin Seeds with Alcalase Enzyme to Achieve Maximum Antioxidant and Nitric Oxide Inhibition Activity

In this research, optimization of pumpkin (Cucurbita pepo) hydrolysis condition was investigated in order to achieve maximum 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and nitric oxide inhibition activity using Design Expert software and response surface method. For this purpose, hydrolysis conditions including concentration of alcalase enzyme were selected within 0.7-3.30%, temperature 32-58 °C and hydrolysis time 30-290 min as independent variable. The results showed that optimum hydrolysis conditions to achieve the maximum DPPH radical scavenging and nitric oxide inhibition activity were temperature of 44 °C, hydrolysis time of 260 min, and enzyme to substrate concentration of 3% which under this condition the DPPH radical scavenging and nitric oxide inhibition activity of hydrolyzate was 72.03 and 89.34%, respectively that was largely similar to the results proposed by the software (75.33 and 84.71%). According to the results, pumpkin seed protein hydrolyzate showed high antioxidant and nitric oxide inhibitory properties and can be used as a suitable ingredient in food formulations.