Optimization of the Process for Obtaining Antioxidant Protein Hydrolysates from Pumpkin Seed Oil Cake Using Response Surface Methodology

Abstract

Pumpkin seed cake, a byproduct of cold-pressed oil production, represents a food waste material with a great potential for valorization. The objective of the present study is to optimize the papain enzymatic hydrolysis process of pumpkin seed cold-pressed oil cake (CPC) to obtain protein hydrolysates with the highest antioxidant activity. Box–Behnken Response Surface Methodology (RSM) was used to optimize the simultaneous effects of an enzyme concentration of papain, a temperature, and a reaction time on the process of enzymatic hydrolysis on pumpkin seed cold-pressed oil cake (CPC). For these three input factors, different values are used—1, 2, and 3% for papain concentration, 20, 30, and 40 °C for temperature, and 60, 120, and 180 min for hydrolysis time. Thus, the design generated a total of 21 experimental runs. The aim is to obtain protein hydrolysates with the highest antioxidant activity. The responses DPPH and ABTS were calculated and the determined regression models were statistically analyzed and validated. The results revealed that optimal conditions included a papain concentration of 1.0%, a temperature of 40 °C, and a hydrolysis time of 60 min to retrieve the highest level of bioactive compounds.