Optimization of Protease Treatment Conditions for Protein Extraction and Investigation of Its Potential as an Alternative Protein Source.

This study aimed to determine enzymes that effectively extract proteins and optimize the processing conditions using response surface methods. Furthermore, the potential of enzymatically hydrolyzed protein extract (CPE) as a substitute protein source was investigated. The enzymatic hydrolysis conditions for protein extraction were optimized using single-factor analysis and a response surface methodology-Box-Behnken design. The R value of the optimized model was 0.9270, indicating the reliability of the model, and the optimal conditions were as follows: a hydrolysis temperature of 45.56 °C, pH 9.1, and a hydrolysis time of 49.85 min. The amino acid composition of CPE was compared to that of powder (CP), which was found to have a higher content of essential amino acids (EAA). The electrophoretic profiles of CP and CPE confirmed that CPE has a low molecular weight. Furthermore, CPE showed higher antioxidant activity and phenol content than CP, with ABTS and DPPH radical scavenging abilities of 69.40 ± 1.61% and 19.27 ± 3.16%, respectively. CPE had high EAA content, antioxidant activity, and phenol content, indicating its potential as an alternative protein source. Overall, in this study, we developed an innovative, ecofriendly, and gentle enzymatic hydrolysis strategy for the extraction and refinement of Chlorella proteins.