Dual-enzyme synergistic preparation and characterization of antioxidant peptide from perilla meal and its alleviation on cellular oxidative injury

To improve the comprehensive utilization of perilla meal, dual-enzyme hydrolysis was optimized to prepare antioxidant peptides (PAP). The optimal conditions were pH 8.0, 50°C, 6 h, solid-liquid ratio of 1:22 g mL −1 , 1:1 compound enzymes and 6% enzyme dosage. The dual-enzyme product presented higher hydrolysis degree and antioxidant capacity than single-enzyme groups, with excellent solubility and processing stability against food additives, metal ions and food ingredients. The ultrafiltration screened active fraction PAP-SF-4 effectively alleviated oxidative damage in H 2 O 2 -stressed 4T1 cells by regulating ROS, MDA and endogenous antioxidant indexes. A key antioxidant peptide GDF was identified, which tightly bound to Keap1 through hydrogen bonds and hydrophobic interactions. This study provides a theoretical basis for the development and application of perilla-derived bioactive peptides.