Novel approach to binary protein construction via pulsed electric field-induced unfolding: Comprehensive enhancement of chickpea protein properties

Developing functional, nutritionally rich plant-based protein resources is crucial for global food production. Here, we present a novel physical strategy to construct high-performance binary proteins by utilizing pulsed electric fields (PEF) to induce protein unfolding and subsequent co-folding. Under optimized conditions (30 kV/cm; chickpea protein (CP): bovine serum albumin (BSA) ratio 1:0.1, w/w), the resulting binary complex (C/B-P) exhibited a 228.3 % increase in solubility relative to native CP, accompanied by a significant reduction in aggregation. Multi-scale structural analyses and molecular dynamics simulations confirmed that PEF treatment unfolded the rigid CP structure, exposing an internal hydrophobic cavity (radius increased by 52 %). This newly accessible cavity serves as a high-affinity binding site (binding energy: −69.3 kcal/mol) for BSA, which stabilizes the unfolded state through co-folding. Consequently, C/B-P demonstrated superior functional properties, with foaming capacity soaring from 10 % to 90 % and markedly improved emulsion stability. Nutritionally, C/B-P possesses a complete amino acid profile, meeting FAO/WHO recommendations for both adults and preschool children. This PEF-based strategy not only effectively enhances CP's properties but also provides a powerful, rapid, and chemical-free paradigm for designing next-generation high-performance plant proteins for food applications.