Design, fabrication and molecular dynamics simulations of luteolin pickering emulsion using phosphorylated and succinylated lactoferrin nanocomplex.

Abstract

Chemically modified protein-based nanocomplexes are widely used as food-grade carriers, but their molecular mechanisms remain unclear. This study developed Pickering emulsions (PEs) composed of phosphorylated (PLF) or succinylated (SLF) lactoferrin (LF) complexed with luteolin (LUT) via a pH-driven method, integrating experimental and computational approaches. SLF-LUT PEs showed superior physicochemical properties, including smaller droplet sizes, encapsulation efficiency up to 80% under neutral conditions, and enhanced storage stability at pH 8 over 30 days. SLF-LUT also significantly inhibited lipid oxidation, with reduced primary and secondary oxidation products. Multi-ligand docking and 100 ns molecular dynamics simulations revealed that LUT binds multiple flaxseed oil fatty acids within lactoferrin's hydrophobic pocket, positioning it for radical scavenging at the oil-water interface. The LF-LUT complex remained stable in a lipid-rich environment, with key residues (LEU320, PRO251, SER252) critical for stabilization. Network pharmacology further linked these interactions to antioxidant pathways (KEAP1/NRF2, FoxO, AGE-RAGE). These findings highlight the potential of nanocomplex-based oil-in-water PEs for delivering hydrophobic bioactives in food and pharmaceutical applications.