Effects of homogeneous and ultrasonic treatment on casein/phosphatidylcholine complex-emulsions: Stability and bioactivity insights

•Ultrasonic treatment changed solubility and surface hydrophobicity of casein.•Ultrasonic treatment enhanced storage stability.•Phosphatidylcholine protected casein structure under high temperature through hydrogen bonding.•Ultrasonic treatment improved digestibility of casein/phosphatidylcholine emulsion. Casein (CAS), a typical protein emulsifier, has functional properties limited by its chemical structure in practical production applications. This study aimed to combine phosphatidylcholine (PC) and casein to form a stable complex (CAS/PC) and improve its functional properties through physical modification (homogeneous and ultrasonic treatment). To date, few studies have explored the effects of physical modification on the stability and biological activity of CAS/PC. Interface behavior analysis showed that compared to homogeneous treatment, PC addition and ultrasonic treatment could decrease the mean particle size (130.20 ± 3.96 nm) and increase the zeta potential (−40.13 ± 1.12 mV), indicating the emulsion is more stable. The chemical structural analysis of CAS showed that PC addition and ultrasonic treatment promoted changes in its sulfhydryl content and surface hydrophobicity, exposing more free sulfhydryl groups and hydrophobic binding sites, thereby enhancing its solubility and improving the stability of the emulsion. Additionally, storage stability analysis revealed that the incorporation of PC with ultrasonic treatment could improve the root mean square deviation value and radius of gyration value of CAS. These modifications resulted in an increase the binding free energy between CAS and PC (−238.786 kJ/mol) at 50 °C, leading to an improvement in the thermal stability of the system. Furthermore, digestive behavior analysis indicated that PC addition and ultrasonic treatment could increase the total FFA release from 667.44 ± 22.33 μmol to 1250.33 ± 21.56 μmol. In conclusion, the study underscores the effectiveness of PC addition and ultrasonic treatment in enhancing the stability and bioactivity of CAS, offering novel ideas for designing stable and healthy emulsifiers.