Protein-flavonoid conjugation is considered to effectively enhance the functionality of proteins, although how different binding modes affect the conformation and antioxidative properties of these conjugates has yet to be revealed. Herein, myofibrillar protein (MP)-luteolin (Lut) conjugates were noncovalently and covalently constructed using equivalent amounts of Lut (10.00, 20.11, and 69.60 μmol/g protein). Fluorescence quenching confirmed that hydrophobic interactions were the main forces in noncovalent MP-Lut conjugates and that the binding was entropy-driven. Liquid chromatography-tandem mass spectrometry results confirmed that Lut could be covalently grafted with MP after alkaline treatment. Proteomics analysis identified that most graft sites were located on the myosin subunits. Intriguingly, in vitro results showed that the antioxidant activity was barely affected by the MP-Lut binding modes. This work provides a theoretical basis for the application of MP-Lut noncovalent/covalent complexes as functional components.
Keywords: antioxidant activity; covalent modification; flavonoid protein conjugates; noncovalent modification.