Molecular Mechanism of Rice Protein Amyloid Fibrils in Modulating Gel Properties of Northern Pike (Esox lucius) Muscle Protein

) myofibrillar protein (MP) forms inherently weak gels due to endogenous proteolytic activity and the low thermal stability of fish myosin, limiting its application in surimi products. This study investigated the reinforcing effect and underlying mechanism of rice protein amyloid fibrils (RFs) on pike MP gels. Dynamic rheology revealed that RFs increased both the storage and loss moduli in a concentration-dependent manner, with the 5% group exhibiting an approximately threefold increase in the G' at 100 rad/s relative to the control. The gel strength, hardness, and chewiness increased progressively with the RF content, whereas the water-holding capacity peaked at 1-3% RFs and declined sharply at 5% RFs. Microstructural imaging showed that moderate RF levels promoted a dense, homogeneous network architecture, while excessive RFs induced phase separation and structural heterogeneity. Hydrophobic interactions and hydrogen bonds were strengthened via RF incorporation, while disulfide bonds decreased monotonically with the increasing fibril concentration. FTIR spectroscopy revealed an α-helix-to-β-sheet transition, with the β-sheet content reaching a maximum of 49.37% at 3% RFs, and SDS-PAGE confirmed that the RF-MP interactions were predominantly non-covalent in nature. These results demonstrate that RFs reinforce pike MP gels through a molecular mechanism involving rigid fibrils acting as structural scaffolds within the protein network and a progressive shift from disulfide-mediated covalent crosslinking toward non-covalent stabilization via hydrophobic interactions and hydrogen bonding. The 1-3% RF range delivers the most balanced gel properties, while excessive fibril loading at 5% induces over-aggregation and impairs water retention. These findings establish amyloid fibrils as effective structural modifiers for freshwater fish gel products and provide a mechanistic basis for their application in surimi processing.