The Reconstruction of Sesame Protein-Derived Amyloid Fibrils Alleviates the Gastric Digestion Instability of β-Carotene Nanoparticles

In this study, the structural changes and reconstruction mechanism of sesame protein-derived amyloid fibrils under varied digestive parameters (pepsin concentration, digestive pH and ionic strength) during gastric digestion were investigated, and the effect of fibril reconstruction on the gastric digestion stability of β-carotene nanoparticles was also explored. The results demonstrated that amyloid fibrils underwent a three-stage dynamic process of enzymatic hydrolysis, regeneration and degradation during gastric digestion. The pepsin concentration of 2 mg/mL was found to promote the balance between fibril hydrolysis and regeneration. The fibrils displayed a pronounced regenerative capacity at pH values of 1.5 and 2.5, whereas at pH 3.5, which was proximal to the isoelectric point of protein, aggregation and precipitation were observed. Furthermore, it was found that 10 mM NaCl exerted minimal influence on fibril stability, whereas the higher concentrations of salt ions were shown to obstruct regeneration and promote aggregation. Analyses through SDS-PAGE, GPC, and MALDI-TOF-MS revealed a gradual reduction in the molecular weight of the fibrils during gastric digestion, with certain fragments reaggregating to form new fibril structures. The fibril-based delivery system formed a stable protective structure for β-carotene nanoparticles, which not only prevented their aggregation but also facilitated their release in the small intestine.