In this study, glycosylated gluten was modified to form a glycosylated gluten covalent complex. The covalent complex then forms nanoparticles through self-assembly under heating to stabilize the Pickering emulsion. The emulsion was loaded with β-carotene to investigate its protective effects on carotene. The research findings showed the thermal and storage stability of Pickering emulsion was the best when the concentration of glycosylated gluten nanoparticles was 4 % and oil-water volume was 40 %. Under these conditions, the smallest and most uniform distribution of emulsion droplets was observed using scanning electron microscopy (SEM) and particle size analysis. Confocal laser scanning microscopy (CLSM) confirmed that the glycosylated gluten nanoparticles formed a three-dimensional network through their interactions at the oil-water interface, inhibited droplet aggregation, and increased the stability of the emulsion. Through simulated digestion in vitro, confirmed β-carotene can be well protected by the prepared Pickering emulsion.