Facile and moderate immobilization of proteases on SPS nanospheres for the active collagen peptides

Although collagen peptides have been proved to possess wide applications in functional foods, cosmetics, medical materials and pharmaceuticals, the production of collagen peptides are deeply affected by proteases and substrate. In this study, the scalable-synthesis sulfonated polystyrene (SPS) nanospheres were utilized as accessible supports for efficient subtilisin immobilization. Detailed characterizations through SEM-EDS, TEM, TGA and FT-IR confirmed the undamaged formation of the SPS-subtilisin. Owing to the moderate hydrophobic effect and electrostatic interaction, the SPS-subtilisin could achieve 397.15 mg/g enzyme loading and 77.3% activity recovery. The tilapia skin collagen, as a resource-rich raw material, was hydrolyzed by the prepared immobilized subtilisin. The antioxidant activity of the attained peptides was verified. With the mass spectrometry and molecular docking analysis of product peptides sequences, representative peptides were synthesized and their anti-oxidation capacity and mechanism were affirmed, which further verified the undiminished catalytic ability of immobilized subtilisin.