Valorization of Schizophyllum commune mycelia through green extraction and hydrolysate preparation: Proteomics-informed antioxidant potential and Caco-2 cell-based assessment.

Schizophyllum commune mycelia obtained from submerged fermentation were investigated as a sustainable source of bioactive ingredients using an integrated workflow involving green extraction, proteomics, in silico peptide-motif prediction, and Caco-2 cellular assays. Hot-water extraction (HWE) and ultrasound-assisted extraction (UAE) were compared at different biomass loads and extraction times. Protein solubilization increased strongly with biomass load regardless of extraction method, whereas polysaccharide recovery depended mainly on extraction time. In crude extracts, UAE gave higher phenolic content than HWE. Shotgun proteomics identified 312 high-confidence proteins, from which selected proteins were used as candidate precursor sequences for antioxidant-peptide motif prediction. The hydrolysate, which contained peptides together with polysaccharides and phenolic compounds, maintained >90% Caco-2 viability and significantly reduced H2O2-induced intracellular reactive oxygen species in a dose-dependent manner. These findings provide preliminary support for the further development of the hydrolysate as a multicomponent antioxidant ingredient for functional food applications.