Chemical and Proteomic Characterization of Bioactive Protein-Enriched Fraction from Schizophyllum commune with Anticancer Activity in HeLa Cells

Abstract

Schizophyllum commune, commonly known as the split gill mushroom, is an edible fungus recognized as a source of bioactive compounds with potential applications in the food and biotechnology industries. In this study, protein-enriched fractions were isolated from the fruiting bodies of S. commune using ion-exchange chromatography coupled with a bioassay-guided approach. Cytotoxic activity was evaluated in HeLa cells using MTT and colony formation assays. Among the fractions, the protein-enriched fraction F2 exhibited antiproliferative activity with an IC50 value of 0.37 μg/mL and significantly inhibited colony formation, while showing minimal effects on normal HaCaT cells. Flow cytometry and fluorescence staining further supported the involvement of apoptosis in F2-treated HeLa cells. Chemical and proteomic characterization of the protein-enriched fraction F2 by SDS-PAGE and LC–MS/MS identified 64 protein groups. Gene ontology analysis revealed proteins involved in diverse biological processes, while physicochemical profiling demonstrated a heterogeneous protein composition with isoelectric points ranging from 4 to 7. These findings provide insight into the chemical and proteomic composition of protein-enriched fraction F2. The results support further investigation of S. commune-derived protein-enriched fractions as a source of bioactive proteins, particularly in bioactivity-guided fractionation studies.