Abstract
Edible mushrooms are increasingly recognized as sources of bioactive compounds with potential health benefits, yet comprehensive characterization of recently cultivated species remains limited. Herein, multi-platform metabolomics and in vitro fermentation were employed to compare their chemical composition and prebiotic potential. GC-MS and UHPLC-HRMS identified over 4800 differential metabolites between RP and AM. Targeted amino acid profiling revealed significantly higher levels of free and acid-extractable amino acids, including essential amino acids, in RP compared to AM, indicating distinct amino acid profiles between the two species. Lipidomics identified over 1500 differential lipids, providing chemotaxonomic markers for species authentication. In vitro fermentation demonstrated that RP crude polysaccharides significantly modulated gut microbiota, enriching beneficial Lachnospiraceae and Ruminococcaceae while suppressing Proteobacteria. Notably, RP and AM polysaccharides exhibited distinct fermentation signatures, suggesting species-specific prebiotic applications. These findings establish RP as a nutritionally superior edible mushroom with promising prebiotic potential.