Metabolomic Revelations into the Dynamic Transformations Across Various Developmental Stages of Coprinus comatus Through UHPLC-Q-Orbitrap-HRMS Analysis

<b>Background</b>: Dietary supplements and functional foods derived from mushrooms have gained increasing popularity. Among these, <i>Coprinus comatus</i> stands out due to its excellent flavor and high nutritional value. However, its susceptibility to autolysis and short shelf life significantly limits its utilization. Although a few studies have attempted to elucidate the autolysis mechanism of <i>C. comatus</i>, only few research has been conducted on the detailed metabolic changes occurring during its growth cycle. <b>Objectives</b>: By conducting a dynamic metabolic profiling analysis of <i>C. comatus</i> metabolites across different developmental stages and tissue parts, this study aims to elucidate the variations in its metabolic composition. <b>Methods</b>: In this study, fruiting bodies of <i>C. comatus</i> were cultivated and collected at four distinct developmental stages. These samples were further divided into cap and gills (CG) and stipe (ST) tissues. Subsequently, UHPLC-Q-Orbitrap was employed for non-targeted dynamic metabolomics analysis of <i>C. comatus</i> samples. The identification of analytes was performed using Compound Discovery 3.3. Then, differential accumulated metabolites (DAMs) between CG and ST at the same stage and CG or ST between adjacent stages were identified. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was conducted to identify potential contributors to the observed metabolic changes. In addition, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity of samples was determined. <b>Results</b>: A total of 490 metabolites were annotated, and the most abundant metabolite groups were lipids, alkaloids, amino acids and their derivatives. It revealed that the metabolites of the ST remained relatively stable across the four growth stages, whereas autolysis induced significant alterations in the metabolites of the CG. KEGG pathway analysis indicated that these changes were primarily linked to lipid and amino acid biosynthesis and metabolic pathways. Furthermore, DPPH assays demonstrated a significant increase in the free radical scavenging activity of CG following autolysis. <b>Conclusions</b>: The metabolites of <i>C. comatus</i> exhibit dynamic variations across different growth stages and tissue locations. The significant morphological changes in CG induced by autolysis are mirrored by corresponding alterations in its metabolic profile. The enhanced DPPH free radical scavenging activity observed in the autolyzed samples, along with the distribution patterns of bioactive components, provides valuable insights for the efficient utilization of <i>C. comatus</i>.