H NMR and LC-MS/MS were used to evaluate taste metabolites and peptide profiles. Among the identified peptides, LPGVAE, LDELEK, DVELSK, LPDEAR, and TTLPDK with high umami scores which threshold in the range of 0.091-0.371 mmol/L were screened by iUmami-SCM and BIOPEP-UWM, and further verified by sensory evaluation. The results of molecular docking suggested that Ser148, Asn150, Ser276, Ser278 of T1R1 and Asn68, Val277, Ala302, Ser306 of T1R3 played a key role in the umami peptides docking. The study revealed continuous enzymolysis of L.edodes could obtain more umami substances and umami peptides, which laid a foundation for researching flavor substances and developing flavor products from L.edodes.