Tricholoma matsutake, a rare fungus, is highly valued for its remarkable nutritional content, making it a sought-after biomass resource in both the cosmetics and food sectors. However, its scarcity and restricted natural growth impede large-scale utilization. An endophytic fungus, Umbelopsis sp. TM01, was successfully isolated from the fruiting body of T. matsutake. Research on the biological activity and cytotoxicity of the extract from Umbelopsis sp. TM01 (UFE) and the extract from T. matsutake (TME) showed that UFE outperformed TME in biological activity and had lower cytotoxicity. In the cosmetics-relevant bio-functions, UFE exhibited more potent anti-tyrosinase activity, greater anti-wrinkle efficacy and comparable wound-healing effect to that of TME. UFE was safe for human dermal fibroblasts even at 10% concentration. Metabolomic analysis revealed UFE had diverse secondary metabolites. All in all, Umbelopsis sp. TM01 has great potential as a substitute for T. matsutake in the cosmetics industry.