Triterpenoid compounds are important bioactivity materials. Morchella is an abundant medicinal fungi found worldwide. In this study, we optimised the isolation and purification of triterpenoid compounds from Morchella mycelium fermentation. The results showed that the triterpenoid compounds yield was 35.22 mg/g, and we also identified two triterpenoid compounds using high-performance liquid chromatography. In addition, we evaluated the anti-tumour and antioxidant activity of the products, and the results showed that triterpenoid compounds from Morchella mycelium fermentation showed good bioactivity. The IC values of four cancer cell lines treated with the triterpenoid compounds for 48 h were 7.20, 14.96, 4.41, and 13.43 mg/mL, respectively. Morphological changes associated with the apoptosis of PC-3 cells were observed using confocal scanning laser microscopy after treatment with triterpenoid compounds for 48 and 72 h. The triterpenoid compounds also exhibited DPPH radical, hydroxyl, and ABTS-free radical scavenging activities in vitro. These results suggest that triterpenoid compounds from Morchella mycelium fermentation, which are found in functional foods and used in the field of pharmacology, might be excellent products for the treatment of cancer and age-related illnesses.