This study was performed to know the potentialities of the fruits of Opuntia ficus-indica, as a medium for mushroom mycelial culture. Five mushroom mycelial (Agrocus blazei, Grifola frondosa, Hericium erinaceum, Innonotus obliquus, Phellinus linteus) were frown on the malt extract broth (MEB) and the cactus broth medium (CB). The submerged culture mixtures were extracted using equal volume of ethyl acetate, and their extract yields, total polyphenol contents, and some physiological activities were compared with each other Each extract from mycelial culture grown on CB medium showed remarkable enhancement in physiological activities compared with each counterpart grown on MEB. Among five mycelial cultures grown on CB medium, the extract yield and polyphenyl content were highest in the extract from Grifola frondosa (extract yield, 0.4 g/L and polyphenol content, 22.7%). Also, the extracts from Grifola frondosa showed the highest physiological activities, such as DPPH radical scavenging ($IC_{50}=362.9{\mu}g/ml$), xanthine oxidase inhibition (about 80% at $500{\mu}g/ml$), and superoxide radical scavenging (about 80% at $500{\mu}g/ml$), and NO production inhibition ($IC_{50}=43.1{\mu}g/ml$) in LPS-stimulated RAW 264.7 cells. This result suggests that the fruit of Opuntia ficus-indica can be used as a culture medium for improving the functional properties of various mushroom mycelia.