This study analyzed the physicochemical characteristics of Astragalus membranaceus (AM) fermented with seven different mushroom mycelia. Physicochemical characteristics, such as contents of moisture, pH, total reducing sugars, free sugar, and isoflavonoid, were investigated. The moisture content was increased in most of the samples. The pH values of AM fermented with Phellinus linteus and Flammulina velutipes were increased, while the pH of other samples were similar to that of non-fermented AM. The reducing sugar content was in the range of 211.69~391.74 mg/100 g. The extraction yield using water was higher than that when extracted with 80% ethanol. The free sugar content was increased through fermentation with mushroom mycelia. However, the glucose contents of the 80% ethanol and water extracts were decreased. Finally, the calycosin and formononetin contents in 80% ethanol and water extracts of AM fermented with Phellinus linteus were 2,549.24 mg/g, and 827.66 mg/g for calycosin, and 1,366.69 mg/g and 221.28 mg/g for formononetin, respectively. These results suggest that fermentation with mushroom mycelia could be used to increase the bioactivity of AM. The mycelium-fermented AM might be a valuable source of functional material and edible resource for industry.