The major active components of Astragalus membranaceus (AM) are isoflavones, which exist in the form of various glycosides. Nuruk is a traditional fermentation starter in Korea, and is suitable for the biotransformation of isoflavone glycosides because it contains various microorganisms and enzymes. This study was performed to determine changes in the isoflavones and antioxidant properties of AM fermented (AF) with nuruk over 24 hours. AF was sampled after 0, 2, 4, 6, 12, 18, and 24 h of fermentation, and calycosin 7-glucoside, ononin, calycosin, and formononetin content, and the antioxidant properties of AF were analyzed. The total phenolic content increased with fermentation time, and the ABTS radical scavenging activity increased until 6 h of fermentation and then decreased. During fermentation, the isoflavone glycosides decreased significantly as fermentation time increased. The contents of calycosin and formononetin, which are aglycons of calycosin-7-glucoside and ononin, increased from 100.54 μg/g to 276.84 μg/g and from 56.29 μg/g to 123.04 μg/g, respectively, at 18 h of fermentation. Significant correlations were observed between fermentation time, isoflavone content, and antioxidant properties. The results of this study showed that fermentation with nuruk is suitable for the biotransformation of isoflavones in AM.