In this study, ethanol extracts from 2-day fermented black soybean milk (FBE) by immobilized Rhizopus oligosporus NTU5 have been evaluated for both antioxidant and cytotoxic activities. The results reveal that a 2-day FBE had strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging effect (76 %). The extracts were further fractionated by silica gel column chromatography and an unknown compound, FBE5-A, was obtained, which exhibited strong antioxidant activity. IC50 of the DPPH scavenging effect of FBE5-A was 7.5 mg/mL, which is stronger than a commonly used antioxidant, vitamin E (a-tocopherol; 17.4 mg/mL), and similar to vitamin C (ascorbic acid; 7.6 mg/mL). The cytotoxic test demonstrated that extracts of 2-day fermented broth exhibited selective cytotoxic activity towards human carcinoma cells, Hep 3B (IC50=150.2 mg/mL), and did not affect normal human lung fibroblasts, MRC-5 (p<0.05). The results indicate the potential applications of fermented black soybean milk as functional food, pharmaceutical or cancer therapy formula.