This study was carried out to investigate the physiological activities of the ethanol extract from Gymnopilus spectabilis mycelium (EGM) and of the supernatant obtained from fermentation broth (SGB). The contents of polysaccharides, phenol compounds and total β-glucans of EGM were found to be 80.14%, 3.5 mg/ml and 5.91%, respectively and those for SGB were 78.68%, 3.32 mg/ml and 3.28%, respectively. Both EGM and SGB exhibited dose-dependent nitrate-scavenging abilities at pH 1.2. In addition, both EGM and SGB on the autoxidation rate of the linoleic acid demonstrated powerful antioxidant activities at 1 mg/ml level. With respect to fibrolytic activity, EGM showed 1,180 unit/g, which was the same activity as streptokinase, while SGB was 1,011 unit/g. The angiotensin converting enzyme inhibition activity of EMG determined by both the normal and pretreatment methods were estimated to be 8.2% and 10.2%, respectively. However, SGB showed no corresponding activity. The growth inhibitory effects of EGM on AGS, A549, HeLa and NCTC cells were over 58.88%, respectively. And the growth inhibitory effects of the SGB on HeLa and NCTC cells were 44.92 and 76.76%, respectively. Also, EGM and SGB activated the components of the alternative complement pathway from 51 and 62% at the concentration of 100 mg/ ml. The xanthine oxidase inhibition activities of EGM and SGB (1 mg/ml) were 9.53 and 16.92%, respectively.