Polyporus umbellatus is a medicinal fungus, has been used in traditional Chinese medicine for thousands years for treatment of edema, scanty urine, vaginal discharge, jaundice and diarrhea. The structure of a soluble polysaccharide (named PUP80S1), purified from the sclerotia of Polyporus umbellatus was elucidated by gas chromatography (GC), GC-mass spectrometry and nuclear magnetic resonance spectroscopy. PUP80S1 is a branched polysaccharide containing approximately 8.5% uronic acid and having an average molecular weight of 8.8kDa. Atomic force microscopy of PUP80S1 reveals a globular chain conformation in water. Antioxidant tests, Oxygen radical absorption capacity and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assays indicate that PUP80S1 possesses significant antioxidant activity. But the related polysaccharide, PUP60S2, which contains more uronic acid residues and a higher level of branching, shows better antioxidant activity. These results suggest that structure features of polysaccharides play an important role in their physiological functions.