The paper dealt with the characterization of polysaccharide of Paecilomyces lilacinus NH-PL-03 strain. First, we extracted and purified exude polysaccharide from the fungal fermentation broth by ethanol depositing method. Second, the proteins were removed by the Sevage method from the crude polysaccharide. Third, the purified polysaccharide (EP-1) was obtained after Superdex G-75 column separation. The results of UV-spectrometer and Sephacryl S-200 HR chromatography experiments showed that the EP-1 was a homogeneous pure polysaccharide with molecular weight of 35.2 kDa. Tested by paper chromatography analysis using the complete hydrolysis by sulfuric acid, we found that the EP-1 comprise single component as glucose. The chemical structure of EP-1 was confirmed as a kind of linear glucan linked by beta-(1,3) linkage. The Congo red reaction performed that EP-1 probable presented a triple-helical conformation in the dilute alkali.