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Pleurotus djamor var. roseus is an edible mushroom, contain rich source of nutrients and is popularly cultivated for edible purpose world wide. However, the medicinal properties of P. djamor var. roseus are not well documented. Hence, the present study was focused on isolation, purification and characterization of medicinally important polysaccharide.1. P. djamor var. roseus was cultivated using paddy straw as substrate and 600 g of fresh fruiting bodies were harvested with biological efficiency of 120%.2. The total sugar and protein content of P. djamor var. roseus was estimated as 51% and 39% respectively.3. Partially purified polysaccharide from P. djamor var. roseus was extracted by hot water extraction and precipitation by ice cold ethanol. It was further purified by DEAE cellulose column followed by Sepharose 6B column. 30mg of purified polysaccharide was obtained from 200 g of dried powder of P. djamor var. roseus.4. The hydrolyzed purified polysaccharide exhibited three spots which correspond to lactose, glucose and xylose.5. FT-IR spectrum of purified polysaccharide confirmed the presence of a polysaccharide moiety, a xylopentose type of compound.6.