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Three polysaccharides (WZP1, WZP2, WZP3) and their Se-enriched products (SeWZP1, SeWZP2 and SeWZP3) were obtained from Pleurotus ostreatus using a simple, rapid method and HNO 3 -Na 2 SeO 3 method, respectively. The molecular weight distribution profiles of all samples except SeWZP2 showed double peaks. The average molecular weights (Mw) of WZP1–3 were 48.6 kDa, 20.2 kDa and 11.8 kDa, respectively, and of SeWZP1–3 were 19.6 kDa, 37.7 kDa, 14.5 kDa, respectively. The complexity of monosaccharide composition of WZP1–3 was inversely proportional to the ethanol concentration used in the ethanol precipitation process. Additionally, the results of biological activity tests indicated that α-glucosidase inhibitory activity of WZP1–3 was related to the molecular weight and the monosaccharide composition complexity. The selenized modification can improve the α-glucosidase-inhibiting, hydroxyl radical-scavenging activity of P. ostreatus polysaccharides. Therefore, by improving their bioactivities by selenization, the polysaccharides of P. ostreatus could be utilized as a natural health food supplement. Unlabelled Image • Three different polysaccharides were obtained from Pleurotus ostreatus using a simple and rapid method. • Se-enrich polysaccharides were successfully obtained using the HNO 3 -Na 2 SeO 3 method. • The selenized modification triggered potential the glucose to be isomerized into galactose. • Selenized modification could obviously elevate the α-glucosidase inhibitory, hydroxyl radical scavenging activities of original polysaccharides. • Se-enrich products could be explored as novel antidiabetic healthcare products. [ABSTRACT FROM AUTHOR]