Mushroom polysaccharides are important bioactive compounds derived from mushrooms with various beneficial properties. In this study, the chemical characterization and bioactivities of polysaccharide extracts from four different edible mushrooms, Clavariadelphus truncatus Donk, Craterellus tubaeformis (Fr.) Quél., Hygrophorus pudorinus (Fr.) Fr., and Macrolepiota procera (Scop.) Singer were studied. Glucose (13.24-56.02%), galactose (14.18-64.05%), mannose (2.18-18.13%), fucose (1.21-5.78%), and arabinose (0.04-5.43%) were identified in all polysaccharide extracts by GC-MS (gas chromatography-mass spectrometry). FT-IR (Fourier transform infrared spectroscopy) confirmed the presence of characteristic carbohydrate patterns. 1H NMR suggested that all polysaccharide extracts had α- and β-d-mannopyranose, d-glucopyranose, d-galactopyranose, α-l-arabinofuranose, and α-l-fucopyranose residues. Approximate molecular weights of polysaccharide extracts were determined by HPLC (high-performance liquid chromatography). The best antioxidant activity was found in M. procera polysaccharide extract in DPPH• (1,1-diphenyl-2-picrylhydrazyl) scavenging (39.03% at 800 μg/mL), CUPRAC (cupric reducing antioxidant capacity) (A0.50: 387.50 μg/mL), and PRAP (phosphomolybdenum reducing antioxidant power) (A0.50: 384.08 μg/mL) assays. C. truncatus polysaccharide extract showed the highest antioxidant activity in ABTS•+ scavenging (IC50: 734.09 μg/mL), β-carotene-linoleic acid (IC50: 472.16 μg/mL), and iron chelating (IC50: 180.35 μg/mL) assays. Significant anticancer activity was found in C. truncatus polysaccharide extract on HT-29 (IC50: 46.49 μg/mL) and HepG2 (IC50: 48.50 μg/mL) cell lines and H. pudorinus polysaccharide extract on the HeLa cell line (IC50: 51.64 μg/mL). Also, H. pudorinus polysaccharide extract possessed prominent AChE (acetylcholinesterase) inhibition activity (49.14% at 200 μg/mL).