The activities of xylanase extracted from spent mushroom composts (SMCs) of Coprinus comatus, Auricularia auricular, Pleurotus ostreatus, Pleurotus citrinopileatus, Agrocybe cylindracea, Hericium erinaceus, Hypsizygus marmoreus, and Tremella fuciformis were investigated. The crude extract from T. fuciformis SMC showed high xylanase activity with a value of 255.2 U/mg. Furthermore, this xylanase was purified using a combination of ammonium sulfate precipitation, diethylaminoethyl-cellulose (DEAE-cellulose), and gel filtration column chromatography. The enzyme was purified 20.7-fold with a yield of 43.1% and activity of 5293.8 U/mg. The purified xylanase showed maximum activity at 50 °C and pH 6, retained 80% activity after 1 h incubation at 50 °C, and sustained stability over a wide range of pH values (2 to 10). Under the optimal conditions, the enzyme exhibited a Km value of 2.5 mg/mL towards birchwood xylan. The activity of xylanase was enhanced in the presence of Mg2+, Ca2+, Ba2+, NH4+, and Tween 80, while some metal ions, particularly Fe3+, inhibited its activity. The saccharification of several biomass wastes using the crude xylanase enzyme was studied. The results showed the potential for saccharification of alkaline-pretreated wheat bran solution where 75% saccharification was achieved.