An a-L-fucosidase [EC 3.2.1. -] has been purified from Bacillus fulminans by ammonium sulfate fractionation and subsequent column chromatography on Sephadex G-200, on DEAE-cellulose and on hydroxylapatite. Free fucose liberated by enzyme action was determined by microdiffusion method. The enzyme has a molecular weight of 70,000 to 80,000, hydrolyzes specifically a terminal, α-(l→2)-fucosidic bond of glyco-proteins, glycopeptides and oligosaccharides and fails to cleave (1→3)- and (l-→4)-linkages of ar-L-fucosides in lacto-N-fucopentaose III and lacto-N-fucopentaose II, respectively, as well as -nitrophenyl a-L-fucoside. It has apparent Km values of 3.3×10−4M, 1.6×10−3M, 2.0×10−3M, and 2.5×10−3M for bound fucose in desialyzed PSM, glycopeptide of desialyzed PSM, lacto-N-fucopentaose I and 2′-fucosyllactose, respectively.