Glycobiology, 2002, Vol. 12, No. 4 273-282
© 2002 Oxford University Press
Characterization of a new 
-L-fucosidase isolated from the marine mollusk Pecten maximus that catalyzes the hydrolysis of -L-fucose from algal fucoidan (Ascophyllum nodosum)
2Laboratoire de Recherches sur les Macromolécules, UMR 7540 CNRS, Université Paris 13, avenue J.-B. Clément, 93430 Villetaneuse, France; 3Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, UMR 8601 CNRS, Université René Descartes, 45 rue des Saints-Pères, 75270 Paris, France; and 4Laboratoire de Chimie Structurale Organique et Biologique, UMR 7613 CNRS, Université Paris 6, 4 place Jussieu, 75252 Paris, France
Algal fucoidan is an 
-L-fucose-based polysaccharide endowed with important biological properties for which the structure has not yet been fully elucidated. In an attempt to implement new enzymatic tools for structural study of this polysaccharide, we have found a fucosidase activity in the digestive glands of the common marine mollusk Pecten maximus, which is active on a fucoidan extracted from the brown algae Ascophyllum nodosum. We now report the purification and characterization of this -L-fucosidase (EC 3.2.1.51). The enzyme was purified by three chromatographic steps, including an essential affinity chromatography based on the glycosidase inhibitor analog 6-amino-deoxymannojirimycin as the ligand. The purified -L-fucosidase is a tetrameric glycoprotein of 200 kDa that hydrolyzes the synthetic substrate p-nitrophenyl -L-fucopyranoside with a Km value of 650 µM. This enzyme has high catalytic activity (85 µmol · min–1 · mg–1) compared with the other known fucosidases and also possesses an unusual thermal stability. The purified -L-fucosidase is a retaining glycosidase. The activity of the purified fucosidase was determined on two structurally different fucoidans of the brown algae A. nodosum and Fucus vesiculosus to delineate glycosidic bond specificity. This report is to our knowledge the first demonstration of a fucosidase that can efficiently release -L-fucose from fucoidan.
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