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Basidiomycetes constitute a natural source of biologically active metabolites. Many basidiomycetes have been classified by the National Cancer Institute of the United States as antitumor agents exhibiting an immunomodulatory activity.1 The therapeutic activity is mainly related to polysaccharides or protein-bound polysaccharides, such as glucans, heterogalactans, and glucanproteins, which are present either in the mycelium or in the fruit body.2–6 Among these polysaccharides are b-D-glucans, which are of particular interest because of their pharmacological properties. Most of the b-D-glucans exhibiting a biological activity have been extracted from Grifola frondosa, Ganoderma lucidum, Trametes versicolor, Schizophyllum commune, Lentinula edodes, and Flammulina velutipes | ELSEVIER Available online at www.sciencedirect.com Carbohydrate RESEARCH Carbohydrate Research 339 2004 1129-1134 NMR characterization of the polysaccharidic fraction from Lentinuỉa edodes grown on olive mill waste waters Umberto Tomati a Monica Belardinelli a Emanuela Galli a Valentina Iori a Donatella Capitani b Luisa Mannina Stephane Vielb c and Annalaura Segreb aIstituto di Bioỉogia Agroambientaỉe e Forestaỉe CNR Area deỉỉa Ricerca di Roma I-00016 Monterotondo Scaỉo Rome Itaỉy Istituto di Metodoỉogie Chimiche CNR Area deỉỉa Ricerca di Roma I-00016 Monterotondo Scaỉo Rome Itaỉy cFacoỉta di Agraria Dipartimento S.T.A.A.M Università degỉi Studi deỉ Moỉise I-86100 Campobasso Itaỉy Received 25 July 2003 received in revised form 11 February 2004 accepted 14 February 2004 Abstract A high-field NMR study of the polysaccharidic fraction extracted from Lentinuỉa edodes mycelium grown on olive mill waste waters is reported. Diffusion-ordered NMR spectroscopy DOSY was applied to the polysaccharidic fraction. The results showed the presence of two polysaccharides of different sizes whose structures were revealed using one- and two-dimensional NMR techniques. These two polysaccharides were identified as xylan and lentinan. 2004 Elsevier Ltd. All rights reserved. Keywords Lentinula edodes Olive mill waste waters Lentinan Xylan NMR DOSY 1. Introduction Basidiomycetes constitute a natural source of biologically active metabolites. Many basidiomycetes have been classified by the National Cancer Institute of the United States as antitumor agents exhibiting an immunomodulatory activity.1 The therapeutic activity is mainly related to polysaccharides or protein-bound polysaccharides such as glucans heterogalactans and glucanproteins which are present either in the mycelium or in the fruit body.2-6 Among these polysaccharides are p-D-glucans which are of particular interest because of their pharmacological properties. Most of the p-D-glu-cans exhibiting a biological activity have .