Production and consume of mushrooms have grown in the world, and beside these, the nutritional properties and biological active components of fungi have received more attention by researchers. Considering these, a mannofucogalactan was isolated from Flammulina velutipes, and characterized using C-13 and H-1 (obs.), C-13 HMQC nuclear magnetic resonance spectroscopy and methylation analysis. The monosaccharide composition of this polymer was determined by GC-MS and showed Fucp, Manp, and Galp in the molar ratio 20:16:64, respectively. C-13 NMR and H-1 (obs.), C-13 HMQC indicated an anomeric region containing signals (C-1/H-1) at delta 102.9/5.19, 102.0/5.16, and 98.8/5.05 corresponding, sequentially, to non-reducing end of alpha-D-Manp, 3-O-substituted alpha-L-Fucp, and 6-O- and 2,6-di-O-substituted alpha-D-Galp units. Along with methylation analysis, these data showed a structure with a main chain composed of 6-O-substituted Galp units, partially substituted at O-2 by 3-O-D-mannopyranosyl-L-fucopyranosyl, alpha-D-mannopyranosyl, and in a minor proportion, alpha-L-fucopyranosyl groups. Furthermore, some nutritional values of this edible mushroom were evaluated, like amino acid and mineral nutrient contents. (C) 2007 Elsevier Ltd. All rights reserved.