Separation, purification, structures and anticoagulant activities of fucosylated chondroitin sulfates from Holothuria scabra

In this study, a new fucosylated chondroitin sulfate (HsG) with an average molecular weight of 69.1 kDa was isolated from sea cucumber Holothuria scabra. We investigated the structure of the HsG by adopting monosaccharide composition analysis, disaccharide composition analysis, IR,1H and13C NMR spectra and methylation analysis. According to methylation results of desulfated/carboxyl-reduced polysaccharides and the analysis of unsaturated disaccharides generated through the enzymolysis of the defucosed polysaccharides, it is shown that each branch is formed by one fucopyranosyl residue, wherein 55.7% of the fucopyranosyl residues are linked to the O-6 position of the N-acetylgalactosamine moiety, 21.2% of the fucopyranosyl residues are linked to the O-3 position of β-d-glucuronic acid, 13.0% of the fucopyranosyl residues are linked to the O-4 positions of the N-acetylgalactosamine moiety, and 10.1% of the fucopyranosyl residues are not linked to sulfate groups on the backbone. The backbone →4)GlcUAβ(1 → 3)GalNAcβ(1→ and sulfated fucose branches were composed of the β-d-glucuronic acid, N-acetyl-β-d- galactosamine, α-l-fucose and sulfate groups by the molar ratio of 1:1.72:2.34:3.29. The anticoagulant activities of the HsG was evaluated and compared with heparin. The result showed that the HsG could prolong the activated partial thromboplastin time.