Purification, structure identification and immune activity of a neutral polysaccharide from Cynanchum Auriculatum
The crude polysaccharides CAPS and CAP of Cynanchum Auriculatum , which were prepared by degrading starch by single-enzymatic method (α-amylase) and double-enzymatic method (α-amylase and glucoamylase) respectively, were compared. CAP had good water solubility and higher non-starch polysaccharide content. A homogeneous neutral polysaccharide CAP-W, with the degree of acetylation about 17 %, was obtained from CAP by anion exchange column chromatography. Its detailed structure was identified by various methods. CAP-W, with the weight average molecular weight of 8.4 kDa, was composed of mannose, glucose, galactose, xylose, and arabinose in a molar ratio of 1.27:1.00:0.25:0.10:1.16. The backbone included β -1,4-Man p , β -1,4,6-Man p , β -1,4-Glc p and β -1,4,6-Glc p residues, with branches at the O-6 position of β -1,4,6-Man p and β -1,4,6-Glc p residues, consisting of α -T-Ara f , α -1,5-Ara f , α -1,2,5-Ara f , α -1,3,5-Ara f , T-Xyl p ,1,4-Xyl p , β -T-Man p and β -T-Gal p residues. In vitro immunological experiments suggested that CAP-W improved the phagocytic ability of macrophages, stimulated the release of NO, TNF-α and IL-6 from RAW264.7 cells, promoted the expression of NF-κB and caused nuclear translocation of NF-κB p65.
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