Macrophage stimulating polysaccharide purified from peels of grape (Vitis labrusca)

To characterize the macrophage stimulating polysaccharide in grape (Vitis labrusca) peels, the active crude polysaccharide (VL-3) has been fractionated from the hot-water extract of grape peels. A macrophage stimulating polysaccharide-rich fraction (VL-3IIb-1-1) was purified from VL-3 by 3 successive column chromatographies on DEAE-Sepharose CL-6B, Sepharose CL-6B, and Sephacryl S-300. VL-3IIb-1-1 was eluted as a single peak on high performance liquid chromatography (HPLC) and its molecular weight was estimated to be 194 kDa. VL-3IIb-1-1 consisted mainly of Ara and Gal in addition to uronic acid (GalA+GlcA) (molar ratio 1.00:0.81:0.72). Methylation analysis indicated that VL-3IIb-1-1 consisted mainly of terminal Araf, 4- or 5-linked Ara, 2,4-branched Rha, 6- or 3,4- or 3,6-branched Gal, and 3,4,6-branched Glc. Single radial gel diffusion also indicated that VL-3IIb-1-1 showed an intermediate reactivity with beta-glucosyl-Yariv antigen. In addition, oral administration of VL-3IIb enhanced the stimulatory responses of macrophage stimulating activity ex vivo. Therefore, VL-3IIb-1-1 purified from grape peels is suggested to be pectic polysaccharide with arabino-3,6-galactan, and it is assumed that VL-3IIb-1-1 plays an important role for expression of its activity.