Arabino-3,6-galactan (AMA-1-b-PS2), an intestinal immunomodulatory compound, was purified from the above-ground portion of Astragalus membranaceus (Bunge). Its structure was characterized using sequential enzymatic digestion with exo-α-L-arabinofuranosidase (AFase) and exo-β-D-(1 → 3)-galactanase (GNase), producing small amounts of intermediate-sized and shorter oligosaccharide (AF-PS2-G2 and AF-PS2-G3) fractions, and a large GNase-resistant fraction (AF-PS2-G1). Simultaneous AFase and GNase digestion of the enzyme-resistant fraction produced two long fragments (AF3-PS2-G1-1-1 and AF3-PS2-G1-1-2). Products of GNase digestion of the upper fractions showed decreased intestinal immunomodulatory activity; the GNase-resistant fraction (AF-PS2-G1) retained significant activity. Sugar component, methylation, and FAB-MS analyses indicated that the oligosaccharides consisted of hexosyl tri- to hexa-decasaccharides and hexosyl di- to hepta-saccharides mainly comprising 6-linked Gal(f) and Gal(p); some were partially mono- or di-arabinosylated. These oligosaccharide fractions were attached to the non-reducing terminus of the β-D-(1 → 3)-galactan backbone as side chains at position 6. AMA-1-b-PS2 likely modulates both the systemic and gastric mucosal immune systems.
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