Structural features of a pectic arabinogalactan with immunological activity from the leaves of Diospyros kaki

A water-soluble acidic heteroglycan, DL-3Bb, isolated from the leaves of Diospyros kaki, had [alpha](D)(20) - 19.9degrees (c 0.30, water), and contained rhamnose, arabinose, xylose, galactose and galacturonic acid in the molar ratio of 1.0:4.5:0.7:1.5:1.0. About 44% of the galacturonic acid existed as its methyl ester, and O-acetyl groups (approx 5.7%) were also identified. Its molecular weight was determined to be 9.0 x 10(5) Da by high-performance gel-permeation chromatography. Its structural features were elucidated by a combination of methylation analysis, periodate oxidation, two steps of partial acid hydrolysis, and H-1 and C-13 NMR spectroscopy and ESI mass spectrometry. The data obtained indicated that DL-3Bb possessed a backbone of a disaccharide of [--> 4)-alpha-GalAp- (1 --> 2)-alpha-Rhap -(1 -->], with approx 58.7% substitution at O-4 of the rhamnopyranosyl residues by beta-(1 --> 4)-linked xylopyranosyl residues, and by beta-(1 --> 3) and beta-(1 --> 6)-linked galactopyranosyl (galactan) residues. The side chains were further substituted by arabinofuranosyl residues at O-2 by beta-(1 --> 4)-linked xylopyranosyl residues and at O-3 by beta-(1 --> 6)-linked galactopyranosyl residues. Preliminary tests in vitro revealed that it could stimulate LPS-induced B lymphocyte proliferation, but not for ConA-induced T lymphocyte proliferation. It was proposed that the acid-labile arabinofuranosyl residues in the side chains would not be needed for the expression of the enhancement of the immunological activity, and that the presence of GalAp in the backbone has an important, but not crucial effect on the expression of the activity. (C) 2003 Elsevier Science Ltd. All rights reserved.