Qun Dong

Structural characterization of a water-soluble β-d-glucan from fruiting bodies of Agaricus blazei Murr

A beta-D-glucan, Ab2-2N, was isolated from the hot-water extract of fruiting bodies of Agaricus blazei Murr by ethanol precipitation, anion-exchange and gel-permeation chromatography. Its structure was investigated by composition analysis, methylation analysis, Smith degradation, mild hydrolysis, and NMR spectroscopy. It contains a (1-->6)-linked beta-D-glucopyranosyl backbone, with one side chain consisting of terminal and 3-substituted beta-D-glucopyranosyl residues attached at O-3 for every three backbone residues.

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) -19.9 degrees (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 9.0x10(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 1H and 13C 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.

Structural elucidation of a new arabinogalactan from the leaves of Nerium indicum.

A polysaccharide fraction, NIB-2, was obtained from the 3% aqueous sodium carbonate extract of Nerium indicum leaves using anion-exchange chromatography and gel-permeation chromatography. It was found to be composed of rhamnose, arabinose, galactose, in the ratios of 1.0:10.4:4.4, along with 4% of galacturonic acid. The results of methylation analysis, periodate oxidation, partial acid hydrolysis, pectinase treatment, and 13C and 1H NMR spectroscopy indicate that it is mainly an arabinogalactan having a backbone of 1,6-linked beta-Galp, with branches at O-3, consisting of terminal, 1,5-, and 1,3,5-linked arabinofuranosyl residues, and a small proportion of galactosyl residues at the termini. Rhamnose and galacturonic acid arose from a contaminating rhamnogalacturonan.

Structural Characterization of an Arabinogalactan from Platycodon grandiflorum Roots and Antiangiogenic Activity of Its Sulfated Derivative

A water-soluble polysaccharide, PGAW1, with an average molecular mass of 9.2 kDa determined by high performance gel permeation chromatography (HPGPC), was isolated from radix of Platycodon grandiflorum . Monosaccharide composition analysis indicated that PGAW1 contains Ara and Gal in the molar ratio of 1.42:1.0. Using methylation analysis, partial hydrolysis, endo-1,4-β-d-galactanase digestion, NMR and ESI-MS, PGAW1 was determined to possess a backbone consisting of 1,4- and 1,6-linked galactopyranosyl residues, with branches attached to O-3 of 1,6-linked galactose residues. By the chlorosulfonic acid-pyridine method we produced a sulfated derivative of PGAW1, Sul-w1, with a substitution degree of 1.52. The substitution was at O-6 on 1,4-linked Gal residues according to the (13)C NMR spectra. Bioactivity test showed that Sul-w1 could inhibit tube formation by human microvascular endothelial cells (HMEC) in a dose-dependent manner.

Structure and potential immunological activity of a pectin from Centella asiatica (L.) Urban.

S3A was a RG-I pectin isolated from Centella asiatica that contained Rha, Ara, Gal, Glc and GalA in molar ratio of 1.0:0.6:1.5:0.2:1.1 and had been found to have a backbone composed mainly of the disaccharide repeat unit, -->4)-alpha-D-GalpA-(1-->2)-alpha-L-Rhap-(1-->. Based on methylation analysis, NaIO4 oxidation, partial acid hydrolysis and lithium-treatment, the structural features were elucidated. Side chains of S3A were predominantly linked to O-4 of 1,2,4-linked alpha-L-Rhap. The side chains are comprised of arabinosyl chains, galactosyl chains, arabinogalactosyl chains and short glucosyl chains. A total of 45% Rhap in the backbone was substituted by side chains. The arabinosyl residues were mostly distributed in the arabinosyl side chains. According to the immunological results of S3A and its degraded derivatives, S3A had no immunological activity, but its derivatives had immuno-stimulating activities to some extent.

Structural characterization of a pectic polysaccharide from Nerium indicum flowers.

A polysaccharide fraction, J6, was isolated from the hot-water extract of flowers of oleander Nerium indicum Mill., using ethanol precipitation, cetyltrimethylammonium bromide (CTAB) complexing, anion-exchange chromatography and gel permeation chromatography. J6 was found to contain L-rhamnose, L-arabinose, D-galactose, and D-galacturonic acid, in the ratio of 10.1:49.8:30.1:10.0. Its structure was investigated by methylation analysis, periodate oxidation, Smith degradation, partial acid hydrolysis, electrospray ionization mass spectrometry and NMR spectroscopic methods. It was found that J6 is an RG-I type polysaccharide, which contains a rhamnogalacturonan backbone, with various branches attached to O-4 of L-rhamnose. The branches probably involve (1-->4)-beta-D-galactan, branched L-arabino-(1-->3)(1-->6)-beta-D-galactan, and (1-->5)-alpha-L-arabinan. J6 stimulated NO production of macrophage RAW264.7 cells in a preliminary test.

Structural characterization of a polysaccharide from Chrysanthemum morifolium flowers and its antioxidant activity.

The flowers of Chrysanthemum morifolium were extracted by boiling water, and a water-soluble polysaccharide (CMJA0S2) with a molecular weight of 6.5 kDa was isolated by anion-exchange chromatography on a DEAE-cellulose column and gel permeation chromatography on a Sephacryl S-300 HR column. Monosaccharide composition analysis indicated that CMJA0S2 was composed of galactose, glucose, mannose and arabinose in molar ratio of 4.1: 3.3: 1.0: 2.3. According to linkage analysis, partial acid hydrolysis and NMR spectra, the backbone was shown to contain 1, 4-linked β-Galp, 1, 4-linked β-Glcp and 1, 4-linked β-Manp, with branches substituted at C-6 of 1, 4-linked β-Manp by 1, 6-linked β-Galp and at O-6 of partial 1, 4-linked β-Galp substituted by T-linked α-Glcp. About 40% of 1, 6-linked β-Galp with T-linked α-Glcp was substituted at O-3 by α-Araf-(1→[5)-α-Araf-(1]3. The anti-oxidative analysis showed that CMJA0S2 could scavenge the DPPH radicals and relieve the damage of PC12 cells caused by H2O2, thus it could be regarded as a potential natural antioxidant.

Inducement of cytokine release by GFPBW2, a novel polysaccharide from fruit bodies of Grifola frondosa , through dectin-1 in macrophages.

Polysaccharides, especially β-glucans isolated from various species of mushrooms, are considered as biological response modifiers (BRMs) to be widely used in the treatment of cancer, especially due to their immunostimulatory activity. We herein characterized the structure of a novel water-soluble homogeneous polysaccharide (GFPBW2) from the fruit bodies of mushroom Grifola frondosa and investigated its immunomodulatory activity in vitro. GFPBW2 was purified from the alkali-extracted fractions by stepwise elution with a molecular weight of 26.2 kDa. On the basis of infrared and NMR spectroscopy, methylation and monosaccharide composition analysis, partial acid hydrolysis, and Smith degradation, its structure was elucidated to possess a backbone consisting of β-d-1,3- and β-d-1,4-linked glucopyranosyl residues, with branches attached to O-6 of β-d-1,3-linked glucopyranosyl residues. Functionally, it is an effective inducer of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) secretion in murine resident peritoneal macrophages. Using quartz crystal microbalance (QCM) analysis, we found that GFPBW2 could bind dendritic cell-associated C-type lectin-1 (Dectin-1) with an affinity constant (Kd) value of 1.08 × 10 (-7) M, while it could activate Syk and enhance TNF-α production in RAW264.7 cells overexpressing wild type but not mutant Dectin-1. Furthermore, Syk, NF-κB signaling, and cytokine release in resident peritoneal macrophages induced by GFPBW2 could be significantly inhibited by a specific Dectin-1 blocking reagent, Laminarin. These data suggested that GFPBW2 might be a potential ligand of Dectin-1, and the potential of GFPBW2 to activate macrophage through triggering cytokine secretion might be attributed, at least in part, to the involvement of Dectin-1.

Structural characterization and effect on anti-angiogenic activity of a fucoidan from Sargassum fusiforme.

A fucoidan FP08S2 was isolated from the boiling-water extract of Sargassum fusiforme, purified by CaCl2 precipitation and chromatography on DEAE-cellulose and Sephacryl S-300. FP08S2 contained fucose, xylose, galactose, mannose, glucuronic acid, and 20.8% sulfate. The sulfate groups were attached to diverse positions of fucose, xylose, mannose, and galactose residues. The backbone of FP08S2 consisted of alternate 1,2-linked α-D-Manp and 1,4-linked β-D-GlcpA. Sugar composition analysis and ESI-MS revealed that the oligosaccharides from branches contained fucose, xylose, galactose, glucuronic acid and sulfate. FP08S2 could significantly inhibit tube formation and migration of human microvascular endothelial cells (HMEC-1) dose-dependently. These results suggested that the fucoidan FP08S2 from brown seaweeds S. fusiforme could be a potent anti-angiogenic agent.

A novel water-soluble β-d-glucan isolated from the spores of Ganoderma lucidum

Ganoderma lucidum is an edible and medicinal mushroom used widely in East Asia. In recent years, its spores have been used as a supplement in combination with other forms of antitumor therapies. The cell wall of Ganoderma lucidum spores contains a high amount of polysaccharides. In this study, a neutral polysaccharide, GLSA50-1B, was isolated from sporoderm-broken spores of Ganoderma lucidum, by hot-water extraction, graded ethanol precipitation, anion-exchange chromatography, and gel permeation chromatography. Using sugar compositional analysis, methylation analysis, partial acid hydrolysis, acetolysis, and NMR and ESI-MS spectroscopy, GLSA50-1B was elucidated to be a novel β-D-glucan featured by a 1,6-linked β-D-Glcp backbone with different length of branches consisting of terminal and 1,4-linked Glcp residues, attached to O-4 of alternative Glc residues in the backbone.