Galp Residues Research Articles

Structural characterization and prebiotic activity of oligo-galacturonic acids from Osmunda japonica Thunb

An oligosaccharide, WOJP-AOS-b, with a molecular size of 1.8 kDa, was obtained through enzymatic hydrolysis of Osmunda japonica Thunb polysaccharides (WOJP-A). The structural characteristics of WOJP-AOS-b were elucidated using Fourier transform-infrared spectroscopy (FT-IR), Ultraviolet (UV) spectroscopy, Nuclear magnetic resonance (NMR) spectroscopy, and Eelectrospray ionization tandem mass spectrometry (ESI-MS/MS). Structural analysis revealed that galacturonic acid (GalpA) was the predominant monosaccharide component of WOJP-AOS-b, accounting for 90.91% of the total composition. FT-IR and NMR analyses confirmed that WOJP-AOS-b possesses a linear structure comprised of α-1,4-linked GalpA residues. MS/MS results indicated that WOJP-AOS-b consists of α-1,4-linked oligo-galacturonic acids with degrees of polymerization ranging from 2 to 6 (DP 2–6). Moreover, in vitro probiotic activity assays demonstrated that WOJP-AOS-b exerted beneficial effects on intestinal microbiota, particularly Bifidobacterium adolescentis ATCC 15703 and Bifidobacterium breve 689b. WOJP-AOS-b enhanced the optical density, short-chain fatty acid (SCFA) content, and lactic acid content in the culture medium. Furthermore, 16S rRNA analysis suggested that WOJP-AOS-b exhibits potential for treating obesity and inhibiting pathogenic bacteria, indicating its promising applications as a prebiotic.

Fucoidan from brown seaweed Tubinaria decurrens: Structure and structure - anticancer activity relationship

The aims of this study are to determine the structure of a fucoidan from brown seaweed Turbinaria decurrens, to investigate its anticancer activity and structure–activity relationship. SEC-MALLS, IR, ESI-MS and NMR spectra analysis indicated that dominant structure of the fucoidan, with a Mw 122.6 KDa, has a backbone of (1 → 3)– and (1 → 4)–α–L–Fucp residues, branched at C–4, sulfate groups are attached at C–2, C–3 and C–4; branches are (1 → 4)–β–D–Galp residues and sulfated at C–2. The fucoidan was hydrolyzed by HCl aqueous solution to obtain hydrolyzed fucoidans. It is assumed that native and hydrolyzed fucoidans have a rod-like conformation in solution with cross–sectional radius of gyration (Rgc) ranged from 0.53 to 1.52 nm as estimated from SAXS measurements. The fucoidans show great anticancer activity against HT29 human colon cancer cell line with IC50 ranging from 5.41 ± 0.36 to 73.52 ± 2.54 μg/mL. Anticancer activity of the fucoidan could be significantly improved by lowering molecular weight, furthermore, fucoidan required small molecular weight, small molecular weight distribution and rod–like structure with a short branch length for high anticancer activity.

Characterization and biological activity of acidic sugarcane leaf polysaccharides by microwave-assisted hot alkali extraction

As a waste of sugarcane, the added value of sugarcane leaves has not been fully utilized, and sugarcane leaf polysaccharide is an important active component of sugarcane leaves. Using new extraction methods to study the structure and biological activity of polysaccharides from sugarcane leaves is of great significance for the development and application of sugarcane leaves. To determine the structural characteristics and biological activities of different acidic sugarcane leaf polysaccharides (ASLPs) extracted by microwave-assisted hot alkali extraction (MALE), three new acidic sugarcane leaf polysaccharides (ASLP-50, ASLP-70 and ASLP-90) were precipitated by ethanol. The structure of ASLPs extracted by MALE contains protein, sulfate and uronic acid. ASLP-50 (195.88 kDa), ASLP-70 (94.62 kDa) and ASLP-90 (16.37 kDa) were acidic polysaccharides composed of mannose, galacturonic acid, glucose, arabinose and fucose. The results of XRD and NMR showed that ASLPs were amorphous polysaccharides composed of α-L-Araf-(1→ main chain, and its branching points were composed of Arap and GalpA residues. In addition, ASLPs showed significant antioxidant, hypoglycemic, hypolipidemic and antibacterial activities in vitro. The research and application of ASLPs provide a reference for the recycling of sugarcane leaves, which is conducive to the development of food, agriculture and pharmaceutical industries.

Structural characterization of pectic polysaccharides from Amaranth caudatus leaves and the promotion effect on hippocampal glucagon-like peptide-1 level

In this study, decolorized pectic polysaccharides (D-ACLP) with molecular weight (Mw) distribution of 3483– 2,023,656 Da were prepared from Amaranth caudatus leaves. Purified polysaccharides (P-ACLP) with the Mw of 152,955 Da were further isolated from D-ACLP through gel filtration. The structure of P-ACLP was analyzed by 1D and 2D NMR spectra. P-ACLP were identified as rhamnogalacturonan-I (RG-I) containing dimeric arabinose side chains. The main chain of P-ACLP was composed of →4)-α-GalpA-(1→, →2)-β-Rhap-(1→, →3)-β-Galp-(1→ and →6)-β-Galp-(1→. There was a branched chain of α-Araf-(1→2)-α-Araf-(1→ connected to the O-6 position of →3)-β-Galp-(1→. The GalpA residues were partially methyl esterified at O-6 and acetylated at O-3. The 28-day consecutive gavage of D-ALCP (400 mg/kg) significantly elevated the hippocampal glucagon-like peptide-1 (GLP-1) levels in rats. The concentrations of butyric acid and total short chain fatty acids in the cecum contents also increased significantly. Moreover, D-ACLP could significantly increase the gut microbiota diversity and dramatically up-regulated the abundance of Actinobacteriota (phylum) and unclassified Oscillospiraceae (genus) in intestinal bacteria. Taking together, D-ACLP might promote the hippocampal GLP-1 level through the beneficial regulation of butyric acid-producing bacteria in gut microbiota. This study contributed to making full use of Amaranth caudatus leaves for cognitive dysfunction intervention in food industry.

Comparative Studies on the Structure and Biological Activities of Two New Polysaccharides from Tricholoma sinoportentosum (TS-P) and Termitomyces albuminosus (TA-P).

Polysaccharides are important active ingredients of living organisms. In this study, two new polysaccharides, Tricholoma sinoportentosum polysaccharide (TS-P) and Termitomyces albuminosus (TA-P), were extracted and purified using anion exchange column chromatography. The structure of each polysaccharide was identified by HPGPC, FT-IR, HPLC, GC-MS and NMR, and the biological activities were also investigated. The results of the structure identification showed that TS-P was composed of arabinose, mannose, glucose and galactose at a ratio of 1:1:3:2 and its main chain was composed of (1→4)-Arap residues, (1→4,6)-D-Manp residues and two (1→6)-Galp residues. The TA-P was composed of arabinose, glucose and galactose at a ratio of 2:4:8. Its main chain was composed of two (1→4)-β-L-Arap residues, one (1→4)-Glcp residues, three (1→2,6)-Galp residues and five (1→6)-Galp residues. The immunoassay showed that TS-P and TA-P could significantly promote the proliferation of T cells, B cells and RAW264.7 cells. The cell cycle results showed that for B cells and macrophages, TS-P and TA-P mainly affected the G0/G1 phases of the cell cycle; for T cells, TS-P affected G2/M phase, while TA-P mainly affected the G0/G1 phases. TS-P could significantly promote B cells to secrete IgA, IgG and IgD (p < 0.01), while TA-P could significantly promote the secretion of IgA and IgG (p < 0.01). The chemical structure and biological activity of TS-P and TA-P were first studied and compared to lay a theoretical foundation for the application of fungal polysaccharide.

Structural characterization and immunomodulatory activity of an arabinogalactan from Jasminum sambac (L.) Aiton tea processing waste

An arabinogalactan named JSP-1a was isolated from Jasmine tea processing waste by DEAE Sepharose FF and Sephacryl S-200 HR chromatography. Polysaccharide JSP-1a, with an average molecular weight of 87.5 kDa, was composed of galactose (59.60 %), arabinose (33.89 %), mannose (4.81 %), and rhamnose (1.70 %). JSP-1a was found to be a type II arabinogalactan comprising the main backbone of 1, 6-linked Galp residues, and the side chain containing α-T-Araf, α-1,5-Araf, β-T-Galp, β-1,3-Galp, and β-1,4-Manp residues was attached to the O-3 position of β-1,3,6-Galp residues. Evidence from bioactivity assays indicated that JSP-1a possessed potent immunomodulatory effects on RAW264.7 macrophages: treatment with JSP-1a increased phagocytosis, activated NF-κB p65 translocation, and promoted the production of NO, reactive oxygen species (ROS), the tumor necrosis factor (TNF)-α, and interleukin (IL)-6. Furthermore, inhibition of Toll-like receptor 4 caused the suppression of NO release and cytokines secretion, which indicated that TLR-4/NF-κB pathway might play a significant role in JSP-1a-induced macrophages' immune response. The results of this study could provide a theoretical basis of JSP-1a as a safe immunostimulatory functional foods or a treatment for immunological diseases.

Pectin from leaves of birch (Betula pendula Roth.): Results of NMR experiments and hypothesis of the RG-I structure

We report that in birch leaf pectin, rhamnogalacturonan-I (RG-I) and galacturonan (HG) were found as separate polymers rather than domains of a complex macromolecule. RG-I and HG were separated by anion-exchange and size-exclusion chromatography and studied by using NMR spectroscopy. NMR spectra showed that methyl-esterified D-galactosyluronic acid residues were located only in HG. Oligosaccharides of similar structure to the backbone, but without terminal reducing residues in the NMR spectra, were found in RG-I. We hypothesize, these oligosaccharides and RG-I backbone can be covalently bound due to its co-eluted of from DEAE-cellulose and Sepharose CL-4B. This result differs from the classical RG-I model, which assumes that all Rhap and GalpA residues are located only in the RG-I backbone. In the heteronuclear multiple bond correlation (HMBC) and rotating frame Overhauser effect spectroscopy (ROESY) spectra, the correlation peaks confirming the substitution of 2,4-rhamnose residues at O-4 by only single D-galactose residues were identified.

Galactofucoidans from Sargassum fusiforme and their antagonistic effects against the proliferation-inhibition of RAW264.7 macrophage induced by culture supernatants of melanoma cells

Two purified galactofucoidans, SFA1B-1 and SFA1C-2, with apparent molecular weights of 1.6 × 105 and 8.9 × 104, respectively, were isolated from Sargassum fusiforme via ion-exchange and gel-filtration column chromatography. SFA1B-1 and SFA1C-2 were composed of large amounts of fucose (Fuc), minor amounts of galactose (Gal), and small amounts of xylose and glucuronic acid. The backbone of these galactofucoidans consisted of α-(1→3)-linked Fucp residues sulfated mostly at the C-2 and/or C-4 positions. The backbone was primarily branched at the C-2 and/or C-4 positions probably by side chains comprising t-Fucp residues and (1→4)-Galp residues. Bioactive tests preliminarily showed that SFA1B-1 and SFA1C-2 could strongly not only antagonize against but also reverse the proliferation-inhibition of RAW264.7 cells induced by B16F10-CS. This finding suggests that SFA1B-1 and SFA1C-2 may antagonize the immunosuppressed function of macrophages. However, their possible effects on immunosuppressed peritoneal macrophages from mice or human with tumors and the underlying antagonistic mechanism warrant further investigation.

The Lipid A from the Lipopolysaccharide of the Phototrophic Bacterium Rhodomicrobium vannielii ATCC 17100 Revisited.

The structure of lipid A from lipopolysaccharide (LPS) of Rhodomicrobium vannielii ATCC 17100 (Rv) a phototrophic, budding bacterium was re-investigated using high-resolution mass spectrometry, NMR, and chemical degradation protocols. It was found that the (GlcpN)-disaccharide lipid A backbone was substituted by a GalpA residue that was connected to C-1 of proximal GlcpN. Some of this GalpA residue was β-eliminated by alkaline de-acylation, which indicated the possibility of the presence of another so far unidentified substituent at C-4 in non-stoichiometric amounts. One Manp residue substituted C-4′ of distal GlcpN. The lipid A backbone was acylated by 16:0(3-OH) at C-2 of proximal GlcpN, and by 16:0(3-OH), i17:0(3-OH), or 18:0(3-OH) at C-2′ of distal GlcpN. Two acyloxy-acyl moieties that were mainly formed by 14:0(3-O-14:0) and 16:0(3-O-22:1) occupied the distal GlcpN of lipid A. Genes that were possibly involved in the modification of Rv lipid A were compared with bacterial genes of known function. The biological activity was tested at the model of human mononuclear cells (MNC), showing that Rv lipid A alone does not significantly stimulate MNC. At low concentrations of toxic Escherichia coli O111:B4 LPS, pre-incubation with Rv lipid A resulted in a substantial reduction of activity, but, when higher concentrations of E. coli LPS were used, the stimulatory effect was increased.

Structural characteristics, molecular properties and immunostimulatory effects of sulfated polysaccharide from freshwater Myriophyllum spicatum L

Water-soluble polysaccharides were isolated from a freshwater plant, Myriophyllum spicatum L., at elevated temperature and purified using DEAE Sepharose FF column to elucidate their structural properties and immunopotentiating effects. Polysaccharides were composed of neutral sugars (73.9–78.2%), sulfate esters (12.8–16.8%) and uronic acids (3.1–4.9%). Weight average molecular weight (Mw) of polysaccharides varied between 217.4 and 529.0 × 103 g/mol. Polysaccharide structures were predominantly consisted of glucose (31.9–59.1%) and galactose (16.6–34.5%) with lower amounts of mannose, xylose, arabinose and rhamnose. As the most active fraction, MSF1 induced RAW264.7 murine macrophage cells to release nitric oxide and initiate the expression of TNF-α, IL-1β, IL-6 and IL-12 cytokines. Besides, MSF1 activated NK-92 cells to produce TNF-α and INF-γ cytokines and initiate cell-mediated cytotoxicity via perforin/granzyme B, NKG2D receptor and FasL pathways. Stimulation of RAW264.7 and NK-92 cells with MSF1 resulted in high expressions of phosphorylated NF-κB, ERK, JNK and p38 proteins revealing the involvement of NF-κB and MAPKs signaling pathways. MSF1 possessed a highly-branched structure consisting a combination of (1 → 4)-Glcp, (1 → 6)-Glcp, (1 → 4,6)-Glcp, (1 → 3,4)-Galp and (1 → 2,3)-Galp residues. Altogether, the present findings suggested the potential application of sulfated-polysaccharides from M. spicatum L. for immunostimulation and thus the necessity for further biomedical research.

Structural characterization of three polysaccharides from the roots of Codonopsis pilosula and their immunomodulatory effects on RAW264.7 macrophages.

Three polysaccharides were isolated from the roots of Codonopsis pilosula by ion-exchange and gel-filtration chromatography. They were named RCNP, RCAP-1, and RCAP-2, and had apparent molecular weights of 1.14 × 104, 5.09 × 104, and 2.58 × 105, respectively. Their structures were characterized by HPGPC, chemical derivative analysis, GC-MS and NMR analyses. Results showed that RCNP contained arabinan and arabinogalactan regions. The arabinan region had a main chain comprising (1 → 5)-linked Araf residues, and the side chains branched at the O-3 position by the single Araf residues. The arabinogalactan region comprised alternating (1 → 4)-, (1 → 6)- or (1 → 3)-linked Galp along with small amounts of branches mainly at the O-3 position of the (1 → 6)-linked Galp or O-6 position of the (1 → 3)-linked Galp residues by terminally linked Araf residues. RCAP-1 and RCAP-2 were highly methyl-esterified pectin-type polysaccharides with long homogalacturonan regions interrupted by a short rhamnogalacturonan I (RG-I) region. The side chains of the RG-I region consisted of (1 → 2)-linked Rha residues attached to the position O-4 of rhamnose. Their degrees of methyl-esterification were approximately 60.6% and 68.1%, respectively. Bioactivity tests showed that RCAP-1 and RCAP-2 exerted a significant immunostimulatory effect based on NO production from RAW264.7 macrophages. These results suggested that these two pectin-type polysaccharides were potential immunostimulation agents.

Comparison and structural characterization of polysaccharides from natural and artificial Se-enriched green tea.

Se-enriched green tea has been widely used as a functional food and disease prevention. In this study, two kinds of homogeneous polysaccharides namely ASe-TPS2 and NSe-TPS2 were obtained from artificial and natural Se-enriched teas with the molecular weights of 6.73 × 103 Da and 2.44 × 105 Da. The structures of ASe-TPS2 and NSe-TPS2 were characterized by monosaccharide composition analysis, partial acid hydrolysis, methylation, FT-IR spectroscopy, NMR spectroscopy, SEM and TGA analysis. It showed that ASe-TPS2 and NSe-TPS2 were acidic polysaccharides containing high amount of uronic acid. The structure of the ASe-TPS2 was mainly composed of β‑D‑(1 → 3)‑Glcp, α‑D‑(1 → 4)‑GalpA, (1 → 4)‑Glcp, α‑L‑(1 → 2)‑Rhap and α‑D‑(1 → 4)‑GalpA, and the non-reducing ends were mainly composed of Araf and Xylp. However, the NSe-TPS2 was mainly composed of β‑D-(1 → 4)‑Glcp and α‑D‑(1 → 4)-GalpA, and the branches were mainly composed of β‑L‑(1 → 2)‑Araf, α‑D‑(1 → 3)‑Galp and β‑L‑(1 → 2)‑Rhap whereas the non-reducing ends were mainly composed of Glcp and Galp residues. These results suggested that the distinction of selenylation methods could present different polysaccharide chain structures.

Characterizations and immunostimulatory activities of a polysaccharide from Arnebia euchroma (Royle) Johnst. roots

A polysaccharide from Arnebia euchroma (Royle) Johnst. named ARP, was obtained and purified by the hot water extraction, ethanol precipitation and deproteinization of TCA. The molecular weight of the polysaccharide fraction of ARP was calculated to be 1.23 × 104 Da from a calibration curve obtained with dextran standards. Monosaccharide composition analysis revealed that ARP was composed of Gal, Ara, Glu, Man, Rha and Fuc at a molar ratio of 53.8:21.3:11.7:6.8:4.3:2.2. Methylation analysis suggested that ARP was likely an arabinogalactan and that its backbone mainly consisted of Galp residues of 1,6‑linkages and Ara residues of 1,5‑ or 1,3‑linkages. The in vitro experiment indicated that ARP enhanced B- and T-lymphocyte proliferation. A dose-dependent relationship was observed, and a dose of 200 μg/mL resulted in the highest cell viability. In addition, ARP significantly stimulated the production of the cytokine, interferon-γ (IFN-γ), and enhanced B- and T-lymphocyte proliferation. Meanwhile, ARP had little effect on interleukin-2 (IL-2) production. The experiments of the effect of ARP on the activation of macrophage in vitro indicated that ARP significantly enhanced the production of TNF-α, IL-6 and IL-1β which suggested the polysaccharide induced the functional activation of macrophage.

An acidic polysaccharide from Ziziphus Jujuba cv. Muzao: Purification and structural characterization

A novel acidic polysaccharide fraction (PZMP2-2) was isolated from Ziziphus Jujuba cv. Muzao fruit via hot water extraction and purified by DEAE-Sepharose FF and Sephacryl S-300 column chromatography. PZMP2-2 consisted of rhamnose, arabinose, xylose, galactose, and galacturonic acid in a ratio of 1.18: 5.23: 0.22: 2.68: 2.20, with a molecular weight of 62.73 kDa. Structural features of PZMP2-2 were investigated using FT-IR, SEM, AFM, GC–MS analysis and NMR spectroscopy. Methylation and GC–MS analysis revealed that the main glycosidic bonds in PZMP2-2 comprised 1-linked Galp, 1,3-linked Araf, 1,2,4-linked Rhap, 1,3-linked Galp, 1,4-linked GalpA and 1,3,5-linked Araf. NMR analysis indicate that PZMP 2–2 has a linear backbone of (1 → 4)-linked GlcpA and (1 → 2,4)-linked Rhap residues, with branches at the O-4 position, consisting of Araf and Galp residues.

Structural characterization of polysaccharides from Saposhnikovia divaricata and their antagonistic effects against the immunosuppression by the culture supernatants of melanoma cells on RAW264.7 macrophages

Two purified native polysaccharides, namely, SDNP-1 and SDNP-2 with apparent molecular weight of 67.9×103 and 5.2×103, separately, were isolated from the water extract of Saposhnikovia divaricata by combined ion-exchange and gel permeation chromatography. The structures of these polysaccharides were characterized by high-performance gel permeation chromatography, chemical derivative analysis, GC–MS, FT-IR spectroscopy, and NMR spectroscopy. Results showed that SDNP-1 and SDNP-2 were composed of arabinose and galactose with molar ratio of approximately 1:1. The polysaccharides were deduced to be AGII-type arabinogalactans with distinct molecular weights and backbone chains. The backbone chain of SDNP-2 mainly consisted of (1→3)-linked and (1→6)-linked Galp residues with an approximate molar ratio of 1:3. Approximately half of the (1→6)-linked Galp residues were substituted at O-3, and small amounts of (1→6)-linked Galp were substituted at O-3 and O-4 by branches that mainly consisted of terminal and (1→5)-linked Araf residues. Bioactivity tests showed that SDNP-2 exhibited significant antagonistic effect against immunosuppression as shown by the cell viability of the culture supernatants of melanoma cells on RAW264.7 macrophages. However, SDNP-1 showed no effect. Thus, molecular weight and backbone chain may be the key factors for the antagonistic effects against immunosuppression.

Structural and rheological studies of a polysaccharide mucilage from lacebark leaves (Hoheria populnea A. Cunn.)

A water-soluble mucilage extracted from the leaves of Hoheria populnea was chemically and physically studied. Monosaccharide composition and linkages were determined by high performance anion exchange chromatography (HPAEC), gas chromatography–mass spectrometry (GC–MS) and nuclear magnetic resonance (NMR) spectroscopy. Lacebark mucilage was composed of rhamnose, galactose, galacturonic acid and glucuronic acid (2:1:2:1). Proton and 13C NMR spectroscopy, and linkage analysis, revealed a predominantly rhamnogalacturonan I-type (RG I-type) structure comprising of a backbone of →4]-α-D-GalpA-[1→2]-α-L-Rhap-[1→. Data indicated the mucilage likely comprises of a polymer containing several structurally discrete domains or possibly more than one discrete polymer. One domain contains a RG I-type backbone with branching at O-3 of GalpA residues to terminal β-D-GlcpA residues, another similarly contains a RG I-type backbone but is branched at O-4 of the Rhap residues to terminal GalpA residues or oligosaccharides containing α-linked 4-Galp and terminal GalpA residues. A possible third domain contains contiguous 2-Rhap residues, some branched at O-3. Hydrated mucilage exhibited pseudoplastic flow behaviour and viscoelastic properties of an entangled biopolymer network. These rheological behaviours were only slightly affected by pH and may prove advantageous in potential end-product applications including oral pharmaceuticals or as a food ingredient.

Structural characterization and macrophage activation of a hetero-galactan isolated from Flammulina velutipes

We isolated and purified a new polysaccharide (WFVP-N-b1) with a molecular weight of 20 kDa from Flammulina velutipes. Results showed that WFVP-N-b1 is composed of an α (1 → 6)-linked D-galactan backbone and branched at the O-2 of its Galp residues by an α-D-(1 → 6)-linked Manp attached to t-β-D-Glcp or t-α-D-Fucp side chains. WFVP-N-b1 can significantly induce cytokines secretion and release of toxic molecules. On a cellular level, WFVP-N-b1 is recognized by Toll-like receptor 4 (TLR4). Thereby, the hetero-galactan increased the phosphorylation of mitogen-activated protein kinases (MAPKs) and Akt, promoted degradation of IκB-α and the nuclear translocation of the NF-κB p65 subunit. Importantly, our results indicate that WFVP-N-b1 activated macrophage is mediated by autophagy, as blockade of WFVP-N-b1-induced autophagy by Baf-A1 significantly decreases macrophage activation. This is the first report that hetero-galactan-induced macrophage activation is mediated by autophagy. Collectively, WFVP-N-b1 activated RAW264.7 cells through MAPKs, autophagy, and Akt/NF-κB signaling pathways via TLR4 receptor.

Structural analysis and immunomodulatory effect of polysaccharide from Atractylodis macrocephalae Koidz. on bovine lymphocytes

RAMP60, RAMP70, RAMP80 and RAMPtp extracted from the rhizome of Atractylodis macrocephalae Koidz. were chemically analyzed and tested for their lymphocyte stimulation. Among them, only RAMPtp was found to induce proliferation of lymphocytes from bovine supramammary lymph node (SMLN). It was found that RAMPtp was homogeneous polysaccharide with Mn of 1.867×103 by GPC analysis, composed of glucose, mannose, rhamnose, arabinose and galactose, with mass percentages of 60.67%, 14.99%, 10.61%, 8.83% and 4.90%, respectively by HPLC analysis, connected by 1, 3-linked β-D Galp and 1, 6-linked β-D Galp residues by NMR analysis and was amorphous in structure by SEM analysis. When SMLN lymphocytes were cultured with RAMPtp in vitro, increased [Ca2+]i, more cells in S and G2/M phases and upregulated IFN-γ and IL-17A while downregulated IL-4 mRNA expressions as well as the binding between SMLN lymphocytes and RAMPtp were observed. These findings might explain the lymphocyte-stimulating property of RAMPtp.

Structural elucidation of a pectin–type polysaccharide from Hovenia dulcis peduncles and its proliferative activity on RAW264.7 cells

In this paper, an acidic polysaccharide HDP3A was isolated from Hovenia dulcis peduncles with 45.9% of uronic acid content. Structure of HDP3A was elucidated by chemical and spectroscopic analysis. HDP3A was mainly contained of galacturonic acid, galactose, rhamnose and arabinose with a small quantity of xylose, fucose, mannose, glucose. The structural analysis indicated that HDP3A was a pectin-type polysaccharide with HG and RG-I regions and hairy regions. HG was linear homopolymer with repeating units of (1→4)-α-d-galactopyranosyl uronic (GalA) residues. The GalpA residues in HG domain may be acetylated and/or methyl-esterified. The backbone of RG-I contains repeating units of [→α-GalpA-1,2)-α-Rhap-1,4→]n and the hairy regions attached in the Rha at position O-4. Hairy regions were mainly composed of galactans, arabinans, arabinogalactans, glucans, mannans and xylans. The immunomodulatory activities of HDP3A and its reduced (HDP3A-R) and hydrolyzed product (HDP3A-0.1) in vitro on macrophages were determined. The three polysaccharide fractions all stimulated the proliferation of RAW264.7 cells and the order of the proliferative effects was HDP3A>HDP3A-R>HDP3A-0.1, indicating that the immune effects of pectin-type polysaccharides were related to branches and molecular weight. Consequently, HDP3A could be as a potential source for the proliferative activity.

Structural determination of the K14 capsular polysaccharide from an ST25 Acinetobacter baumannii isolate, D46

The structure of the capsular polysaccharide (CPS) recovered from D46, an extensively antibiotic resistant ST25 Acinetobacter baumannii clinical isolate, was elucidated. The structure was resolved on the basis of NMR spectroscopy and chemical analyses, and was found to contain a branched neutral pentasaccharide with a backbone composed of GalpNAc and Galp residues, all d configured, and a d-Glcp side group. The KL14 gene cluster found in the D46 genome includes genes for four glycosyltransferases but no modules for synthesis of complex sugars, and this is consistent with the structure of K14. The K14 structure and KL14 sequence clarify the relationship between the structure and K locus sequence for A. nosocomialis isolate LUH5541. The identity of the first sugar of the K14 repeat unit (K unit), and the functions of the four encoded glycosyltransferases and Wzy polymerase were predicted.