Acidic Heteropolysaccharide Research Articles

Extraction, characterization, and biological activity of polysaccharides from Sophora flavescens Ait.

Four water-soluble polysaccharides, designated as SF1, SF2, SF3 and SF4, were efficiently extracted from the roots of Sophora flavescens by mechanochemistry under the conditions of rotational speed of 400rpm, grinding time of 10min, powder to ball weight ratio of 1:20, and Na2CO3 loading of 7wt%. The results obtained indicated that all of these four acid heteropolysaccharides are composed of rhamnose, arabinose, xylose, mannose, glucose and galactose, with the average molecular weights of 400.9, 98.6, 99.3, 42.7kDa, respectively. In vitro, SF4 showed the most significant scavenging activity on superoxide radical, ABTS, and DPPH radical, while SF3 had the most significant scavenging activity on hydroxyl radical. Immunological tests demonstrated that SF1, SF2, SF3 and SF4 significantly stimulated nitric oxide production without cytotoxicity in macrophages and promoted splenocyte proliferation. These data suggest that the four polysaccharides fractions have the potential as novel natural sources of antioxidative and immunopotentiating agents.

Fractionation, physicochemical characteristics and biological activities of polysaccharides from Pueraria lobata roots

In this paper, three water-soluble polysaccharides (PS-D1, PS-D2, and PS-D3) isolated and purified from Pueraria lobata roots were characterized and their biological activities were evaluated. Results showed that PS-D1 was a neutral heteropolysaccharide comprised glucose and fructose in a molar ratio of 24.4:1.0 with a weight-average molecular weight (Mw) of 62kDa, while PS-D2 and PS-D3 were both acidic heteropolysaccharides consisting of glucose, fructose, arabinose and glucose, fructose, galactose, arabinose in a molar ratio of 54.5:1.0:1.0 and 61.0:1.0:2.7:2.4 with Mws of 159 and 385kDa, respectively. Amongst of them, PS-D3 had the highest uronic acid content (∼12.4%), carbohydrate content (∼88.9%) and Mw. All the three polysaccharides exhibited as random coils or large aggregates in aqueous solution. In vitro antioxidant activity tests indicated that PS-D3 showed the strongest scavenging capacities on hydroxyl (79.6%) and DPPH (60%) radicals at 3.0mg/ml, and exhibited the highest antioxidant activity with a Trolox equivalent antioxidant capacity of 53.8µmol Trolox/g and a ferric reducing ability of plasma of 56.9µmol Fe2+/g. Moreover, the inhibitory activity on advanced glycation end-products (AGEs) formation of PS-D3 was 59.1% at 0.8mg/ml. These results indicated that PS-D3 with prominent antioxidant capability showed pronounced non-enzymatic glycation inhibition activity in vitro.

Structural characterization and in vitro antitumor activity of an acidic polysaccharide from Angelica sinensis (Oliv.) Diels

A water-soluble polysaccharide extracted from the roots of Angelica sinensis (Oliv.) Diels, which is a traditional Chinese medicine herb, was fractioned and purified by Sephadex G-50 gel filtration chromatography. The structural characterization and antitumor activities of the purified polysaccharide fraction, named as ASP, were evaluated in the present study. ASP, which molecular weight was determined to be 80kDa by high-performance gel-permeation chromatography, is an acidic heteropolysaccharide consisting of glucuronic acid, glucose, arabinose and galactose in ratio of 1.00:1.70:1.85:5.02. It has a backbone composed of (1→3)-linked Galp, (1→6)-linked Galp and 2-OMe-(1→6)-linked Galp with three branches attached to O-3 of 2-OMe-(1→6)-linked Galp and terminated with GlcpA and Araf, and all of Araf and the majority of Glcp are distributed in branches. Moreover, all of GlcpA were presented as (1→)-linked GlcpA in branches. In in vitro antitumor assays, ASP displayed cytotoxicity against HepG2 cells (34.32±3.50% at the concentration of 1mg/mL) and MCF-7 cells (28.90±1.50% at the concentration of 1mg/mL) in a dose-dependent manner, and ASP also showed mild inhibitory activity against A549 cells.

Structural characterization of low molecular weight polysaccharide from Astragalus membranaceus and its immunologic enhancement in recombinant protein vaccine against systemic candidiasis

Structure and immunologic enhancement of low molecular weight polysaccharide (LMW-ASP) isolated from the root of Astragalus membranaceus (Fisch) Bge. Were detected in recombinant protein vaccine. Structure analysis of LMW-ASP revealed that LMW-ASP (Mw=5.6kDa) was an acid heteropolysaccharide, which consisted of Glc, Gal, Ara, Xyl and GalA in ratio of 10.0：1.3：1.7：1.0：0.9. Recombinant protein (rP-HSP90C) contained epitope C (LKVIRK) from heat shock protein 90 (HSP90) of Candida albicans was used as a vaccine. The results indicated that LMW-ASP significantly promoted specific antibody titers IgG, IgG1, IgG2b, and IL-2, IL-4, IL-10, IL-12 in sera of mice immunized with rP-HSP90C (p<0.05). It was also found LMW-ASP improved DTH response in HSP90C-injceted mice. More importantly, the mice immunized with rP-HSP90C/LMW-ASP had fewer CFU (colony forming unites) in the kidneys compared to the mice immunized with rP-HSP90C (p<0.05). Therefore, LMW-ASP could be exploited into the novel adjuvant to enhance the efficacy of recombinant protein vaccine.

Supercharged fluorescent protein as a versatile probe for the detection of glycosaminoglycans in vitro and in vivo.

Glycosaminoglycans (GAGs) are linear acidic heteropolysaccharides that are ubiquitously expressed in animal tissues and participate in various life processes. To date, the detection and visualization of GAGs in complex biological samples and living organisms remain a challenge because of the lack of powerful biocompatible probes. In this study, a superpositively charged green fluorescent protein (ScGFP) was shown great potential in GAG detection for the first time. First, on the basis of the phenomenon of GAGs dose-dependently inhibiting the fluorescence quenching of ScGFP by graphene oxide, a simple and highly sensitive signal-on homogeneous platform was established for detecting and quantifying GAGs, even in complex samples such as heparin in citrated plasma and oversulfated chondroitin sulfate in heparin. Furthermore, ScGFP with excellent stability and biocompatibility could be easily used as a highly sensitive and selective probe to visualize different types of GAGs in vitro and in vivo through combination with specific GAG-degrading enzymes. This study introduces a versatile probe for GAG detection, which is easy to prepare and which shows a high practical value in basic research and medical applications.

Acid heteropolysaccharides with potent antileishmanial effects

The present study investigated in vitro the effects of sulphated heterorhamnan (Go3), iota-/nu-carrageenans (G3d and EHW-I) and arabinogalactan (ARAGAL) polysaccharides on macrophage activation and inhibition of intracellular amastigotes of Leishmania (L.) amazonensis. All the sulphated polysaccharides (Go3, G3d and EHW-I) promoted increased nitric oxide production varying from 71 to 110%. The leishmanicidal activity of all compounds was compared to the inhibition effect of Meglumine Antimoniate at 300μg/mL (∼79%), used as positive control. Inhibition of Leishmania (L.) amazonensis growth was 55% with 5μg/mL of Go3, 50% and 98% to G3d and EHW-I, respectively at 10μg/mL, and 88% with 10μg/mL of ARAGAL. The superoxide anion scavenging activity for the sulphated polysaccharides varied from approximately 30–55% at 10μg/mL. In conclusion, the results of the present study indicate the promising potential of these polysaccharides for the development of new alternative therapeutic agents against leishmaniasis.

Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits

A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. RCP-II was an acidic heteropolysaccharide and the characteristic structure of polysaccharide was determined. The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013Da, based on dextran standards. RCP-II presented high scavenging activity toward DPPH•, HO•, O2•− in a concentration-dependent manner. The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases.

The marine bacteria Cobetia marina DSMZ 4741 synthesizes an unexpected K-antigen-like exopolysaccharide

We have studied the exopolysaccharide produced by Cobetia marina DSMZ 4741, a marine bacterium isolated from coastal seawater. This strain is able to produce a polysaccharide in presence of carbon sources as glucose, mannitol and alginate. The maximum production occurs in aerobic condition, during the end of the exponential phase. The polymer is a non-viscous, acidic heteropolysaccharide of 270kDa constituted of a repeating unit of:→2)-β-d-Ribf-(1→4)-[7,8-O-(Pyr)]-α-d-KDOp-(2→This kind of chemical structure is generally related to K-antigen polysaccharide of pathogenic Escherichia coli strains. This is the first time this type of EPS is described from a marine bacterium. Moreover the polysaccharide exhibits a pyruvate substitution on its 3-deoxy-d-manno-oct-2-ulosonic acid (KDO) residue never encountered before. The discovery of such an unexpected EPS with high biotechnological potential is a new incentive for a better exploration of bioactive marine resources.

Water-soluble polysaccharide isolated with alkali from the stem of Physalis alkekengi L.: structural characterization and immunologic enhancement in DNA vaccine.

A water-soluble polysaccharide (WSPA) was isolated with alkali and purified from the mature stem of Physalis alkekengi L. WSPA (Mw=31kDa) was an acid heteropolysaccharide, which consisted of Rha, Ara, Xyl, Gal, Glc and GalA in ratio of 1.0:2.5:0.8:2.7:4.4:1.4. The results from structural analysis indicated backbone and branches of WSPA were composed of (1→3)-linked Glc, (1→3)-linked Gal, (1→2)-linked Xyl, (1→2)-linked Ara, and (1→2)-linked Rha residues. However, GalA was distributed only in the backbone of WSPA. All branches of WSPA were at O-2 of (1→6)-linked Gal and terminated with Glc. More importantly, it was found that WSPA significantly enhanced specific antibody IgG response with higher titers of IgG1 as well as IgG2b (p<0.05) in mice immunized with DNA vaccine. Therefore, WSPA can be considered as a potential adjuvant candidate in DNA vaccine.

Purification, Physicochemical Characterization, and Bioactivities of Polysaccharides from Puerh Tea

Two fractions of polysaccharides, named PTPS-1 and PTPS-2, was extracted and purified from puerh tea by Sephacryl S-300 column chromatography. The physicochemical properties of these two polysaccharides were investigated by high-performance liquid chromatography (HPLC), Fourier Transform IR spectra, scanning electron microscope (SEM) and atomic force microscope (AFM). Analysis of chemical compositions (protein, neutral sugars, uronic acid and monosaccharide composition) suggested that they were both kinds of acid heteropolysaccharides bound with protein, and contained seven monosaccharides with different molar ratio. Meanwhile, evaluation of antioxidant activities by in vitro assays of DPPH, ABTS and FRAP showed that PTPS-1 demonstrated stronger antioxidant ability than PTPS-2. Similarly, PTPS-1 exhibited remarkable inhibitory potential on α-glycosidase in vitro, significantly stronger than that of PTPS-2 and acarbose. Moreover, the results from animal test indicated PTPS-1 possessed significant inhibition on postprandial hyperglycemia in diabetic mice compared with the model group.

Characterization and hepatoprotective effect of polysaccharides from Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou sarcocarp

The polysaccharides from Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou sarcocarp (PWJS) is evaluated for the chemical composition, antioxidant activity and hepatoprotective effect. The characteristics of PWJS were determined by FT-IR spectral and HPLC analysis. The antioxidant activity was investigated using in vitro systems. An in vivo study of PWJS against CCl4 induced liver injury was also conducted. HPLC analysis showed that PWJS is an acidic heteropolysaccharide, rich in glucose (38.59%), arabinose (23.16%), galacturonic acid (17.64%) and galactose (10.44%). PWJS displayed strong antioxidant activity in vitro. In the in vivo study, PWJS treatment lowered the serum levels of ALT and AST in CCl4-intoxicated mice. Additionally, levels of antioxidant enzymes (SOD and CAT) and GSH were elevated in liver damage mice by PWJS intervention, while content of MDA was lessened. Meanwhile, PWJS reverses the suppression of Nrf2 nuclear translocation, and increases the protein expression of HO-1, GSTα and NQO1 in liver damage mice. Hematoxylin-eosin staining showed that the condition of liver damage was mitigated. This study demonstrates that PWJS reverses hepatotoxicity in CCl4-intoxicated mice through the mechanisms of antioxidant activity, as well as an augmentation of the Nrf2 pathway in liver tissue.

Structural elucidation and antioxidant activity of a novel polysaccharide (TAPB1) from Tremella aurantialba

A novel water-soluble acidic heteropolysaccharide (designated as TAPB1), with a mean molecular weight of 7.6 × 105 Da, was obtained from fruiting bodies of Tremella aurantialba. The heteropolysaccharide had a carbohydrate content of ∼97.6% and a negative response to the Bradford test. Monosaccharide composition analysis showed that TAPB1 comprised d-mannose, d-xylose, and d-glucuronic acid in the ratio 3.1:2.9:1.2, with trace amounts of d-galactose, d-glucose, and d-galacturonic acid. Methylation data and nuclear magnetic resonance spectral analysis indicated that TAPB1 contained an α-(1 → 3)-linked mannopyranosyl backbone, partially substituted at the O-4 positions with a side chain composed of two xyloses and at the O-2 position with a side chain comprising xylose and glucuronic acid. Antioxidant assays showed that TAPB1 exerted a significant scavenging effect on superoxide radicals and H2O2 in a dose-dependent manner.

Purification, characterization and immunomodulatory effects of Plantago depressa polysaccharides

Four purified polysaccharide fractions from seeds of Plantago depressa (PDSP-1, PDSP-2, PDSP-3 and PDSP-4) were obtained by isolation and purification using DEAE-52 cellulose and Sephacryl S-400 HR chromatography. Basic physicochemical properties including molecular weight, chemical composition, FT-IR and glycosidic linkage of these fractions were investigated. They seemed to be homogeneous acidic protein-bound heteropolysaccharides with high molecular weight of over 1000kDa and contained a lot more β-type glycosidic linkages than α-type. PDSP-3 mainly contained mannose, arabinose and fucose, and the others were rich in arabinose, fucose and galacturonic acid. The immunomodulatory effects of them were assessed by splenocyte proliferation index and production of NO and TNF-α from macrophages. They all showed significant immunomodulatory activities, and PDSP-3 presented the strongest effect. Their observed differences in biological activities were probably due to their structure differences. And monosaccharide compositions, linkage types and molecular weight may affect their immunomodulatory activities.

Screening and comparison of antioxidant activities of polysaccharides from Coriolus versicolor

Six polysaccharide fractions (Coriolus versicolor polysaccharides: CVPS-1, CVPS-2, CVPS-3, CVPS-4, CVPS-5 and CVPS-6) were isolated and purified from the fruiting bodies of C. versicolor by ion exchange chromatography and gel chromatography. Their chemical and physical characteristics were determined by chemical methods, high performance liquid chromatography, and high-performance gel-permeation chromatography. Finally, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical assay, superoxide radical assay, and hydroxyl radical assay were carried out to test the antioxidant activities of CVPS in vitro. The results indicated that the six CVPS fractions were acidic heteropolysaccharides, composed of mannose, rhamnose, glucuronic acid, glucose and fructose with different ratios. The molecular weights of CVPS-1, CVPS-2, CVPS-3, CVPS-4, CVPS-5 and CVPS-6 were 1740, 1480, 568, 880, 1260 and 1840kDa and the protein contents were 4.2%, 6.4%, 8.5%, 7.8%, 6.5% and 3.9%, respectively. Among the six fractions, CVPS with lower molecular weight, higher protein content and larger uronic acid amount, basically exhibited higher radical scavenging effects at the same concentration. Compared with other fractions, CVPS-3 exhibited the highest antioxidant activities. The effects of the molecular weight, protein content and uronic acid amount of the polysaccharides appeared to be significant on the improvement of the bioactivities.

Preventive effects of jujube polysaccharides on fructose-induced insulin resistance and dyslipidemia in mice.

High fructose intake is associated with adverse metabolic syndromes. This study was designed to investigate whether the polysaccharides derived from Zizyphus jujube cv. Shaanbeitanzao (ZSP) could alleviate high fructose-induced insulin resistance and dyslipidemia in mice. ZSP was identified by capillary zone electrophoresis as an acidic heteropolysaccharide with l-arabinose, d-galactose and d-galacturonic acid being the main component monosaccharides. Mice were provided with 20% high-fructose water and ZSP was administered intragastrically at doses of 0, 200 or 400 mg kg(-1) BW for 4 weeks. Fructose-treated mice showed hyperglycemia, hyperinsulinemia and dyslipidemia with impaired insulin sensitivity (p < 0.05). Administration of ZSP at a dose of 400 mg kg(-1) BW significantly reduced the serum levels of glucose, insulin, TC, TG, LDL-C, and VLDL-C (p < 0.01). ZSP also markedly improved the HDL-C level, homeostasis model assessment for insulin resistance (HOMA-IR) and β-cell function (HOMA-β), and decreased the atherogenic index (AI) of the mice exposed to high-fructose water. Histopathological test with H&E and oil red O staining confirmed liver steatosis induced by a high-fructose diet and the hepatoprotective effect of ZSP. These findings indicate that the jujube polysaccharides may ameliorate insulin resistance and dyslipidemia in fructose-treated mice.

Antioxidant activity and characterization of antioxidant polysaccharides from pine needle (Cedrus deodara)

A novel antioxidant polysaccharide (APC) was isolated and characterized from pine needles of Cedrus deodara with the evaluation of its in vitro antioxidant activity. According to gel filtration chromatography, high performance size exclusion chromatography, gas chromatography–mass spectrometry, partial acid hydrolysis, periodic acid oxidation, Smith degradation and methylation analysis, APC was observed to be an acidic heteropolysaccharide (composed of glucose, arabinose, mannose and xylose in a molar ratio of 45.84:1:2.35:1.73) with the molecular weight of 1.53×104Da, and the backbone was mainly composed by glucose, mannose and xylose in the form of (1→4) linked. Meanwhile, APC exhibited the remarkable antioxidant activity to scavenge free radicals and inhibit the oxidative injury of DNA and cells. The present results suggested that APC could be a potential antioxidant agent for preparing functional foods and nutraceuticals applied in food and pharmaceutical industries.

Comparison on polysaccharide content and PMP-HPLC fingerprints of polysaccharide in stems and leaves of Dendrobium officinale

In order to provide scientific basics for exploitation and sufficient application of Dendrobium officinale leaves resources, the phenol-sulfuric acid method was applied to determine the polysaccharide content. The monosaccharides were derivated by PMP and the derivatives were identified by HPLC-DAD-ESI-MS(n) and the contents of mannose and glucose were determined simultaneously. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A) was employed to generate the mean chromatogram and similarity analysis of the samples was carried out. The results demonstrated that polysaccharide content, monosaccharide compositions and composition ratio had an obvious difference between stems and leaves. The polysaccharide content of stems was higher than that of leaves. Monosaccharide composition in leaf was significantly different from that in stem. The polysaccharide from stems was composed of mannose and glucose, however the polysaccharide of leaves was acid heteropolysaccharide and was mainly composed of five monosaccharides, including mannose, galacturonic acid, glucose, galactose and arabinose. The similarity value of the 14 batches was above 0.9, indicating that similarity of fingerprints among different samples was high. The study can provide evidence for expanding the medicinal parts of D. officinale.

Analysis of monosaccharide compositions in polysaccharides from exopleura of Ginkgo biloba

A high-performance liquid chromatography method of pre-column derivatization with 1-phenyl-3-methyl-5 -pyrazolone (PMP) has been established for determination of 6 kinds of monosaccharides simultaneously. A special Agilent HC-C18 column (4. 6 mm x 250 mm, 5 microm), optimized for the separation of PMP derivatives, was used at ambient temperature of 40 degrees C. The PMP derivatives elution was performed with a mixture of 0.1 mol x L(-1) phosphate buffer (pH 6. 8) and acetonitrile in a ratio of 84: 16 at a flow rate of 1 mL x min(-1), and UV absorbance of the effluent was monitored at 245 nm. The results showed that the polysaccharides from exopleura of Ginkgo biloba were acidic heteropolysaccharides mainly containing mannose, rhamnose, D-galacturonic acid, glucose, galactose, arabinose, with the molar ratio of 0.032: 0.14: 0.296: 0.403:0.106: 0.046.

Physicochemical characterization of puerh tea polysaccharides and their antioxidant and α-glycosidase inhibition

Puerh tea is one of the most popular (or healthiest) beverages in Southeast Asia, because of its higher antioxidant, hypolipidemic and antidiabetic effects. One of the important active ingredients, tea polysaccharides (TPS) has long been recognized to possess immune modulatory and antidiabetic effects. In the present work, polysaccharides were isolated from puerh tea (PTPS) with aging (mild fermentation) for 1year (PTPS-1), 3years (PTPS-3) and 5years (PTPS-5), respectively. The results obtained indicated that all of these three kinds of PTPS are acid heteropolysaccharides bound with proteins, with differences in molecular weights and in monosaccharide compositions. The longer aging process not only produced a higher yield of PTPS but also had higher contents of proteins and uronic acids which proportionally correlated with their antioxidant and α-glycosidase inhibitory activities (PTPS-5>PTPS-3>PTPS-1). The anti-α-glycosidase activities of PTPS-5 and PTPS-3 were found to be three times more and equally potent than that of the antidiabetic drug acarbose. All of these findings firstly indicate that the aging process not only induced changes in chemical compositions and proteins conjugation in PTPS, but also markedly enhanced antioxidant and anti-α-glycosidase activities. PTPS-5 exhibited potential antidiabetic effect and merits further studies.

Purification, characterization and immunomodulating activity of a pectic polysaccharide isolated from Korean mulberry fruit Oddi (Morus alba L.)

A water-soluble polysaccharide (JS-MP-1) was isolated and purified from the Korean mulberry fruits Oddi (Morus alba L.) by crushing the fresh fruits then performing ethanol precipitation and DEAE-cellulose ion exchange chromatography. The neutral monosaccharide composition of the purified JS-MP-1 was determined to be composed mainly of galactose (37.6%, in mole percent), arabinose (36.3%), and rhamnose (18.4%), while other major sugars such as glucose, xylose, mannose, and fucose were present as minor components. HPLC analysis revealed that JS-MP-1 contains both galacturonic acid (GalA) and glucuronic acid (GlcA) at approximately 4:1 in mole percent. Monosaccharide composition, Fourier-transform infrared (FTIR) analysis, biochemical analysis, and elemental analysis suggested that JS-MP-1 is an acidic heteropolysaccharide, most likely a rhamnoarabinogalacturonan type plant pectic polysaccharide, with an apparent molecular mass of 1600 kDa containing no, or if any, negligible level of sulfate esters and proteins. Enzyme-Linked Immunosorbent Assay and RT-PCR analysis demonstrated that JS-MP-1 significantly stimulates murine RAW264.7 macrophage cells to release chemokines (RANTES and MIP-1α) and proinflammatory cytokines (TNF-α and IL-6) and to induce the expression of iNOS and COX-2, which are responsible for the production of NO and prostaglandin PGE2, respectively. These results suggest that the mulberry fruit-derived polysaccharide JS-MP-1 can act as a potent immunomodulator, and these observations may support the applicability of this polysaccharide as an immunotherapeutic adjuvant or the water extracts of the mulberry fruit as a beneficial health food.