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PubMed1983

Different polysaccharides in the external layers (capsule and slime) of the cell envelope of Rhodopseudomonas capsulata Sp11.

Two different acidic polysaccharides (I and II) were detected in the external cell envelope layers (slime and capsule) of Rhodopseudomonas capsulata Sp11. Polysaccharide I contains rhamnose, fucose, glucosamine and an unknown acidic sugar, it represents the slime material of the strain. Polysaccharide II contains rhamnose, galactose, 3-amino-3,6-dideoxygalactose, an unknown amino sugar and galacturonic acid, it represents very likely the capsule of R. capsulata Sp11. Polysaccharide I has a serological specificity different from that of polysaccharide II as shown by immunoprecipitation using antisera against living cells. Polysaccharide II, but not polysaccharide I, reacts in antiserum against heat-treated cells (100 degrees C, 2.5 h). Whole cells are agglutinated in the antisera against living but not in those against heat-treated cells.

Cell Membrane↗
PubMed2000

Characterisation of extracellular polysaccharides from suspension cultures of members of the poaceae.

Microscopic examination of suspension- cultured cells of Phleum pratense L., Panicum miliaceum L., Phalarisaquatica L. and Oryza sativa L. showed that they were comprised of numerous root primordia. Polysaccharides secreted by these suspension cultures contained glycosyl linkages consistent with the presence of high proportions of root mucilage-like polysaccharides. In contrast, suspension-cultured cells of Hordeum vulgare L. contained mostly undifferentiated cells more typical of plant cells in suspension culture. The polysaccharides secreted by H. vulgare cultures contained mostly linkages consistent with the presence of glucuronoarabinoxylan. The soluble polymers secreted by cell-suspension cultures of Phleum pratense contained 70% carbohydrate, 14% protein and 6% inorganic material. The extracellular polysaccharides were separated into four fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7.0. From glycosyl-linkage analyses, five polysaccharides were identified: an arabinosylated xyloglucan (comprising 20% of the total polysaccharide), a glucomannan (6%), a type-II arabinogalactan (an arabinogalactan-protein; 7%), an acidic xylan (3%), and a root-slime-like polysaccharide, which contained features of type-II arabinogalactans and glucuronomannans (65%).

Cell Wall↗
PubMed2005

Three types of acidic polysaccharides associated with coccolith of Pleurochrysis haptonemofera: comparison with Pleurochrysis carterae and analysis using fluorescein-isothiocyanate-labeled lectins.

In the coccolithophorid microalgae acidic polysaccharides are considered to be involved in the formation of the calcified scale, coccolith. Characteristics of the acidic polysaccharides extracted from the cell surface of the coccolithophorid Pleurochrysis haptonemofera were analyzed. The acidic polysaccharides on the cell surface can be detected by measuring fluorescence of cells after fluorescein-isothiocyanate-labeled lectin staining by flow cytometry. Flow cytometric analyses revealed that the acidic polysaccharides remained on the cell surface even after CaCO3 in the coccolith was dissolved by lowering pH, but they were extracted by subsequent EDTA or EGTA treatment, suggesting that they are bound not into the CaCO3 crystals of the coccolith, but onto the surface via Ca2+. Analyses of the acidic polysaccharides by anion exchange chromatography, colloidal precipitation with divalent cations, and polyacrylamide gel electrophoresis (PAGE) revealed that P. haptonemofera has 3 types of acidic polysaccharides (Ph-PS-l, -2, and -3). The PAGE patterns suggested that Ph-PS-2 has a repeated structure with a broad range of molecular weight, as in Pleurochrysis carterae, while Ph-PS-1 and -3 contain several minor components in addition to a major component, respectively. The minor components in Ph-PS-1 and -3 that have not been found in P. carterae might be characteristic of P. haptonemofera. Analyses of both the cell surface treated by various concentrations of EDTA and EGTA and the extracts suggested that Ph-PS-2, which is distinguishable by a higher affinity to concanavalin A, is bound onto the coccolith surface more intensely than the other two types of acidic polysaccharides.

Calcium↗
PubMed2005

Polysaccharide-producing bacteria isolated from paper machine slime deposits.

Development of novel enzymatic methods for slime deposit control in paper mills requires knowledge of polysaccharide-producing organisms and the polysaccharide structures present in deposits. In this work, 27 polysaccharide-producing bacteria were isolated from slime samples collected from different parts of a paper machine. Most of the isolates produced polysaccharides in liquid culture and nine of them were selected for production of polysaccharides for characterisation. The selected isolates belonged to seven different genera: Bacillus, Brevundimonas, Cytophaga, Enterobacter, Klebsiella, Paenibacillus and Starkeya. Using ribotyping, partial 16S rDNA sequencing, physiological tests and fatty acid analysis, four of the nine isolates: Bacillus cereus, Brevundimonas vesicularis, K. pneumoniae and P. stellifer were identified to the species level. Production of polysaccharides by the selected isolates varied between 0.07 and 1.20 g L(-1), the highest amount being produced by B. vesicularis. The polysaccharides were heteropolysaccharides with varying proportions of galactose, glucose mannose, rhamnose fucose and uronic acids.

Bacteria↗
PubMed1984

Rate, mechanism, and immunochemical studies of lactonisation in serogroup B and C polysaccharides of Neisseria meningitidis.

Meningococcal Serogroup B polysaccharide and colominic acid, which are (2----8)-alpha-linked homopolymers of sialic acid, undergo lactonisation at low pH at a rate which is dependent upon the molecular size and upon the salt form (Na+ or Ca2+). Meningococcal Serogroup C polysaccharide, a (2----9)-alpha-linked homopolymer of sialic acid with acetyl groups present at O-7 and/or O-8, reacts with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to give an O-acylisourea. The rate of formation of O-acylisourea does not differ substantially between O-acetylated (O-Ac+) C, non-O-acetylated (O-Ac-) C, and B polysaccharide. 13C-N.m.r. spectroscopy shows that, in the absence of O-acetyl groups, the majority of the activated carboxyl groups of C polysaccharide condense with an adjacent HO-8 to form a delta-lactone. Immunochemical studies show that the antigenicity of B polysaccharide is markedly reduced on lactonisation of less than 20%, as measured by a radioimmunoassay using an anti-B monoclonal antibody, and that low-molecular-weight colominic acid is poorly antigenic both before and after lactonisation, suggesting the presence of conformational determinants on B polysaccharide. In contrast, lactonisation and/or formation of O-acylisourea groups in the (O-Ac+)-C polysaccharide does not cause a significant decrease in the antigenicity, which is consistent with a sequential (structural) determinant on the molecule.

Hydrogen-Ion Concentration↗
PubMed1984

Conformational and dynamic differences between N. meningitidis serogroup B and C polysaccharides, using n.m.r. spectroscopy and molecular mechanics calculations.

1H-N.m.r. spectroscopy has been used to determine the conformation in aqueous solution of the sialic acid residues of the N. meningitidis serogroup B and non-O-acetylated (O-Ac-)-C polysaccharides, and of N-acetylneuraminic acid (NeuNAc). In all cases, the sugar adopts the 2C5 conformation. The side-chain of NeuNAc adopts a conformation such that H-7 and H-8 are approximately anti-periplanar. This conformation is also found in the (O-Ac-)-C polysaccharide, whereas H-7 and H-8 are gauche in the B polysaccharide. Molecular mechanics calculations have been used to probe the conformational preferences of the variously linked sialic acid residues, and the results are in general agreement with those based on the 1H-n.m.r. data. The 13C-n.m.r. spin-lattice relaxation-times have been interpreted in terms of the molecular dynamics of the B and (O-Ac-)-C polysaccharides. Molecular correlation times have been calculated and details of internal rotational or segmental motion elucidated. The C polysaccharide is characterised by internal or segmental motion in the C-7 to C-9 side-chain of the sialic acid repeating-unit, whereas the B polysaccharide has little or no such movement and tumbles in solution as a rigid species with internal rotation of only the pendant C-9 group. The conformational differences suggest a substantially different three-dimensional structure in solution for these polysaccharides.

Magnetic Resonance Spectroscopy↗
PubMed1987

Structural investigation of the capsular polysaccharide from Escherichia coli O9:K37 (A 84a).

The structure of the capsular polysaccharide from E. coli O9:K37 (A 84a) has been studied, using methylation analysis, Smith degradation, and graded acid hydrolysis. The configurations at the anomeric centres were assigned by 1H-n.m.r. spectroscopy of the polysaccharide and its derivatives and oligosaccharide fragments. The polysaccharide has the following trisaccharide repeating-unit which is unique in the E. coli series of capsular polysaccharides in possessing a 1-carboxyethylidene group as the sole acidic function. (Formula: see text) E. coli capsular polysaccharides have been classified into seventy-four serotypes. The structures of about twenty of these polysaccharides have been elucidated, one of which, K29, has been reported to contain a 1-carboxyethylidene group. In continuation of a programme aimed at establishing the structural basis for the serology and immunochemistry of the E. coli capsular antigens, we now report on the structure of the capsular polysaccharide from E. coli O9:K37.

Carbohydrate Conformation↗
PubMed1987

O-polysaccharide-protein conjugates induce high levels of specific antibodies to Pseudomonas aeruginosa immunotype 3 lipopolysaccharide.

A semi-synthetic vaccine against Pseudomonas aeruginosa immunotype 3 was prepared by chemical coupling of P. aeruginosa immunotype 3 O-polysaccharide to tetanus toxoid. The O-polysaccharide was obtained by mild acid hydrolysis of immunotype 3 lipopolysaccharide, and purified by gel permeation chromatography. The purification was evaluated by high performance liquid chromatography. Additional analyses revealed a high grade of purity of the O-polysaccharide, and an at least 1000-fold reduction of endotoxic activity as compared to homologous lipopolysaccharide. O-Polysaccharide was conjugated to tetanus toxoid, using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as coupling reagent. Antigenic determinants of both O-polysaccharide and tetanus toxoid were retained after conjugation, as tested in a sandwich enzyme-linked immunosorbent assay. Immunization of mice revealed that O-polysaccharide was nonimmunogenic in mice, while the O-specific part of the conjugate was able to induce high levels of IgG antibodies reacting with immunotype 3 lipopolysaccharide in an enzyme-linked immunosorbent assay. By immunoblotting it was shown that the antibodies were directed to high molecular weight lipopolysaccharide only, demonstrating specificity for its O-polysaccharide moiety.

Animals↗
PubMed1991

Isolation and purification of cell wall polysaccharide of Bacillus anthracis (delta Sterne).

A polysaccharide fraction was isolated form sodium-dodecyl-sulfate (SDS) treated cell walls of Bacillus anthracis (delta Sterne) by hydrofluoric acid (HF) hydrolysis and ethanolic precipitation. The polysaccharide fraction was subsequently purified by several washings with absolute ethanol. Purity of the isolated polysaccharide was tested using the anthrone assay and amino acid analyzer. The molecular mass of the polysaccharide fraction as determined by gel filtration chromatography was about 12000 Da. Preliminary analyses of the polysaccharide was done using thin layer chromatography and amino acid analyzer, and results obtained from these analyses were further confirmed by gas liquid chromatography and 13C-NMR spectroscopy. Results showed that the polysaccharide moiety contained galactose, N-acetylglucosamine, and N-acetylmannosamine in an approximate molar ratio of 3:2:1. This moiety was devoid of muramic acid, alanine, diaminopimelic acid, glutamic acid, and lipid, thus indicating that the isolated polysaccharide was of pure quality.

Acetylglucosamine↗
PubMed2004

Characterization and immunomodulating activities of polysaccharide from Lentinus edodes.

The polysaccharide L-II was isolated and purified from the fruiting body of Lentinus edodes, which consisted of d-glucopyranose and had the molecular weight of 2.03 x 10(5) Da. We evaluated the effects of the polysaccharide L-II on the cellular immune response of Sarcoma 180-bearing mice. Mice were treated with three doses of the polysaccharide L-II (1, 5, and 10 mg/kg body weight) for 10 days. Tumor weight, relative spleen and thymus weight, delayed-type hypersensitivity (DTH) response, phagocytosis of macrophage, splenocytes proliferation were studied. Concentration of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) in mice serum were measured in control and polysaccharide groups. At the dose of 1, 5 and 10 mg/kg, a significant increase (p<0.05) in relative spleen and thymus weight, DTH, phagocytosis of macrophage was observed, as well as a significant decrease in tumor formation. The concentration of TNF-alpha, IFN-gamma in serum increased significantly in the polysaccharide groups compared with the model control group, but IL-2 not. Moreover, the polysaccharide L-II could increase NO production and catalase activity in macrophages. Results of these studies demonstrated the antitumor activity of the polysaccharide L-II on mice-transplanted sarcoma 180 was mediated by immunomodulation in inducing T-cells and macrophage-dependent immune system responses.

Animals↗
PubMed2006

Effect of the Lycium barbarum polysaccharides on age-related oxidative stress in aged mice.

Oxidative damage of biomolecules increases with age and is postulated to be a major causal factor of various physiological function disorders. Consequently, the concept of anti-age by antioxidants has been developed. Lycium barbarum fruits have been used as a traditional Chinese herbal medicine and the data obtained in in vitro models have clearly established the antioxidant potency of the polysaccharides isolated from the fruits. In the present study, the age-dependent changes in the antioxidant enzyme activity, immune function and lipid peroxidation product were investigated and effect of Lycium barbarum polysaccharides on age-induced oxidative stress in different organs of aged mice was checked. Lycium barbarum polysaccharides (200, 350 and 500 mg/kg b.w. in physiological saline) were orally administrated to aged mice over a period of 30 days. Aged mice receiving vitamin C served as positive control. Enzymatic and non-enzymatic antioxidants, lipid peroxides in serum and tested organs, and immune function were measured. Result showed that increased endogenous lipid peroxidation, and decreased antioxidant activities, as assessed by superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and total antioxidant capacity (TAOC), and immune function were observed in aged mice and restored to normal levels in the polysaccharides-treated groups. Antioxidant activities of Lycium barbarum polysaccharides can be compable with normal antioxidant, vitamin C. Moreover, addition of vitamin C to the polysaccharides further increased the in vivo antioxidant activity of the latter. It is concluded that the Lycium barbarum polysaccharides can be used in compensating the decline in TAOC, immune function and the activities of antioxidant enzymes and thereby reduces the risks of lipid peroxidation accelerated by age-induced free radical.

Aging↗
PubMed2005

Interaction of antimicrobial peptides with bacterial polysaccharides from lung pathogens.

The interaction of two cathelicidin antimicrobial peptides, LL-37 and SMAP-29, with three bacterial polysaccharides, respectively, produced by Pseudomonas aeruginosa, Burkholderia cepacia and Klebsiella pneumoniae, was investigated to identify possible mechanisms adopted by lung pathogens to escape the action of innate immunity effectors. In vitro assays indicated that the antibacterial activity of both peptides was inhibited to a variable extent by the three polysaccharides. Circular dichroism experiments showed that these induced an alpha-helical conformation in the two peptides, with the polysaccharides from K. pneumoniae and B. cepacia showing, respectively, the highest and the lowest effect. Fluorescence measurements also indicated the presence of peptide-polysaccharide interactions. A model is proposed in which the binding of peptides to the polysaccharide molecules induces, at low polysaccharide to peptide ratios, a higher order of aggregation, due to peptide-peptide interactions. Overall, these results suggest that binding of the peptides by the polysaccharides produced by lung pathogens can contribute to the impairment of peptide-based innate defenses of airway surface.

Antigens, Bacterial↗
PubMed2005

Characterization of cell wall polysaccharides from the medicinal plant Panax notoginseng.

Panax notoginseng is a commonly used medicinal plant in south-western China. Recent studies indicate that wall polysaccharides are responsible for some of the immunostimulatory activity. Fractionation of the P. notoginseng root powder alcohol insoluble residue (AIR) and its compositional analysis enabled us to deduce the polysaccharide and protein composition of the root cell walls. P. notoginseng walls are composed primarily of polysaccharide (approximately 97% w/w) and some protein. The polysaccharides include pectic polysaccharides (neutral Type I 4-galactan (21%), arabinan (5%), acidic rhamnogalacturonan I (RG I, 2%) and homogalacturonan (HGA, 24%), non-cellulosic polysaccharides (heteroxylan, 3%), xyloglucan (XG, 3%) and heteromannan (1%)) and cellulose (24%). The root AIR also contains Type II AG/AGPs (5% w/w) typically associated with the plasma membrane and extracellular matrix. Thus, P. notoginseng roots contain polysaccharides typical of Type I primary cell walls but are distinguished by their very high levels of Type I 4-galactans and low levels of XGs. The major amino acids in the AIR were Leu (14 mol%), Asx (16 mol%), Glx (10 mol%), Ala (9 mol%), Thr (9 mol%) and Val (9 mol%).

Cell Wall↗
PubMed2003

The nontoxic mushroom Auricularia auricula contains a polysaccharide with anticoagulant activity mediated by antithrombin.

An acidic polysaccharide with anticoagulant activity was isolated from the edible mushroom Auricularia auricula using water, alkali or acid extracts. The alkali extract showed the highest anticoagulant activity and was thereby further purified using gel filtration chromatography. Specific anticoagulant activity of the purified polysaccharide was 2 IU/mg and its average mass was approximately 160 kDa. The polysaccharide from this species of mushroom contains mainly mannose, glucose, glucuronic acid and xylose but no sulfate esters. Its anticoagulant activity was due to catalysis of thrombin inhibition by antithrombin but not by heparin cofactor II. Inhibition of Factor Xa by antithrombin was not catalyzed by the polysaccharide. The glucuronic acid residues were essential for the anticoagulant action of the mushroom polysaccharide since the activity disappeared after reduction of its carboxyl groups. In ex vivo tests using rats orally fed with the polysaccharide, we observed an inhibitory effect on platelet aggregation as observed with aspirin, a well-known antiplatelet agent. The polysaccharides from these mushrooms may constitute a new source of compounds with action on coagulation, platelet aggregation and, perhaps, on thrombosis.

Animals↗
PubMed1978

Isolation, partial characterization, and biological properties of polysaccharides from crude papain.

Two polysaccharides have been isolated from crude papain by precipitation of papain with ammonium sulfate, further precipitation of other proteins with trichloroacetic acid, and chromatography of the supernatant on DEAE-cellulose. The first polysaccharide to be eluted, designated PP-I, contained D-glucuronic acid, D-glucose, D-galactose, L-arabinose, and L-rhamnose, in the approximate molar ratios of 4:1:12:10:4. The other (PP-II), eluted at a higher salt-concentration, contained the same sugars (with about one-third less glucose and more uronic acid) in the approximate molar ratios of 13:1:40:26:12. Reduction of the uronic acid groups of PP-II produced a polysaccharide (PP-II-R) containing the same sugars in the approximate molar ratios of 2:11:37:28:12. Hydrolysis of a mixture of the two polysaccharides yielded an aldobiouronic acid, D-glucosyluronic acid-D-galactose. Neither polysaccharide preparation contained protein. These polysaccharides dramatically affected aggregation and alignment of normal human fibroblasts but had no effect on a mouse embryo fibroblast aneuploid cell-line that does not exhibit contact inhibition of growth or movement. In aggregating cells, these polysaccharides caused the cells to behave as contact-inhibited cells, that is, cell division and nuclear area were decreased.

Adult↗
PubMed1996

Full 1H NMR assignment and detailed O-acetylation patterns of capsular polysaccharides from Neisseria meningitidis used in vaccine production.

We report essentially complete 1H NMR assignments for the capsular polysaccharides from Neisseria meningitidis serotypes A, C, W-135, and Y. These polysaccharides are components of current polysaccharide vaccines against meningococcal infection and of the polysaccharide-protein conjugate vaccines under development. From these NMR data the pattern of O-acetylation was determined. O-Acetylation of the W-135 polysaccharide is reported for the first time. We also show that, for the Types C and W-135 polysaccharides a migration of O-acetyl groups occurs during storage in solution, and demonstrate that high field 1H NMR represents a simple and sensitive method to define the O-acetylation pattern of individual batches of these polysaccharides.

Acetylation↗
PubMed2003

A polysaccharide isolated from Cordyceps sinensis, a traditional Chinese medicine, protects PC12 cells against hydrogen peroxide-induced injury.

Cordyceps sinensis, a well-known traditional Chinese medicine, possesses activities in anti-tumour, anti-oxidation and stimulating the immune system; however, the identity of active component(s) is not determined. By using anti-oxidation activity-guided fractionation, a polysaccharide of molecular weight approximately 210 kDa was isolated from cultured Cordyceps mycelia by ion-exchange and sizing chromatography. The isolated polysaccharide, having strong anti-oxidation activity, contains glucose, mannose and galactose in a ratio of 1 : 0.6 : 0.75. The pre-treatment of isolated polysaccharide on the cultured rat pheochromocytoma PC12 cells shows strong protective effect against hydrogen peroxide (H(2)O(2))-induced insult. Treatment of the cells with the isolated polysaccharide at 100 microg/ml prior to H(2)O(2) exposure significantly elevated the survival of PC12 cells in culture by over 60%. In parallel, the H(2)O(2)-induced production of malondialdehyde in cultured cells was markedly reduced by the polysaccharide treatment. Moreover, the pre-treatment of the isolated polysaccharide significantly attenuated the changes of glutathione peroxidase and superoxide dismutase activities in H(2)O(2)-treated cells in a dose-dependent manner. This is the first report in identifying a polysaccharide from Cordyceps, which protects against the free radical-induced neuronal cell toxicity.

Animals↗
PubMed1987

The persistence of antibodies induced by meningococcal polysaccharides of groups A and C in human volunteers.

Ninety-nine college students (58 males and 41 females) aged 17 to 23 years were each injected subcutaneously with 50 micrograms of meningococcal polysaccharides A and C. Titrations of antisera obtained at various time intervals during the two subsequent years were made by passive haemagglutination microtitration using human O Rh-negative red blood cells from a single source. The percentage of responders (those who developed a fourfold increase in titre) to polysaccharide A was 97.9% and that to polysaccharide C was 94.8%. Pre-immunization titres of 16 to polysaccharide A and 8 to polysaccharide C were considered to be threshold values above which the response might be impaired. There was a significant (P less than 0.01) difference in the geometric mean HA titre between vaccinees and control groups after vaccination at each time interval studied. The majorities of both the vaccinees and the controls had a higher peak titre to polysaccharide A than to polysaccharide C.

Adolescent↗