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Sample records for polysaccharide copolymerase protein

  1. Pivotal roles of the outer membrane polysaccharide export and polysaccharide copolymerase protein families in export of extracellular polysaccharides in gram-negative bacteria.

    PubMed

    Cuthbertson, Leslie; Mainprize, Iain L; Naismith, James H; Whitfield, Chris

    2009-03-01

    Many bacteria export extracellular polysaccharides (EPS) and capsular polysaccharides (CPS). These polymers exhibit remarkably diverse structures and play important roles in the biology of free-living, commensal, and pathogenic bacteria. EPS and CPS production represents a major challenge because these high-molecular-weight hydrophilic polymers must be assembled and exported in a process spanning the envelope, without compromising the essential barrier properties of the envelope. Emerging evidence points to the existence of molecular scaffolds that perform these critical polymer-trafficking functions. Two major pathways with different polymer biosynthesis strategies are involved in the assembly of most EPS/CPS: the Wzy-dependent and ATP-binding cassette (ABC) transporter-dependent pathways. They converge in an outer membrane export step mediated by a member of the outer membrane auxiliary (OMA) protein family. OMA proteins form outer membrane efflux channels for the polymers, and here we propose the revised name outer membrane polysaccharide export (OPX) proteins. Proteins in the polysaccharide copolymerase (PCP) family have been implicated in several aspects of polymer biogenesis, but there is unequivocal evidence for some systems that PCP and OPX proteins interact to form a trans-envelope scaffold for polymer export. Understanding of the precise functions of the OPX and PCP proteins has been advanced by recent findings from biochemistry and structural biology approaches and by parallel studies of other macromolecular trafficking events. Phylogenetic analyses reported here also contribute important new insight into the distribution, structural relationships, and function of the OPX and PCP proteins. This review is intended as an update on progress in this important area of microbial cell biology.

  2. Functional and structural characterization of polysaccharide co-polymerase proteins required for polymer export in ATP-binding cassette transporter-dependent capsule biosynthesis pathways.

    PubMed

    Larue, Kane; Ford, Robert C; Willis, Lisa M; Whitfield, Chris

    2011-05-13

    Neisseria meningitidis serogroup B and Escherichia coli K1 bacteria produce a capsular polysaccharide (CPS) that is composed of α2,8-linked polysialic acid (PSA). Biosynthesis of PSA in these bacteria occurs via an ABC (ATP-binding cassette) transporter-dependent pathway. In N. meningitidis, export of PSA to the surface of the bacterium requires two proteins that form an ABC transporter (CtrC and CtrD) and two additional proteins, CtrA and CtrB, that are proposed to form a cell envelope-spanning export complex. CtrA is a member of the outer membrane polysaccharide export (OPX) family of proteins, which are proposed to form a pore to mediate export of CPSs across the outer membrane. CtrB is an inner membrane protein belonging to the polysaccharide co-polymerase (PCP) family. PCP proteins involved in other bacterial polysaccharide assembly systems form structures that extend into the periplasm from the inner membrane. There is currently no structural information available for PCP or OPX proteins involved in an ABC transporter-dependent CPS biosynthesis pathway to support their proposed roles in polysaccharide export. Here, we report cryo-EM images of purified CtrB reconstituted into lipid bilayers. These images contained molecular top and side views of CtrB and showed that it formed a conical oligomer that extended ∼125 Å from the membrane. This structure is consistent with CtrB functioning as a component of an envelope-spanning complex. Cross-complementation of CtrA and CtrB in E. coli mutants with defects in genes encoding the corresponding PCP and OPX proteins show that PCP-OPX pairs require interactions with their cognate partners to export polysaccharide. These experiments add further support for the model of an ABC transporter-PCP-OPX multiprotein complex that functions to export CPS across the cell envelope.

  3. PssP2 Is a Polysaccharide Co-Polymerase Involved in Exopolysaccharide Chain-Length Determination in Rhizobium leguminosarum

    PubMed Central

    Marczak, Małgorzata; Matysiak, Paulina; Kutkowska, Jolanta; Skorupska, Anna

    2014-01-01

    Production of extracellular polysaccharides is a complex process engaging proteins localized in different subcellular compartments, yet communicating with each other or even directly interacting in multicomponent complexes. Proteins involved in polymerization and transport of exopolysaccharide (EPS) in Rhizobium leguminosarum are encoded within the chromosomal Pss-I cluster. However, genes implicated in polysaccharide synthesis are common in rhizobia, with several homologues of pss genes identified in other regions of the R. leguminosarum genome. One such region is chromosomally located Pss-II encoding proteins homologous to known components of the Wzx/Wzy-dependent polysaccharide synthesis and transport systems. The pssP2 gene encodes a protein similar to polysaccharide co-polymerases involved in determination of the length of polysaccharide chains in capsule and O-antigen biosynthesis. In this work, a mutant with a disrupted pssP2 gene was constructed and its capabilities to produce EPS and enter into a symbiotic relationship with clover were studied. The pssP2 mutant, while not altered in lipopolysaccharide (LPS), displayed changes in molecular mass distribution profile of EPS. Lack of the full-length PssP2 protein resulted in a reduction of high molecular weight EPS, yet polymerized to a longer length than in the RtTA1 wild type. The mutant strain was also more efficient in symbiotic performance. The functional interrelation between PssP2 and proteins encoded within the Pss-I region was further supported by data from bacterial two-hybrid assays providing evidence for PssP2 interactions with PssT polymerase, as well as glycosyltransferase PssC. A possible role for PssP2 in a complex involved in EPS chain-length determination is discussed. PMID:25268738

  4. Proteins Among the Polysaccharides

    PubMed Central

    Wen, Fushi; Curlango-Rivera, Gilberto

    2007-01-01

    Charles Darwin recognized the power of the root cap as a model for plant signalling and behavior, and used it to explore the ways plants sense and respond to diverse stimuli. Over ensuing decades, various groups have reported tantalizing clues regarding the role of a complex extracellular matrix that ensheaths the tip region housing the apical and root cap meristems. In the course of characterizing root tip resistance to infection and injury and the role border cells play in this phenomenon, we confirmed and extended early- and mid-20th century studies reporting enzyme activities secreted from the root cap. Multidimensional protein analysis revealed, in fact, that >100 proteins are actively synthesized and secreted from the root cap and border cells. This ‘root cap secretome’ appears to be a critical component of root tip resistance to infection. We have developed a microscopic assay to quantify the protein-based extracellular response to dynamic changes in environmental conditions including hydroponic culture, and present the results here. This tool provides a simple, direct measure that can be used to explore the ways border cells may function in the manner of white blood cells to trap, immobilize and neutralize threats to the growing root tip. PMID:19704617

  5. Rheology of interfacial protein-polysaccharide composites

    NASA Astrophysics Data System (ADS)

    Fischer, P.

    2013-05-01

    The morphology and mechanical properties of protein adsorption layers can significantly be altered by the presence of surfactants, lipids, particles, other proteins, and polysaccharides. In food emulsions, polysaccharides are primarily considered as bulk thickener but can under appropriate environmental conditions stabilize or destabilize the protein adsorption layer and, thus, the entire emulsion system. Despite their ubiquitous usage as stabilization agent, relatively few investigations focus on the interfacial rheology of composite protein/polysaccharide adsorption layers. The manuscript provides a brief review on both main stabilization mechanisms, thermodynamic phase separation and electrostatic interaction and discusses the rheological response in light of the environmental conditions such as ionic strength and pH.

  6. Influence of polysaccharides on wine protein aggregation.

    PubMed

    Jaeckels, Nadine; Meier, Miriam; Dietrich, Helmut; Will, Frank; Decker, Heinz; Fronk, Petra

    2016-06-01

    Polysaccharides are the major high-molecular weight components of wines. In contrast, proteins occur only in small amounts in wine, but contribute to haze formation. The detailed mechanism of aggregation of these proteins, especially in combination with other wine components, remains unclear. This study demonstrates the different aggregation behavior between a buffer and a model wine system by dynamic light scattering. Arabinogalactan-protein, for example, shows an increased aggregation in the model wine system, while in the buffer system a reducing effect is observed. Thus, we could show the importance to examine the behavior of wine additives under conditions close to reality, instead of simpler buffer systems. Additional experiments on melting points of wine proteins reveal that only some isoforms of thaumatin-like proteins and chitinases are involved in haze formation. We can confirm interactions between polysaccharides and proteins, but none of these polysaccharides is able to prevent haze in wine.

  7. Designing Whey Protein-Polysaccharide Particles for Colloidal Stability.

    PubMed

    Wagoner, Ty; Vardhanabhuti, Bongkosh; Foegeding, E Allen

    2016-01-01

    Interactions between whey proteins and polysaccharides, in particular the formation of food-grade soluble complexes, are of interest because of potential functional and health benefits. A specific application that has not received much attention is the use of complexes for enhanced colloidal stability of protein sols, such as protein-containing beverages. In beverages, the primary goal is the formation of complexes that remain dispersed after thermal processing and extended storage. This review highlights recent progress in the area of forming whey protein-polysaccharide soluble complexes that would be appropriate for beverage applications. Research in this area indicates that soluble complexes can be formed and stabilized that are reasonably small in size and possess a large surface charge that would predict colloidal stability. Selection of specific proteins and polysaccharides can be tailored to desired conditions. The principal challenges involve overcoming restrictions on protein concentration and ensuring that protein remains bioavailable.

  8. The specificity of interactions between proteins and sulfated polysaccharides.

    PubMed

    Mulloy, Barbara

    2005-12-01

    Sulfated polysaccharides are capable of binding with proteins at several levels of specificity. As highly acidic macromolecules, they can bind non-specifically to any basic patch on a protein surface at low ionic strength, and such interactions are not likely to be physiologically significant. On the other hand, several systems have been identified in which very specific substructures of sulfated polysaccharides confer high affinity for particular proteins; the best-known example of this is the pentasaccharide in heparin with high affinity for antithrombin, but other examples may be taken from the study of marine invertebrates: the importance of the fine structure of dermatan sulfate (DS) to its interaction with heparin cofactor II (HCII), and the involvement of sea urchin egg-jelly fucans in species specific fertilization. A third, intermediate, kind of specific interaction is described for the cell-surface glycosaminoglycan heparan sulfate (HS), in which patterns of sulfate substitution can show differential affinities for cytokines, growth factors, and morphogens at cell surfaces and in the intracellular matrix. This complex interplay of proteins and glycans is capable of influencing the diffusion of such proteins through tissue, as well as modulating cellular responses to them. PMID:16341442

  9. The specificity of interactions between proteins and sulfated polysaccharides.

    PubMed

    Mulloy, Barbara

    2005-12-01

    Sulfated polysaccharides are capable of binding with proteins at several levels of specificity. As highly acidic macromolecules, they can bind non-specifically to any basic patch on a protein surface at low ionic strength, and such interactions are not likely to be physiologically significant. On the other hand, several systems have been identified in which very specific substructures of sulfated polysaccharides confer high affinity for particular proteins; the best-known example of this is the pentasaccharide in heparin with high affinity for antithrombin, but other examples may be taken from the study of marine invertebrates: the importance of the fine structure of dermatan sulfate (DS) to its interaction with heparin cofactor II (HCII), and the involvement of sea urchin egg-jelly fucans in species specific fertilization. A third, intermediate, kind of specific interaction is described for the cell-surface glycosaminoglycan heparan sulfate (HS), in which patterns of sulfate substitution can show differential affinities for cytokines, growth factors, and morphogens at cell surfaces and in the intracellular matrix. This complex interplay of proteins and glycans is capable of influencing the diffusion of such proteins through tissue, as well as modulating cellular responses to them.

  10. Protein/Polysaccharide Electrostatic Complexes and Their Applications in Stabilizing Oil-in-Water Emulsions.

    PubMed

    Ai, Wenjia; Fang, Yapeng; Xiang, Shengping; Yao, Xiaolin; Nishinari, Katsuyoshi; Phillips, Glyn O

    2015-01-01

    Consumers are becoming increasingly fastidious in demanding food products with improved quality and functionality. This largely relies on rational design of food structures. As the two key food ingredients, protein and polysaccharides play important roles in food structuring. The combination of protein and polysaccharide provides rich opportunities for food structure and function designs through molecular interaction and assembly. This paper provides a brief review on the formation and characterization of protein/polysaccharide electrostatic complexes and their applications in stabilizing oil-in-water emulsions, particularly those containing polyunsaturated fatty acids. PMID:26598842

  11. [Comparative Analysis of Spectrophotometric Methods of the Protein Measurement in the Pectic Polysaccharide Samples].

    PubMed

    Ponomareva, S A; Golovchenko, V V; Patova, O A; Vanchikova, E V; Ovodov, Y S

    2015-01-01

    For the assay to reliability of determination of the protein content in the pectic polysaccharide samples by absorbance in the ultraviolet and visible regions of the spectrum a comparison of the eleven techniques called Flores, Lovry, Bradford, Sedmak, Rueman (ninhydrin reaction) methods, the method of ultraviolet spectrophotometry, the method Benedict's reagent, the method Nessler's reagent, the method with amide black, the bicinchoninic reagent and the biuret method was carried out. The data obtained show that insufficient sensitivity of the seven methods from the listed techniques doesn't allow their usage for determination of protein content in pectic polysaccharide samples. But the Lowry, Bradford, Sedmak methods, and the method Nessler's reagent may be used for determination of protein content in pectic polysaccharide samples, and the Bradford method is advisable for protein contaminants content determination in pectic polysaccharide samples in case protein content is less than 15%, and the Lowry method--for samples is more than 15%. PMID:26165122

  12. Bioinspired matrices assembled by polysaccharide-protein interactions

    NASA Astrophysics Data System (ADS)

    Zhang, Le

    Bioinspired matrices assembled on the basis of noncovalent interactions between proteins and polysaccharides have been proved suitable to deliver therapeutically relevant proteins or DNAs. Our initial efforts were dedicated to the relationship between mechanical properties of hydrogels assembled based on specific interactions between low molecular weight heparin (LMWH) and heparin binding peptides (HBPs) such as HIP, ATIII, and PF4ZIP peptides. The measured differences in affinity and kinetics for LMWH-HBP binding likely lead to observed differences in the phase separation behavior of the poly (ethylene glycol) (PEG)-LMWH/PEG-HIP hydrogels versus the PEG-LMWH/PEG-ATIII hydrogels. More attention has been given to the PF4ZIP peptide employed for the noncovalent assembly of heparinized hydrogels. Multifunctional star PEG-PF4ZIP bioconjugates complexed with star PEG-LMWH form hydrogels that exhibit increasing elastic moduli with increasing mole ratio of PEG-PF4ZIP. The viscoelastic properties of the hydrogels can be controlled via alterations in the ratio between LMWH and PF4ZIP peptide, and comparisons with other PEG-LMWH/PEG-HBP hydrogels suggest the importance of both LMWH/HBP binding kinetics and the binding capacity of LMWH in determining rheological properties in these hydrogels. Characterization of the PEG-LMWH/PEG-PF4ZIP hydrogels suggests that useful moduli for soft tissue engineering applications are obtained at physiological temperatures and after applying high shear. Furthermore, in the basic fibroblast growth factor (bFGF) release, bFGF/vascular endothelial growth factor (VEGF) co-release, and hydrogel erosion results, the combination of growth factor (GF) release profiles and hydrogel erosion profiles suggests that GF delivery from the assembled hydrogels is mainly an erosion-controlled process that may permit co-release of GF with PEG-LMWH and may therefore also improve the bioactivity of GF delivered from these matrices. Hydrogels with such engineered

  13. Protein/polysaccharide interactions and their impact on haze formation in white wines.

    PubMed

    Dufrechou, Marie; Doco, Thierry; Poncet-Legrand, Céline; Sauvage, François-Xavier; Vernhet, Aude

    2015-11-18

    Proteins in white wines may aggregate and form hazes at room temperature. This was previously shown to be related to pH-induced conformational changes and to occur for pH <3.5. The aim of the present work was to study the impact of wine polysaccharides on pH-induced haze formation by proteins but also the consequences of their interactions with these proteins on the colloidal stability of white wines. To this end, model systems and purified global pools of wine proteins and polysaccharides were used first. Kinetics of aggregation, proteins involved, and turbidities related to final hazes were monitored. To further identify the impact of each polysaccharide, fractions purified to homogeneity were used in a second phase. These included two neutral (mannoprotein and arabinogalactan) and two negatively charged (rhamnogalacturonan II dimer (RG-II) and arabinogalactan) polysaccharides. The impact of major wine polysaccharides on wine protein aggregation at room temperature was clearly less marked than those of the pH and the ionic strength. Polysaccharides modulated the aggregation kinetics and final haziness, indicating that they interfere with the aggregation process, but could not prevent it.

  14. Blend-modification of soy protein/lauric acid edible films using polysaccharides.

    PubMed

    Pan, Hongyang; Jiang, Bo; Chen, Jie; Jin, Zhengyu

    2014-05-15

    Different types of polysaccharides (propyleneglycol alginate (PGA), pectin, carrageenan and aloe polysaccharide) were incorporated into soy protein isolate (SPI)/lauric acid (La) films using a co-drying process or by direct addition to form biodegradable composite films with modified water vapour permeability (WVP) and mechanical properties. The WVP of SPI/La/polysaccharide films decreased when polysaccharides were added using the co-drying process, regardless of the type of polysaccharide. The tensile strength of SPI/La film was increased by the addition of polysaccharides, and the percentage elongation at break was increased by incorporating PGA using the co-drying process. Regarding oxygen-barrier performance, no notable differences were observed between the SPI/La and SPI/La/polysaccharide films. The most significant improvement was observed by blending PGA, with the co-dried preparation exhibiting better properties than the direct-addition preparation. Scanning electron microscopy (SEM) revealed that the microstructures of the films are the basis for the differences in the barrier and mechanical properties of the modified blends of SPI, polysaccharides and La.

  15. Maillard reaction products of rice protein hydrolysates with mono-, oligo- and polysaccharides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rice protein, a byproduct of rice syrup production, is abundant but, its lack of functionality prevents its wide use as a food ingredient. Maillard reaction products of (MRPs) hydrolysates from the limited hydrolysis of rice protein (LHRP) and various mono-, oligo- and polysaccharides were evaluat...

  16. Role for Golgi reassembly and stacking protein (GRASP) in polysaccharide secretion and fungal virulence

    PubMed Central

    Kmetzsch, Lívia; Joffe, Luna S.; Staats, Charley C.; de Oliveira, Débora L.; Fonseca, Fernanda L.; Cordero, Radames J. B.; Casadevall, Arturo; Nimrichter, Leonardo; Schrank, Augusto; Vainstein, Marilene H.; Rodrigues, Marcio L.

    2011-01-01

    Secretion of virulence factors is a critical mechanism for the establishment of cryptococcosis, a disease caused by the yeast pathogen Cryptococcus neoformans. One key virulence strategy of C. neoformans is the release of glucuronoxylomannan (GXM), a capsule-associated immune-modulatory polysaccharide that reaches the extracellular space through secretory vesicles. Golgi reassembly and stacking protein (GRASP) is required for unconventional protein secretion mechanisms in different eukaryotic cells, but its role in polysaccharide secretion is unknown. This study demonstrates that a C. neoformans functional mutant of a GRASP ortholog had attenuated virulence in an animal model of cryptococcosis, in comparison to wild type (WT) and reconstituted cells. Mutant cells manifested altered Golgi morphology, failed to produce typical polysaccharide capsules and showed a reduced ability to secrete GXM both in vitro and during animal infection. Isolation of GXM from cultures of WT, reconstituted or mutant strains revealed that the GRASP ortholog mutant produced polysaccharides with reduced dimensions. The mutant was also more efficiently associated to and killed by macrophages than WT and reconstituted cells. These results demonstrate that GRASP, a protein involved in unconventional protein secretion, is also required for polysaccharide secretion and virulence in C. neoformans. PMID:21542865

  17. Role for Golgi reassembly and stacking protein (GRASP) in polysaccharide secretion and fungal virulence.

    PubMed

    Kmetzsch, Lívia; Joffe, Luna S; Staats, Charley C; de Oliveira, Débora L; Fonseca, Fernanda L; Cordero, Radames J B; Casadevall, Arturo; Nimrichter, Leonardo; Schrank, Augusto; Vainstein, Marilene H; Rodrigues, Marcio L

    2011-07-01

    Secretion of virulence factors is a critical mechanism for the establishment of cryptococcosis, a disease caused by the yeast pathogen Cryptococcus neoformans. One key virulence strategy of C. neoformans is the release of glucuronoxylomannan (GXM), a capsule-associated immune-modulatory polysaccharide that reaches the extracellular space through secretory vesicles. Golgi reassembly and stacking protein (GRASP) is required for unconventional protein secretion mechanisms in different eukaryotic cells, but its role in polysaccharide secretion is unknown. This study demonstrates that a C. neoformans functional mutant of a GRASP orthologue had attenuated virulence in an animal model of cryptococcosis, in comparison with wild-type (WT) and reconstituted cells. Mutant cells manifested altered Golgi morphology, failed to produce typical polysaccharide capsules and showed a reduced ability to secrete GXM both in vitro and during animal infection. Isolation of GXM from cultures of WT, reconstituted or mutant strains revealed that the GRASP orthologue mutant produced polysaccharides with reduced dimensions. The mutant was also more efficiently associated to and killed by macrophages than WT and reconstituted cells. These results demonstrate that GRASP, a protein involved in unconventional protein secretion, is also required for polysaccharide secretion and virulence in C. neoformans.

  18. Seminal fluid from sea urchin (Lytechinus variegatus) contains complex sulfated polysaccharides linked to protein.

    PubMed

    Cinelli, Leonardo P; Vilela-Silva, Ana-Cristina E S; Mourão, Paulo A S

    2009-09-01

    The eggs of sea urchins are covered by a jelly coat, which contains high concentrations of sulfated polysaccharides. These carbohydrates show species-specificity in inducing the sperm acrosome reaction. Several studies about the egg jelly of sea urchins have been published, but there is no information about the composition of the seminal fluid of these echinoderms. Here we report for the first time the occurrence of complex sulfated polysaccharides in the seminal fluid of the sea urchin Lytechinus variegatus. These polysaccharides occur as three fractions that differ mostly in their carbohydrate/protein ratios. The native molecular masses of the polymers are very high (> or = 200 kDa) but, after digestion with papain the size decreases to approximately 8 kDa. All fractions have a similar carbohydrate composition, containing mostly galactose, glucosamine and mannose. The heterogeneous sulfated polysaccharides differ from vertebrate glycosaminoglycans and also from all previously described polysaccharides from invertebrates. The physiological role of the sulfated carbohydrates from seminal fluid is not yet determined. However, by analogy with the effects proposed for some glycoproteins found in vertebrate seminal fluid, it may be possible that the sulfated polysaccharides from invertebrate are also involved in fertilization process.

  19. Translocation of capsular polysaccharides in pathogenic strains of Escherichia coli requires a 60-kilodalton periplasmic protein.

    PubMed Central

    Silver, R P; Aaronson, W; Vann, W F

    1987-01-01

    An 11.6-kilobase (kb) region of a 34-kb fragment of Escherichia coli DNA that encodes the K1 capsular polysaccharide genes is necessary for translocation of the K1 polysaccharide to the bacterial cell surface. This 11.6-kb region contains a gene, kpsD, encoding a 60-kilodalton protein. The kpsD gene was localized to a 2.4-kb PstI-BamHI fragment. Cells harboring a Tn1000 insertion in kpsD did not synthesize the 60-kilodalton protein and did not express polysaccharide on the cell surface. Immunodiffusion and rocket immunoelectrophoresis of cell extracts, however, demonstrated that K1 polysaccharide was synthesized by these cells. We present evidence that the kpsD gene product is synthesized as a precursor and that the processed form is located in the periplasmic space. Analysis of alkaline phosphatase activity of a kpsD-phoA fusion demonstrated that kpsD expression was under positive regulation. A 260-base-pair AluI fragment located within the kpsD coding sequence was used as a probe and was found to hybridize to chromosomal DNA from E. coli that synthesizes the K2, K5, K7, K12, and K13 capsular polysaccharides but not K3 and K100. These results suggest that the kpsD gene product may be required for export not only of K1 but for other K antigens as well. Images PMID:3119565

  20. A dry platform for separation of proteins from biomass-containing polysaccharides, lignin, and polyphenols.

    PubMed

    Barakat, Abdellatif; Jérôme, François; Rouau, Xavier

    2015-04-13

    License to mill: Proteins were continuously extracted from polysaccharides, lignin, and polyphenol by combining ultrafine milling with electrostatic separation. Such a fractionation process does not involve any solvent, catalyst, or external source of heating. In addition, this dry process is compatible with downstream enzymatic reactions, thus opening an attractive route for producing valuable chemicals from biomass. PMID:25760796

  1. Improved conjugation and purification strategies for the preparation of protein-polysaccharide conjugates.

    PubMed

    Suárez, N; Massaldi, H; Franco Fraguas, L; Ferreira, F

    2008-12-12

    A glycoconjugate constituted by the Streptococcus pneumoniae serotype 14 capsular polysaccharide (CPS14) and bovine serum albumin (BSA) was prepared, and the unique properties of Sephadex LH-20 were used to separate the conjugate from the unconjugated material. The strength of this approach consists in its capacity to produce pure polysaccharide-protein conjugate in good yield and free from unconjugated material, a common residual contaminant of this type of immunobiologicals. The CPS14-BSA conjugate prepared via an improved 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP)-activation technique was characterized chemically and its immunogenicity was evaluated in mice. The purified conjugate, unlike the corresponding polysaccharide, produced a T-cell-dependent response in this species. PMID:18992885

  2. Improved conjugation and purification strategies for the preparation of protein-polysaccharide conjugates.

    PubMed

    Suárez, N; Massaldi, H; Franco Fraguas, L; Ferreira, F

    2008-12-12

    A glycoconjugate constituted by the Streptococcus pneumoniae serotype 14 capsular polysaccharide (CPS14) and bovine serum albumin (BSA) was prepared, and the unique properties of Sephadex LH-20 were used to separate the conjugate from the unconjugated material. The strength of this approach consists in its capacity to produce pure polysaccharide-protein conjugate in good yield and free from unconjugated material, a common residual contaminant of this type of immunobiologicals. The CPS14-BSA conjugate prepared via an improved 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP)-activation technique was characterized chemically and its immunogenicity was evaluated in mice. The purified conjugate, unlike the corresponding polysaccharide, produced a T-cell-dependent response in this species.

  3. Structural and immunological characterization of Burkholderia pseudomallei O-polysaccharide-flagellin protein conjugates.

    PubMed Central

    Brett, P J; Woods, D E

    1996-01-01

    The O-polysaccharide moiety of Burkholderia pseudomallei 319a lipopolysaccharide was covalently linked to flagellin protein isolated from the same strain. A glycoconjugate incorporating adipic acid dihydrazide as a spacer molecule elicited high-titer immunoglobulin G responses to both the protein and carbohydrate components of the construct. This immunoglobulin G was capable of protecting diabetic rats from challenge with a heterologous B. pseudomallei strain. PMID:8698517

  4. Polysaccharide-free nucleic acids and proteins of Abelmoschus esculentus for versatile molecular studies.

    PubMed

    Manoj-Kumar, A; Reddy, K N; Manjulatha, M; Blanco, L

    2012-01-01

    Abelmoschus esculentus (okra) is one of the polysaccharide rich crop plants. The polysaccharides interfere with nucleic acids and protein isolation thereby affecting the downstream molecular analysis. So, to understand the molecular systematics of okra, high quality DNA, RNA and proteins are essential. In this study we present a method for extracting genomic DNA, RNA and proteins from polysaccharide rich okra tissues. The conventional extraction procedures were integrated with purification treatments with pectinase, RNase and proteinase K, which improved the quality and quantity of DNA as well. Using SDS, additional washes with CIA and NaCl precipitation improved the RNA isolation both quantitatively and qualitatively. Finally, ammonium acetate mediated protein precipitation and re-solubilization increased the quality of total protein extracts from the okra leaves. All of the methods above not only eliminated the impurities but also improved the quality and quantity of nucleic acids and proteins. Further, we subjected these samples to versatile downstream molecular analyses such as restriction endonuclease digestion, RAPD, Southern, reverse transcription-PCR and Western analysis and were proved to be successful.

  5. Proteins, peptides, polysaccharides, and nucleotides with inhibitory activity on human immunodeficiency virus and its enzymes.

    PubMed

    Ng, Tzi Bun; Cheung, Randy Chi Fai; Wong, Jack Ho; Chan, Wai Yee

    2015-12-01

    Human immunodeficiency virus (HIV), the causative agent of acquired immune deficiency syndrome, has claimed innumerable lives in the past. Many biomolecules which suppress HIV replication and also other biomolecules that inhibit enzymes essential to HIV replication have been reported. Proteins including a variety of milk proteins, ribosome-inactivating proteins, ribonucleases, antifungal proteins, and trypsin inhibitors; peptides comprising cathelicidins, defensins, synthetic peptides, and others; polysaccharides and polysaccharopeptides; nucleosides, nucleotides, and ribozymes, demonstrated anti-HIV activity. In many cases, the mechanism of anti-HIV action has been elucidated. Strategies have been devised to augment the anti-HIV potency of these compounds.

  6. Moderate PEGylation of the carrier protein improves the polysaccharide-specific immunogenicity of meningococcal group A polysaccharide conjugate vaccine.

    PubMed

    Zhang, Tingting; Yu, Weili; Wang, Yanfei; Hu, Tao

    2015-06-22

    Neisseria meningitidis can cause severe and fulminant diseases such as meningitis. Meningococcal capsular polysaccharide (PS) is a key virulence determinant that is not able to induce immunological memory. Conjugation of PS to a carrier protein can significantly increase the immunogenicity of PS and induce immunological memory. Due to the classically described carrier-induced epitopic suppression (CIES) mechanisms, a strong immune response against the carrier protein could suppress the immune response to PS after coadministration of free carrier protein with the conjugate vaccine. However, it was not clear whether suppressing or enhancing the protein-specific immunogenicity could improve the PS-specific immunogenicity of the conjugate vaccine. Thus, moderate PEGylation, extensive PEGylation and oligomerization were used to regulate the immunogenicity of tetanus toxoid (TT) in the conjugate vaccine (PS-TT). Moderate PEGylation led to a 2.7-fold increase in the PS-specific IgG titers elicited by PS-TT. In contrast, extensive PEGylation and oligomerization of TT led to 1.4-fold and 1.6-fold decrease in the PS-specific IgG titers elicited by PS-TT, respectively. The PS-specific immunogenicity of PS-TT can be increased by moderate PEGylation through mild suppression of the TT-specific immunogenicity. The PS-specific immunogenicity of PS-TT was decreased through significant suppression or enhancement of the TT-specific immunogenicity. Thus, our study contributes to understand the CIES mechanisms and improve the PS-specific immunogenicity of a meningococcal PS conjugate vaccine.

  7. Cellular response to poly(vinyl alcohol) nanofibers coated with biocompatible proteins and polysaccharides

    NASA Astrophysics Data System (ADS)

    Lee, So Young; Jang, Da Hyun; Kang, Yun Ok; Kim, O. Bok; Jeong, Lim; Kang, Hyun Ki; Lee, Seung Jin; Lee, Chong-Heon; Park, Won Ho; Min, Byung-Moo

    2012-07-01

    A PVA nanofibrous matrix was prepared by electrospinning an aqueous 10 wt% PVA solution. The mean diameter of the PVA nanofibers electrospun from the aqueous PVA solution was 240 nm. The water resistance of the as-spun PVA nanofibrous matrix was improved by physically crosslinking the PVA nanofibers by heat treatment at 150 °C for 10 min. In addition, the heat-treated PVA nanofibrous matrix was coated with biocompatible polysaccharides (chitosan (CHI) or hyaluronic acid (HA)) and proteins (collagen (COL) or silk fibroin (SF)) to construct biomimetic nanofibrous scaffolds. The coating of proteins or polysaccharides on the PVA nanofibrous matrix was confirmed by ATR-IR spectra, and the degree of coating was determined by elemental analysis based on nitrogen content. The coated PVA matrices exhibited less hydrophilicity, except for the HA coating, and better tensile properties than the pure PVA nanofibrous matrix. The increase in tensile properties was due to interfiber bonds formed by the coating. The effect of protein and polysaccharide coating on normal human keratinocytes (NHEKs) and fibroblasts (NHEFs) was examined by cytocompatibility assessment in vitro. Among the CHI-, COL-, HA- and SF-coated PVA matrices, the SF-coated PVA nanofibrous matrix was found to be the most promising scaffold for the attachment and spreading of NHEKs and NHEFs as compared to the pure PVA matrix. This approach to controlling the surface properties of nanofibrous structures with SF may be useful in the design and tailoring of novel matrices for skin regeneration.

  8. Effect of Protein, Polysaccharide, and Oxygen Concentration Profiles on Biofilm Cohesiveness▿

    PubMed Central

    Ahimou, Francois; Semmens, Michael J.; Haugstad, Greg; Novak, Paige J.

    2007-01-01

    It is important to control biofilm cohesiveness to optimize process performance. In this study, a membrane-aerated biofilm reactor inoculated with activated sludge was used to grow mixed-culture biofilms of different ages and thicknesses. The cohesions, or cohesive energy levels per unit volume of biofilm, based on a reproducible method using atomic force microscopy (F. Ahimou, M. J. Semmens, P. J. Novak, and G. Haugstad, Appl. Environ. Microbiol. 73:2897-2904, 2007), were determined at different locations within the depths of the biofilms. In addition, the protein and polysaccharide concentrations within the biofilm depths, as well as the dissolved oxygen (DO) concentration profiles within the biofilms, were measured. It was found that biofilm cohesion increased with depth but not with age. Level of biofilm cohesive energy per unit volume was strongly correlated with biofilm polysaccharide concentration, which increased with depth in the membrane-aerated biofilm. In a 12-day-old biofilm, DO also increased with depth and may therefore be linked to polysaccharide production. In contrast, protein concentration was relatively constant within the biofilm and did not appear to influence cohesion. PMID:17337565

  9. The derivatization of oxidized polysaccharides for protein immobilization and affinity chromatography.

    PubMed

    Junowicz, E; Charm, S E

    1976-03-25

    The present report describes the preparation of modified polysaccharides matrices useful for the synthesis of affinity adsorbents and immobilized proteins. Hydrazido-matrices were synthesized by condensing an excess of the bifunctional reagent, adipic acid dihydrazide, with periodate oxidized cellulose paper, Sephadex, or Sepharose matrices. Ribonucleotide dialdehyde cofactors, glyceraldehyde 3-phosphate, pyridoxal 5'-phosphate and oxidized DNAase B were separately bound to the hydrazido-polymers. Azido-matrices obtained by modification of the hydrazido-derivatives were coupled to specific amino ligands such as amino acids and proteins. Several adsorbents were prepared and used as models for affinity chromatography. PMID:1260016

  10. Bacteriophage Tailspikes and Bacterial O-Antigens as a Model System to Study Weak-Affinity Protein-Polysaccharide Interactions.

    PubMed

    Kang, Yu; Gohlke, Ulrich; Engström, Olof; Hamark, Christoffer; Scheidt, Tom; Kunstmann, Sonja; Heinemann, Udo; Widmalm, Göran; Santer, Mark; Barbirz, Stefanie

    2016-07-27

    Understanding interactions of bacterial surface polysaccharides with receptor protein scaffolds is important for the development of antibiotic therapies. The corresponding protein recognition domains frequently form low-affinity complexes with polysaccharides that are difficult to address with experimental techniques due to the conformational flexibility of the polysaccharide. In this work, we studied the tailspike protein (TSP) of the bacteriophage Sf6. Sf6TSP binds and hydrolyzes the high-rhamnose, serotype Y O-antigen polysaccharide of the Gram-negative bacterium Shigella flexneri (S. flexneri) as a first step of bacteriophage infection. Spectroscopic analyses and enzymatic cleavage assays confirmed that Sf6TSP binds long stretches of this polysaccharide. Crystal structure analysis and saturation transfer difference (STD) NMR spectroscopy using an enhanced method to interpret the data permitted the detailed description of affinity contributions and flexibility in an Sf6TSP-octasaccharide complex. Dodecasaccharide fragments corresponding to three repeating units of the O-antigen in complex with Sf6TSP were studied computationally by molecular dynamics simulations. They showed that distortion away from the low-energy solution conformation found in the octasaccharide complex is necessary for ligand binding. This is in agreement with a weak-affinity functional polysaccharide-protein contact that facilitates correct placement and thus hydrolysis of the polysaccharide close to the catalytic residues. Our simulations stress that the flexibility of glycan epitopes together with a small number of specific protein contacts provide the driving force for Sf6TSP-polysaccharide complex formation in an overall weak-affinity interaction system. PMID:27045683

  11. A Review of Pneumococcal Vaccines: Current Polysaccharide Vaccine Recommendations and Future Protein Antigens

    PubMed Central

    Daniels, Calvin C.; Rogers, P. David

    2016-01-01

    This review describes development of currently available pneumococcal vaccines, provides summary tables of current pneumococcal vaccine recommendations in children and adults, and describes new potential vaccine antigens in the pipeline. Streptococcus pneumoniae, the bacteria responsible for pneumonia, otitis media, meningitis and bacteremia, remains a cause of morbidity and mortality in both children and adults. Introductions of unconjugated and conjugated pneumococcal polysaccharide vaccines have each reduced the rate of pneumococcal infections caused by the organism S. pneumoniae. The first vaccine developed, the 23-valent pneumococcal polysaccharide vaccine (PPSV23), protected adults and children older than 2 years of age against invasive disease caused by the 23 capsular serotypes contained in the vaccine. Because PPSV23 did not elicit a protective immune response in children younger than 2 years of age, the 7-valent pneumococcal conjugate vaccine (PCV7) containing seven of the most common serotypes from PPSV23 in pediatric invasive disease was developed for use in children younger than 2 years of age. The last vaccine to be developed, the 13-valent pneumococcal conjugate vaccine (PCV13), contains the seven serotypes in PCV7, five additional serotypes from PPSV23, and a new serotype not contained in PPSV23 or PCV7. Serotype replacement with virulent strains that are not contained in the polysaccharide vaccines has been observed after vaccine implementation and stresses the need for continued research into novel vaccine antigens. We describe eight potential protein antigens that are in the pipeline for new pneumococcal vaccines. PMID:26997927

  12. Ultrastructural localization of acid protein polysaccharides and calcium in the vacuoles of the chicken thrombocyte.

    PubMed

    Daimon, T; Uchida, K; Mizuhira, V

    1977-04-20

    The coexistence of acid protein polysaccharides and calcium in the vacuoles of chicken thrombocytes were studied by means of ultrahistochemical methods and electron probe X-ray microanalysis. The thrombocytes possessed large vacuoles of a surface connected membrane system. After both ruthenium red staining and tannic acid fixation the innersurface coat of vacuoles was always strongly and continously visualized. Electron microscopic X-ray microprobe analysis of antimonate precipitates in thrombocytes fixed in K-antimonate-OsO4 revealed calcium localization on the innersurface of vacuoles. From these facts it seems likely that the vacuoles of the surface connected membrane system may take part in the pool or the transport of calcium.

  13. In vitro protein-polysaccharide conjugation: tyrosinase-catalyzed conjugation of gelatin and chitosan.

    PubMed

    Chen, Tianhong; Embree, Heather D; Wu, Li-Qun; Payne, Gregory F

    2002-09-01

    The enzyme tyrosinase was used for the in vitro conjugation of the protein gelatin to the polysaccharide chitosan. Tyrosinases are oxidative enzymes that convert accessible tyrosine residues of proteins into reactive o-quinone moieties. Spectrophotometric and dissolved oxygen studies indicate that tyrosinase can oxidize gelatin and we estimate that 1 in 5 gelatin chains undergo reaction. Oxidized tyrosyl residues (i.e., quinone residues) can undergo nonenzymatic reactions with available nucleophiles such as the nucleophilic amino groups of chitosan. Ultraviolet/visible, (1)H-NMR, and ir provided chemical evidence for the conjugation of oxidized gelatin with chitosan. Physical evidence for conjugation was provided by dynamic viscometry, which indicated that tyrosinase catalyzes the sol-to-gel conversion of gelatin/chitosan mixtures. The gels formed from tyrosinase-catalyzed reactions were observed to differ from gels formed by cooling gelatin. In contrast to gelatin gels, tyrosinase-generated gels had different thermal behavior and were broken by the chitosan-hydrolyzing enzyme chitosanase. These results demonstrate that tyrosinase can be exploited for the in vitro formation of protein-polysaccharide conjugates that offer interesting mechanical properties.

  14. The role of the plant-specific ALTERED XYLOGLUCAN9 protein in Arabidopsis cell wall polysaccharide O-acetylation.

    PubMed

    Schultink, Alex; Naylor, Dan; Dama, Murali; Pauly, Markus

    2015-04-01

    A mutation in the ALTERED XYLOGLUCAN9 (AXY9) gene was found to be causative for the decreased xyloglucan acetylation phenotype of the axy9.1 mutant, which was identified in a forward genetic screen for Arabidopsis (Arabidopsis thaliana) mutants. The axy9.1 mutant also exhibits decreased O-acetylation of xylan, implying that the AXY9 protein has a broad role in polysaccharide acetylation. An axy9 insertional mutant exhibits severe growth defects and collapsed xylem, demonstrating the importance of wall polysaccharide O-acetylation for normal plant growth and development. Localization and topological experiments indicate that the active site of the AXY9 protein resides within the Golgi lumen. The AXY9 protein appears to be a component of the plant cell wall polysaccharide acetylation pathway, which also includes the REDUCED WALL ACETYLATION and TRICHOME BIREFRINGENCE-LIKE proteins. The AXY9 protein is distinct from the TRICHOME BIREFRINGENCE-LIKE proteins, reported to be polysaccharide acetyltransferases, but does share homology with them and other acetyltransferases, suggesting that the AXY9 protein may act to produce an acetylated intermediate that is part of the O-acetylation pathway.

  15. The role of the plant-specific ALTERED XYLOGLUCAN9 protein in Arabidopsis cell wall polysaccharide O-acetylation.

    PubMed

    Schultink, Alex; Naylor, Dan; Dama, Murali; Pauly, Markus

    2015-04-01

    A mutation in the ALTERED XYLOGLUCAN9 (AXY9) gene was found to be causative for the decreased xyloglucan acetylation phenotype of the axy9.1 mutant, which was identified in a forward genetic screen for Arabidopsis (Arabidopsis thaliana) mutants. The axy9.1 mutant also exhibits decreased O-acetylation of xylan, implying that the AXY9 protein has a broad role in polysaccharide acetylation. An axy9 insertional mutant exhibits severe growth defects and collapsed xylem, demonstrating the importance of wall polysaccharide O-acetylation for normal plant growth and development. Localization and topological experiments indicate that the active site of the AXY9 protein resides within the Golgi lumen. The AXY9 protein appears to be a component of the plant cell wall polysaccharide acetylation pathway, which also includes the REDUCED WALL ACETYLATION and TRICHOME BIREFRINGENCE-LIKE proteins. The AXY9 protein is distinct from the TRICHOME BIREFRINGENCE-LIKE proteins, reported to be polysaccharide acetyltransferases, but does share homology with them and other acetyltransferases, suggesting that the AXY9 protein may act to produce an acetylated intermediate that is part of the O-acetylation pathway. PMID:25681330

  16. Ultrastructural localization of capsules, cell wall polysaccharide, cell wall proteins, and F antigen in pneumococci.

    PubMed Central

    Skov Sørensen, U B; Blom, J; Birch-Andersen, A; Henrichsen, J

    1988-01-01

    The localization of pneumococcal capsular and cell wall antigens was examined by immunoelectron microscopy. C polysaccharide (C-Ps), a common component of all pneumococci, was uniformly distributed on both the inside and outside of the cell walls. The thickness of the C-Ps varied with the strain. Encapsulated strains were covered by varied amounts of capsular polysaccharide concealing the C-Ps of the bacteria so as to render it inaccessible to anti-C-Ps antibodies. In addition to C-Ps, protein antigens were demonstrable on the surface of nonencapsulated pneumococci. The proteins were not masked by the C-Ps layer. An extra layer on the cell walls was conspicuous on electron micrographs of both rough and encapsulated pneumococci. The nature of this extra layer has not been disclosed. F antigen, another common antigen of pneumococci, was uniformly distributed on the surface of the plasma membranes. During the course of the experimental work a reproducible method of gold labeling immunoglobulins was developed. Images PMID:3397179

  17. Physico-chemical characterization of protein-associated polysaccharides extracted from sugar beet pulp.

    PubMed

    Fishman, Marshall L; Chau, Hoa K; Qi, Phoebe X; Hotchkiss, Arland T; Yadav, Madhav P

    2013-02-15

    We have solubilized and separated polysaccharides from sugar beet pulp (SBP) into three fractions with steam assisted flash extraction (SAFE). For pectin, recovery ranged from 8 to 14%, degree of methy-esterification 66-73%, crude protein 1.3-1.7%, M(w) 262-318 kDa, η(w) 0.22-0.23 dL/g, Rg(z) 36-39 nm and Rh(z) 41-42 nm. For alkaline soluble polysaccharides, (ASP I) recovery ranged from 4.0 to 6.5%, crude protein 1.2-4.8%, weight average molar mass (M(w)) 66-68 kDa, weight average intrinsic viscosity (η(w)) 0.27-0.30 dL/g, z-average radius of gyration (Rg(z)) 25-29 nm and z-average hydrated radius (Rh(z)) 10-11 nm. ASP II recovery ranged from 2.0 to 8.6%, crude protein 1.2-4.8%, M(w) 299-339 kDa, η(w) 0.22-0.33 dL/g, Rg(z) 33-34 nm and Rh(z) 30-34 nm. Recovery of the residue mainly cellulose, ranged from 20.3 to 22.3%. The cellulose in this fraction was converted to carboxymethyl cellulose (CMC). The CMC fraction contained 0.33-0.43 crude protein and had an M(w) ranging from 127 to 263 kDa, η(w) 3.6-8.0 dL/g, Rg(z) 35-45 nm and Rh(z) 27-40 nm.

  18. Microanalysis characterization of bioactive protein-bound polysaccharides produced by Amanita ponderosa cultures.

    PubMed

    Salvador, Cátia; Martins, M Rosário; Caldeira, A Teresa

    2015-02-01

    Different compounds of edible mushrooms are responsible for their bioactivity. The ability to synthesize polysaccharides, namely protein-polysaccharide (PPS) complexes, is related to the antioxidant capacity of these compounds and present great interest in preventing a number of diseases, including cancer, cardiovascular and auto-immune diseases, and accelerated aging. Amanita ponderosa are wild edible mushrooms that grow in Mediterranean "montado" areas [Portuguese name given to cork oak (Quercus suber) and holm oak (Quercus ilex) forests]. The aim of this study was to evaluate the production of PPS complexes obtained from A. ponderosa cultures using a new microanalytical approach to quickly and easily monitor the production process. Microanalysis using Fourier-transform infrared using attenuated total reflection and Raman spectroscopy of PPS samples showed spectra compatible with identification of this type of compound in culture extracts. PPS separated by size-exclusion chromatography showed seven main complexes. Molecular weights of the main PPS complexes isolated from cultures ranged between 1.5 and 20 kDa and did not present toxicity against Artemia salina, demonstrating the potential of A. ponderosa as a source of biologically active compounds with nutraceutical value. Application of this microanalytical approach to monitoring the production of PPS compounds can be successfully applied in biotechnological processes.

  19. Structural correlates of carrier protein recognition in tetanus toxoid-conjugated bacterial polysaccharide vaccines.

    PubMed

    Lockyer, Kay; Gao, Fang; Derrick, Jeremy P; Bolgiano, Barbara

    2015-03-10

    An analysis of structure-antibody recognition relationships in nine licenced polysaccharide-tetanus toxoid (TT) conjugate vaccines was performed. The panel of conjugates used included vaccine components to protect against disease caused by Haemophilus influenzae type b, Neisseria meningitidis groups A, C, W and Y and Streptococcus pneumoniae serotype 18C. Conformation and structural analysis included size exclusion chromatography with multi-angle light scattering to determine size, and intrinsic fluorescence spectroscopy and fluorescence quenching to evaluate the protein folding and exposure of Trp residues. A capture ELISA measured the recognition of TT epitopes in the conjugates, using four rat monoclonal antibodies: 2 localised to the HC domain, and 2 of which were holotoxoid conformation-dependent. The conjugates had a wide range of average molecular masses ranging from 1.8×10(6) g/mol to larger than 20×10(6) g/mol. The panel of conjugates were found to be well folded, and did not have spectral features typical of aggregated TT. A partial correlation was found between molecular mass and epitope recognition. Recognition of the epitopes either on the HC domain or the whole toxoid was not necessarily hampered by the size of the molecule. Correlation was also found between the accessibility of Trp side chains and polysaccharide loading, suggesting also that a higher level of conjugated PS does not necessarily interfere with toxoid accessibility. There were different levels of carrier protein Trp side-chain and epitope accessibility that were localised to the HC domain; these were related to the saccharide type, despite the conjugates being independently manufactured. These findings extend our understanding of the molecular basis for carrier protein recognition in TT conjugate vaccines.

  20. Structural correlates of carrier protein recognition in tetanus toxoid-conjugated bacterial polysaccharide vaccines.

    PubMed

    Lockyer, Kay; Gao, Fang; Derrick, Jeremy P; Bolgiano, Barbara

    2015-03-10

    An analysis of structure-antibody recognition relationships in nine licenced polysaccharide-tetanus toxoid (TT) conjugate vaccines was performed. The panel of conjugates used included vaccine components to protect against disease caused by Haemophilus influenzae type b, Neisseria meningitidis groups A, C, W and Y and Streptococcus pneumoniae serotype 18C. Conformation and structural analysis included size exclusion chromatography with multi-angle light scattering to determine size, and intrinsic fluorescence spectroscopy and fluorescence quenching to evaluate the protein folding and exposure of Trp residues. A capture ELISA measured the recognition of TT epitopes in the conjugates, using four rat monoclonal antibodies: 2 localised to the HC domain, and 2 of which were holotoxoid conformation-dependent. The conjugates had a wide range of average molecular masses ranging from 1.8×10(6) g/mol to larger than 20×10(6) g/mol. The panel of conjugates were found to be well folded, and did not have spectral features typical of aggregated TT. A partial correlation was found between molecular mass and epitope recognition. Recognition of the epitopes either on the HC domain or the whole toxoid was not necessarily hampered by the size of the molecule. Correlation was also found between the accessibility of Trp side chains and polysaccharide loading, suggesting also that a higher level of conjugated PS does not necessarily interfere with toxoid accessibility. There were different levels of carrier protein Trp side-chain and epitope accessibility that were localised to the HC domain; these were related to the saccharide type, despite the conjugates being independently manufactured. These findings extend our understanding of the molecular basis for carrier protein recognition in TT conjugate vaccines. PMID:25640334

  1. Maternal immunization of mice with group B streptococcal type III polysaccharide-beta C protein conjugate elicits protective antibody to multiple serotypes.

    PubMed Central

    Madoff, L C; Paoletti, L C; Tai, J Y; Kasper, D L

    1994-01-01

    Group B streptococcal infection is a major cause of neonatal mortality. Antibody to the capsular polysaccharide protects against invasive neonatal disease, but immunization with capsular polysaccharides fails to elicit protective antibody in many recipients. Conjugation of the polysaccharide to tetanus toxoid has been shown to increase immune response to the polysaccharide. In animal models, C proteins of group B streptococci are also protective determinants. We examined the ability of the beta C protein to serve in the dual role of carrier for the polysaccharide and protective immunogen. Type III polysaccharide was covalently coupled to beta C protein by reductive amination. Immunization of rabbits with the polysaccharide-protein conjugate elicited high titers of antibody to both components, and the serum induced opsonophagocytic killing of type III, Ia/C, and Ib/C strains of group B streptococci. Female mice were immunized with the conjugate vaccine and then bred; 93% of neonatal pups born to these dams vaccinated with conjugate survived type III group B streptococcal challenge and 76% survived type Ia/C challenge, compared with 3% and 8% survival, respectively, in controls (P < 0.001). The beta C protein acted as an effective carrier for the type III polysaccharide while simultaneously induced protective immunity against beta C protein--containing strains of group B streptococci. Images PMID:7518832

  2. Noncovalent Dispersion and Functionalization of Cellulose Nanocrystals with Proteins and Polysaccharides.

    PubMed

    Fang, Wenwen; Arola, Suvi; Malho, Jani-Markus; Kontturi, Eero; Linder, Markus B; Laaksonen, Päivi

    2016-04-11

    Native cellulose nanocrystals (CNCs) are valuable high quality materials with potential for many applications including the manufacture of high performance materials. In this work, a relatively effortless procedure was introduced for the production of CNCs, which gives a nearly 100% yield of crystalline cellulose. However, the processing of the native CNCs is hindered by the difficulty in dispersing them in water due to the absence of surface charges. To overcome these difficulties, we have developed a one-step procedure for dispersion and functionalization of CNCs with tailored cellulose binding proteins. The process is also applicable for polysaccharides. The tailored cellulose binding proteins are very efficient for the dispersion of CNCs due to the selective interaction with cellulose, and only small fraction of proteins (5-10 wt %, corresponds to about 3 μmol g(-1)) could stabilize the CNC suspension. Xyloglucan (XG) enhanced the CNC dispersion above a fraction of 10 wt %. For CNC suspension dispersed with carboxylmethyl cellulose (CMC) we observed the most long-lasting stability, up to 1 month. The cellulose binding proteins could not only enhance the dispersion of the CNCs, but also functionalize the surface. This we demonstrated by attaching gold nanoparticles (GNPs) to the proteins, thus, forming a monolayer of GNPs on the CNC surface. Cryo transmission electron microscopy (Cryo-TEM) imaging confirmed the attachment of the GNPs to CNC solution conditions. PMID:26907991

  3. A new family of β-helix proteins with similarities to the polysaccharide lyases

    DOE PAGESBeta

    Close, Devin W.; D'Angelo, Sara; Bradbury, Andrew R. M.

    2014-09-27

    Microorganisms that degrade biomass produce diverse assortments of carbohydrate-active enzymes and binding modules. Despite tremendous advances in the genomic sequencing of these organisms, many genes do not have an ascribed function owing to low sequence identity to genes that have been annotated. Consequently, biochemical and structural characterization of genes with unknown function is required to complement the rapidly growing pool of genomic sequencing data. A protein with previously unknown function (Cthe_2159) was recently isolated in a genome-wide screen using phage display to identify cellulose-binding protein domains from the biomass-degrading bacterium Clostridium thermocellum. Here, the crystal structure of Cthe_2159 is presentedmore » and it is shown that it is a unique right-handed parallel β-helix protein. Despite very low sequence identity to known β-helix or carbohydrate-active proteins, Cthe_2159 displays structural features that are very similar to those of polysaccharide lyase (PL) families 1, 3, 6 and 9. Cthe_2159 is conserved across bacteria and some archaea and is a member of the domain of unknown function family DUF4353. This suggests that Cthe_2159 is the first representative of a previously unknown family of cellulose and/or acid-sugar binding β-helix proteins that share structural similarities with PLs. More importantly, these results demonstrate how functional annotation by biochemical and structural analysis remains a critical tool in the characterization of new gene products.« less

  4. A new family of β-helix proteins with similarities to the polysaccharide lyases.

    PubMed

    Close, Devin W; D'Angelo, Sara; Bradbury, Andrew R M

    2014-10-01

    Microorganisms that degrade biomass produce diverse assortments of carbohydrate-active enzymes and binding modules. Despite tremendous advances in the genomic sequencing of these organisms, many genes do not have an ascribed function owing to low sequence identity to genes that have been annotated. Consequently, biochemical and structural characterization of genes with unknown function is required to complement the rapidly growing pool of genomic sequencing data. A protein with previously unknown function (Cthe_2159) was recently isolated in a genome-wide screen using phage display to identify cellulose-binding protein domains from the biomass-degrading bacterium Clostridium thermocellum. Here, the crystal structure of Cthe_2159 is presented and it is shown that it is a unique right-handed parallel β-helix protein. Despite very low sequence identity to known β-helix or carbohydrate-active proteins, Cthe_2159 displays structural features that are very similar to those of polysaccharide lyase (PL) families 1, 3, 6 and 9. Cthe_2159 is conserved across bacteria and some archaea and is a member of the domain of unknown function family DUF4353. This suggests that Cthe_2159 is the first representative of a previously unknown family of cellulose and/or acid-sugar binding β-helix proteins that share structural similarities with PLs. Importantly, these results demonstrate how functional annotation by biochemical and structural analysis remains a critical tool in the characterization of new gene products.

  5. A new family of β-helix proteins with similarities to the polysaccharide lyases

    SciTech Connect

    Close, Devin W.; D'Angelo, Sara; Bradbury, Andrew R. M.

    2014-09-27

    Microorganisms that degrade biomass produce diverse assortments of carbohydrate-active enzymes and binding modules. Despite tremendous advances in the genomic sequencing of these organisms, many genes do not have an ascribed function owing to low sequence identity to genes that have been annotated. Consequently, biochemical and structural characterization of genes with unknown function is required to complement the rapidly growing pool of genomic sequencing data. A protein with previously unknown function (Cthe_2159) was recently isolated in a genome-wide screen using phage display to identify cellulose-binding protein domains from the biomass-degrading bacterium Clostridium thermocellum. Here, the crystal structure of Cthe_2159 is presented and it is shown that it is a unique right-handed parallel β-helix protein. Despite very low sequence identity to known β-helix or carbohydrate-active proteins, Cthe_2159 displays structural features that are very similar to those of polysaccharide lyase (PL) families 1, 3, 6 and 9. Cthe_2159 is conserved across bacteria and some archaea and is a member of the domain of unknown function family DUF4353. This suggests that Cthe_2159 is the first representative of a previously unknown family of cellulose and/or acid-sugar binding β-helix proteins that share structural similarities with PLs. More importantly, these results demonstrate how functional annotation by biochemical and structural analysis remains a critical tool in the characterization of new gene products.

  6. Stratification structure of polysaccharides and proteins in activated sludge with different aeration in membrane bioreactor.

    PubMed

    Zhang, Haifeng; Yu, Haihuan; Zhang, Lanhe; Song, Lianfa

    2015-09-01

    The effect of distribution pattern of polysaccharides (PS) and proteins (PN) in activated sludge (AS) stratification with different aeration rates on membrane fouling and rejection efficiency were investigated. During high aeration, PN and PS concentrations increased in supernatant, the dominant fraction (84% of PN and 73% of PS) was small molecules (<1 kDa). Less slime and loose bound extracellular polymeric substances (LB-EPS), more tight bound EPS (TB-EPS) were observed compared with low aeration. The decrease in PN/PS ratio and Ca(2+) concentration within EPS deteriorated AS flocculation ability. At slow trans-membrane pressure (TMP) rise stage, fouling rate under high aeration was 41% lower than low aeration due to lower PN within EPS outer. Low PS rejection rate (about 23%) leaded to higher PS in effluent at this stage. High PS rejection rate (about 94%) at rapid TMP rise stage resulted in about 2.2-time higher fouling rate than that low aeration.

  7. Effects of various protein- and polysaccharide-based clarification agents on antioxidative compounds and colour of pomegranate juice.

    PubMed

    Erkan-Koç, Betül; Türkyılmaz, Meltem; Yemiş, Oktay; Özkan, Mehmet

    2015-10-01

    The effects of natural sedimentation and clarification agents [protein-based (albumin, casein and gelatin) and polysaccharide-based (chitosan and xanthan gum)] on total phenolics, hydrolysable tannins, anthocyanins and antioxidant activity of pomegranate juice were investigated. While protein-based agents led to higher reductions in the contents of total phenolics (7.2-17.2%), hydrolysable tannins (16.7-59.5%) and anthocyanins (11.7-23.7%) (p<0.05) than natural sedimentation, polysaccharide-based agents had similar effects on these compounds, like natural sedimentation (p>0.05). The stabilities of β-punicalagin and α-punicalagin (the major hydrolysable tannins) against clarification were similar. Moreover, antioxidant activities of the samples clarified with polysaccharide-based agents were higher (2.4-26.6%) than those of the protein-based agents. For both high total phenolic, hydrolysable tannin and anthocyanins contents, as well as high antioxidant activity, we suggest that polysaccharide-based agents be used for the clarification of pomegranate juice. In fact, chitosan is especially recommended for this purpose since chitosan (10.3NTU) led to lower turbidity as compared to xanthan gum (20.0 NTU).

  8. Solution conformation and flexibility of capsular polysaccharides from Neisseria meningitidis and glycoconjugates with the tetanus toxoid protein

    PubMed Central

    Abdelhameed, Ali Saber; Morris, Gordon A.; Almutairi, Fahad; Adams, Gary G.; Duvivier, Pierre; Conrath, Karel; Harding, Stephen E.

    2016-01-01

    The structural integrity of meningococcal native, micro-fluidized and activated capsular polysaccharides and their glycoconjugates – in the form most relevant to their potential use as vaccines (dilute solution) - have been investigated with respect to their homogeneity, conformation and flexibility. Sedimentation velocity analysis showed that the polysaccharide size distributions were generally bimodal with some evidence for higher molar mass forms at higher concentration. Weight average molar masses Mw where lower for activated polysaccharides. Conjugation with tetanus toxoid protein however greatly increased the molar mass and polydispersity of the final conjugates. Glycoconjugates had an approximately unimodal log-normal but broad and large molar mass profiles, confirmed by sedimentation equilibrium “SEDFIT MSTAR” analysis. Conformation analysis using HYDFIT (which globally combines sedimentation and viscosity data), “Conformation Zoning” and Wales-van Holde approaches showed a high degree of flexibility – at least as great as the unconjugated polysaccharides, and very different from the tetanus toxoid (TT) protein used for the conjugation. As with the recently published finding for Hib-TT complexes, it is the carbohydrate component that dictates the solution behaviour of these glycoconjugates, although the lower intrinsic viscosities suggest some degree of compaction of the carbohydrate chains around the protein. PMID:27782149

  9. Protein translation inhibition by Stachybotrys chartarum conidia with and without the mycotoxin containing polysaccharide matrix.

    PubMed

    Karunasena, Enusha; Cooley, J Danny; Straus, Douglas; Straus, David C

    2004-07-01

    Recent studies have correlated the presence of Stachybotrys chartarum in structures with SBS. S. chartarum produces mycotoxins that are thought to produce some of the symptoms reported in sick-building syndrome (SBS). The conidia (spores) produced by Stachybotrys species are not commonly found in the air of buildings that have been found to contain significant interior growth of this organism. This could be due in part to the large size of the Stachybotrys spores, or the organism growing in hidden areas such as wall cavities. However, individuals in buildings with significant Stachybotrys growth frequently display symptoms that may be attributed to exposure to the organism's mycotoxins. In addition, Stachybotrys colonies produce a "slime" or polysaccharide (carbohydrate) matrix that coats the hyphae and the spores. The intent of this project was to determine whether the carbohydrate matrix and the mycotoxins embedded in it could be removed from the spores by repeated washings with either aqueous or organic solvents. The results demonstrated that the process of spore washing removed compounds that were toxic in a protein translation assay as compared to spores that were washed with an organic solution, however a correlation between carbohydrate removal during the washing process and the removal of mycotoxins from the spore surface was not observed. These data demonstrated that mycotoxins are not likely to be found exclusively in the carbohydrate matrix of the spores. Therefore, mycotoxin removal from the spore surface can occur without significant loss of polysaccharide. We also showed that toxic substances may be removed from the spore surface with an aqueous solution. These results suggest that satratoxins are soluble in aqueous solutions without being bound to water-soluble moieties, such as the carbohydrate slime matrix. PMID:15487326

  10. Evidence for a role of the polysaccharide capsule transport proteins in pertussis pathogenesis.

    PubMed

    Hoo, Regina; Lam, Jian Hang; Huot, Ludovic; Pant, Aakanksha; Li, Rui; Hot, David; Alonso, Sylvie

    2014-01-01

    Polysaccharide (PS) capsules are important virulence determinants for many bacterial pathogens. Bordetella pertussis, the agent of whooping cough, produces a surface associated microcapsule but its role in pertussis pathogenesis remained unknown. Here we showed that the B. pertussis capsule locus is expressed in vivo in murine lungs and that absence of the membrane-associated protein KpsT, involved in the transport of the PS polymers across the envelope, but not the surface-exposed PS capsule itself, affects drastically B. pertussis colonization efficacy in mice. Microarray analysis revealed that absence of KpsT in B. pertussis resulted in global down-regulation of gene expression including key virulence genes regulated by BvgA/S, the master two-component system. Using a BvgS phase-locked mutant, we demonstrated a functional link between KpsT and BvgA/S-mediated signal transduction. Whereas pull-down assays do not support physical interaction between BvgS sensor and any of the capsule locus encoded proteins, absence of KpsT impaired BvgS oligomerization, necessary for BvgS function. Furthermore, complementation studies indicated that instead of KpsT alone, the entire PS capsule transport machinery spanning the cell envelope likely plays a role in BvgS-mediated signal transduction. Our work thus provides the first experimental evidence of a role for a virulence-repressed gene in pertussis pathogenesis.

  11. Evidence for a Role of the Polysaccharide Capsule Transport Proteins in Pertussis Pathogenesis

    PubMed Central

    Hoo, Regina; Lam, Jian Hang; Huot, Ludovic; Pant, Aakanksha; Li, Rui; Hot, David; Alonso, Sylvie

    2014-01-01

    Polysaccharide (PS) capsules are important virulence determinants for many bacterial pathogens. Bordetella pertussis, the agent of whooping cough, produces a surface associated microcapsule but its role in pertussis pathogenesis remained unknown. Here we showed that the B. pertussis capsule locus is expressed in vivo in murine lungs and that absence of the membrane-associated protein KpsT, involved in the transport of the PS polymers across the envelope, but not the surface-exposed PS capsule itself, affects drastically B. pertussis colonization efficacy in mice. Microarray analysis revealed that absence of KpsT in B. pertussis resulted in global down-regulation of gene expression including key virulence genes regulated by BvgA/S, the master two-component system. Using a BvgS phase-locked mutant, we demonstrated a functional link between KpsT and BvgA/S-mediated signal transduction. Whereas pull-down assays do not support physical interaction between BvgS sensor and any of the capsule locus encoded proteins, absence of KpsT impaired BvgS oligomerization, necessary for BvgS function. Furthermore, complementation studies indicated that instead of KpsT alone, the entire PS capsule transport machinery spanning the cell envelope likely plays a role in BvgS-mediated signal transduction. Our work thus provides the first experimental evidence of a role for a virulence-repressed gene in pertussis pathogenesis. PMID:25501560

  12. Influence of pea protein aggregates on the structure and stability of pea protein/soybean polysaccharide complex emulsions.

    PubMed

    Yin, Baoru; Zhang, Rujing; Yao, Ping

    2015-01-01

    The applications of plant proteins in the food and beverage industry have been hampered by their precipitation in acidic solution. In this study, pea protein isolate (PPI) with poor dispersibility in acidic solution was used to form complexes with soybean soluble polysaccharide (SSPS), and the effects of PPI aggregates on the structure and stability of PPI/SSPS complex emulsions were investigated. Under acidic conditions, high pressure homogenization disrupts the PPI aggregates and the electrostatic attraction between PPI and SSPS facilitates the formation of dispersible PPI/SSPS complexes. The PPI/SSPS complex emulsions prepared from the PPI containing aggregates prove to possess similar droplet structure and similar stability compared with the PPI/SSPS emulsions produced from the PPI in which the aggregates have been previously removed by centrifugation. The oil droplets are protected by PPI/SSPS complex interfacial films and SSPS surfaces. The emulsions show long-term stability against pH and NaCl concentration changes. This study demonstrates that PPI aggregates can also be used to produce stable complex emulsions, which may promote the applications of plant proteins in the food and beverage industry.

  13. Effects of drying methods on physicochemical and immunomodulatory properties of polysaccharide-protein complexes from litchi pulp.

    PubMed

    Huang, Fei; Guo, Yajuan; Zhang, Ruifen; Yi, Yang; Deng, Yuanyuan; Su, Dongxiao; Zhang, Mingwei

    2014-01-01

    Dried litchi pulp has been used in traditional remedies in China for many years to treat various diseases, and the therapeutic activity has been, at least partly, attributed to the presence of bioactive polysaccharides. Polysaccharide-protein complexes from vacuum freeze-(VF), vacuum microwave-(VM) and heat pump (HP) dried litchi pulp, which were coded as LP-VF, LP-VM and LP-HP, were comparatively studied on the physicochemical and immunomodulatory properties. LP-HP had a predominance of galactose, while glucose was the major sugar component in LP-VF and LP-VM. Compared with LP-VF and LP-VM, LP-HP contained more aspartate and glutamic in binding protein. LP-HP also exhibited a stronger stimulatory effect on splenocyte proliferation at 200 μg/mL and triggered higher NO, TNF-α and IL-6 secretion from RAW264.7 macrophages. Different drying methods caused the difference in physicochemical properties of polysaccharide-protein complexes from dried litchi pulp, which resulted in significantly different immunomodulatory activity. HP drying appears to be the best method for preparing litchi pulp to improve its immunomodulatory properties. PMID:25140451

  14. Preparation, characterization, and immunological properties in mice of Escherichia coli O157 O-specific polysaccharide-protein conjugate vaccines.

    PubMed Central

    Konadu, E; Robbins, J B; Shiloach, J; Bryla, D A; Szu, S C

    1994-01-01

    Escherichia coli O157 causes severe enteritis and the extraintestinal complication of hemolytic-uremic syndrome, with their highest incidence occurring in children. We postulated that serum immunoglobulin G (IgG) antibodies to the O-specific polysaccharide of lipopolysaccharide (LPS) may confer protective immunity to enteric pathogens by inducing bactericidal reactions against the ingested organisms in the jejunum (J. B. Robbins, C. Chu, and R. Schneerson, Clin. Infect. Dis. 15:346-361, 1992; S. C. Szu, R. Gupta, and J. B. Robbins, p. 381-394, in I. K. Wachsmuth, P. A. Blake, and O. Olsvik, ed., Vibrio cholerae, 1994). Because polysaccharide-protein conjugates induce serum IgG antibodies in infants, we bound the O-specific polysaccharide of E. coli O157 to proteins. E. coli O157 LPS, treated with acetic acid or hydrazine, was derivatized with adipic acid dihydrazide and bound to proteins by carbodiimide-mediated condensation. Conjugates of these adipic hydrazide derivative were prepared with bovine serum albumin, formalin-treated exotoxin C of Clostridium welchii (Pig Bel toxoid), or Pseudomonas aeruginosa recombinant exoprotein A. The conjugates had low levels of endotoxin and elicited serum antibodies with bactericidal activity to the O157 LPS. The largest increase in LPS antibodies was of the IgG class. Clinical evaluation of E. coli O157-toxoid conjugates is planned. Images PMID:7927787

  15. Structural and Functional Properties of Soy Protein Isolates Modified by Soy Soluble Polysaccharides.

    PubMed

    Xu, Yan-Teng; Liu, Ling-Ling

    2016-09-28

    Aiming to achieve the modification to soy protein isolate (SPI) by soy soluble polysaccharides (SSPS), electrically driven complex systems were first established in the environment of pH 3.0, and then reconstituted SPI particles with different SPI-SSPS ratios were obtained under freeze-drying process. Through this treatment, the structures of SPI particles were partly unfolded and adsorbed SSPS mainly via hydrophobic interactions and hydrogen bonding with larger particle sizes. The adherence of SSPS decreased the surface hydrophobicity of reconstituted SPI particles, but exerted not much influence on the emulsifying and foaming activities and increased the corresponding stabilities due to enhancing the unfolded extent of structure and improving the conformation flexibility. Reconstituted SPI-SSPS particles might rearrange and link each other due to the presence of SSPS on the air-water interface to better stabilize these systems. At SPI-SSPS ratio of 10:1, lower temperature was required to form gels with lower gel intensity and porous structure. The findings provide a further comprehension to the relationship between structures and functional properties of SPI modified by SSPS and the feasibility of applying these reconstituted particles to needed areas. PMID:27608266

  16. Protection of mice against Salmonella typhimurium with an O-specific polysaccharide-protein conjugate vaccine.

    PubMed Central

    Watson, D C; Robbins, J B; Szu, S C

    1992-01-01

    Serious infections with salmonellae remain a threat in many human populations. Despite extensive study of salmonella infections in animals and clinical experience with killed cellular vaccines, there are no vaccines against serotypes other than Salmonella typhi licensed for human use. Serum antibodies to the O-specific polysaccharide (O-SP) of salmonellae protect mice against invasive infection. In order to render it immunogenic, we have conjugated the O-SP of Salmonella typhimurium to carrier proteins by various schemes. O-SP conjugated to tetanus toxoid (O-SP-TT) elicited antibodies in outbred mice after three subcutaneous injections without adjuvant. The O-SP alone elicited no detectable antibody. The antibody response to O-SP-TT was boosted by successive doses and consisted of immunoglobulin G (IgG) and IgM. Most mice only produced antibodies specific for the abequose (O:4 factor) region of the O-SP. Occasional animals also produced antibodies to the core oligosaccharide. Immunized mice were protected against intraperitoneal challenge with S. typhimurium, demonstrating a 160-fold increase in the 50% lethal dose. Passive immunization with conjugate-induced IgM or IgG also protected against challenge. These results indicate that an O-SP-TT conjugate, when given by a route and formulation acceptable for human use, protects mice against challenge with S. typhimurium. Images PMID:1383154

  17. The polysaccharides from porphyra yezoensis suppress the denaturation of bighead carp myofibrillar protein.

    PubMed

    Jiang, Long-Fa

    2014-07-01

    In this study, investigated was the effect of the porphyra yezoensis polysaccharides (PPs) on the denaturation of bighead carp (Aristichthys nobilis) myofibrillar protein (Mf) during frozen storage at -18°C for 90d. The PPs (2.5%, 5%, and 7.5%, respectively) was added to 100g of Mf. The changes in the Ca-adenylpyrophosphatase (ATPase) activity and unfrozen water content in Mf were examined to evaluate he denaturation of Mf during frozen storage. Ca-ATPase activity decreased gradually during frozen storage at -18°C upon addition of PPs. By contrast, Ca-ATPase activity in the control group dropped drastically during the first 45d of storage and then further decreased gradually for up to 90d of storage, indicating a biphasic denaturation pattern. PPs addition significantly increased sulfhydryl contents in the Mf of the treatment groups compared with that of the control group (p<0.05) during frozen storage at -18°C. PMID:24769211

  18. Understanding low-lipid algae hydrothermal liquefaction characteristics and pathways through hydrothermal liquefaction of algal major components: crude polysaccharides, crude proteins and their binary mixtures.

    PubMed

    Yang, Wenchao; Li, Xianguo; Li, Zihui; Tong, Chenhong; Feng, Lijuan

    2015-11-01

    Crude polysaccharides and proteins extracted from algae were chosen as model materials to investigate the hydrothermal liquefaction (HTL) characteristics and pathways of low-lipid algae. Liquefaction behavior of the two individuals and their binary mixtures with different mass ratios were evaluated under different temperatures. Formation pathways of bio-oil from crude polysaccharides/proteins were proposed. Results showed that polysaccharides had a small contribution to bio-oil (<5%) and approximately 60% distributed in aqueous phase, while proteins played a crucial role on bio-oil formation (maximum 16.29%). Bio-oil from polysaccharides mainly contained cyclic ketones and phenols and from proteins composed of pyrazines, pyrroles and amines. Interaction between polysaccharides and proteins forming polycyclic nitrogenous compounds had a negative effect on bio-oil yield at 220 and 260°C. However, their further decomposition caused increase of bio-oil yield at 300°C. Mixture liquefaction obtained the highest higher heating value (HHV) of bio-oil and energy recovery than polysaccharides/proteins liquefaction at 300°C.

  19. Molecularly imprinted polymer grafted on polysaccharide microsphere surface by the sol-gel process for protein recognition.

    PubMed

    Li, Feng; Li, Jing; Zhang, Shusheng

    2008-02-15

    An interfacial organic-inorganic hybridization concept was applied to the preparation of a new spherical imprinted material for protein recognition. The functional biopolymer chitosan (CS), shaped as microsphere and high-density cross-linked, constituted of the polysaccharide core for surface imprinting. After the model template protein, bovine serum albumin, was covalently immobilized by forming imine bonds with the functional amine groups of CS, two kinds of organic siloxane (3-aminopropyltrimethoxysiloxane: APTMS, and tetraethoxysiloxane: TEOS) assembled and polymerized on the polysaccharide-protein surface via sol-gel process in aqueous solution at room temperature. After template removal, the protein-imprinted sol-gel surface exhibited a prevalent preference for the template protein in adsorption experiments, as compared with four contrastive proteins. Bioinformatics methods were also employed to investigate the imprinting process and the recognition effect. The influence of siloxane type, pH, siloxane/water ratio on template removal and recognition selectivity was assessed. Under optimized imprinting conditions, a large quantity of well-distributed pores was observed on the immobilized-template imprinted surface. The surface-imprinted adsorbent offered a fast kinetics for template re-adsorption and could be reused. Compared with the imprinted material prepared with free-template, material prepared with immobilized-template possessed higher adsorption capacity towards template protein. Easy preparation of the described imprinted material, high affinity and good reusability make this approach attractive and broadly applicable in biotechnology for down-stream processing and biosensor. PMID:18371777

  20. Vi capsular polysaccharide-protein conjugates for prevention of typhoid fever. Preparation, characterization, and immunogenicity in laboratory animals.

    PubMed

    Szu, S C; Stone, A L; Robbins, J D; Schneerson, R; Robbins, J B

    1987-11-01

    The Vi has proven to be a protective antigen in two double masked, controlled clinical trials in areas with high rates of typhoid fever (approximately 1% per annum). In both studies the protective efficacy of the Vi was approximately 70%. Approximately 75% of subjects in these areas responded with a fourfold or greater rise of serum Vi antibodies. In contrast, the Vi elicited a fourfold or greater rise in 95-100% of young adults in France and the United States. Methods were devised, therefore, to synthesize Vi-protein conjugates in order to both enhance the antibody response and confer T-dependent properties to the Vi (and theoretically increase its protective action in populations at high risk for typhoid fever). We settled on a method that used the heterobifunctional crosslinking reagent, N-succinimidyl-3-(2-pyridyldithio)-propionate (SPDP), to bind thiol derivatives of the Vi to proteins. This synthetic scheme was reproducible, provided high yields of Vi-protein conjugates, and was applicable to several medically relevant proteins such as diphtheria and tetanus toxoids. The resultant conjugates were more immunogenic in mice and juvenile Rhesus monkeys than the Vi alone. In contrast to the T-independent properties of the Vi, conjugates of this polysaccharide with several medically relevant proteins induced booster responses in mice and in juvenile Rhesus monkeys. Clinical studies with Vi-protein conjugates are planned. This scheme is also applicable to synthesize protein conjugates with other polysaccharides that have carboxyl functions.

  1. Response to Pneumococcal Polysaccharide and Protein-Conjugate Vaccines Singly or Sequentially in Adults who have Recovered from Pneumococcal Pneumonia

    PubMed Central

    Musher, Daniel M.; Rueda, Adriana M.; Nahm, Moon H.; Graviss, Edward A.; Rodriguez-Barradas, Maria C.

    2008-01-01

    Background Controversy persists over the benefits of pneumococcal polysaccharide vaccine (PPV) in at-risk adults. We studied PPV, protein-conjugate pneumococcal vaccine (PCV), or immunologic ‘priming’ with PCV followed by ‘boosting’ with PPV in adults who recovered from pneumococcal pneumonia. Methods Subjects received PPV followed in 6 months by PCV, or vice-versa. IgG to capsular polysaccharide and opsonophagocytic killing activity (OPK) were studied at baseline, 4–8 weeks and 6 months after each vaccination. Results PPV and PCV stimulated similar IgG levels and OPK at 4–8 weeks. Six months post-PPV, antibody declined to baseline but remained modestly elevated post-PCV. PCV given 6 months post-PPV stimulated modest IgG increases that failed to reach post-PPV peaks. In contrast, PPV 6 months after PCV caused dramatic increases in IgG and OPK to all polysaccharides, consistent with a booster effect. Six months after the second vaccination, however, IgG and OPK in all patients fell precipitously, returning toward original baseline levels. Conclusions In high-risk subjects, the effect of PPV is short-lived; PCV stimulates a more prolonged response. PPV as a booster following PCV causes early antibody rises, but IgG declines rapidly thereafter, consistent with induction of suppressor cells or tolerance. Protein vaccines may be needed for high-risk adults. PMID:18710324

  2. A review of research on the protein-bound polysaccharide (polysaccharopeptide, PSP) from the mushroom Coriolus versicolor (Basidiomycetes: Polyporaceae).

    PubMed

    Ng, T B

    1998-01-01

    1. Protein-bound polysaccharides, designated as PSK and PSP, have been isolated from the CM-101 strain and the COV-1 strain, respectively, of the mushroom Coriolus versicolor. This article aims at summarizing existing research findings about PSP since information on PSK is well documented. 2. PSP possesses a molecular weight of approximately 100 kDa. Glutamic and aspartic acids are abundant in its polypeptide component, whereas its polysaccharide component is made up of monosaccharides with alpha-1,4 and beta-1,3 glucosidic linkages. The presence of fucose in PSK and rhamnose and arabinose in PSP distinguishes the two protein-bound polysaccharides, which are otherwise chemically similar. 3. PSP is classified as a biological response modifier. It induces, in experimental animals, increased gamma-interferon production, interleukin-2 production, and T-cell proliferation. It also counteracts the depressive effect of cyclophosphamide on white blood cell count, interleukin-2 production and delayed-type hypersensitivity reaction. Its antiproliferative activity against tumor cell lines and in vivo antitumor activity have been demonstrated. A small peptide with a molecular weight of 16-18 kDa originating from PSP has been produced with antiproliferative and antitumor activities. 4. PSP administered to patients with esophageal cancer, gastric cancer and lung cancer, and who are undergoing radiotherapy or chemotherapy, helps alleviate symptoms and prevents the decline in immune status.

  3. A unique capsular polysaccharide structure from the psychrophilic marine bacterium Colwellia psychrerythraea 34H that mimics antifreeze (glyco)proteins.

    PubMed

    Carillo, Sara; Casillo, Angela; Pieretti, Giuseppina; Parrilli, Ermenegilda; Sannino, Filomena; Bayer-Giraldi, Maddalena; Cosconati, Sandro; Novellino, Ettore; Ewert, Marcela; Deming, Jody W; Lanzetta, Rosa; Marino, Gennaro; Parrilli, Michelangelo; Randazzo, Antonio; Tutino, Maria L; Corsaro, M Michela

    2015-01-14

    The low temperatures of polar regions and high-altitude environments, especially icy habitats, present challenges for many microorganisms. Their ability to live under subfreezing conditions implies the production of compounds conferring cryotolerance. Colwellia psychrerythraea 34H, a γ-proteobacterium isolated from subzero Arctic marine sediments, provides a model for the study of life in cold environments. We report here the identification and detailed molecular primary and secondary structures of capsular polysaccharide from C. psychrerythraea 34H cells. The polymer was isolated in the water layer when cells were extracted by phenol/water and characterized by one- and two-dimensional NMR spectroscopy together with chemical analysis. Molecular mechanics and dynamics calculations were also performed. The polysaccharide consists of a tetrasaccharidic repeating unit containing two amino sugars and two uronic acids bearing threonine as substituent. The structural features of this unique polysaccharide resemble those present in antifreeze proteins and glycoproteins. These results suggest a possible correlation between the capsule structure and the ability of C. psychrerythraea to colonize subfreezing marine environments.

  4. Bypassing the need for subcellular localization of a polysaccharide export-anchor complex by overexpressing its protein subunits

    PubMed Central

    Javens, June; Wan, Zhe; Hardy, Gail G.; Brun, Yves V.

    2013-01-01

    Summary Subcellular protein localization is thought to promote protein-protein interaction by increasing the effective concentration and enabling spatial coordination and proper segregation of proteins. We found that protein overexpression allowed the assembly of a productive polysaccharide biosynthesis-export-anchoring complex in the absence of polar localization in Caulobacter crescentus. Polar localization of the holdfast export protein, HfsD, depends on the presence of the other export proteins, HfsA, and HfsB, and on the polar scaffold protein PodJ. The holdfast deficiency of hfsB and podJ mutants is suppressed by the overexpression of export proteins. Restored holdfasts are randomly positioned and co-localize with a holdfast anchor protein in these strains, indicating that functional complexes can form at non-polar sites. Therefore, overexpression of export proteins surpasses a concentration threshold necessary for holdfast synthesis. Restoration of holdfast synthesis at non-polar sites reduces surface adhesion, consistent with the need to spatially coordinate the holdfast synthesis machinery with the flagellum and pili. These strains lack the cell-specific segregation of the holdfast, resulting in the presence of holdfasts in motile daughter cells. Our results highlight the fact that multiple facets of subcellular localization can be coupled to improve the phenotypic outcome of a protein assembly. PMID:23714375

  5. A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate.

    PubMed

    Abdelhameed, Ali Saber; Adams, Gary G; Morris, Gordon A; Almutairi, Fahad M; Duvivier, Pierre; Conrath, Karel; Harding, Stephen E

    2016-02-26

    Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyribosylribitol (PRP) polysaccharides extracted from Haemophilus influenzae type b (Hib), and the corresponding glycoconjugate made by conjugating this with the tetanus toxoid (TT) protein have been characterized and compared using a combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge with viscometry. The weight average molar mass of the activated material was considerably reduced (Mw ~ 0.24 × 10(6) g.mol(-1)) compared to the native (Mw ~ 1.2 × 10(6) g.mol(-1)). Conjugation with the TT protein yielded large polydisperse structures (of Mw ~ 7.4 × 10(6) g.mol(-1)), but which retained the high degree of flexibility of the native and activated polysaccharide, with frictional ratio, intrinsic viscosity, sedimentation conformation zoning behaviour and persistence length all commensurate with highly flexible coil behaviour and unlike the previously characterised tetanus toxoid protein (slightly extended and hydrodynamically compact structure with an aspect ratio of ~3). This non-protein like behaviour clearly indicates that it is the carbohydrate component which mainly influences the physical behaviour of the glycoconjugate in solution.

  6. A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate

    PubMed Central

    Abdelhameed, Ali Saber; Adams, Gary G.; Morris, Gordon A.; Almutairi, Fahad M.; Duvivier, Pierre; Conrath, Karel; Harding, Stephen E.

    2016-01-01

    Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyribosylribitol (PRP) polysaccharides extracted from Haemophilus influenzae type b (Hib), and the corresponding glycoconjugate made by conjugating this with the tetanus toxoid (TT) protein have been characterized and compared using a combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge with viscometry. The weight average molar mass of the activated material was considerably reduced (Mw ~ 0.24 × 106 g.mol−1) compared to the native (Mw ~ 1.2 × 106 g.mol−1). Conjugation with the TT protein yielded large polydisperse structures (of Mw ~ 7.4 × 106 g.mol−1), but which retained the high degree of flexibility of the native and activated polysaccharide, with frictional ratio, intrinsic viscosity, sedimentation conformation zoning behaviour and persistence length all commensurate with highly flexible coil behaviour and unlike the previously characterised tetanus toxoid protein (slightly extended and hydrodynamically compact structure with an aspect ratio of ~3). This non-protein like behaviour clearly indicates that it is the carbohydrate component which mainly influences the physical behaviour of the glycoconjugate in solution. PMID:26915577

  7. A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate.

    PubMed

    Abdelhameed, Ali Saber; Adams, Gary G; Morris, Gordon A; Almutairi, Fahad M; Duvivier, Pierre; Conrath, Karel; Harding, Stephen E

    2016-01-01

    Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyribosylribitol (PRP) polysaccharides extracted from Haemophilus influenzae type b (Hib), and the corresponding glycoconjugate made by conjugating this with the tetanus toxoid (TT) protein have been characterized and compared using a combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge with viscometry. The weight average molar mass of the activated material was considerably reduced (Mw ~ 0.24 × 10(6) g.mol(-1)) compared to the native (Mw ~ 1.2 × 10(6) g.mol(-1)). Conjugation with the TT protein yielded large polydisperse structures (of Mw ~ 7.4 × 10(6) g.mol(-1)), but which retained the high degree of flexibility of the native and activated polysaccharide, with frictional ratio, intrinsic viscosity, sedimentation conformation zoning behaviour and persistence length all commensurate with highly flexible coil behaviour and unlike the previously characterised tetanus toxoid protein (slightly extended and hydrodynamically compact structure with an aspect ratio of ~3). This non-protein like behaviour clearly indicates that it is the carbohydrate component which mainly influences the physical behaviour of the glycoconjugate in solution. PMID:26915577

  8. Construction of a single-axis molecular puller for measuring polysaccharide and protein mechanics by atomic force microscopy.

    PubMed

    Rabbi, Mahir; Marszalek, Piotr E

    2007-12-01

    INTRODUCTIONPolysaccharides and proteins are frequently subjected to mechanical forces in vivo. Because these forces affect a wide range of biological activities, it is important to develop methods that directly investigate the mechanical properties of these molecules. Recent progress in techniques that allow the mechanical manipulation of biopolymers at a single-molecule level has revealed the complex nature of the elasticity of proteins and polysaccharides. The atomic force microscope (AFM) is an excellent force spectrometer for probing the mechanical properties (e.g., length and tension) of individual polysaccharides and proteins. The following protocol describes the basic design and construction of an AFM (a single-axis molecular puller) that has four parts: a head, a base, electronics, and software. Those with a background in mechanical engineering, basic knowledge of electronics and data acquisition techniques, and some computer programming skills (e.g., with LabView, Matlab, or Igor) should be able to construct this instrument. It is advisable to inspect commercial AFMs before constructing one from scratch.

  9. Further Studies on the Immunogenicity of Haemophilus influenzae Type b and Pneumococcal Type 6A Polysaccharide-Protein Conjugates

    PubMed Central

    Chu, Chiayung; Schneerson, Rachel; Robbins, John B.; Rastogi, Suresh C.

    1983-01-01

    Conjugates were prepared by carbodiimide-mediated coupling of adipic acid hydrazide derivatives of Haemophilus influenzae type b (Hib), Escherichia coli K100, and pneumococcal 6A (Pn6A) polysaccharides with tetanus toxoid (TT), as an example of a “useful” carrier, and horseshoe crab hemocyanin (HCH), as an example of a “nonsense” carrier. These conjugates were injected into NIH mice, and their serum antibody responses to the polysaccharides and proteins were characterized. As originally reported, Hib conjugates increased the immunogenicity of the capsular polysaccharide and elicited greater than the estimated protective levels of anti-Hib antibodies in most recipients after one injection and in all after the third injection (Schneerson et al., J. Exp. Med. 152:361-376, 1980). Both Hib conjugates induced similar anti-Hib responses. The K100-HCH conjugate was more immunogenic than the K100-TT conjugate and elicited anti-Hib responses similar to the Hib conjugates after the third injection. Simultaneous injection of the K100 and the Hib conjugates did not enhance the anti-Hib response. The Pn6A-TT conjugate induced low levels of anti-Hib antibodies; when injected simultaneously with the Hib conjugates, the anti-Hib response was enhanced, as all mice responded after the first injection and with higher levels of anti-Hib than observed with the Hib conjugates alone (P < 0.05). The Pn6A conjugates were not as immunogenic as the Hib conjugates. Pn6A-TT was more effective than was Pn6A-HCH; it elicited anti-Pn6A (>100 ng of antibody nitrogen per ml) in 6 of 10 mice after the third injection. The addition of the Hib-HCH conjugate to the Pn6A-TT conjugate increased the anti-Pn6A response with a higher geometric mean antibody titer, and 9 of 10 mice responded after the third injection. A preparation of diphtheria toxoid, TT, and pertussis vaccine increased the anti-Hib antibody levels after the first injection only in mice receiving Hib-TT, but not in mice receiving

  10. Examination of an aloe vera galacturonate polysaccharide capable of in situ gelation for the controlled release of protein therapeutics

    NASA Astrophysics Data System (ADS)

    McConaughy, Shawn David

    A therapeutic delivery platform has been investigated with the ultimate goal of designing a sustained protein release matrix utilizing an in-situ gelling, acidic polysaccharide derived from the Aloe vera plant. The Aloe vera polysaccharide (AvP) has been examined in order to determine how chemical composition, structure, molecular weight and solution behavior affect gelation and protein/peptide delivery. Correlations are drawn between structural characteristics and solution behavior in order to determine the impact of polymer conformation and solvation on gel formation under conditions designed to simulate nasal applications. Steady state and dynamic rheology, classic and dynamic light scattering, zeta potential, pulse field gradient nuclear magnetic resonance and fluorescence spectroscopy have been employed to gain insight into the effects of galacturonic acid content, degree of methylation, entanglement and ionic strength on both solution behavior and the hydrogel state which ultimately governs protein/peptide release. This dissertation is divided into two sections. In the first section, a series of Aloe vera polysaccharides (AvP), from the pectin family have been structurally characterized indicating high galacturonic acid (GalA) content, low degree of methylester substitution (DM), low numbers of rhamnose residues and high molecular weight with respect to pectins extracted from traditional sources. The behavior of AvP was examined utilizing dilute solution, low-shear rheological techniques for specific molecular weight samples at selected conditions of ionic strength. From these dilute aqueous solution studies, the Mark-Houwink-Sakurada (MHS) constants (K and alpha), persistence length (Lp) and inherent chain stiffness (B parameter) were determined, indicating an expanded random coil in aqueous salt solutions. The critical concentration for transition from dilute to concentrated solution, C e, was determined by measuring both the zero shear viscosity and

  11. The Role of the Plant-Specific ALTERED XYLOGLUCAN9 Protein in Arabidopsis Cell Wall Polysaccharide O-Acetylation1[OPEN

    PubMed Central

    Schultink, Alex; Naylor, Dan; Dama, Murali; Pauly, Markus

    2015-01-01

    A mutation in the ALTERED XYLOGLUCAN9 (AXY9) gene was found to be causative for the decreased xyloglucan acetylation phenotype of the axy9.1 mutant, which was identified in a forward genetic screen for Arabidopsis (Arabidopsis thaliana) mutants. The axy9.1 mutant also exhibits decreased O-acetylation of xylan, implying that the AXY9 protein has a broad role in polysaccharide acetylation. An axy9 insertional mutant exhibits severe growth defects and collapsed xylem, demonstrating the importance of wall polysaccharide O-acetylation for normal plant growth and development. Localization and topological experiments indicate that the active site of the AXY9 protein resides within the Golgi lumen. The AXY9 protein appears to be a component of the plant cell wall polysaccharide acetylation pathway, which also includes the REDUCED WALL ACETYLATION and TRICHOME BIREFRINGENCE-LIKE proteins. The AXY9 protein is distinct from the TRICHOME BIREFRINGENCE-LIKE proteins, reported to be polysaccharide acetyltransferases, but does share homology with them and other acetyltransferases, suggesting that the AXY9 protein may act to produce an acetylated intermediate that is part of the O-acetylation pathway. PMID:25681330

  12. Contributions of polysaccharide and lipid regions of lipopolysaccharide to the recognition by spike G protein of bacteriophage phi X174.

    PubMed

    Kawaura, Tomoko; Inagaki, Minoru; Tanaka, Akiyoshi; Kato, Muneharu; Nishikawa, Shiro; Kashimura, Naoki

    2003-04-01

    A histidine-tagged G protein of bacteriophage phi X174 (HisG) bound strongly with lipopolysaccharide (LPS) of Escherichia coli C, one of a phi X174-sensitive Ra strain. The dissociation constant, Kd, was measured to be 0.16 +/- 0.04 microM by fluorometric titration. HisG showed slightly less affinity to LPSs of the insensitive Rc and Rd 2 strains having shorter R-core polysaccharide sequences than that of the sensitive Ra strains. The difference between the two types of LPS was demonstrated by CD spectra; LPSs of the sensitive strains increased the signal intensity for beta-sheet, while the insensitive strains decreased it. The chemically degraded LPS derivatives lacking a hydrophobic lipid region showed much less affinity to HisG, indicating the importance of the lipid region of LPS for strong binding with HisG. On the other hand, since the degraded derivatives increased the intensity of CD spectra, the polysaccharide region is thought to contribute to the conformation change of the protein.

  13. Bismuth Dimercaptopropanol (BisBAL) Inhibits the Expression of Extracellular Polysaccharides and Proteins by Brevundimonas diminuta: Implications for Membrane Microfiltration

    SciTech Connect

    Badireddy, Appala R.; Chellam, Shankararaman; Yanina, Svetlana; Gassman, Paul L.; Rosso, Kevin M.

    2008-02-15

    A 2:1 molar ratio preparation of bismuth with a lipophilic dithiol (3-dimercapto-1-propanol, BAL)significantly reduced extracellular polymeric substances (EPS) expression by Brevundimonas diminuta in suspended cultures at levels just below the minimum inhibitory concentration (MIC). Total polysaccharides and proteins secreted by B. diminuta decreased by approximately 95% over a 5-day period when exposed to the bismuth-BAL chelate (BisBAL) at near MIC (12 μM). Fourier-transform infrared spectroscopy (FTIR) suggested that a possible mechanism of biofilm disruption by BisBAL is the inhibition of carbohydrate Oacetylation. FTIR also revealed extensive homology between EPS samples with and without BisBAL treatment, with proteins, polysaccharides, and peptides varying predominantly only in the amount expressed. EPS secretion decreased following BisBAL treatment as verified by atomic force microscopy and scanning electron microscopy. Without BisBAL treatment, a slime-like EPS matrix secreted by B. diminuta resulted in biofouling and inefficient hydrodynamic backwashing of microfiltration membranes.

  14. Physical Stability of Octenyl Succinate-Modified Polysaccharides and Whey Proteins for Potential Use as Bioactive Carriers in Food Systems.

    PubMed

    Puerta-Gomez, Alex F; Castell-Perez, M Elena

    2015-06-01

    The high cost and potential toxicity of biodegradable polymers like poly(lactic-co-glycolic)acid (PLGA) has increased the interest in natural and modified biopolymers as bioactive carriers. This study characterized the physical stability (water sorption and state transition behavior) of selected starch and proteins: octenyl succinate-modified depolymerized waxy corn starch (DWxCn), waxy rice starch (DWxRc), phytoglycogen, whey protein concentrate (80%, WPC), whey protein isolate (WPI), and α-lactalbumin (α-L) to determine their potential as carriers of bioactive compounds under different environmental conditions. After enzyme modification and particle size characterization, glass transition temperature and moisture isotherms were used to characterize the systems. DWxCn and DWxRc had increased water sorption compared to native starch. The level of octenyl succinate anhydrate (OSA) modification (3% and 7%) did not reduce the water sorption of the DWxCn and phytoglycogen samples. The Guggenheim-Andersen-de Boer model indicated that native waxy corn had significantly (P < 0.05) higher water monolayer capacity followed by 3%-OSA-modified DWxCn, WPI, 3%-OSA-modified DWxRc, α-L, and native phytoglycogen. WPC had significantly lower water monolayer capacity. All Tg values matched with the solid-like appearance of the biopolymers. Native polysaccharides and whey proteins had higher glass transition temperature (Tg) values. On the other hand, depolymerized waxy starches at 7%-OSA modification had a "melted" appearance when exposed to environments with high relative humidity (above 70%) after 10 days at 23 °C. The use of depolymerized and OSA-modified polysaccharides blended with proteins created more stable blends of biopolymers. Hence, this biopolymer would be suitable for materials exposed to high humidity environments in food applications.

  15. Role of the Apt1 Protein in Polysaccharide Secretion by Cryptococcus neoformans

    PubMed Central

    Rizzo, Juliana; Oliveira, Débora L.; Joffe, Luna S.; Hu, Guanggan; Gazos-Lopes, Felipe; Fonseca, Fernanda L.; Almeida, Igor C.; Frases, Susana; Kronstad, James W.

    2014-01-01

    Flippases are key regulators of membrane asymmetry and secretory mechanisms. Vesicular polysaccharide secretion is essential for the pathogenic mechanisms of Cryptococcus neoformans. On the basis of the observations that flippases are required for polysaccharide secretion in plants and the putative Apt1 flippase is required for cryptococcal virulence, we analyzed the role of this enzyme in polysaccharide release by C. neoformans, using a previously characterized apt1Δ mutant. Mutant and wild-type (WT) cells shared important phenotypic characteristics, including capsule morphology and dimensions, glucuronoxylomannan (GXM) composition, molecular size, and serological properties. The apt1Δ mutant, however, produced extracellular vesicles (EVs) with a lower GXM content and different size distribution in comparison with those of WT cells. Our data also suggested a defective intracellular GXM synthesis in mutant cells, in addition to changes in the architecture of the Golgi apparatus. These findings were correlated with diminished GXM production during in vitro growth, macrophage infection, and lung colonization. This phenotype was associated with decreased survival of the mutant in the lungs of infected mice, reduced induction of interleukin-6 (IL-6) cytokine levels, and inefficacy in colonization of the brain. Taken together, our results indicate that the lack of APT1 caused defects in both GXM synthesis and vesicular export to the extracellular milieu by C. neoformans via processes that are apparently related to the pathogenic mechanisms used by this fungus during animal infection. PMID:24337112

  16. Role of the Apt1 protein in polysaccharide secretion by Cryptococcus neoformans.

    PubMed

    Rizzo, Juliana; Oliveira, Débora L; Joffe, Luna S; Hu, Guanggan; Gazos-Lopes, Felipe; Fonseca, Fernanda L; Almeida, Igor C; Frases, Susana; Kronstad, James W; Rodrigues, Marcio L

    2014-06-01

    Flippases are key regulators of membrane asymmetry and secretory mechanisms. Vesicular polysaccharide secretion is essential for the pathogenic mechanisms of Cryptococcus neoformans. On the basis of the observations that flippases are required for polysaccharide secretion in plants and the putative Apt1 flippase is required for cryptococcal virulence, we analyzed the role of this enzyme in polysaccharide release by C. neoformans, using a previously characterized apt1Δ mutant. Mutant and wild-type (WT) cells shared important phenotypic characteristics, including capsule morphology and dimensions, glucuronoxylomannan (GXM) composition, molecular size, and serological properties. The apt1Δ mutant, however, produced extracellular vesicles (EVs) with a lower GXM content and different size distribution in comparison with those of WT cells. Our data also suggested a defective intracellular GXM synthesis in mutant cells, in addition to changes in the architecture of the Golgi apparatus. These findings were correlated with diminished GXM production during in vitro growth, macrophage infection, and lung colonization. This phenotype was associated with decreased survival of the mutant in the lungs of infected mice, reduced induction of interleukin-6 (IL-6) cytokine levels, and inefficacy in colonization of the brain. Taken together, our results indicate that the lack of APT1 caused defects in both GXM synthesis and vesicular export to the extracellular milieu by C. neoformans via processes that are apparently related to the pathogenic mechanisms used by this fungus during animal infection.

  17. Identification and functional characterization of the putative polysaccharide biosynthesis protein (CapD) of Enterococcus faecium U0317.

    PubMed

    Ali, Liaqat; Spiess, Meike; Wobser, Dominique; Rodriguez, Marta; Blum, Hubert E; Sakιnç, Türkân

    2016-01-01

    Most bacterial species produce capsular polysaccharides that contribute to disease pathogenesis through evasion of the host innate immune system and are also involved in inhibiting leukocyte killing. In the present study, we identified a gene in Enterococcus faecium U0317 with homologies to the polysaccharide biosynthesis protein CapD that is made up of 336 amino acids and putatively catalyzes N-linked glycosylation. A capD deletion mutant was constructed and complemented by homologous recombination that was confirmed by PCR and sequencing. The mutant revealed different growth behavior and morphological changes compared to wild-type by scanning electron microscopy, also the capD mutant showed a strong hydrophobicity and that was reversed in the reconstituted mutant. For further characterization and functional analyses, in-vitro cell culture and in-vivo a mouse infection models were used. Antibodies directed against alpha lipotechoic acid (αLTA) and the peptidyl-prolyl cis-trans isomerase (αPpiC), effectively mediated the opsonophagocytic killing in the capD knock-out mutant, while this activity was not observed in the wild-type and reconstituted mutant. By comparison more than 2-fold decrease was seen in mutant colonization and adherence to both T24 and Caco2 cells. However, a significant higher bacterial colonization was observed in capD mutant during bacteremia in the animal model, while virulence in a mouse UTI (urinary tract infection) model, there were no obvious differences. Further studies are needed to elucidate the function of capsular polysaccharide synthesis gene clusters and its involvement in the disease pathogenesis with the aim to develop targeted therapies to treat multidrug-resistant E. faecium infections.

  18. Salmonella enterica serovar Typhi O:1,9,12 polysaccharide-protein conjugate as a diagnostic tool for typhoid fever.

    PubMed

    Zuñiga, Jessica; Lillo, Luis; Shin, Junghee J; Machavarapu, Rajya L; Quiroga, Teresa; Goycoolea, Manuela; Matsuhiro, Betty; Aron-Hott, L; Godfrey, Henry P; Cabello, Felipe C

    2005-09-01

    Serologic tests play an important role in diagnosis of typhoid fever. In an effort to develop a more defined reagent for these tests, purified Salmonella enterica serovar Typhi (ST) O:1,9,12 polysaccharide was conjugated to human serum albumin (HSA), and the conjugate was purified chromatographically to yield a reagent with 2 moles ST O polysaccharide per mole HSA. In 40 patients with bacteriologically confirmed typhoid fever, significant dot immunobinding titers (> or =20,000) were present in 28 (70%) tested with 100 ng of ST O antigen-HSA (ST O-HSA) conjugate, in 38 (95%) tested with 100 ng of ST lipopolysaccharide, and in 16 (40%) tested with purified unconjugated ST O chains. In sera from 22 patients with other nontyphoid fevers, 2 (9.1%) had such reactivities with 100 ng of ST O-HSA, 1 (4.5%) had such reactivity with 100 ng of ST lipopolysaccharide (4.5%), and none reacted with 100 ng of unconjugated ST O chains. None of the 17 healthy-control sera reacted significantly with any of the ST reagents. None of the patient or control sera reacted with unconjugated HSA. The sensitivity of dot immunobinding for typhoid fever was 70% with 100 ng of ST O-HSA, somewhat lower than that with 100 ng of ST lipopolysaccharide (95%) but similar to that of the Widal H agglutination test with a > or =1/160 cutoff (74%). Specificities of these tests were 91%, 95%, and 86%, respectively. These preliminary results suggest that ST O polysaccharide-protein conjugates could provide a nontoxic, easily quality-controlled synthetic reagent for analysis of human immune responses to ST as well as for the development of new diagnostics and vaccines for typhoid fever.

  19. Alpha C Protein as a Carrier for Type III Capsular Polysaccharide and as a Protective Protein in Group B Streptococcal Vaccines

    PubMed Central

    Gravekamp, Claudia; Kasper, Dennis L.; Paoletti, Lawrence C.; Madoff, Lawrence C.

    1999-01-01

    The alpha C protein, a protective surface protein of group B streptococci (GBS), is present in most non-type III GBS strains. Conjugate vaccines composed of the alpha C protein and type III capsular polysaccharide (CPS) might be protective against most GBS infections. In this study, the type III CPS was covalently coupled to full-length, nine-repeat alpha C protein (resulting in III-α9r conjugate vaccine) or to two-repeat alpha C protein (resulting in III-α2r conjugate vaccine) by reductive amination. Initial experiments with the III-α9r vaccine showed that it was poorly immunogenic in mice with respect to both vaccine antigens and was suboptimally efficacious in providing protection in mice against challenge with GBS. Therefore, modified vaccination protocols were used with the III-α2r vaccine. Female mice were immunized three times with 0.5, 5, or 20 μg of the III-α2r vaccine with an aluminum hydroxide adjuvant and bred. Ninety-five percent of neonatal mice born to dams immunized with the III-α2r vaccine survived challenge with GBS expressing type III CPS, and 60% survived challenge with GBS expressing wild-type (nine-repeat) alpha C protein; 18 and 17%, respectively, of mice in the negative control groups survived (P, <0.0001). These protection levels did not differ significantly from those obtained with the type III CPS-tetanus toxoid conjugate vaccine and the unconjugated two-repeat alpha C protein, which protected 98 and 58% of neonates from infection with GBS expressing type III CPS or the alpha C protein, respectively. Thus, the two-repeat alpha C protein in the vaccine was immunogenic and simultaneously enhanced the immunogenicity of type III CPS. III-α vaccines may be alternatives to GBS polysaccharide-tetanus toxoid vaccines, eliciting additional antibodies protective against GBS infection. PMID:10225912

  20. The impact of protein-conjugate polysaccharide vaccines: an endgame for meningitis?

    PubMed Central

    Maiden, Martin C. J.

    2013-01-01

    The development and implementation of conjugate polysaccharide vaccines against invasive bacterial diseases, specifically those caused by the encapsulated bacteria Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae, has been one of the most effective public health innovations of the last 25 years. These vaccines have resulted in significant reductions in childhood morbidity and mortality worldwide, with their effectiveness due in large part to their ability to induce long-lasting immunity in a range of age groups. At the population level this immunity reduces carriage and interrupts transmission resulting in herd immunity; however, these beneficial effects can be counterbalanced by the selection pressures that immunity against carriage can impose, potentially promoting the emergence and spread of virulent vaccine escape variants. Studies following the implementation of meningococcal serogroup C vaccines improved our understanding of these effects in relation to the biology of accidental pathogens such as the meningococcus. This understanding has enabled the refinement of the implementation of conjugate polysaccharide vaccines against meningitis-associated bacteria, and will be crucial in maintaining and improving vaccine control of these infections. To date there is little evidence for the spread of virulent vaccine escape variants of the meningococcus and H. influenzae, although this has been reported in pneumococci. PMID:23798695

  1. Protecting the herd: the remarkable effectiveness of the bacterial meningitis polysaccharide-protein conjugate vaccines in altering transmission dynamics.

    PubMed

    Stephens, David S

    2011-01-01

    Interrupting human-to-human transmission of the agents (Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae) of bacterial meningitis by new capsular polysaccharide-protein conjugate vaccines (PPCVs) has proven to be a remarkable (and unanticipated) contributor to vaccine effectiveness. Herd immunity accounts for ∼50% of the protection by meningococcal serogroup C PPCVs, pneumococcal PPCV7, and H. influenzae b PPCVs. Nasopharyngeal carriage can be reduced ≥75% for vaccine serotypes; the decrease in carriage is correlated with disease reduction in unvaccinated individuals, and the impact of herd immunity lasts for years. Based on these data, models for using herd immunity in vaccine-based prevention strategies are underway for control of meningitis in sub-Saharan Africa. Although the immunologic basis of herd immunity and impact on microbial biology need more study, protecting the unvaccinated by altering pathogen transmission dynamics is a powerful effect of PPCVs and increasingly important in vaccine introduction, implementation, and evaluation strategies.

  2. Polysaccharide protein complex isolated from mushroom Phellinus rimosus (berk.) Pilat alleviates γ radiation-induced toxicity in mice.

    PubMed

    Joseph, Jini; Smina, Thozhuthum Parambil Pathmanabhan; Janardhanan, Kainoor Krishnankutty

    2011-06-01

    Ionizing radiations generate reactive oxygen species in irradiated tissue that induces several pathophysiological changes in the body. Radiotherapy induced toxicity is a major dose-limiting factor in anticancer treatments. Radioprotective agents are of significant importance in medical, industrial, environmental, military, and space applications. Radioprotective effect of polysaccharide protein complex (PPC-Pr) isolated from mushroom, Phellinus rimosus, was evaluated in Swiss albino mice. PPC-Pr (5 and 10 mg/kg bwt, i.p.) significantly increased leukocyte count, bone marrow cellularity, glutathione content, and activities of antioxidant enzymes such as catalase, superoxide dismutase, and glutathione peroxidase in blood as well as intestinal mucosa when compared with the irradiated control group. Histopathological observation of intestinal jejunal mucosa revealed the tissue protective effects of PPC-Pr. Further radioprotective activity of PPC-Pr was in a dose-dependent manner. The findings suggest potential radioprotective efficacy of PPC-Pr.

  3. Effect of conjugation methodology on the immunogenicity and protective efficacy of meningococcal group C polysaccharide-P64k protein conjugates.

    PubMed

    Carmenate, Tania; Canaán, Leonardo; Alvarez, Anabel; Delgado, Maité; González, Sonia; Menéndez, Tamara; Rodés, Lorenzo; Guillén, Gerardo

    2004-04-01

    Neisseria meningitidis serogroup C polysaccharide (CCPS) was conjugated to the carrier protein P64k using two different conjugation procedures, condensation mediated by carbodiimide with adipic acid dihydrazide as spacer and the reductive amination method. BALB/c mice were immunized with the resultant polysaccharide-protein conjugates and the immune response was evaluated. All conjugates assayed generated at least 10-fold higher antibody titers than the free polysaccharide. The reductive amination method rendered the best conjugate (CCPS-P64kR) that was able to elicit antibody titers statistically higher than the titer elicited by the plain CCPS (P<0.001). The sera of the group immunized with CCPS-P64kR showed a three-fold higher bactericidal response than the sera of the group immunized with the plain CCPS and they were able to protect against challenge with meningococci in the infant rat protection model. In addition, three different conjugates were obtained from polysaccharides with molecular relative sizes of 2000-4000 Da, 4000-10,000 Da or 10,000-50,000 Da, but no differences were detected in the immune response obtained against the three conjugates. Our experiments demonstrate that it is possible to generate a protective, T-cell-dependent response against CCPS using the P64k protein as carrier. PMID:15039094

  4. Effect of conjugation methodology, carrier protein, and adjuvants on the immune response to Staphylococcus aureus capsular polysaccharides.

    PubMed

    Fattom, A; Li, X; Cho, Y H; Burns, A; Hawwari, A; Shepherd, S E; Coughlin, R; Winston, S; Naso, R

    1995-10-01

    Conjugate vaccines were prepared with S. aureus type 8 capsular polysaccharide (CP) using three carrier proteins: Pseudomonas aeruginosa exotoxin A (ETA), a non-toxic recombinant ETA (rEPA), and diphtheria toxoid (DTd). Adipic acid dihydrazide (ADH) or N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) was used as a spacer to link the CP to carrier protein. All conjugates gave a high immune response with a boost after the second immunization. Conjugates prepared with ADH gave higher antibody titers than conjugates prepared with SPDP. IgG1 was the primary subclass elicited by all conjugates regardless of the carrier protein or the conjugation method used to prepare the vaccines. The non-immunogenic CP and the conjugates were formulated with either monophosphoryl lipid A (MPL), QS21, or in Novasomes and evaluated in mice. While the adjuvants failed to improve the immunogenicity of the nonconjugated CP, a more than fivefold increase in the antibody levels was observed when these adjuvants were used with the conjugates. Significant rises in IgG2b and IgG3 were observed with all formulations. The enhancement of the immunogenicity and the IgG subclass shift, as seen with some adjuvants, may prove to be important in immunocompromised patients. PMID:8585282

  5. Fat body in some genera of leaf-cutting ants (Hymenoptera: Formicidae). Proteins, lipids and polysaccharides detection.

    PubMed

    Roma, Gislaine Cristina; Mathias, Maria Izabel Camargo; Bueno, Odair Corrêa

    2006-01-01

    The comparative histochemical analysis of the fat body of workers belonging to the basal species Cyphomyrmex rimosus and Mycetarotes parallelus and to derived species Acromyrmex disciger and Atta laevigata revealed that this tissue is constituted mainly by cells denominated trophocytes and oenocytes. The trophocytes of all species studied here were characterized mainly by the proteins and lipids synthesis and storage, being the derived species the ones who have presented higher quantity of lipids in the trophocytes when compared to the trophocytes of basal species. In workers M. parallelus and A. laevigata, besides proteins and lipids, there has being observed the presence of polysaccharides, however, in C. rimosus and A. disciger these elements were detected in lower quantities. The histochemical studies of the oenocytes of basal and derived species revealed significant presence of proteins as well as lipids in these cells. In the oenocytes of derived species A. disciger and A. laevigata a higher quantity of lipidic inclusions has being observed, when compared to the basal species.

  6. POLYSACCHARIDE OF COCCIDIOIDES IMMITIS

    PubMed Central

    Pappagianis, D.; Putman, E. W.; Kobayashi, G. S.

    1961-01-01

    Pappagianis, D. (University of California, Berkeley), E. W. Putman, and G. S. Kobayashi. Polysaccharide of Coccidioides immitis. J. Bacteriol. 82:714–723. 1961.—Soluble polysaccharide from mycelia or culture filtrates of Coccidioides immitis was found to consist mainly of mannose, but also included small quantities of galactose and another reducing sugar. Isolation of the polysaccharide by ethanol precipitation provided accompanying nitrogenous material. There was 3 to 4% nitrogen, present in amino acids, but in a nondialyzable (possibly protein or peptide) form. The average molecular weight of the complex was 31,700. Attempts to separate the nitrogenous and polysaccharide materials by chemical methods or by moving boundary electrophoresis were unsuccessful; ultracentrifugation showed a single peak. Despite these findings, there appeared to be several antigenic species present, as indicated by multiple lines of precipitation in double diffusion and quantitative precipitin tests. In the latter, polysaccharide-containing fractions gave precipitates without measurable carbohydrate, suggesting separability of nitrogenous and polysaccharide components. Images PMID:14483753

  7. Safety and Immunogenicity of Improved Shigella O-Specific Polysaccharide-Protein Conjugate Vaccines in Adults in Israel

    PubMed Central

    Passwell, Justen H.; Harlev, Efrat; Ashkenazi, Shai; Chu, Chiayung; Miron, Dan; Ramon, Reut; Farzan, Naheed; Shiloach, Joseph; Bryla, Dolores A.; Majadly, Fathy; Roberson, Robin; Robbins, John B.; Schneerson, Rachel

    2001-01-01

    Data suggest that the O-specific polysaccharide (O-SP) domain of the lipopolysaccharide (LPS) of Shigella species is both an essential virulence factor and a protective antigen and that a critical level of serum immunoglobulin G (IgG) to this antigen will confer immunity to shigellosis. Because covalent attachment of polysaccharides to proteins increases their immunogenicity, especially in infants and in young children, the O-SP of Shigella species were bound to medically useful proteins, and the safety and immunogenicity of the resultant conjugates were confirmed in adults and 4- to 7-year-old children. Succinylation of the carrier protein improved the immunogenicity of Shigella conjugates in mice and increased their yield. Based on these results, a clinical trial of O-SP conjugates of Shigella sonnei and Shigella flexneri 2a bound to succinylated mutant Pseudomonas aeruginosa exotoxin A (rEPAsucc) or native or succinylated Corynebacterium diphtheriae toxin mutant (CRM9 or CRM9succ) was conducted in healthy adults. The conjugates were safe and immunogenic. S. sonnei-CRM9, S. sonnei-CRM9succ, and S. sonnei-rEPAsucc elicited significant rises of geometric mean (GM) IgG anti-LPS within 1 week of injection (P < 0.001). At 26 weeks, the GM anti-LPS levels elicited by these three conjugates were similar and higher than their prevaccination levels (P < 0.0001). GM IgG anti-LPS levels elicited by S. flexneri 2a-rEPAsucc were significantly higher than those elicited by S. flexneri 2a-rCRM9succ at all intervals after injection. At 26 weeks, the levels of IgG anti-LPS in vaccinees were higher than their prevaccination levels (P < 0.0001). The serum antibody responses were specific, as there was no significant rise of anti-LPS to the heterologous O-SP in any vaccinee. Both conjugates elicited statistically significant rises of serum antibodies to the injected carrier protein. At 6 months, these five Shigella conjugates elicited higher fold rises than similar conjugates (D. N

  8. Examination of an aloe vera galacturonate polysaccharide capable of in situ gelation for the controlled release of protein therapeutics

    NASA Astrophysics Data System (ADS)

    McConaughy, Shawn David

    A therapeutic delivery platform has been investigated with the ultimate goal of designing a sustained protein release matrix utilizing an in-situ gelling, acidic polysaccharide derived from the Aloe vera plant. The Aloe vera polysaccharide (AvP) has been examined in order to determine how chemical composition, structure, molecular weight and solution behavior affect gelation and protein/peptide delivery. Correlations are drawn between structural characteristics and solution behavior in order to determine the impact of polymer conformation and solvation on gel formation under conditions designed to simulate nasal applications. Steady state and dynamic rheology, classic and dynamic light scattering, zeta potential, pulse field gradient nuclear magnetic resonance and fluorescence spectroscopy have been employed to gain insight into the effects of galacturonic acid content, degree of methylation, entanglement and ionic strength on both solution behavior and the hydrogel state which ultimately governs protein/peptide release. This dissertation is divided into two sections. In the first section, a series of Aloe vera polysaccharides (AvP), from the pectin family have been structurally characterized indicating high galacturonic acid (GalA) content, low degree of methylester substitution (DM), low numbers of rhamnose residues and high molecular weight with respect to pectins extracted from traditional sources. The behavior of AvP was examined utilizing dilute solution, low-shear rheological techniques for specific molecular weight samples at selected conditions of ionic strength. From these dilute aqueous solution studies, the Mark-Houwink-Sakurada (MHS) constants (K and alpha), persistence length (Lp) and inherent chain stiffness (B parameter) were determined, indicating an expanded random coil in aqueous salt solutions. The critical concentration for transition from dilute to concentrated solution, C e, was determined by measuring both the zero shear viscosity and

  9. A polysaccharide from Huaier induced apoptosis in MCF-7 breast cancer cells via down-regulation of MTDH protein.

    PubMed

    Luo, Zhiyong; Hu, Xiaopeng; Xiong, Hua; Qiu, Hong; Yuan, Xianglin; Zhu, Feng; Wang, Yihua; Zou, Yanmei

    2016-10-20

    In this study, one homogeneous polysaccharide (SP1), with a molecular weight of 56kDa, was isolated from the Huaier fruiting bodies. It had a backbone consisting of 1,4-linked-β-d-Galp and 1,3,6-linked-β-d-Galp residues, which was terminated with 1-linked-α-d-Glcp and 1-linked-α-l-Araf terminal at O-3 position of 1,3,6-linked-β-d-Galp unit along the main chain in the ratio of 1.1:2.0:1.1:1.1. MTT assay showed that shMTDH or SP1 (100, 200 and 400μg/ml) was able to suppress the proliferation of MCF-7 cells, due to a significant increase in the number of apoptotic cells as determined by flow cytometric analysis. Furthermore, Western blot analysis revealed that SP1 or shMTDH treatment led to a rise of ratio between proapoptotic Bax and antiapoptotic Bcl-2 protein in MCF-7 cells. In addition, carcinogene MTDH protein expression in MCF-7 cells received SP1 (100, 200 and 400μg/mL) or shMTDH treatment was also repressed after 48h incubation. Taken together, these findings indicated that SP1 has anticancer potential in the treatment of human breast cancer. PMID:27474651

  10. Polysaccharides: Occurrence, Significance, and Properties

    NASA Astrophysics Data System (ADS)

    Bemiller, James N.

    Polysaccharides are properties present significance in all living organisms where they carry out one or more of their diverse functions. While there is no specific category or definition of a complex polysaccharide, most are structurally complex. Polysaccharides contain 1-5 different monosaccharide (sugar) units. The different sugar units may have different anomeric configurations and/or be joined by different glycosidic linkages. Polysaccharides may be linear or branched. Branches may be short saccharide units on a linear backbone or the molecule may have a branch-on-branch structure; in either case, the branches may be isolated or clustered. Polysaccharides may contain non-carbohydrate groups. Esters or cyclic acetal groups, when present, can be removed by appropriate treatments. All polysaccharides are polydisperse, i. e., are present in a range of molecular weights rather than having a single molecular weight. Most are polymolecular, i. e., differ in fine structure from molecule to molecule. So most polysaccharides can be said to be structurally complex. They may be attached to protein molecules or to other polysaccharide molecules. They are solvated by water. Most dissolve in aqueous systems, especially if they are alkaline. Polysaccharides can be depolymerized by acids and heat, specific enzymes, and high pH systems following oxidation. Their hydroxyl groups can be esterified (acylated), etherified (alkylated), and oxidized. Amino groups can be acylated (and deacylated). Carboxyl groups can be converted into esters, amides, and amines. Structural modification makes the molecules even more complex and polymolecular and, perhaps, polydisperse.

  11. Food Protein-polysaccharide Conjugates Obtained via the Maillard Reaction: A Review.

    PubMed

    de Oliveira, Fabíola Cristina; Coimbra, Jane Sélia Dos Reis; de Oliveira, Eduardo Basílio; Zuñiga, Abraham Damian Giraldo; Rojas, Edwin E Garcia

    2016-05-18

    The products formed by glycosylation of food proteins with carbohydrates via the Maillard reaction, also known as conjugates, are agents capable of changing and improving techno-functional characteristics of proteins. The Maillard reaction uses the covalent bond between a group of a reducing carbohydrates and an amino group of a protein. This reaction does not require additional chemicals as it occurs naturally under controlled conditions of temperature, time, pH, and moisture. Moreover, there is growing interest in modifying proteins for industrial food applications. This review analyses the current state of art of the Maillard reaction on food protein functionalities. It also discusses the influence of the Maillard reaction on the conditions and formulation of reagents that improve desirable techno-functional characteristics of food protein.

  12. In vitro evaluation of the effects of protein-polyphenol-polysaccharide interactions on (+)-catechin and cyanidin-3-glucoside bioaccessibility.

    PubMed

    Oliveira, Ana; Pintado, Manuela

    2015-11-01

    The bioaccessibility of cyanidin-3-glucoside and (+)-catechin in model solutions when β-lactoglobulin (β-LG) and pectin/chitosan are present was investigated using an in vitro model simulating gastrointestinal conditions. In the mouth, the free cyanidin content increased (+) 90 and 14% while the (+)-catechin content decreased (-) 23 and 13%, respectively for mixtures with -pectin and -β-LG-pectin. Under gastric conditions, the cyanidin content decreased 85 and 28% for mixtures with -pectin and -β-LG-pectin. On the contrary, after gastric digestion, (+)-catechin bioaccessibility increased and exhibited values similar to the original samples for all the systems tested. The transition to the intestinal environment induced a significant alteration on both polyphenols and this effect was more marked for cyanidin. Systems with pectin allowed obtaining a higher content of bioaccessible cyanidin. The gastric conditions promoted an increase in the antioxidant capacity, followed by a decrease of it in the intestine. The free (+)-catechin and cyanidin-3-glucoside contents decreased when exposed to the gastrointestinal tract conditions. However, when incorporated in food matrix components, the gastrointestinal tract may act positively on the extraction of polyphenols, since they are progressively released from protein and polysaccharide bonds, being available for the absorption and to exert their biological effects.

  13. Influence of Hydrocolloids (Dietary Fibers) on Lipid Digestion of Protein-Stabilized Emulsions: Comparison of Neutral, Anionic, and Cationic Polysaccharides.

    PubMed

    Qin, Dingkui; Yang, Xiaojun; Gao, Songran; Yao, Junhu; McClements, David Julian

    2016-07-01

    The impact of dietary fibers on lipid digestion within the gastrointestinal tract depends on their molecular and physicochemical properties. In this study, the influence of the electrical characteristics of dietary fibers on their ability to interfere with the digestion of protein-coated lipid droplets was investigated using an in vitro small intestine model. Three dietary fibers were examined: cationic chitosan; anionic alginate; neutral locust bean gum (LBG). The particle size, ζ-potential, microstructure, and apparent viscosity of β-lactoglobulin stabilized oil-in-water emulsions containing different types and levels of dietary fiber were measured before and after lipid digestion. The rate and extent of lipid digestion depended on polysaccharide type and concentration. At relatively low dietary fiber levels (0.1 to 0.2 wt%), the initial lipid digestion rate was only reduced by chitosan, but the final extent of lipid digestion was unaffected by all 3 dietary fibers. At relatively high dietary fiber levels (0.4 wt%), alginate and chitosan significantly inhibited lipid hydrolysis, whereas LBG did not. The impact of chitosan on lipid digestion was attributed to its ability to promote fat droplet aggregation through bridging flocculation, thereby retarding access of the lipase to the droplet surfaces. The influence of alginate was mainly ascribed to its ability to sequester calcium ions and promote depletion flocculation.

  14. Sulfated polysaccharide-protein complex sensitizes doxorubicin-induced apoptosis of breast cancer cells in vitro and in vivo

    PubMed Central

    Wang, Jie; Wu, Hua Jian; Zhou, Chao Zhu; Wang, Hao

    2016-01-01

    The present study aimed to investigate the effect of sulfated polysaccharide-protein complex (SPPC) on the antitumor effect of doxorubicin (Dox) on MDA-MB-231 breast cancer cells in vitro and in vivo. MTT and Annexin V/propidium iodide staining assays demonstrated that SPPC selectively sensitized MDA-MB-231 cells to Dox-induced cytotoxicity. The half maximal inhibitory concentration of Dox against MDA-MB-231 cells was decreased from 5.3 to 1.5 µM when it was used concomitantly with 5 µM SPPC. SPPC potentiated Dox-induced apoptosis in breast cancer cells via the mitochondrial apoptosis signaling pathway by activating caspase-3 and caspase-9. Notably, the caspase inhibitor Z-VAD-fmk diminished the effect of SPPC on Dox-mediated apoptosis. Furthermore, combination treatment with SPPC and Dox markedly reduced the growth of breast cancer xenografts in mice. The present study demonstrated that SPPC was able to enhance the antitumor effect of Dox on breast cancer cells, thus suggesting that SPCC may be used to reduce the cumulative dose of Dox and its associated toxicities in the chemotherapy of breast cancer and other types of cancer.

  15. In vitro evaluation of the effects of protein-polyphenol-polysaccharide interactions on (+)-catechin and cyanidin-3-glucoside bioaccessibility.

    PubMed

    Oliveira, Ana; Pintado, Manuela

    2015-11-01

    The bioaccessibility of cyanidin-3-glucoside and (+)-catechin in model solutions when β-lactoglobulin (β-LG) and pectin/chitosan are present was investigated using an in vitro model simulating gastrointestinal conditions. In the mouth, the free cyanidin content increased (+) 90 and 14% while the (+)-catechin content decreased (-) 23 and 13%, respectively for mixtures with -pectin and -β-LG-pectin. Under gastric conditions, the cyanidin content decreased 85 and 28% for mixtures with -pectin and -β-LG-pectin. On the contrary, after gastric digestion, (+)-catechin bioaccessibility increased and exhibited values similar to the original samples for all the systems tested. The transition to the intestinal environment induced a significant alteration on both polyphenols and this effect was more marked for cyanidin. Systems with pectin allowed obtaining a higher content of bioaccessible cyanidin. The gastric conditions promoted an increase in the antioxidant capacity, followed by a decrease of it in the intestine. The free (+)-catechin and cyanidin-3-glucoside contents decreased when exposed to the gastrointestinal tract conditions. However, when incorporated in food matrix components, the gastrointestinal tract may act positively on the extraction of polyphenols, since they are progressively released from protein and polysaccharide bonds, being available for the absorption and to exert their biological effects. PMID:26289110

  16. Influence of Hydrocolloids (Dietary Fibers) on Lipid Digestion of Protein-Stabilized Emulsions: Comparison of Neutral, Anionic, and Cationic Polysaccharides.

    PubMed

    Qin, Dingkui; Yang, Xiaojun; Gao, Songran; Yao, Junhu; McClements, David Julian

    2016-07-01

    The impact of dietary fibers on lipid digestion within the gastrointestinal tract depends on their molecular and physicochemical properties. In this study, the influence of the electrical characteristics of dietary fibers on their ability to interfere with the digestion of protein-coated lipid droplets was investigated using an in vitro small intestine model. Three dietary fibers were examined: cationic chitosan; anionic alginate; neutral locust bean gum (LBG). The particle size, ζ-potential, microstructure, and apparent viscosity of β-lactoglobulin stabilized oil-in-water emulsions containing different types and levels of dietary fiber were measured before and after lipid digestion. The rate and extent of lipid digestion depended on polysaccharide type and concentration. At relatively low dietary fiber levels (0.1 to 0.2 wt%), the initial lipid digestion rate was only reduced by chitosan, but the final extent of lipid digestion was unaffected by all 3 dietary fibers. At relatively high dietary fiber levels (0.4 wt%), alginate and chitosan significantly inhibited lipid hydrolysis, whereas LBG did not. The impact of chitosan on lipid digestion was attributed to its ability to promote fat droplet aggregation through bridging flocculation, thereby retarding access of the lipase to the droplet surfaces. The influence of alginate was mainly ascribed to its ability to sequester calcium ions and promote depletion flocculation. PMID:27300319

  17. Sulfated polysaccharide-protein complex sensitizes doxorubicin-induced apoptosis of breast cancer cells in vitro and in vivo

    PubMed Central

    Wang, Jie; Wu, Hua Jian; Zhou, Chao Zhu; Wang, Hao

    2016-01-01

    The present study aimed to investigate the effect of sulfated polysaccharide-protein complex (SPPC) on the antitumor effect of doxorubicin (Dox) on MDA-MB-231 breast cancer cells in vitro and in vivo. MTT and Annexin V/propidium iodide staining assays demonstrated that SPPC selectively sensitized MDA-MB-231 cells to Dox-induced cytotoxicity. The half maximal inhibitory concentration of Dox against MDA-MB-231 cells was decreased from 5.3 to 1.5 µM when it was used concomitantly with 5 µM SPPC. SPPC potentiated Dox-induced apoptosis in breast cancer cells via the mitochondrial apoptosis signaling pathway by activating caspase-3 and caspase-9. Notably, the caspase inhibitor Z-VAD-fmk diminished the effect of SPPC on Dox-mediated apoptosis. Furthermore, combination treatment with SPPC and Dox markedly reduced the growth of breast cancer xenografts in mice. The present study demonstrated that SPPC was able to enhance the antitumor effect of Dox on breast cancer cells, thus suggesting that SPCC may be used to reduce the cumulative dose of Dox and its associated toxicities in the chemotherapy of breast cancer and other types of cancer. PMID:27698706

  18. Protein-bound polysaccharide-K (PSK) induces apoptosis and inhibits proliferation of promyelomonocytic leukemia HL-60 cells.

    PubMed

    Hirahara, Noriyuki; Fujioka, Masaki; Edamatsu, Takeo; Fujieda, Ayako; Sekine, Fujio; Wada, Tsutomu; Tanaka, Tsuneo

    2011-09-01

    Protein-bound polysaccharide-K (PSK) is extracted from Coriolus versicolor (CM101), and is clinically used in combination therapy for gastrointestinal cancer and small cell lung carcinoma. PSK is a biological response modifier (BRM), and its mechanism of action is partly mediated, by modulating host immune systems, such as the activation of immune effector cells and the neutralization of transforming growth factor-beta (TGFβ) activity. Direct inhibition of tumor cell proliferation has been reported as another mechanism, but how PSK induces such an effect remains to be elucidated. Here, the anti-proliferative activity of PSK was examined using seven different human malignant cell lines (WiDr, HT29, SW480, KATOIII, AGS, HL60 and U937), and PSK was found to inhibit the proliferation of HL-60 cells most profoundly. Therefore, HL-60 cells were used to clarify the mechanism of anti-proliferative activity. Caspase-3 activation followed by apoptosis are involved at least in part in the PSK-induced anti-proliferative activity against HL-60 cells. PMID:21868514

  19. Short communication: Changes in fluorescence intensity induced by soybean soluble polysaccharide-milk protein interactions during acidification.

    PubMed

    Li, Y H; Wang, W J; Xu, X J; Meng, Y C; Zhang, L W; Chen, J; Qiu, R

    2015-12-01

    Interactions between stabilizer and milk protein are believed to influence the stabilizing behavior of the milk system. We investigated changes in fluorescence intensity induced by interactions of soybean soluble polysaccharide (SSPS) and milk protein (Mp) during acidification. The fluorescence intensity (If) of Mp increased as pH decreased from 6.8 to 5.2. Compared with Mp alone, If of SSPS-Mp mixtures increased as the pH decreased from 6.8 to 5.2. We found that the If of the SSPS-Mp mixture decreased in a pH range from 5.2 to 3.6, which indicated a change in the polarity microenvironment around the Trp residues. We also found that the maximum emission wavelength (λmax) shifted from 337 to 330nm as pH decreased from 6.8 to 3.6, in further support of SSPS interacting with the polar portion of Mp during acidification. Furthermore, an excited monomeric molecule (pyrene exciplex) was found as a ground-state pyrene formed and a broad band was shown at about 450nm. The intensity ratio of the first peak to the third peak (I1:I3) of Mp increased slightly, and the ratio of intensity of pyrene exciplex to monomer (Ie:Im) decreased because pyrene molecules were located in a less hydrophobic microenvironment during acidification. However, the ratio of I1:I3 decreased clearly at pH below 5.6 and the ratio of Ie:Im showed the opposite trend in the SSPS-Mp mixture. Changes in intrinsic and exogenous fluorescence intensity confirmed that interactions of SSPS and Mp could change the polarity of the microenvironment and that SSPS probably interacted with the polar portion of Mp. These results could give insight into the behavior of stabilizers in acid milk products. PMID:26476946

  20. A structural overview of GH61 proteins – fungal cellulose degrading polysaccharide monooxygenases

    PubMed Central

    Leggio, Leila Lo; Welner, Ditte; De Maria, Leonardo

    2012-01-01

    Recent years have witnessed a spurt of activities in the elucidation of the molecular function of a class of proteins with great potential in biomass degradation. GH61 proteins are of fungal origin and were originally classified in family 61 of the glycoside hydrolases. From the beginning they were strongly suspected to be involved in cellulose degradation because of their expression profiles, despite very low detectable endoglucanase activities. A major breakthrough came from structure determination of the first members, establishing the presence of a divalent metal binding site and a similarity to bacterial proteins involved in chitin degradation. A second breakthrough came from the identification of cellulase boosting activity dependent on the integrity of the metal binding site. Finally very recently GH61 proteins were demonstrated to oxidatively cleave crystalline cellulose in a Cu and reductant dependant manner. This mini-review in particular focuses on the contribution that structure elucidation has made in the understanding of GH61 molecular function and reviews the currently known structures and the challenges remaining ahead for exploiting this new class of enzymes to the full. PMID:24688660

  1. Alr5068, a Low-Molecular-Weight protein tyrosine phosphatase, is involved in formation of the heterocysts polysaccharide layer in the cyanobacterium Anabaena sp. PCC 7120.

    PubMed

    Tan, Hui; Wan, Shuang; Liu, Pi-Qiong; Wang, Li; Zhang, Cheng-Cai; Chen, Wen-Li

    2013-10-01

    The filamentous cyanobacterium Anabaena sp. PCC 7120 forms nitrogen-fixing heterocysts after deprivation of combined nitrogen. Under such conditions, vegetative cells provide heterocysts with photosynthate and receive fixed nitrogen from the latter. Heterocyst envelope contains a glycolipid layer and a polysaccharide layer to restrict the diffusion of oxygen into heterocysts. Low-Molecular-Weight protein tyrosine phosphatases (LMW-PTPs) are involved in the biosynthesis of exopolysaccharides in bacteria. Alr5068, a protein from Anabaena sp. PCC 7120, shows significant sequence similarity with LMW-PTPs. In this study we characterized the enzymatic properties of Alr5068 and showed that it can dephosphorylate several autophosphorylated tyrosine kinases (Alr2856, Alr3059 and All4432) of Anabaena sp. PCC 7120 in vitro. Several conserved residues among LMW-PTPs are shown to be essential for the phosphatase activity of Alr5068. Overexpression of alr5068 results in a strain unable to survive under diazotrophic conditions, with the formation of morphologically mature heterocysts detached from the filaments. Overexpression of an alr5068 allele that lost phosphatase activity led to the formation of heterocyst with an impaired polysaccharide layer. The alr5068 gene was upregulated after nitrogen step-down and its mutation affected the expression of hepA and hepC, two genes necessary for the formation of the heterocyst envelope polysaccharide (HEP) layer. Our results suggest that Alr5068 is associated with the production of HEP in Anabaena sp. PCC 7120.

  2. Alr5068, a Low-Molecular-Weight protein tyrosine phosphatase, is involved in formation of the heterocysts polysaccharide layer in the cyanobacterium Anabaena sp. PCC 7120.

    PubMed

    Tan, Hui; Wan, Shuang; Liu, Pi-Qiong; Wang, Li; Zhang, Cheng-Cai; Chen, Wen-Li

    2013-10-01

    The filamentous cyanobacterium Anabaena sp. PCC 7120 forms nitrogen-fixing heterocysts after deprivation of combined nitrogen. Under such conditions, vegetative cells provide heterocysts with photosynthate and receive fixed nitrogen from the latter. Heterocyst envelope contains a glycolipid layer and a polysaccharide layer to restrict the diffusion of oxygen into heterocysts. Low-Molecular-Weight protein tyrosine phosphatases (LMW-PTPs) are involved in the biosynthesis of exopolysaccharides in bacteria. Alr5068, a protein from Anabaena sp. PCC 7120, shows significant sequence similarity with LMW-PTPs. In this study we characterized the enzymatic properties of Alr5068 and showed that it can dephosphorylate several autophosphorylated tyrosine kinases (Alr2856, Alr3059 and All4432) of Anabaena sp. PCC 7120 in vitro. Several conserved residues among LMW-PTPs are shown to be essential for the phosphatase activity of Alr5068. Overexpression of alr5068 results in a strain unable to survive under diazotrophic conditions, with the formation of morphologically mature heterocysts detached from the filaments. Overexpression of an alr5068 allele that lost phosphatase activity led to the formation of heterocyst with an impaired polysaccharide layer. The alr5068 gene was upregulated after nitrogen step-down and its mutation affected the expression of hepA and hepC, two genes necessary for the formation of the heterocyst envelope polysaccharide (HEP) layer. Our results suggest that Alr5068 is associated with the production of HEP in Anabaena sp. PCC 7120. PMID:23827083

  3. Proteins among the polysaccharides: a new perspective on root cap slime.

    PubMed

    Wen, Fushi; Curlango-Rivera, Gilberto; Hawes, Martha C

    2007-09-01

    Charles Darwin recognized the power of the root cap as a model for plant signalling and behavior, and used it to explore the ways plants sense and respond to diverse stimuli. Over ensuing decades, various groups have reported tantalizing clues regarding the role of a complex extracellular matrix that ensheaths the tip region housing the apical and root cap meristems. In the course of characterizing root tip resistance to infection and injury and the role border cells play in this phenomenon, we confirmed and extended early- and mid-20(th) century studies reporting enzyme activities secreted from the root cap. Multidimensional protein analysis revealed, in fact, that >100 proteins are actively synthesized and secreted from the root cap and border cells. This 'root cap secretome' appears to be a critical component of root tip resistance to infection. We have developed a microscopic assay to quantify the protein-based extracellular response to dynamic changes in environmental conditions including hydroponic culture, and present the results here. This tool provides a simple, direct measure that can be used to explore the ways border cells may function in the manner of white blood cells to trap, immobilize and neutralize threats to the growing root tip.

  4. Dynamic contact angle analysis of protein adsorption on polysaccharide multilayer's films for biomaterial reendothelialization.

    PubMed

    Benni, Safiya; Avramoglou, Thierry; Hlawaty, Hanna; Mora, Laurence

    2014-01-01

    Atherosclerosis is a major cardiovascular disease. One of the side effects is restenosis. The aim of this work was to study the coating of stents by dextran derivates based polyelectrolyte's multilayer (PEM) films in order to increase endothelialization of injured arterial wall after stent implantation. Films were composed with diethylaminoethyl dextran (DEAE) as polycation and dextran sulphate (DS) as polyanion. One film was composed with 4 bilayers of (DEAE-DS)4 and was labeled D-. The other film was the same as D- but with an added terminal layer of DEAE polycation: (DEAE-DS)4-DEAE (labeled D+). The dynamic adsorption/desorption of proteins on the films were characterized by dynamic contact angle (DCA) and atomic force microscopy (AFM). Human endothelial cell (HUVEC) adhesion and proliferation were quantified and correlated to protein adsorption analyzed by DCA for fibronectin, vitronectin, and bovine serum albumin (BSA). Our results showed that the endothelial cell response was optimal for films composed of DS as external layer. Fibronectin was found to be the only protein to exhibit a reversible change in conformation after desorption test. This behavior was only observed for (DEAE-DS)4 films. (DEAE-DS)4 films could enhance HUVEC proliferation in agreement with fibronectin ability to easily change from conformation.

  5. Dynamic Contact Angle Analysis of Protein Adsorption on Polysaccharide Multilayer's Films for Biomaterial Reendothelialization

    PubMed Central

    Benni, Safiya; Mora, Laurence

    2014-01-01

    Atherosclerosis is a major cardiovascular disease. One of the side effects is restenosis. The aim of this work was to study the coating of stents by dextran derivates based polyelectrolyte's multilayer (PEM) films in order to increase endothelialization of injured arterial wall after stent implantation. Films were composed with diethylaminoethyl dextran (DEAE) as polycation and dextran sulphate (DS) as polyanion. One film was composed with 4 bilayers of (DEAE-DS)4 and was labeled D−. The other film was the same as D− but with an added terminal layer of DEAE polycation: (DEAE-DS)4-DEAE (labeled D+). The dynamic adsorption/desorption of proteins on the films were characterized by dynamic contact angle (DCA) and atomic force microscopy (AFM). Human endothelial cell (HUVEC) adhesion and proliferation were quantified and correlated to protein adsorption analyzed by DCA for fibronectin, vitronectin, and bovine serum albumin (BSA). Our results showed that the endothelial cell response was optimal for films composed of DS as external layer. Fibronectin was found to be the only protein to exhibit a reversible change in conformation after desorption test. This behavior was only observed for (DEAE-DS)4 films. (DEAE-DS)4 films could enhance HUVEC proliferation in agreement with fibronectin ability to easily change from conformation. PMID:25276808

  6. Dynamic contact angle analysis of protein adsorption on polysaccharide multilayer's films for biomaterial reendothelialization.

    PubMed

    Benni, Safiya; Avramoglou, Thierry; Hlawaty, Hanna; Mora, Laurence

    2014-01-01

    Atherosclerosis is a major cardiovascular disease. One of the side effects is restenosis. The aim of this work was to study the coating of stents by dextran derivates based polyelectrolyte's multilayer (PEM) films in order to increase endothelialization of injured arterial wall after stent implantation. Films were composed with diethylaminoethyl dextran (DEAE) as polycation and dextran sulphate (DS) as polyanion. One film was composed with 4 bilayers of (DEAE-DS)4 and was labeled D-. The other film was the same as D- but with an added terminal layer of DEAE polycation: (DEAE-DS)4-DEAE (labeled D+). The dynamic adsorption/desorption of proteins on the films were characterized by dynamic contact angle (DCA) and atomic force microscopy (AFM). Human endothelial cell (HUVEC) adhesion and proliferation were quantified and correlated to protein adsorption analyzed by DCA for fibronectin, vitronectin, and bovine serum albumin (BSA). Our results showed that the endothelial cell response was optimal for films composed of DS as external layer. Fibronectin was found to be the only protein to exhibit a reversible change in conformation after desorption test. This behavior was only observed for (DEAE-DS)4 films. (DEAE-DS)4 films could enhance HUVEC proliferation in agreement with fibronectin ability to easily change from conformation. PMID:25276808

  7. Noncovalent association of protein and capsular polysaccharide on bacteria-sized latex beads as a model for polysaccharide-specific humoral immunity to intact gram-positive extracellular bacteria.

    PubMed

    Colino, Jesus; Duke, Leah; Snapper, Clifford M

    2013-09-15

    Intact Streptococcus pneumoniae expressing type 14 capsular polysaccharide (PPS14) and type III S. agalactiae containing a PPS14 core capsule identical to PPS14 exhibit noncovalent associations of PPS14 and bacterial protein, in contrast to soluble covalent conjugates of these respective Ags. Both bacteria and conjugates induce murine PPS14-specific IgG responses dependent on CD4⁺ T cells. Further, secondary immunization with conjugate and S. agalactiae, although not S. pneumoniae, results in a boosted response. However, in contrast to conjugate, PPS14-specific IgG responses to bacteria lack affinity maturation use the 44.1-idiotype and are dependent on marginal zone B cells. To better understand the mechanism underlying this dichotomy, we developed a minimal model of intact bacteria in which PPS14 and pneumococcal surface protein A (PspA) were stably attached to 1 μm (bacteria-sized) latex beads, but not directly linked to each other, in contrast to PPS14-PspA conjugate. Beads coated simultaneously with PPS14+[PspA], similar to conjugate, induced in mice boosted PPS14-specific IgG secondary responses, dependent on T cells and ICOS-dependent costimulation, and in which priming could be achieved with PspA alone. In contrast to conjugate, but similar to intact bacteria, the primary PPS14-specific IgG response to beads coated simultaneously with PPS14+[PspA] peaked rapidly, with the secondary response highly enriched for the 44.1-idiotype and lacking affinity maturation. These results demonstrate that noncovalent association in a particle, of polysaccharide and protein, recapitulates essential immunologic characteristics of intact bacteria that are distinct from soluble covalent conjugates of these respective Ags.

  8. Effects of the conjugation of whey proteins with gellan polysaccharides on surfactant-induced competitive displacement from the air-water interface.

    PubMed

    Cai, B; Ikeda, S

    2016-08-01

    Whey proteins can be used to stabilize foams and emulsions against coalescence because of their ability to form viscoelastic films at the interface that resist film rupture on collision between colloidal particles. However, whey proteins are competitively displaced from the interface if small-molecule surfactants are added, leading to destabilization of the entire system. This is because surfactants are more effective in molecular packing at the interface, and they lower interfacial tension to a greater degree than whey proteins do, but their interfacial films are poor in viscoelasticity. We hypothesized that whey proteins would become more resistant to surfactant-induced competitive displacement if they were conjugated with network-forming polysaccharides. The protein moiety of the conjugate would be expected to enable its adsorption to the interface, and the polysaccharide moiety would be expected to form self-assembled networks, strengthening the interfacial film as a whole. In this study, whey proteins were conjugated with gellan polysaccharides using the Maillard reaction. Atomic force microscopy images of interfacial films formed by the whey protein-gellan conjugate at the air-water interface and transferred onto mica sheets using the Langmuir-Blodgett method revealed that gellan did form self-assembled networks at the interface and that interfacial films also contained a large number of unconjugated whey protein molecules. Following the addition of a small-molecule surfactant (Tween 20) to the sub-phase, surface pressure increased, indicating spontaneous adsorption of surfactants to the interface. Atomic force microscopy images showed decreases in interfacial area coverage by whey proteins as surface pressure increased. At a given surface pressure, the interfacial area coverage by whey protein-gellan conjugates was greater than coverage by unconjugated whey proteins, confirming that whey proteins became more resistant to surfactant-induced displacement after

  9. Effect of polysaccharides extract of rhizoma atractylodis macrocephalae on thymus, spleen and cardiac indexes, caspase-3 activity ratio, Smac/DIABLO and HtrA2/Omi protein and mRNA expression levels in aged rats.

    PubMed

    Guo, Ling; Sun, Yong Le; Wang, Ai Hong; Xu, Chong En; Zhang, Meng Yuan

    2012-10-01

    This study was designed to determine the possible protective effect of polysaccharides extract of rhizoma atractylodis macrocephalae on heart function in aged rats. Polysaccharides extract of rhizoma atractylodis macrocephalae was administered to aged rats. Results showed that thymus, spleen and cardiac indexs were significantly increased, whereas caspase-3 activity ratio, Smac/DIABLO and HtrA2/Omi protein expression, Smac/DIABLO and HtrA2/Omi mRNA expression levels were markedly reduced. It can be concluded that polysaccharides extract of rhizoma atractylodis macrocephalae may enhance immunity and improve heart function in aged rats.

  10. Protein-bound polysaccharide-K induces apoptosis via mitochondria and p38 mitogen-activated protein kinase-dependent pathways in HL-60 promyelomonocytic leukemia cells.

    PubMed

    Hirahara, Noriyuki; Edamatsu, Takeo; Fujieda, Ayako; Fujioka, Masaki; Wada, Tsutomu; Tajima, Yoshitsugu

    2013-07-01

    Protein-bound polysaccharide-K (PSK) is extracted from Coriolus versicolor (CM101). PSK is a biological response modifier (BRM), and its mechanism of action is partly mediated by modulating host immune systems; however, recent studies showed antiproliferative activity of PSK. Therefore, we examined the mechanism underlying the antiproliferative activity of PSK using seven different human malignant cell lines (WiDr, HT29, SW480, KATOIII, AGS, HL-60 and U937), and PSK was found to inhibit the proliferation of HL-60 cells most profoundly. Therefore, HL-60 cells were used to elucidate the mechanism of the antiproliferative activity. Western blotting was performed to detect phosphorylated p38 mitogen-activated protein kinase (MAPK). A p38 MAPK inhibitor, SB203580, was used to examine the roles in PSK-induced apoptosis and growth inhibition. Flow cytometry was performed for mitochondrial membrane potential detection. PSK activated caspase-3 and induced p38 MAPK phosphorylation. Co-treatment with SB203580 blocked PSK-induced apoptosis, caspase-3 activation and growth inhibition. PSK induced apoptosis via the mitochondrial pathway. The depolarization of mitochondria induced by PSK was reversed by co-treatment with SB203580. The present study revealed that PSK induced apoptosis in HL-60 cells via a mitochondrial and p38 MAPK-dependent pathway. PMID:23604455

  11. Lipopolysaccharide and β-1,3-glucan-binding protein (LGBP) bind to seaweed polysaccharides and activate the prophenoloxidase system in white shrimp Litopenaeus vannamei.

    PubMed

    Chen, Yu-Yuan; Chen, Jiann-Chu; Kuo, Yi-Hsuan; Lin, Yong-Chin; Chang, Yu-Hsuan; Gong, Hong-Yi; Huang, Chien-Lun

    2016-02-01

    Lipopolysaccharide and β-1,3-glucan-binding protein (LGBP), important pattern recognition proteins (PRPs), recognize lipopolysaccharide (LPS) and β-1,3-glucan (βG), known as pathogen-associated molecular patterns (PAMPs), and subsequently trigger innate immunity. Several seaweed polysaccharides and seaweed extracts increase immune parameters and resistance to pathogens. Here, we constructed the expression vector pET28b-LvLGBP and transferred it into Escherichia coli BL21 (DE3) for protein expression and to produce the recombinant protein LGBP (rLvLGBP) in white shrimp Litopenaeus vannamei. We examined the binding of rLvLGBP with seaweed-derived polysaccharides including alginate, carrageenan, fucoidan, laminarin, Gracilaria tenuistipitata extract (GTE), and Sargassum duplicatum extract (SDE), and examined the phenoloxidase activity of shrimp haemocytes incubated with a mixture of rLvLGBP and each polysaccharide. We also examined the binding of rLvLGBP with LPS and βG, and the phenoloxidase activity of shrimp haemocytes incubated with a mixture of rLvLGBP and LPS (rLvLGBP-LPS) or a mixture of rLvLGBP and βG (rLvLGBP-βG). An ELISA binding assay indicated that rLvLGBP binds to LPS, βG, alginate, carrageenan, fucoidan, laminarin, GTE, and SDE with dissociation constants of 0.1138-0.1770 μM. Furthermore, our results also indicated that the phenoloxidase activity of shrimp haemocytes incubated with a mixture of rLvLGBP and LPS, βG, alginate, carrageenan, fucoidan, laminarin, GTE, and SDE significantly increased by 328%, 172%, 200%, 213%, 197%, 194%, 191%, and 197%, respectively compared to controls (cacodylate buffer). We conclude that LvLGBP functions as a PRP, recognizes and binds to LPS, βG, alginate, carrageenan, fucoidan, laminarin, GTE, and SDE, and subsequently leads to activating innate immunity in shrimp. PMID:26522339

  12. Microanalysis, Pharmacokinetics and Tissue Distribution of Polysaccharide-Protein Complexes from Longan Pulp in Mice

    PubMed Central

    Min, Ting; Sun, Jie; Yi, Yang; Wang, Hong-Xun; Hang, Fei; Ai, You-Wei; Wang, Li-Mei

    2015-01-01

    A high performance size exclusion-fluorescence detection (HPSEC-FD) method combined with fluorescein isothiocyanate (FITC) prelabeling was established for the microanalysis of polysaccharide–protein complexes from longan pulp (LPP). FITC-labeled LPP (LPPF) was fractionated by gel filtration chromatography. The weight-average molecular weight and FITC substitution degree of LPPF were 39.01 kDa and 0.20%, respectively. The HPSEC-FD calibration curves linear over the range of 1–200 µg/mL in mouse plasma, spleen and lung samples with correlation coefficients greater than 0.995. The inter-day and intra-day precisions of the method were not more than 6.9%, and the relative recovery ranged from 93.7% to 106.4%. The concentration–time curve of LPPF in plasma following intravenous (i.v.) administration at 40 mg/kg body weight well fitted to a two-compartment model. LPPF rapidly eliminated from plasma according to the short half-lives (t1/2α = 2.23 min, t1/2β = 39.11 min) and mean retention times (MRT0–t = 1.15 h, MRT0–∞ = 1.39 h). After administration over 5 to 360 min, the concentration of LPPF in spleen homogenate decreased from 7.41 to 3.68 µg/mL; the concentration in lung homogenate decreased from 9.08 to 3.40 µg/mL. On the other hand, the increasing concentration of LPPF fraction with low molecular weight in heart homogenate was observed. PMID:26501257

  13. Structural characterization and immunomodulatory activity of Grifola frondosa polysaccharide via toll-like receptor 4-mitogen-activated protein kinases-nuclear factor κB pathways.

    PubMed

    Ma, Xiaolei; Meng, Meng; Han, Lirong; Cheng, Dai; Cao, Xiaohong; Wang, Chunling

    2016-06-15

    We isolated a neutral polysaccharide from the fruiting body of a mushroom Grifola frondosa (GFP-A). The aim of this study was to characterize a neutral α-d-polysaccharide derived from G. frondosa and evaluate its immunomodulatory effect on toll-like receptor 4, mitogen-activated protein kinases and nuclear factor κB pathways of protein expression in macrophages. The structural features of GFP-A were characterized by physicochemical and instrumental analyses. Its molecular weight was found to be 8.48 × 10(2) kDa. The main chain of GFP-A consisted of (1 → 4)-linked and (1 → 6)-linked α-d-glucopyranosyl, and (1 → 3,6)-linked α-d-mannopyranosyl residues, which branched at C-3. The branches consisted of (1 → 6)-linked α-d-galactopyranosyl and t-l-rhamnopyranosyl residues. An in vitro immunomodulatory assay for pro-inflammatory cytokines (interleukin-1β, interleukin-2, tumor necrosis factor alpha, etc.) using the macrophage cell line, RAW 264.7, revealed that GFP-A exhibited significant immunomodulatory activity by stimulating the toll-like receptor 4, mitogen-activated protein kinases to nuclear factor κB/pathway. PMID:27220562

  14. Polysaccharide Degradation

    NASA Astrophysics Data System (ADS)

    Stone, Bruce A.; Svensson, Birte; Collins, Michelle E.; Rastall, Robert A.

    An overview of current and potential enzymes used to degrade polysaccharides is presented. Such depolymerases are comprised of glycoside hydrolases, glycosyl transferases, phosphorylases and lyases, and their classification, active sites and action patterns are discussed. Additionally, the mechanisms that these enzymes use to cleave glycosidic linkages is reviewed as are inhibitors of depolymerase activity; reagents which react with amino acid residues, glycoside derivatives, transition state inhibitors and proteinaceous inhibitors. The characterization of various enzymes of microbial, animal or plant origin has led to their widespread use in the production of important oligosaccharides which can be incorporated into food stuffs. Sources of polysaccharides of particular interest in this chapter are those from plants and include inulin, dextran, xylan and pectin, as their hydrolysis products are purported to be functional foods in the context of gastrointestinal health. An alternative use of degraded polysaccharides is in the treatment of disease. The possibility exists to treat bacterial exopolysaccharide with lyases from bacteriophage to produce oligosaccharides exhibiting bioactive sequences. Although this area is currently in its infancy the knowledge is available to investigate further.

  15. Trans-Golgi network localized ECHIDNA/Ypt interacting protein complex is required for the secretion of cell wall polysaccharides in Arabidopsis.

    PubMed

    Gendre, Delphine; McFarlane, Heather E; Johnson, Errin; Mouille, Gregory; Sjödin, Andreas; Oh, Jaesung; Levesque-Tremblay, Gabriel; Watanabe, Yoichiro; Samuels, Lacey; Bhalerao, Rishikesh P

    2013-07-01

    The secretion of cell wall polysaccharides through the trans-Golgi network (TGN) is required for plant cell elongation. However, the components mediating the post-Golgi secretion of pectin and hemicellulose, the two major cell wall polysaccharides, are largely unknown. We identified evolutionarily conserved YPT/RAB GTPase Interacting Protein 4a (YIP4a) and YIP4b (formerly YIP2), which form a TGN-localized complex with ECHIDNA (ECH) in Arabidopsis thaliana. The localization of YIP4 and ECH proteins at the TGN is interdependent and influences the localization of VHA-a1 and SYP61, which are key components of the TGN. YIP4a and YIP4b act redundantly, and the yip4a yip4b double mutants have a cell elongation defect. Genetic, biochemical, and cell biological analyses demonstrate that the ECH/YIP4 complex plays a key role in TGN-mediated secretion of pectin and hemicellulose to the cell wall in dark-grown hypocotyls and in secretory cells of the seed coat. In keeping with these observations, Fourier transform infrared microspectroscopy analysis revealed that the ech and yip4a yip4b mutants exhibit changes in their cell wall composition. Overall, our results reveal a TGN subdomain defined by ECH/YIP4 that is required for the secretion of pectin and hemicellulose and distinguishes the role of the TGN in secretion from its roles in endocytic and vacuolar trafficking. PMID:23832588

  16. Antibody production in vitamin A-depleted rats is impaired after immunization with bacterial polysaccharide or protein antigens.

    PubMed

    Pasatiempo, A M; Kinoshita, M; Taylor, C E; Ross, A C

    1990-05-01

    Vitamin A nutritional status has been implicated as important in maintaining the integrity of immune functions. We have determined the effect of vitamin A (retinol) depletion on the ability of young animals to produce antibodies after challenge with various bacterial antigens. Male Lewis rats raised on vitamin A-free or adequate diets were immunized either near 40 days of age, before signs of vitamin A deficiency were apparent, or near 47 days of age when symptoms of deficiency were beginning to be manifest. For rats immunized with polysaccharide antigens from Streptococcus pneumoniae or Neisseria meningitidis, antibody production did not exceed 0-19% of the response of control rats. Vitamin A depletion also severely compromised the response to two T cell-dependent antigens, tetanus toxoid and sheep red blood cells. In striking contrast, retinol-depleted rats immunized with lipopolysaccharides from Pseudomonas aeruginosa and Serratia marcesens produced an antibody response indistinguishable from retinol-sufficient animals. These lipopolysaccharides could elicit antibodies in rat pups, whereas the capsular polysaccharide antigens could not. This is consistent with the characteristics of type 1 and type 2 antigens, respectively. These studies indicate that retinol status is an important determinant of the humoral immune response to certain types of antigen and suggest that antibody production to capsular polysaccharides and T cell-dependent antigens is particularly dependent on adequate retinol status.

  17. Polysaccharide-Based Micelles for Drug Delivery

    PubMed Central

    Zhang, Nan; Wardwell, Patricia R.; Bader, Rebecca A.

    2013-01-01

    Delivery of hydrophobic molecules and proteins has been an issue due to poor bioavailability following administration. Thus, micelle carrier systems are being investigated to improve drug solubility and stability. Due to problems with toxicity and immunogenicity, natural polysaccharides are being explored as substitutes for synthetic polymers in the development of new micelle systems. By grafting hydrophobic moieties to the polysaccharide backbone, self-assembled micelles can be readily formed in aqueous solution. Many polysaccharides also possess inherent bioactivity that can facilitate mucoadhesion, enhanced targeting of specific tissues, and a reduction in the inflammatory response. Furthermore, the hydrophilic nature of some polysaccharides can be exploited to enhance circulatory stability. This review will highlight the advantages of polysaccharide use in the development of drug delivery systems and will provide an overview of the polysaccharide-based micelles that have been developed to date. PMID:24300453

  18. S-Layer Homology Domain Proteins Csac_0678 and Csac_2722 Are Implicated in Plant Polysaccharide Deconstruction by the Extremely Thermophilic Bacterium Caldicellulosiruptor saccharolyticus

    PubMed Central

    Ozdemir, Inci; Blumer-Schuette, Sara E.

    2012-01-01

    The genus Caldicellulosiruptor contains extremely thermophilic bacteria that grow on plant polysaccharides. The genomes of Caldicellulosiruptor species reveal certain surface layer homology (SLH) domain proteins that have distinguishing features, pointing to a role in lignocellulose deconstruction. Two of these proteins in Caldicellulosiruptor saccharolyticus (Csac_0678 and Csac_2722) were examined from this perspective. In addition to three contiguous SLH domains, the Csac_0678 gene encodes a glycoside hydrolase family 5 (GH5) catalytic domain and a family 28 carbohydrate-binding module (CBM); orthologs to Csac_0678 could be identified in all genome-sequenced Caldicellulosiruptor species. Recombinant Csac_0678 was optimally active at 75°C and pH 5.0, exhibiting both endoglucanase and xylanase activities. SLH domain removal did not impact Csac_0678 GH activity, but deletion of the CBM28 domain eliminated binding to crystalline cellulose and rendered the enzyme inactive on this substrate. Csac_2722 is the largest open reading frame (ORF) in the C. saccharolyticus genome (predicted molecular mass of 286,516 kDa) and contains two putative sugar-binding domains, two Big4 domains (bacterial domains with an immunoglobulin [Ig]-like fold), and a cadherin-like (Cd) domain. Recombinant Csac_2722, lacking the SLH and Cd domains, bound to cellulose and had detectable carboxymethylcellulose (CMC) hydrolytic activity. Antibodies directed against Csac_0678 and Csac_2722 confirmed that these proteins bound to the C. saccharolyticus S-layer. Their cellular localization and functional biochemical properties indicate roles for Csac_0678 and Csac_2722 in recruitment and hydrolysis of complex polysaccharides and the deconstruction of lignocellulosic biomass. Furthermore, these results suggest that related SLH domain proteins in other Caldicellulosiruptor genomes may also be important contributors to plant biomass utilization. PMID:22138994

  19. A major facilitator superfamily protein, HepP, is involved in formation of the heterocyst envelope polysaccharide in the cyanobacterium Anabaena sp. strain PCC 7120.

    PubMed

    López-Igual, Rocío; Lechno-Yossef, Sigal; Fan, Qing; Herrero, Antonia; Flores, Enrique; Wolk, C Peter

    2012-09-01

    Some filamentous cyanobacteria such as Anabaena sp. strain PCC 7120 produce cells, termed heterocysts, specialized in nitrogen fixation. Heterocysts bear a thick envelope containing an inner layer of glycolipids and an outer layer of polysaccharide that restrict the diffusion of air (including O(2)) into the heterocyst. Anabaena sp. mutants impaired in production of either of those layers show a Fox(-) phenotype (requiring fixed nitrogen for growth under oxic conditions). We have characterized a set of transposon-induced Fox(-) mutants in which transposon Tn5-1063 was inserted into the Anabaena sp. chromosome open reading frame all1711 which encodes a predicted membrane protein that belongs to the major facilitator superfamily (MFS). These mutants showed higher nitrogenase activities under anoxic than under oxic conditions and altered sucrose uptake. Electron microscopy and alcian blue staining showed a lack of the heterocyst envelope polysaccharide (Hep) layer. Northern blot and primer extension analyses showed that, in a manner dependent on the nitrogen-control transcription factor NtcA, all1711 was strongly induced after nitrogen step-down. Confocal microscopy of an Anabaena sp. strain producing an All1711-green fluorescent protein (All1711-GFP) fusion protein showed induction in all cells of the filament but at higher levels in differentiating heterocysts. All1711-GFP was located in the periphery of the cells, consistent with All1711 being a cytoplasmic membrane protein. Expression of all1711 from the P(glnA) promoter in a multicopy plasmid led to production of a presumptive exopolysaccharide by vegetative cells. These results suggest that All1711, which we denote HepP, is involved in transport of glycoside(s), with a specific physiological role in production of Hep.

  20. Protein-bound polysaccharides from Coriolus versicolor attenuate LPS-induced synthesis of pro-inflammatory cytokines and stimulate PBMCs proliferation.

    PubMed

    Jędrzejewski, Tomasz; Pawlikowska, Małgorzata; Piotrowski, Jakub; Kozak, Wiesław

    2016-10-01

    Protein-bound polysaccharides (PBP) isolated from Coriolus versicolor (CV) are classified as biological response modifiers capable of exhibiting various biological activities, such as anti-tumour and immunopotentiating activity. Since we have found in vivo studies that the tested PBP induced prolongation of endotoxin fever in rats, the aim of the present study was to investigate the in vitro effect of the PBP on the production of pro-inflammatory cytokines by the lipolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs). The results showed that the PBP affect the immunomodulating properties of the LPS-treated PBMCs by the enhancement of mitogenic activity and attenuation of the LPS-induced production of interleukin (IL)-1β and IL-6. Moreover, the tested polysaccharides peptides themselves also exhibit immunomodulatory properties manifested in the increased cell proliferation and pro-inflammatory cytokine release from PBMCs. The effect of PBP on the both phenomena was time-dependent and occurred in the U-shaped dose response manner. These findings are significant when considering the use of commercially available PBP from CV extract by cancer patients suffering from immunodeficiency, who may experience microbial infections during therapy.

  1. Protein-bound polysaccharides from Coriolus versicolor attenuate LPS-induced synthesis of pro-inflammatory cytokines and stimulate PBMCs proliferation.

    PubMed

    Jędrzejewski, Tomasz; Pawlikowska, Małgorzata; Piotrowski, Jakub; Kozak, Wiesław

    2016-10-01

    Protein-bound polysaccharides (PBP) isolated from Coriolus versicolor (CV) are classified as biological response modifiers capable of exhibiting various biological activities, such as anti-tumour and immunopotentiating activity. Since we have found in vivo studies that the tested PBP induced prolongation of endotoxin fever in rats, the aim of the present study was to investigate the in vitro effect of the PBP on the production of pro-inflammatory cytokines by the lipolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs). The results showed that the PBP affect the immunomodulating properties of the LPS-treated PBMCs by the enhancement of mitogenic activity and attenuation of the LPS-induced production of interleukin (IL)-1β and IL-6. Moreover, the tested polysaccharides peptides themselves also exhibit immunomodulatory properties manifested in the increased cell proliferation and pro-inflammatory cytokine release from PBMCs. The effect of PBP on the both phenomena was time-dependent and occurred in the U-shaped dose response manner. These findings are significant when considering the use of commercially available PBP from CV extract by cancer patients suffering from immunodeficiency, who may experience microbial infections during therapy. PMID:27594322

  2. Antihyperglycaemic mechanisms of an aceteoside polymer from rose flowers and a polysaccharide-protein complex from abalone mushroom.

    PubMed

    Chen, Rong-Rong; Liu, Zhao-Kun; Liu, Fang; Ng, Tzi Bun

    2015-01-01

    Oral administration of an aceteoside polymer from rose Rosa rugosa (P1-b) and a polysaccharide-peptide complex from abalone mushroom Pleurotus abalonus (LB-1b), both with antioxidant activity, produced antihyperglycaemic effects in alloxan-induced diabetic mice. The expression of insulin, superoxide dismutase and pancreas duodenum homeobox factor-1 essential for pancreatic islet function as estimated by real-time PCR was augmented. The reactive oxygen species-scavenging ability of the rose constituent was notably stronger than the mushroom constituent. Thus, the two biomolecules protected the pancreas from oxidative stress, elevated pancreatic insulin expression and lowered circulating glucose level.

  3. Antihyperglycaemic mechanisms of an aceteoside polymer from rose flowers and a polysaccharide-protein complex from abalone mushroom.

    PubMed

    Chen, Rong-Rong; Liu, Zhao-Kun; Liu, Fang; Ng, Tzi Bun

    2015-01-01

    Oral administration of an aceteoside polymer from rose Rosa rugosa (P1-b) and a polysaccharide-peptide complex from abalone mushroom Pleurotus abalonus (LB-1b), both with antioxidant activity, produced antihyperglycaemic effects in alloxan-induced diabetic mice. The expression of insulin, superoxide dismutase and pancreas duodenum homeobox factor-1 essential for pancreatic islet function as estimated by real-time PCR was augmented. The reactive oxygen species-scavenging ability of the rose constituent was notably stronger than the mushroom constituent. Thus, the two biomolecules protected the pancreas from oxidative stress, elevated pancreatic insulin expression and lowered circulating glucose level. PMID:25200621

  4. Gamma-irradiation influence on the structure and properties of calcium caseinate-whey protein isolate based films. Part 2. Influence of polysaccharide addition and radiation treatment on the structure and functional properties of the films.

    PubMed

    Cieśla, Krystyna; Salmieri, Stephane; Lacroix, Monique

    2006-11-15

    The influence of gamma-irradiation (32 kGy) followed by the addition of polysaccharides (potato starch, soluble potato starch, and sodium alginate) and heating on the properties of the films based on calcium caseinate (CC)-whey proteins isolate (WPI) and the gels formed with CaCl(2) was evaluated. Radiation induced an improvement of the mechanical and barrier properties of all films. The polysaccharides' effect on the irradiated and non-irradiated CC-WPI gels could be predicted as the sum of their separate effects on CC and on WPI, apart from the alginate interaction with the irradiated CC-WPI. The better properties of the films achieved after admixing polysaccharides to the formerly irradiated protein solution correspond to the smaller strength of gels. Properties of the films and gels prepared using the irradiated proteins and alginate differed depending on whether alginate was admixed before or after irradiation. Results were related to the protein structure, interaction with polysaccharides, and the film's microstructure.

  5. The combination of oligo- and polysaccharides and reticulated protein for the control of symptoms in patients with irritable bowel syndrome: Results of a randomised, placebo-controlled, double-blind, parallel group, multicentre clinical trial

    PubMed Central

    Alexea, Octavian; Bacarea, Vlad

    2015-01-01

    Background A medical device containing the film-forming agent reticulated protein and a prebiotic mixture of vegetable oligo- and polysaccharides has been developed, recently receiving European approval as MED class III for the treatment of chronic/functional or recidivant diarrhoea due to different causes including irritable bowel syndrome (IBS). In the present paper, we evaluate a protein preparation containing these components in comparison with placebo in adult patients with diarrhoea-predominant IBS. Methods In a randomised, placebo-controlled, double-blind, parallel group, multicentre clinical trial, patients were randomly assigned to receive the combination of oligo- and polysaccharides and reticulated protein and placebo (four oral tablets/day for 56 days). Demographic, clinical and quality of life characteristics and presence and intensity of abdominal pain and flatulence (seven-point Likert scale) were assessed at three study visits (baseline and at 28 and 56 days). Stool emissions were recorded on the diary card using the seven-point Bristol Stool Scale. Results A total of 128 patients were randomised to receive either tablets containing the combination (n = 63) or placebo (n = 65). Treatment with oligo- and polysaccharides and reticulated protein was safe and well tolerated. A significant improvement in symptoms across the study was observed in patients treated with oligo- and polysaccharides and reticulated protein between visit 2 and visit 3 in abdominal pain (p = 0.0167) and flatulence (p = 0.0373). We also detected a statistically significant increase in the quality of life of patients receiving the active treatment from baseline to visit 3 (p < 0.0001). Conclusions Treatment with oligo- and polysaccharides and reticulated protein is safe, improving IBS symptoms and quality of life of patients with diarrhoea-predominant IBS. PMID:27403313

  6. Ultrasonic disruption of fungal mycelia for efficient recovery of polysaccharide-protein complexes from viscous fermentation broth of a medicinal fungus.

    PubMed

    Cheung, Yi-Ching; Liu, Xing-Xun; Wang, Wing-Qiang; Wu, Jian-Yong

    2015-01-01

    High-intensity ultrasound (US) was applied to facilitate the extraction of intracellular and extracellular polysaccharide-protein complexes (PSPs) from the viscous mycelial fermentation broth of a medicinal fungus Cordyceps sinensis Cs-HK1. The US treatment caused the disruption of fungal mycelia, a dramatic reduction of the apparent broth viscosity, and the release of intracellular products into the liquid medium. The degree of mycelium disruption and the rate of intracellular product release were dependent on US power intensity, treatment period and biomass concentration of broth. The extraction or release kinetics of total water-soluble products and PSPs (yield Y versus time t) under the effect of US was fitted closely to the Elovich model Y=Yo+Y1 lnt and parabolic model Y=Yo+Y1t(½), respectively. Another interesting effect of the US treatment was a notable increase in the antioxidant cytoprotective activity of PSP against H2O2 induced cell death.

  7. Preclinical evaluation of group B Neisseria meningitidis and Escherichia coli K92 capsular polysaccharide-protein conjugate vaccines in juvenile rhesus monkeys.

    PubMed Central

    Devi, S J; Zollinger, W D; Snoy, P J; Tai, J Y; Costantini, P; Norelli, F; Rappuoli, R; Frasch, C E

    1997-01-01

    We reported the first use of group B meningococcal conjugate vaccines in a nonhuman primate model (S. J. N. Devi, C. E. Frasch, W. Zollinger, and P. J. Snoy, p. 427-429, in J. S. Evans, S. E. Yost, M. C. J. Maiden, and I. M. Feavers, ed., Proceedings of the Ninth International Pathogenic Neisseria Conference, 1994). Three different group B Neisseria meningitidis capsular polysaccharide (B PS)-protein conjugate vaccines and an Escherichia coli K92 capsular polysaccharide-tetanus toxoid (K92-TT) conjugate vaccine are here evaluated for safety and relative immunogenicities in juvenile rhesus monkeys with or without adjuvants. Monkeys were immunized intramuscularly with either B PS-cross-reactive material 197 conjugate, B PS-outer membrane vesicle (B-OMV) conjugate, or N-propionylated B PS-outer membrane protein 3 (N-pr. B-OMP3) conjugate vaccine with or without adjuvants at weeks 0, 6, and 14. A control group of monkeys received one injection of the purified B PS alone, and another group received three injections of B PS noncovalently complexed with OMV. Antibody responses as measured by enzyme-linked immunosorbent assay varied among individual monkeys. All vaccines except B PS and the K92-TT conjugate elicited a twofold or greater increase in total B PS antibodies after one immunization. All vaccines, including the K92-TT conjugate, elicited a rise in geometric mean B PS antibody levels of ninefold or more over the preimmune levels following the third immunization. Antibodies elicited by N-pr. B-OMP3 and B-OMV conjugates were directed to the N-propionylated or to the spacer-containing B PS antigens as well as to the native B PS complexed with methylated human serum albumin. None of the vaccines caused discernible safety-related symptoms. PMID:9038314

  8. Conjugation of Polysaccharide 6B from Streptococcus pneumoniae with Pneumococcal Surface Protein A: PspA Conformation and Its Effect on the Immune Response

    PubMed Central

    Perciani, Catia T.; Barazzone, Giovana C.; Goulart, Cibelly; Carvalho, Eneas; Cabrera-Crespo, Joaquin; Gonçalves, Viviane M.; Leite, Luciana C. C.

    2013-01-01

    Despite the substantial beneficial effects of incorporating the 7-valent pneumococcal conjugate vaccine (PCV7) into immunization programs, serotype replacement has been observed after its widespread use. As there are many serotypes currently documented, the use of a conjugate vaccine relying on protective pneumococcal proteins as active carriers is a promising alternative to expand PCV coverage. In this study, capsular polysaccharide serotype 6B (PS6B) and recombinant pneumococcal surface protein A (rPspA), a well-known protective antigen from Streptococcus pneumoniae, were covalently attached by two conjugation methods. The conjugation methodology developed by our laboratory, employing 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) as an activating agent through carboxamide formation, was compared with reductive amination, a classical methodology. DMT-MM-mediated conjugation was shown to be more efficient in coupling PS6B to rPspA clade 1 (rPspA1): 55.0% of PS6B was in the conjugate fraction, whereas 24% was observed in the conjugate fraction with reductive amination. The influence of the conjugation process on the rPspA1 structure was assessed by circular dichroism. According to our results, both conjugation processes reduced the alpha-helical content of rPspA; reduction was more pronounced when the reaction between the polysaccharide capsule and rPspA1 was promoted between the carboxyl groups than the amine groups (46% and 13%, respectively). Regarding the immune response, both conjugates induced functional anti-rPspA1 and anti-PS6B antibodies. These results suggest that the secondary structure of PspA1, as well as its reactive groups (amine or carboxyl) involved in the linkage to PS6B, may not play an important role in eliciting a protective immune response to the antigens. PMID:23554468

  9. H-NS Nucleoid Protein Controls Virulence Features of Klebsiella pneumoniae by Regulating the Expression of Type 3 Pili and the Capsule Polysaccharide

    PubMed Central

    Ares, Miguel A.; Fernández-Vázquez, José L.; Rosales-Reyes, Roberto; Jarillo-Quijada, Ma. Dolores; von Bargen, Kristine; Torres, Javier; González-y-Merchand, Jorge A.; Alcántar-Curiel, María D.; De la Cruz, Miguel A.

    2016-01-01

    Klebsiella pneumoniae is an opportunistic pathogen causing nosocomial infections. Main virulence determinants of K. pneumoniae are pili, capsular polysaccharide, lipopolysaccharide, and siderophores. The histone-like nucleoid-structuring protein (H-NS) is a pleiotropic regulator found in several gram-negative pathogens. It has functions both as an architectural component of the nucleoid and as a global regulator of gene expression. We generated a Δhns mutant and evaluated the role of the H-NS nucleoid protein on the virulence features of K. pneumoniae. A Δhns mutant down-regulated the mrkA pilin gene and biofilm formation was affected. In contrast, capsule expression was derepressed in the absence of H-NS conferring a hypermucoviscous phenotype. Moreover, H-NS deficiency affected the K. pneumoniae adherence to epithelial cells such as A549 and HeLa cells. In infection experiments using RAW264.7 and THP-1 differentiated macrophages, the Δhns mutant was less phagocytized than the wild-type strain. This phenotype was likely due to the low adherence to these phagocytic cells. Taken together, our data indicate that H-NS nucleoid protein is a crucial regulator of both T3P and CPS of K. pneumoniae. PMID:26904512

  10. Serogroup quantitation of multivalent polysaccharide and polysaccharide-conjugate meningococcal vaccines from China.

    PubMed

    Cook, Matthew C; Gibeault, Sabrina; Filippenko, Vasilisa; Ye, Qiang; Wang, Junzhi; Kunkel, Jeremy P

    2013-07-01

    The active components of most meningococcal vaccines are four antigenic serogroup capsular polysaccharides (A, C, Y, W135). The vaccines, monovalent or multivalent mixtures of either free polysaccharides or polysaccharides conjugated to antigenic carrier proteins, may be in liquid or lyophilised formulations, with or without excipients. Acid hydrolysis and chromatographic methods for serogroup quantitation, which were previously optimised and qualified using polysaccharide-based standards and a narrow range of real vaccines, are here challenged with multiple lots of a broad assortment of additional multivalent polysaccharide-based meningococcal vaccine products. Centrifugal filtration successfully removed all interfering lactose excipient without loss of polysaccharides to allow for the determination of Y and W135 serogroups. Replicate operations by three different analysts indicated high method reproducibility. Results indicated some lot-to-lot and product-to-product variations. However, all vaccines were within general specifications for each serogroup polysaccharide, with the exception of all lots of one polysaccharide vaccine - which by these methods were found to be deficient in the serogroup A component only. These robust techniques are very useful for the evaluation of antigen content and consistency of manufacture. The deformulation, hydrolysis and chromatographic methods may be adaptable for the evaluation of other types of polysaccharide-based vaccines.

  11. Protein-bound Polysaccharide-K Inhibits Hedgehog Signaling Through Down-regulation of MAML3 and RBPJ Transcription Under Hypoxia, Suppressing the Malignant Phenotype in Pancreatic Cancer.

    PubMed

    Yamasaki, Akio; Onishi, Hideya; Imaizumi, Akira; Kawamoto, Makoto; Fujimura, Akiko; Oyama, Yasuhiro; Katano, Mitsuo

    2016-08-01

    Hedgehog signaling is activated in pancreatic cancer and could be a therapeutic target. We previously demonstrated that recombination signal binding protein for immunoglobulin-kappa-J region (RBPJ) and mastermind-like 3 (MAML3) contribute to the hypoxia-induced up-regulation of Smoothened (SMO) transcription. We have also shown that protein-bound polysaccharide-K (PSK) could be effective for refractory pancreatic cancer that down-regulates SMO transcription under hypoxia. In this study, we evaluated whether the anticancer mechanism of PSK involves inhibiting RBPJ and MAML3 expression under hypoxia. PSK reduced SMO, MAML3 and RBPJ expression in pancreatic cancer cells under hypoxia. PSK also blocked RBPJ-induced invasiveness under hypoxia by inhibiting matrix metalloproteinase expression. Lastly, we showed that PSK attenuated RBPJ-induced proliferation both in vitro and in vivo. These results suggest that PSK suppresses Hedgehog signaling through down-regulation of MAML3 and RBPJ transcription under hypoxia, inhibiting the induction of a malignant phenotype in pancreatic cancer. Our results may lead to development of new treatments for refractory pancreatic cancer using PSK as a Hedgehog inhibitor. PMID:27466498

  12. Preventive Effect of Novel Bacterial Polysaccharide and Animal Splenic Protein as Natural Adjuvants on Animal Model of Asthma

    PubMed Central

    Mirsadraee, Majid; Mohaghegh Hazrati, Saleh; Khakzad, Mohammad Reza; Ghafarzadegan, Kamran; Boskabady, Mohhamad Hosein

    2012-01-01

    Objective(s) Two new adjuvants from natural animal lipids (G2) and bacterial polysaccharide extracts (PC) were previously prepared by our group and showed a reduction in tracheal responsiveness. The aim of this study was to evaluate the preventive effect of recently introduced natural products (G2 and PC) on the development of asthma. Materials and Methods Asthma was induced using a standard method in four groups of BALB/c mice. A non-sensitized control group was also included in order to be compared with treated groups. Three groups were premedicated with novel agents named G2, PC, and a combination of these two for 20 days before starting the induction of asthma. Bronchoalveolar lavage fluid (BALF) was collected and analyzed for inflammatory cells. Interferon-γ, and IL-4 and the histopathological of both lungs were also evaluated. Results In all pretreated groups, the inflammatory cells infiltration especially eosinophils and smooth muscle hyperplasia decreased significantly. BALF cytology also showed significant decrease in eosinophil count in all pretreated groups. There was a significant increase in the BALF and serum INF-γ in all pretreated groups but the combination of G2/PC was more effective. BALF IL-4 decreased significantly in the group pretreated with a combination of G2 and G2/PC (4.11±0.86 and 4.02±0.52 pg/ml in G2 and G2/PC, respectively). Serum IL-4 in the PC group was significantly higher than the sensitized control. Conclusion G2 and PC may effectively prevent asthma development by activation of the type 1 T helper system. PMID:23493252

  13. NMR structure of a lytic polysaccharide monooxygenase provides insight into copper binding, protein dynamics, and substrate interactions

    PubMed Central

    Aachmann, Finn L.; Sørlie, Morten; Skjåk-Bræk, Gudmund; Eijsink, Vincent G. H.; Vaaje-Kolstad, Gustav

    2012-01-01

    Lytic polysaccharide monooxygenases currently classified as carbohydrate binding module family 33 (CBM33) and glycoside hydrolase family 61 (GH61) are likely to play important roles in future biorefining. However, the molecular basis of their unprecedented catalytic activity remains largely unknown. We have used NMR techniques and isothermal titration calorimetry to address structural and functional aspects of CBP21, a chitin-active CBM33. NMR structural and relaxation studies showed that CBP21 is a compact and rigid molecule, and the only exception is the catalytic metal binding site. NMR data further showed that His28 and His114 in the catalytic center bind a variety of divalent metal ions with a clear preference for Cu2+ (Kd = 55 nM; from isothermal titration calorimetry) and higher preference for Cu1+ (Kd ∼ 1 nM; from the experimentally determined redox potential for CBP21-Cu2+ of 275 mV using a thermodynamic cycle). Strong binding of Cu1+ was also reflected in a reduction in the pKa values of the histidines by 3.6 and 2.2 pH units, respectively. Cyanide, a mimic of molecular oxygen, was found to bind to the metal ion only. These data support a model where copper is reduced on the enzyme by an externally provided electron and followed by oxygen binding and activation by internal electron transfer. Interactions of CBP21 with a crystalline substrate were mapped in a 2H/1H exchange experiment, which showed that substrate binding involves an extended planar binding surface, including the metal binding site. Such a planar catalytic surface seems well-suited to interact with crystalline substrates. PMID:23112164

  14. TtsI, a key regulator of Rhizobium species NGR234 is required for type III-dependent protein secretion and synthesis of rhamnose-rich polysaccharides.

    PubMed

    Marie, Corinne; Deakin, William J; Ojanen-Reuhs, Tuula; Diallo, Ericka; Reuhs, Brad; Broughton, William J; Perret, Xavier

    2004-09-01

    Formation of nitrogen-fixing nodules on legume roots by Rhizobium sp. NGR234 requires an array of bacterial factors, including nodulation outer proteins (Nops) secreted through a type III secretion system (TTSS). Secretion of Nops is abolished upon inactivation of ttsI (formerly y4xI), a protein with characteristics of two-component response regulators that was predicted to activate transcription of TTSS-related genes. During the symbiotic interaction, the phenotype of NGR omega ttsI differs from that of a mutant with a nonfunctional secretion machine, however. This indicated that TtsI regulates the synthesis of other symbiotic factors as well. Conserved sequences, called tts boxes, proposed to act as binding sites for TtsI, were identified not only within the TTSS cluster but also in the promoter regions of i) genes predicted to encode homologs of virulence factors secreted by pathogenic bacteria, ii) loci involved in the synthesis of a rhamnose-rich component (rhamnan) of the lipopolysaccharides (LPS), and iii) open reading frames that play roles in plasmid partitioning. Transcription studies showed that TtsI and tts boxes are required for the activation of TTSS-related genes and those involved in rhamnose synthesis. Furthermore, extraction of polysaccharides revealed that inactivation of ttsI abolishes the synthesis of the rhamnan component of the LPS. The phenotypes of mutants impaired in TTSS-dependent protein secretion, rhamnan synthesis, or in both functions were compared to assess the roles of some of the TtsI-controlled factors during symbiosis.

  15. Bactericidal antibody responses of juvenile rhesus monkeys immunized with group B Neisseria meningitidis capsular polysaccharide-protein conjugate vaccines.

    PubMed

    Zollinger, W D; Moran, E E; Devi, S J; Frasch, C E

    1997-03-01

    Reports on the bactericidal activities of antibodies to group B Neisseria meningitidis capsular polysaccharide (B PS) are conflicting. Using three different complement sources, we analyzed the bactericidal activities of sera of juvenile rhesus monkeys immunized with five conjugate vaccines of B PS synthesized by different schemes, an Escherichia coli K92 conjugate, and a noncovalent complex of B PS with group B meningococcal outer membrane vesicles (B+OMV) (S. J. N. Devi, W. D. Zollinger, P. J. Snoy, J. Y. Tai, P. Costantini, F. Norelli, R. Rappuoli, and C. E. Frasch, Infect. Immun. 65:1045-1052, 1997). With rabbit complement, nearly all preimmune sera showed relatively high bactericidal titers, and all vaccines, except the K92 conjugate, induced a fourfold or greater increase in bactericidal titers in most of the monkeys vaccinated. In contrast, with human complement, most prevaccination sera showed no bactericidal activity and in most of the vaccine groups, little or no increase in bactericidal titer was observed. However, the covalent conjugation of P BS and OMV (B-OMV) administered with and without the Ribi adjuvant induced relatively high bactericidal titers which persisted up to 30 weeks. An analysis of the specificities of bactericidal antibodies revealed that absorption with E. coli K1 cells did not change the bactericidal titer with human complement but reduced the titers observed with the rabbit and monkey complements. A significant increase in anti-lipopolysaccharide (LPS) antibodies was elicited by the B-OMV conjugates, and nearly all of the bactericidal activity with human complement could be inhibited with the purified group B meningococcal L3,7,8 LPS. B-OMV covalently coupled via adipic acid dihydrazide elicited significantly elevated levels (P < or = 0.02) of anti-OMV antibodies compared to those of the noncovalently complexed B+OMV. An initial small-scale evaluation of B PS conjugates in adult human males appears feasible, with careful monitoring

  16. Antifungal and anticancer effects of a polysaccharide-protein complex from the gut bacterium Raoultella ornithinolytica isolated from the earthworm Dendrobaena veneta.

    PubMed

    Fiołka, Marta J; Lewtak, Kinga; Rzymowska, Jolanta; Grzywnowicz, Krzysztof; Hułas-Stasiak, Monika; Sofińska-Chmiel, Weronika; Skrzypiec, Krzysztof

    2013-06-21

    The polysaccharide-protein complex (PPC) isolated from metabolites of gut bacteria Raoultella ornithinolytica from Dendrobaena veneta earthworms exhibits activity against Candida albicans, in breast ductal carcinoma (line T47D) and in the endometrioid ovarian cancer line (TOV-112D) in vitro. The action against C. albicans was analyzed using light, SEM, TEM, and AFM microscopes. The changes observed indicated two directions of the action of the complex, that is, disturbance of metabolic activity and cell wall damage. The PPC is an adhesion-promoting complex inducing death of C. albicans cells by necrosis. Owing to its significant effect on C. albicans, the complex is a promising source of antifungal compounds. The PPC showed a minimal cytotoxic effect against human skin fibroblasts; however, the cytotoxicity against the T47D line was determined at 20% and 15% against the TOV-112D line. The action of the PPC against the T47D line exerted a cytopathic effect, whereas in the TOV-112D line, it caused a reduction in the cell number. The PPC induced death of tumor cells by apoptosis and necrosis. In view of the negligible cytotoxicity on fibroblasts, the PPC will be subjected to chemical modifications to increase its antitumor activity for prospective medical applications.

  17. A Synthetic Disaccharide Analogue from Neisseria meningitidis A Capsular Polysaccharide Stimulates Immune Cell Responses and Induces Immunoglobulin G (IgG) Production in Mice When Protein-Conjugated.

    PubMed

    Fallarini, Silvia; Buzzi, Benedetta; Giovarruscio, Sara; Polito, Laura; Brogioni, Giulia; Tontini, Marta; Berti, Francesco; Adamo, Roberto; Lay, Luigi; Lombardi, Grazia

    2015-10-01

    Some new phosphonoester-linked oligomers, stabilized analogues of the corresponding phosphate-bridged oligomers of Neisseria meningitidis A (MenA) capsular polysaccharide (CPS), were conjugated to human serum albumin (HSA), as a protein carrier model, and studied for immunological activities. We determined (i) in vitro, their biocompatibility (CAM test) and activity in inducing both T cell proliferation (CFSE method) and IL-2 release (ELISA), and (ii) in vivo, their ability to stimulate specific IgG antibody production (ELISA). All HSA-conjugated compounds induce T cell proliferation (40% of proliferation at 10(2) μM), whereas only the phosphonodisaccharide was effective (28% of proliferation at 10(2) μM) among the unconjugated forms. IL-2 release confirmed these results. In addition, the HSA-conjugated showed in vivo the capacity of eliciting the production of specific IgG antibodies. In conclusion, we obtained novel biocompatible, water-stable, and immunoactive MenA CPS analogues. A short disaccharide fragment showed the unusual behavior of triggering T cell proliferation in vitro. PMID:27623315

  18. Protective effect of a protein-bound polysaccharide, PSK, on CLP-induced sepsis in mice transplanted orthotopically with colon tumor.

    PubMed

    Ohmura, Yoshio; Matsunaga, Kenichi; Suzuki, Tatsuo

    2006-01-01

    We investigated the effects of a protein-bound polysaccharide, PSK, on the resistance of tumor-bearing mice against sepsis induced by cecal ligation and puncture (CLP). (a) In BALB/c mice that had received intracecal transplantation of colon 26 (C26) tumor, CLP with a 21-gauge needle significantly shortened the survival time, compared with that of non-tumor-bearing mice. Oral administration of PSK to such mice resulted in a significant prolongation of the survival time and increase of the survival rates. The effects were dependent on the timing of PSK administration and the dose. (b) CLP significantly increased the IL-10 level in serum, the IL-10 gene expression by spleen cells, the number of IL-10-producing CD4-positive T cells, and the productivity of IL-10 by spleen of tumor-bearing mice compared with that of non-tumor-bearing mice. PSK administration to such mice suppressed the increase. Further, PSK prevented the reduction of gene expression of IFN-gamma and the number of IFN-gamma-producing CD4-positive T cells and IFN-gamma productivity by spleen cells of tumor-bearing CLP-treated mice. (c) Treatment with anti-IFN-gamma monoclonal antibody before CLP significantly reduced the effects of PSK. These findings suggest that the protective effect of PSK on the CLP-induced sepsis in mice transplanted orthotopically with C26 tumor is possibly mediated by suppression of IL-10 and promotion of IFN-gamma. PMID:16369184

  19. Whey protein/polysaccharide-stabilized oil powders for topical application-release and transdermal delivery of salicylic acid from oil powders compared to redispersed powders.

    PubMed

    Kotzé, Magdalena; Otto, Anja; Jordaan, Anine; du Plessis, Jeanetta

    2015-08-01

    Oil-in-water (o/w) emulsions are commonly converted into solid-like powders in order to improve their physical and chemical stabilities. The aim of this study was to investigate whether whey protein/polysaccharide-stabilized o/w emulsions could be converted into stable oil powders by means of freeze-drying. Moreover, during this study, the effects of pH and polymer type on release and trans(dermal) delivery of salicylic acid, a model drug, from these oil powders were investigated and compared to those of the respective template emulsions and redispersed oil powders. Physical characterization of the various formulations was performed, such as droplet size analysis and oil leakage, and relationships drawn with regards to release and trans(dermal) delivery. The experimental outcomes revealed that the oil powders could be redispersed in water without changing the release characteristics of salicylic acid. pH and polymer type affected the release of salicylic acid from the oil powders, template emulsions, and redispersed powders similarly. Contrary, the transdermal delivery from the oil powders and from their respective redispersed oil powders was differently affected by pH and polymer type. It was hypothesized that the release had been influenced by the electrostatic interactions between salicylic acid and emulsifiers, whereas the transdermal performance could have been determined by the particle or aggregate sizes of the formulations.

  20. Method for producing capsular polysaccharides

    NASA Technical Reports Server (NTRS)

    Kern, Roger G. (Inventor); Petersen, Gene R. (Inventor); Richards, Gil F. (Inventor)

    1994-01-01

    Structurally altered capsular polysaccharides are produced by mutant bacteria. These polysaccharides are isolated by selecting a wild type bacterial strain and a phage producing degradative enzymes that have substrate specificity for the capsular polysaccharides produced by the wild type bacteria. Phage-resistant mutants producing capsular polysaccharides are selected and the structurally altered capsular polysaccharide is isolated therefrom.

  1. Polysaccharides and Proteins Added to Flowing Drinking Water at Microgram-per-Liter Levels Promote the Formation of Biofilms Predominated by Bacteroidetes and Proteobacteria

    PubMed Central

    Sack, Eveline L. W.; van der Kooij, Dick

    2014-01-01

    Biopolymers are important substrates for heterotrophic bacteria in (ultra)oligotrophic freshwater environments, but information about their utilization at microgram-per-liter levels by attached freshwater bacteria is lacking. This study aimed at characterizing biopolymer utilization in drinking-water-related biofilms by exposing such biofilms to added carbohydrates or proteins at 10 μg C liter−1 in flowing tap water for up to 3 months. Individually added amylopectin was not utilized by the biofilms, whereas laminarin, gelatin, and caseinate were. Amylopectin was utilized during steady-state biofilm growth with simultaneously added maltose but not with simultaneously added acetate. Biofilm formation rates (BFR) at 10 μg C liter−1 per substrate were ranked as follows, from lowest to highest: blank or amylopectin (≤6 pg ATP cm−2 day−1), gelatin or caseinate, laminarin, maltose, acetate alone or acetate plus amylopectin, and maltose plus amylopectin (980 pg ATP cm−2 day−1). Terminal restriction fragment length polymorphism (T-RFLP) and 16S rRNA gene sequence analyses revealed that the predominant maltose-utilizing bacteria also dominated subsequent amylopectin utilization, indicating catabolic repression and (extracellular) enzyme induction. The accelerated BFR with amylopectin in the presence of maltose probably resulted from efficient amylopectin binding to and hydrolysis by inductive enzymes attached to the bacterial cells. Cytophagia, Flavobacteriia, Gammaproteobacteria, and Sphingobacteriia grew during polysaccharide addition, and Alpha-, Beta-, and Gammaproteobacteria, Cytophagia, Flavobacteriia, and Sphingobacteriia grew during protein addition. The succession of bacterial populations in the biofilms coincided with the decrease in the specific growth rate during biofilm formation. Biopolymers can clearly promote biofilm formation at microgram-per-liter levels in drinking water distribution systems and, depending on their concentrations, might

  2. Polysaccharides and proteins added to flowing drinking water at microgram-per-liter levels promote the formation of biofilms predominated by bacteroidetes and proteobacteria.

    PubMed

    Sack, Eveline L W; van der Wielen, Paul W J J; van der Kooij, Dick

    2014-04-01

    Biopolymers are important substrates for heterotrophic bacteria in (ultra)oligotrophic freshwater environments, but information about their utilization at microgram-per-liter levels by attached freshwater bacteria is lacking. This study aimed at characterizing biopolymer utilization in drinking-water-related biofilms by exposing such biofilms to added carbohydrates or proteins at 10 μg C liter(-1) in flowing tap water for up to 3 months. Individually added amylopectin was not utilized by the biofilms, whereas laminarin, gelatin, and caseinate were. Amylopectin was utilized during steady-state biofilm growth with simultaneously added maltose but not with simultaneously added acetate. Biofilm formation rates (BFR) at 10 μg C liter(-1) per substrate were ranked as follows, from lowest to highest: blank or amylopectin (≤6 pg ATP cm(-2) day(-1)), gelatin or caseinate, laminarin, maltose, acetate alone or acetate plus amylopectin, and maltose plus amylopectin (980 pg ATP cm(-2) day(-1)). Terminal restriction fragment length polymorphism (T-RFLP) and 16S rRNA gene sequence analyses revealed that the predominant maltose-utilizing bacteria also dominated subsequent amylopectin utilization, indicating catabolic repression and (extracellular) enzyme induction. The accelerated BFR with amylopectin in the presence of maltose probably resulted from efficient amylopectin binding to and hydrolysis by inductive enzymes attached to the bacterial cells. Cytophagia, Flavobacteriia, Gammaproteobacteria, and Sphingobacteriia grew during polysaccharide addition, and Alpha-, Beta-, and Gammaproteobacteria, Cytophagia, Flavobacteriia, and Sphingobacteriia grew during protein addition. The succession of bacterial populations in the biofilms coincided with the decrease in the specific growth rate during biofilm formation. Biopolymers can clearly promote biofilm formation at microgram-per-liter levels in drinking water distribution systems and, depending on their concentrations, might

  3. The distribution of particles characterized by size and free mobility within polydisperse populations of protein-polysaccharide conjugates, determined from two-dimensional agarose electropherograms.

    PubMed

    Tietz, D; Aldroubi, A; Schneerson, R; Unser, M; Chrambach, A

    1991-01-01

    New approaches for the characterization of polydisperse particle populations are presented*. The investigated samples contain virus-sized protein-polysaccharide conjugates which had previously been prepared as immunogens against bacterial meningitis (Hib). The analysis is based on two-dimensional agarose electrophoresis (Serwer-type). This method, like the one of O'Farrell, achieves a separation according to size and charge. It relies on a different principle, however, and is applicable to nondenatured particles which are 100 to more than 1000 times larger in mass than regular uncrosslinked proteins. Data from stained gel patterns are evaluated by the computer program ELPHOFIT, which makes it possible to standardize the gel and to construct a nomogram which defines every position on the gel in terms of particle size and free mobility (related to surface net charge density). The output of ELPHOFIT, consisting of nomogram parameters, is transferred to the image processing program GELFIT. This software is used to evaluate the computer images obtained by digitizing the stained gel patterns: (i) The nomogram is electronically superimposed on the computer image. (ii) The gel pattern is transformed from a curvilinear to a rectangular coordinate system of particle size and free mobility. The center of gravity as well as density maxima are given in coordinates of particle size and free mobility. Ranges of grey levels can be accentuated by adding 16 pseudocolors. (iii) Using surface-stripping techniques, GELFIT provides an estimate for the number of major subpopulations within each preparation. (iv) Numerical values for the distribution of particle size and free mobility are determined. Using program IMAGE, the quantitative physical assessment of a given conjugate preparation is presented in the form of a computer-generated three-dimensional plot, the shape of which serves to identify and characterize the preparation visually. The data analysis based on digitized two

  4. Chemical Modification of Polysaccharides

    PubMed Central

    Cumpstey, Ian

    2013-01-01

    This review covers methods for modifying the structures of polysaccharides. The introduction of hydrophobic, acidic, basic, or other functionality into polysaccharide structures can alter the properties of materials based on these substances. The development of chemical methods to achieve this aim is an ongoing area of research that is expected to become more important as the emphasis on using renewable starting materials and sustainable processes increases in the future. The methods covered in this review include ester and ether formation using saccharide oxygen nucleophiles, including enzymatic reactions and aspects of regioselectivity; the introduction of heteroatomic nucleophiles into polysaccharide chains; the oxidation of polysaccharides, including oxidative glycol cleavage, chemical oxidation of primary alcohols to carboxylic acids, and enzymatic oxidation of primary alcohols to aldehydes; reactions of uronic-acid-based polysaccharides; nucleophilic reactions of the amines of chitosan; and the formation of unsaturated polysaccharide derivatives. PMID:24151557

  5. CHARMM additive all-atom force field for carbohydrate derivatives and its utility in polysaccharide and carbohydrate-protein modeling

    PubMed Central

    Guvench, Olgun; Mallajosyula, Sairam S.; Raman, E. Prabhu; Hatcher, Elizabeth; Vanommeslaeghe, Kenno; Foster, Theresa J.; Jamison, Francis W.; MacKerell, Alexander D.

    2011-01-01

    Monosaccharide derivatives such as xylose, fucose, N-acetylglucosamine (GlcNAc), N-acetylgalactosamine (GlaNAc), glucuronic acid, iduronic acid, and N-acetylneuraminic acid (Neu5Ac) are important components of eukaryotic glycans. The present work details development of force-field parameters for these monosaccharides and their covalent connections to proteins via O-linkages to serine or threonine sidechains and via N-linkages to asparagine sidechains. The force field development protocol was designed to explicitly yield parameters that are compatible with the existing CHARMM additive force field for proteins, nucleic acids, lipids, carbohydrates, and small molecules. Therefore, when combined with previously developed parameters for pyranose and furanose monosaccharides, for glycosidic linkages between monosaccharides, and for proteins, the present set of parameters enables the molecular simulation of a wide variety of biologically-important molecules such as complex carbohydrates and glycoproteins. Parametrization included fitting to quantum mechanical (QM) geometries and conformational energies of model compounds, as well as to QM pair interaction energies and distances of model compounds with water. Parameters were validated in the context of crystals of relevant monosaccharides, as well NMR and/or x-ray crystallographic data on larger systems including oligomeric hyaluronan, sialyl Lewis X, O- and N-linked glycopeptides, and a lectin:sucrose complex. As the validated parameters are an extension of the CHARMM all-atom additive biomolecular force field, they further broaden the types of heterogeneous systems accessible with a consistently-developed force-field model. PMID:22125473

  6. Comparative immunogenicity of conjugates composed of the Staphylococcus aureus type 8 capsular polysaccharide bound to carrier proteins by adipic acid dihydrazide or N-succinimidyl-3-(2-pyridyldithio)propionate.

    PubMed Central

    Fattom, A; Shiloach, J; Bryla, D; Fitzgerald, D; Pastan, I; Karakawa, W W; Robbins, J B; Schneerson, R

    1992-01-01

    Staphylococcus aureus type 8 capsular polysaccharide (CP) was conjugated either to diphtheria toxoid or to Pseudomonas aeruginosa recombinant exoprotein A by using adipic acid dihydrazide (ADH) or N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) as the joining reagent. The polysaccharide/protein ratios of these two pairs of conjugates were similar. The two synthetic schemes bound the linker to the carboxyls of the type 8 CP by carbodiimide-mediated condensation. ADH was bound to the carboxyls of the protein, whereas SPDP reacted with the amino groups of the protein. Intermolecular linking of the carrier protein, caused by the carbodiimide during the conjugation reaction with the type 8 CP derivative, probably accounts for the larger size of the conjugates formed with ADH compared with those formed with SPDP. Both conjugates synthesized with ADH elicited higher levels of CP antibodies, especially after the first immunization, than did those prepared with SPDP. Similar levels of exoprotein A antibodies were elicited by both conjugates. Higher levels of diphtheria toxoid antibodies were elicited by the conjugate prepared with SPDP than by the one prepared with ADH. The basis for the differences in the immunogenicities of these two pairs of S. aureus type 8 CP conjugates is discussed. PMID:1730492

  7. Comparative immunogenicity of conjugates composed of the Staphylococcus aureus type 8 capsular polysaccharide bound to carrier proteins by adipic acid dihydrazide or N-succinimidyl-3-(2-pyridyldithio)propionate.

    PubMed

    Fattom, A; Shiloach, J; Bryla, D; Fitzgerald, D; Pastan, I; Karakawa, W W; Robbins, J B; Schneerson, R

    1992-02-01

    Staphylococcus aureus type 8 capsular polysaccharide (CP) was conjugated either to diphtheria toxoid or to Pseudomonas aeruginosa recombinant exoprotein A by using adipic acid dihydrazide (ADH) or N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) as the joining reagent. The polysaccharide/protein ratios of these two pairs of conjugates were similar. The two synthetic schemes bound the linker to the carboxyls of the type 8 CP by carbodiimide-mediated condensation. ADH was bound to the carboxyls of the protein, whereas SPDP reacted with the amino groups of the protein. Intermolecular linking of the carrier protein, caused by the carbodiimide during the conjugation reaction with the type 8 CP derivative, probably accounts for the larger size of the conjugates formed with ADH compared with those formed with SPDP. Both conjugates synthesized with ADH elicited higher levels of CP antibodies, especially after the first immunization, than did those prepared with SPDP. Similar levels of exoprotein A antibodies were elicited by both conjugates. Higher levels of diphtheria toxoid antibodies were elicited by the conjugate prepared with SPDP than by the one prepared with ADH. The basis for the differences in the immunogenicities of these two pairs of S. aureus type 8 CP conjugates is discussed. PMID:1730492

  8. Preparation and characterization of a nontoxic polysaccharide-protein conjugate that induces active immunity and passively protective antibody against Pseudomonas aeruginosa immunotype 1 in mice.

    PubMed Central

    Tsay, G C; Collins, M S

    1984-01-01

    Acid treatment of Pseudomonas aeruginosa immunotype 1 lipopolysaccharide generated a low-molecular-weight polysaccharide fraction that was detectable in agar gel immunodiffusion but did not induce antibodies or resistance to infection in mice. The polysaccharide was treated with periodate to generate additional aldehyde groups. Oxidized polysaccharide was covalently coupled by reductive amination to 1,4-diaminobutyl-derivatized bovine serum albumin. Physical properties of the conjugate were characterized by gel filtration and high-pressure liquid chromatography. The gelation activity of the conjugate in the Limulus amoebocyte lysate assay was 4,000-fold less than native lipopolysaccharide by weight. Mice immunized with the conjugate resisted challenge with P. aeruginosa immunotype 1 that killed 90% of mice immunized with saline. Immunization with the conjugate vaccine induced humoral immunoglobulin G that passively protected normal and burned mice. These results indicate that conjugation of nonimmunogenic polysaccharide antigen of P. aeruginosa restores immunogenicity similar to that of native lipopolysaccharide without restoring endotoxicity inherent in lipopolysaccharide. PMID:6429046

  9. Tumor necrosis factor-alpha and acute phase proteins in early pregnant ewes after challenge with peptidoglycan-polysaccharide (PG-PS).

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial infection shortly after breeding interferes with establishment and progression of pregnancy. This is particularly relevant in cases of Gram-positive bacterial infection wherein it can be demonstrated that the injection of peptidoglycan-polysaccharide (PG-PS), a component of Gram-positive ...

  10. Escherichia hermannii (ATCC 33651) polysaccharide-protein conjugates: comparison of two conjugation methods for the induction of humoral responses in mice.

    PubMed

    Jacques, I; Dubray, G

    1991-08-01

    Escherichia hermannii (ATCC 33651) LPS O-polysaccharide was covalently linked to a carrier (bovine serum albumin) to form conjugates either directly or with a spacer arm (adipic acid dihydrazide). The immunogenicity of both conjugates at three different doses was tested in mice. Antibodies to the conjugate were produced and were shown to react with free lipopolysaccharide. The directly-coupled conjugate was found to be more immunogenic than the indirect one (i.e. lower dose necessary for a similar response). The antibody response elicited by the directly coupled conjugate (1 microgram/animal) began at 21 days and was sustained for at least 4 months. The mouse model described here may be applicable to the testing of other conjugates composed of bacterial cell wall polysaccharides and LPS O-chains. PMID:1771969

  11. Epitope specificity of rabbit immunoglobulin G (IgG) elicited by pneumococcal type 23F synthetic oligosaccharide- and native polysaccharide-protein conjugate vaccines: comparison with human anti-polysaccharide 23F IgG.

    PubMed Central

    Alonso de Velasco, E; Verheul, A F; van Steijn, A M; Dekker, H A; Feldman, R G; Fernández, I M; Kamerling, J P; Vliegenthart, J F; Verhoef, J; Snippe, H

    1994-01-01

    Streptococcus pneumoniae type 23F capsular polysaccharide (PS23F) consitss of a repeating glycerol-phosphorylated branched tetrasaccharide. The immunogenicities of the following related antigens were investigated: (i) a synthetic trisaccharide comprising the backbone of one repeating unit, (ii) a synthetic tetrasaccharide comprising the complete repeating unit, and (iii) native PS23F (all three conjugated to keyhole limpet hemocyanin [KLH]) and (iv) formalin-killed S. pneumoniae 23F. All antigens except the trisaccharide-KLH conjugate induced relatively high anti-PS23F antibody levels in rabbits. The epitope specificity of such antibodies was then studied by means of an inhibition immunoassay. The alpha(1-->2)-linked L-rhamnose branch was shown to be immunodominant for immunoglobulin G (IgG) induced by tetrasaccharide-KLH, PS23F-KLH, and killed S. pneumoniae 23F: in most sera L-rhamnose totally inhibited the binding of IgG to PS23F. Thus, there appears to be no major difference in epitope specificity between IgG induced by tetrasaccharide-KLH and that induced by antigens containing the polymeric form of PS23F. Human anti-PS23F IgG (either vaccine induced or naturally acquired) had a different epitope specificity: none of the inhibitors used, including L-rhamnose and tetrasaccharide-KLH, exhibited substantial inhibition. These observations suggest that the epitope recognized by human IgG on PS23F is larger than the epitope recognized by rabbit IgG. Both human and rabbit antisera efficiently opsonized type 23F pneumococci, as measured in a phagocytosis assay using human polymorphonuclear leukocytes. PMID:7509318

  12. Bacterial Extracellular Polysaccharides in Biofilm Formation and Function.

    PubMed

    Limoli, Dominique H; Jones, Christopher J; Wozniak, Daniel J

    2015-06-01

    Microbes produce a biofilm matrix consisting of proteins, extracellular DNA, and polysaccharides that is integral in the formation of bacterial communities. Historical studies of polysaccharides revealed that their overproduction often alters the colony morphology and can be diagnostic in identifying certain species. The polysaccharide component of the matrix can provide many diverse benefits to the cells in the biofilm, including adhesion, protection, and structure. Aggregative polysaccharides act as molecular glue, allowing the bacterial cells to adhere to each other as well as surfaces. Adhesion facilitates the colonization of both biotic and abiotic surfaces by allowing the bacteria to resist physical stresses imposed by fluid movement that could separate the cells from a nutrient source. Polysaccharides can also provide protection from a wide range of stresses, such as desiccation, immune effectors, and predators such as phagocytic cells and amoebae. Finally, polysaccharides can provide structure to biofilms, allowing stratification of the bacterial community and establishing gradients of nutrients and waste products. This can be advantageous for the bacteria by establishing a heterogeneous population that is prepared to endure stresses created by the rapidly changing environments that many bacteria encounter. The diverse range of polysaccharide structures, properties, and roles highlight the importance of this matrix constituent to the successful adaptation of bacteria to nearly every niche. Here, we present an overview of the current knowledge regarding the diversity and benefits that polysaccharide production provides to bacterial communities within biofilms. PMID:26185074

  13. Bacterial Extracellular Polysaccharides in Biofilm Formation and Function

    PubMed Central

    Limoli, Dominique H.; Jones, Christopher J.; Wozniak, Daniel J.

    2015-01-01

    Microbes produce a biofilm matrix consisting of proteins, extracellular DNA, and polysaccharides that is integral in the formation of bacterial communities. Historical studies of polysaccharides revealed that their overproduction often alters the colony morphology and can be diagnostic in identifying certain species. The polysaccharide component of the matrix can provide many diverse benefits to the cells in the biofilm, including adhesion, protection, and structure. Aggregative polysaccharides act as molecular glue, allowing the bacterial cells to adhere to each other as well as surfaces. Adhesion facilitates the colonization of both biotic and abiotic surfaces by allowing the bacteria to resist physical stresses imposed by fluid movement that could separate the cells from a nutrient source. Polysaccharides can also provide protection from a wide range of stresses, such as desiccation, immune effectors, and predators such as phagocytic cells and amoebae. Finally, polysaccharides can provide structure to biofilms, allowing stratification of the bacterial community and establishing gradients of nutrients and waste products. This can be advantageous for the bacteria by establishing a heterogeneous population that is prepared to endure stresses created by the rapidly changing environments that many bacteria encounter. The diverse range of polysaccharide structures, properties, and roles highlight the importance of this matrix constituent to the successful adaptation of bacteria to nearly every niche. Here, we present an overview of the current knowledge regarding the diversity and benefits that polysaccharide production provides to bacterial communities within biofilms. PMID:26185074

  14. Polysaccharide structural features that are critical for the binding of sulfated fucans to bindin, the adhesive protein from sea urchin sperm.

    PubMed

    DeAngelis, P L; Glabe, C G

    1987-10-15

    We have investigated the structural features of sulfated fucose-containing polysaccharides which are responsible for their selective binding to Strongylocentrotus purpuratus bindin. The data presented demonstrate that the sulfate esters and a molecular weight in excess of approximately 15,000 are required for high affinity binding of the fucans to bindin. Desulfation destroys the binding activity of the fucans, which can be fully restored by chemical resulfation. Fucan fragments of an average molecular weight of 15,000 were nearly as active as the starting material (Mr 10(6)). The observed IC50 value for fragments of Mr congruent to 10,000 and Mr congruent to 5,000 were 1 and 2 orders of magnitude higher, respectively. The binding of fucoidan to bindin is stable in high salt (50% at 1.2 M NaCl) whereas the binding of fucoidan to DEAE-cellulose or polylysine is inhibited by the concentrations of salt normally found in sea water (50% at 0.2 and 0.5 M NaCl, respectively). This result suggests that the binding mechanism is not a simple ionic interaction and that hydrogen bonding and cooperativity may also be important determinants of the binding mechanism. We also found that polyvinyl sulfate binds to bindin with high affinity and inhibits the bindin-mediated agglutination of sea urchin eggs. The results of these investigations suggest that the spatial orientation of the sulfate esters plays a critical role in determining the selectivity of sulfated polysaccharide binding and that the polysaccharide backbone does not play a direct role in the binding mechanism.

  15. Synthetic trimer and tetramer of 3-beta-D-ribose-(1-1)-D-ribitol-5-phosphate conjugated to protein induce antibody responses to Haemophilus influenzae type b capsular polysaccharide in mice and monkeys.

    PubMed Central

    Peeters, C C; Evenberg, D; Hoogerhout, P; Käyhty, H; Saarinen, L; van Boeckel, C A; van der Marel, G A; van Boom, J H; Poolman, J T

    1992-01-01

    Synthetic oligosaccharides derived from the capsular polysaccharide (PRP) of Haemophilus influenzae type b were conjugated to carrier proteins via a thioether linkage. Conjugates were made of trimeric and tetrameric ribose-ribitol-phosphate and tetanus toxoid or diphtheria toxin. All conjugates elicited anti-PRP antibody responses with an increasing immunoglobulin G/immunoglobulin M ratio in adult mice and monkeys. Trimer conjugates elicited lower anti-PRP antibody responses compared with tetramer conjugates. Adult monkeys responded equally well to the tetrameric oligosaccharide-tetanus toxoid conjugate as to the oligosaccharide-CRM197 conjugate (HbOC), which elicits protective levels of serum antibodies in human infants after two or three injections. PMID:1563770

  16. Immunogenicity of meningococcal B polysaccharide conjugated to tetanus toxoid or CRM197 via adipic acid dihydrazide.

    PubMed

    Bartoloni, A; Norelli, F; Ceccarini, C; Rappuoli, R; Costantino, P

    1995-04-01

    Vaccine development against Group B Neisseria meningitidis is complicated by the nature of the capsular polysaccharide, which is alpha 2-8-linked poly-sialic acid, identical in structure to the poly-sialic acid found in many mammalian tissues during development. To test the feasibility of a vaccine based on this polysaccharide, we synthesized several conjugates of meningococcal B polysaccharide linked to a carrier protein (tetanus toxoid or diphtheria CRM197), via an adipic acid dihydrazide (ADH) spacer. All conjugates induced a strong immune response. However, most of the antibodies were not directed against the Meningococcus B polysaccharide and could not be inhibited by the purified polysaccharide alone. Further investigations showed that the antibodies recognized an epitope composed by the junction between the spacer and the polysaccharide and protein, that is not present in the native polysaccharide and is generated during the coupling reaction. This epitope becomes immunodominant with respect to the poorly immunogenic polysaccharide. While the majority of the immune response is directed against the above epitope, the conjugates induced also an immune response against the Meningococcus B polysaccharide. The anti-Meningococcus B antibodies elicited are of the IgM and IgG class and are inhibitable by the polysaccharide. Moreover, they are bactericidal, thus suggesting that they would induce protection against disease. PMID:7543714

  17. Effect of brefeldin A on the structure of the Golgi apparatus and on the synthesis and secretion of proteins and polysaccharides in sycamore maple (Acer pseudoplatanus) suspension-cultured cells.

    PubMed

    Driouich, A; Zhang, G F; Staehelin, L A

    1993-04-01

    Brefeldin A (BFA), a specific inhibitor of Golgi-mediated secretion in animal cells, has been used to study the organization of the secretory pathway and the function of the Golgi apparatus in plant cells. To this end, we have employed a combination of electron microscopical, immunocytochemical, and biochemical techniques to investigate the effects of this drug on the architecture of the Golgi apparatus as well as on the secretion of proteins and complex cell wall polysaccharides in sycamore maple (Acer pseudoplatanus) suspension-cultured cells. We have used 2.5 and 7.5 micrograms/mL of BFA, which is comparable to the 1 to 10 micrograms/mL used in experiments with animal cells. Electron micrographs of high-pressure frozen and freeze-substituted cells show that although BFA causes swelling of the endoplasmic reticulum cisternae, unlike in animal cells, it does not induce the disassembly of sycamore maple Golgi stacks. Instead, BFA induces the formation of large clusters of Golgi stacks, an increase in the number of trans-like Golgi cisternae, and the accumulation in the cytoplasm of very dense vesicles that appear to be derived from trans Golgi cisternae. These vesicles contain large amounts of xyloglucan (XG), the major hemicellulosic cell wall polysaccharide, as shown by immunocytochemical labeling with anti-XG antibodies. All of these structural changes disappear within 120 min after removal of the drug. In vivo labeling experiments using [3H]leucine demonstrate that protein secretion into the culture medium, but not protein synthesis, is inhibited by approximately 80% in the presence of BFA. In contrast, the incorporation of [3H]fucose into N-linked glycoproteins, which occurs in trans-Golgi cisternae, appears to be affected to a greater extent than the incorporation of [3H]xylose, which has been localized to medial Golgi cisternae. BFA also affects secretion of complex polysaccharides as evidenced by the approximate 50% drop in incorporation of [3H]xylose and

  18. POLYPEPTIDE AND POLYSACCHARIDE PROCESSING IN HYPERTHERMOPHILIC MICROORGANISMS

    SciTech Connect

    KELLY, ROBERT M.

    2008-12-22

    This project focused on the microbial physiology and biochemistry of heterotrophic hyperthermophiles with respect to mechanisms by which these organisms process polypeptides and polysaccharides under normal and stressed conditions. Emphasis is on two model organisms, for which completed genome sequences are available: Pyrococcus furiosus (growth Topt of 98°C), an archaeon, and Thermotoga maritima (growth Topt of 80°C), a bacterium. Both organisms are obligately anaerobic heterotrophs that reduce sulfur facultatively. Whole genome cDNA spotted microarrays were used to follow transcriptional response to a variety of environmental conditions in order to identify genes encoding proteins involved in the acquisition, synthesis, processing and utilization of polypeptides and polysaccharides. This project provided new insights into the physiological aspects of hyperthermophiles as these relate to microbial biochemistry and biological function in high temperature habitats. The capacity of these microorganisms to produce biohydrogen from renewable feedstocks makes them important for future efforts to develop biofuels.

  19. Coprecipitation of Th(4+) and the purified extracellular polysaccharide produced by bacterium Bradyrhizobium (Chamaecytisus) BGA-1.

    PubMed

    Díaz-Marrero, Ana R; Santamaría, Mónica; Hernández, Jairo; Corzo, Javier

    2004-08-01

    The soil bacterium Bradyrhizobium (Chamaecytisus) strain BGA-1 produces an extracellular polymeric substance (EPS) that, in the presence of Fe(3+), Al(3+) or Th(4+) solutions, forms a gel-like precipitate composed of polysaccharide, protein, lipopolysaccharide and the metal. Precipitation of the main component of the EPS, the extracellular polysaccharide, and thorium was studied. The precipitate was stable, but redissolved at pH values below 3.0 or in the presence of 10 mM EDTA. In the precipitate, the ratio thorium/basic repeating unit of the polysaccharide ranged from 0.4 to 0.8 mol/mol. Soluble polysaccharide-thorium complexes were not found, and larger polysaccharide molecules were precipitated in preference to smaller ones. Kinetic studies showed a non-linear dependence of the precipitate on the concentrations of both thorium and polysaccharide. The behaviors of the purified polysaccharide and of whole EPS with the thorium-containing precipitate were compared. The results suggested that EPS components other than polysaccharide are able to modify the precipitating ability of the polysaccharide. Thus, whole EPS is a better substrate than the purified polysaccharide for the removal of thorium from its solutions.

  20. Antitumor polysaccharides from mushrooms: a review on the structural characteristics, antitumor mechanisms and immunomodulating activities.

    PubMed

    Meng, Xin; Liang, Hebin; Luo, Lixin

    2016-04-01

    Mushrooms are popular folk medicines that have attracted considerable attention because of their efficient antitumor activities. This review covers existing research achievements on the mechanisms of isolated mushroom polysaccharides, particularly (1→3)-β-D-glucans. Our review also describes the function in modulating the immune system and potential tumor-inhibitory effects of polysaccharides. The antitumor mechanisms of mushroom polysaccharides are mediated by stimulated T cells or other immune cells. These polysaccharides are able to trigger various cellular responses, such as the expression of cytokines and nitric oxide. Most polysaccharides could bind other conjugate molecules, such as polypeptides and proteins, whose conjugation always possess strong antitumor activities. The purpose of this review is to summarize available information, and to reflect the present situation of polysaccharide research filed with a view for future direction. PMID:26974354

  1. Bioactivities and extraction optimization of crude polysaccharides from the fruits and leaves of Rubus chingii Hu.

    PubMed

    Zhang, Tian-Tian; Lu, Chuan-Li; Jiang, Jian-Guo; Wang, Min; Wang, Dong-Mei; Zhu, Wei

    2015-10-01

    Polysaccharides of Rubus chingii Hu fruit and leaf were extracted to compare their antioxidant, anti-inflammatory, and anticancer activities against breast cancer cells MCF-7 and liver cancer cells Bel-7402. Results showed that all the tested bioactivities of polysaccharides from leaf (L-Ps) were better than those of polysaccharides from fruit (F-Ps). Response surface methodology was then used to optimize the extraction conditions of polysaccharides from leaf. Additionally, polysaccharides from fruit and leaf were characterized and their contents of total sugars, proteins and uronic acid were compared. It was found that polysaccharides from fruit and leaf were similar in IR and UV absorption, but significantly different in contents of total sugars, protein and uronic acid. Their elution profiles of DEAE-Sepharose fast flow column were different too. The main peak of polysaccharides from fruit was eluted with 0.3 mol/l NaCl solution and the main peak of polysaccharides from leaf was eluted with deionized water. The differences between the two polysaccharides may be responsible for their differences in bioactivities. Further studies are required to explore their complete structural characteristics, structure-activity relationship and the mechanism of their activities.

  2. Iodine-Catalyzed Polysaccharide Esterification

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A review is provided of the recent reports to use iodine-catalyzed esterification reaction to produce esters from polysaccharides. The process entails reaction of the polysaccharide with an acid anhydride in the presence of a catalytic level of iodine, and in the absence of additional solvents. T...

  3. Polysaccharides of the red algae.

    PubMed

    Usov, Anatolii I

    2011-01-01

    Red algae (Rhodophyta) are known as the source of unique sulfated galactans, such as agar, agarose, and carrageenans. The wide practical uses of these polysaccharides are based on their ability to form strong gels in aqueous solutions. Gelling polysaccharides usually have molecules built up of repeating disaccharide units with a regular distribution of sulfate groups, but most of the red algal species contain more complex galactans devoid of gelling ability because of various deviations from the regular structure. Moreover, several red algae may contain sulfated mannans or neutral xylans instead of sulfated galactans as the main structural polysaccharides. This chapter is devoted to a description of the structural diversity of polysaccharides found in the red algae, with special emphasis on the methods of structural analysis of sulfated galactans. In addition to the structural information, some data on the possible use of red algal polysaccharides as biologically active polymers or as taxonomic markers are briefly discussed.

  4. Polysaccharides and bacterial plugging

    SciTech Connect

    Fogler, H.S.

    1991-11-01

    Before any successful application of Microbial Enhanced Oil Recovery process can be realized, an understanding of the cells' transport and retentive mechanisms in porous media is needed. Cell transport differs from particle transport in their ability to produce polysaccharides, which are used by cells to adhere to surfaces. Cell injection experiments have been conducted using Leuconostoc cells to illustrate the importance of cellular polysaccharide production as a transport mechanism that hinders cell movement and plugs porous media. Kinetic studies of the Leuconostoc cells, carried out to further understand the plugging rates of porous media, have shown that the cells' growth rates are approximately equal when provided with monosaccharide (glucose and fructose) or sucrose. The only difference in cell metabolism is the production of dextran when sucrose is supplied as a carbon source. Experimentally it has also been shown that the cells' growth rate is weakly dependent upon the sucrose concentration in the media, and strongly dependent upon the concentration of yeast extract. The synthesis of cellular dextran has been found to lag behind cell generation, thus indicating that the cells need to reach maturity before they are capable of expressing the detransucrase enzyme and synthesizing insoluble dextran. Dextran yields were found to be dependent upon the sucrose concentration in the media. 10 refs., 9 figs., 9 tabs.

  5. The molecular basis of polysaccharide cleavage by lytic polysaccharide monooxygenases

    PubMed Central

    Frandsen, Kristian E. H.; Simmons, Thomas J.; Dupree, Paul; Poulsen, Jens-Christian N.; Hemsworth, Glyn R.; Ciano, Luisa; Johnston, Esther M.; Tovborg, Morten; Johansen, Katja S.; von Freiesleben, Pernille; Marmuse, Laurence; Fort, Sébastien; Cottaz, Sylvain; Driguez, Hugues; Henrissat, Bernard; Lenfant, Nicolas; Tuna, Floriana; Baldansuren, Amgalanbaatar; Davies, Gideon J.; Leggio, Leila Lo; Walton, Paul H.

    2016-01-01

    Lytic polysaccharide monooxygenases (LPMOs) are copper-containing enzymes which oxidatively break down recalcitrant polysaccharides such as cellulose and chitin. Since their discovery LPMOs have become integral factors in the industrial utilization of biomass, especially in the sustainable generation of cellulosic bioethanol. We report here the first structural determination of an LPMO–oligosaccharide complex, yielding detailed insights into the mechanism of action of these enzymes. Using a combination of structure and electron paramagnetic resonance spectroscopy, we reveal the means by which LPMOs interact with saccharide substrates. We further uncover electronic and structural features of the enzyme active site, showing how LPMOs orchestrate the reaction of oxygen with polysaccharide chains. PMID:26928935

  6. The molecular basis of polysaccharide cleavage by lytic polysaccharide monooxygenases.

    PubMed

    Frandsen, Kristian E H; Simmons, Thomas J; Dupree, Paul; Poulsen, Jens-Christian N; Hemsworth, Glyn R; Ciano, Luisa; Johnston, Esther M; Tovborg, Morten; Johansen, Katja S; von Freiesleben, Pernille; Marmuse, Laurence; Fort, Sébastien; Cottaz, Sylvain; Driguez, Hugues; Henrissat, Bernard; Lenfant, Nicolas; Tuna, Floriana; Baldansuren, Amgalanbaatar; Davies, Gideon J; Lo Leggio, Leila; Walton, Paul H

    2016-04-01

    Lytic polysaccharide monooxygenases (LPMOs) are copper-containing enzymes that oxidatively break down recalcitrant polysaccharides such as cellulose and chitin. Since their discovery, LPMOs have become integral factors in the industrial utilization of biomass, especially in the sustainable generation of cellulosic bioethanol. We report here a structural determination of an LPMO-oligosaccharide complex, yielding detailed insights into the mechanism of action of these enzymes. Using a combination of structure and electron paramagnetic resonance spectroscopy, we reveal the means by which LPMOs interact with saccharide substrates. We further uncover electronic and structural features of the enzyme active site, showing how LPMOs orchestrate the reaction of oxygen with polysaccharide chains.

  7. Salmonella typhi O:9,12 polysaccharide-protein conjugates: characterization and immunoreactivity with pooled and individual normal human sera, sera from patients with paratyphoid A and B and typhoid fever, and animal sera.

    PubMed

    Aron, L; Di Fabio, J; Cabello, F C

    1993-04-01

    Polysaccharide of O:9,12 specificity purified from Salmonella typhi was conjugated to tetanus toxoid or bovine serum albumin in order to obtain defined antigenic material that would contain O chain free of other S. typhi antigens and that would be suitable for characterizing host humoral response to only S. typhi O-chain antigens. These artificial conjugates were strongly reactive in immunodots with 18 pooled and 3 individual serum samples from patients with typhoid fever and with rabbit anti-Salmonella O antiserum (group D, factors 1, 9, and 12). They reacted weakly with one serum sample from one human with paratyphoid A. These results suggest that the periodate oxidation and the reductive amination used in the conjugation conserved the immunogenicity of the O chain and allowed its absorption to nitrocellulose. They also suggest that the bovine serum albumin conjugate could be used in the diagnosis of S. typhi infections as normal sera may react with the protein molecule of the tetanus toxoid conjugate.

  8. Mediating chemical reactions using polysaccharides

    NASA Astrophysics Data System (ADS)

    Tyler, Lauren E.

    We have studied the NaBH4-mediated hydrogenation of select alkenes catalyzed by polysaccharide-stabilized nanoparticles. We compared the catalytic properties of Ni-based nanoparticles or Au/Co-based nanoparticles on the hydrogenation of cinnamic acid, cinnamide, cinnamyl alcohol, and ethyl cinnamate. We evaluated the possibility that the type of stabilizing polysaccharide surrounding the nanoparticle may affect the selectivity towards the alkene compounds that undergo the hydrogenation reaction. We found that the hydrogenation of cinnamide or ethyl cinnamate proceeded readily to 100% completion independent of the type of polysaccharide stabilizing the nanoparticle. However, the extent of the hydrogenation of cinnamyl alcohol and cinnamic acid varied greatly depending on the type of polysaccharide stabilizing the nanoparticle. In the course of these studies, we observed that some polysaccharides by themselves promoted the hydrolysis of ethyl cinnamate. Thus, we have raised the hypothesis that some polysaccharides may act as "esterases" and explored the interaction between select polysaccharides and a variety of ester compounds.

  9. Effect of aluminum ions on chemical and immunological properties of meningococcal group B polysaccharide.

    PubMed

    Moreno, C; Lifely, M R; Esdaile, J

    1985-09-01

    Soluble salts of aluminum were examined for their capacity to complex with purified meningococcal group B polysaccharide. The formation of the complexes resulted, first, in a markedly reduced rate of internal esterification at acid pH and, consequently, prolonged stability of the antigen as measured by its reactivity with antibody at pH 4 and, second, in an increased resistance to neuraminidase. Al3+ complexes of B polysaccharides were tested for immunogenicity in mice and found to be no better than the purified polysaccharide in the Na+ or Ca2+ form. However, when Neisseria meningitidis type 6 protein (outer membrane) complexed to B polysaccharide was tested, a substantial increase in anti-B titers was detected, whereas antiprotein titers remained unchanged. The possibility of using combinations of metal-polysaccharide-outer membrane protein complexes as vaccines for humans is discussed.

  10. Activation of intrinsic apoptotic signaling pathway in cancer cells by Cymbopogon citratus polysaccharide fractions.

    PubMed

    Thangam, Ramar; Sathuvan, Malairaj; Poongodi, Arasu; Suresh, Veeraperumal; Pazhanichamy, Kalailingam; Sivasubramanian, Srinivasan; Kanipandian, Nagarajan; Ganesan, Nalini; Rengasamy, Ramasamy; Thirumurugan, Ramasamy; Kannan, Soundarapandian

    2014-07-17

    Essential oils of Cymbopogon citratus were already reported to have wide ranging medical and industrial applications. However, information on polysaccharides from the plant and their anticancer activities are limited. In the present study, polysaccharides from C. citratus were extracted and fractionated by anion exchange and gel filtration chromatography. Two different polysaccharide fractions such as F1 and F2 were obtained, and these fractions were found to have distinct acidic polysaccharides as characterized by their molecular weight and sugar content. NMR spectral analysis revealed the presence of (1→4) linked b-d-Xylofuranose moiety in these polysaccharides. Using these polysaccharide fractions F1 and F2, anti-inflammatory and anticancer activities were evaluated against cancer cells in vitro and the mechanism of action of the polysaccharides in inducing apoptosis in cancer cells via intrinsic pathway was also proposed. Two different reproductive cancer cells such as Siha and LNCap were employed for in vitro studies on cytotoxicity, induction of apoptosis and apoptotic DNA fragmentation, changes in mitochondrial membrane potential, and profiles of gene and protein expression in response to treatment of cells by the polysaccharide fractions. These polysaccharide fractions exhibited potential cytotoxic and apoptotic effects on carcinoma cells, and they induced apoptosis in these cells through the events of up-regulation of caspase 3, down-regulation of bcl-2 family genes followed by cytochrome c release.

  11. Depletion of complement has distinct effects on the primary and secondary antibody responses to a conjugate of pneumococcal serotype 14 capsular polysaccharide and a T-cell-dependent protein carrier.

    PubMed

    Test, Samuel T; Mitsuyoshi, Joyce K; Hu, Yong

    2005-01-01

    Complement activation plays a critical role in the immune response to T-cell-dependent and T-cell-independent antigens. However, the effect of conjugation of T-cell-dependent protein carriers to T-cell-independent type 2 antigens on the requirement for complement in the humoral immune response to such antigens remains unknown. We studied the role of complement activation on the antibody response of BALB/c mice immunized with the T-cell-independent type 2 antigen serotype 14 pneumococcal capsular polysaccharide (PPS14), either in unmodified form or conjugated to ovalbumin (OVA). In mice immunized with either PPS14 or PPS14-OVA, depletion of endogenous complement at the time of primary immunization by treatment with cobra venom factor (CVF) diminished serum anti-PPS14 concentrations after primary immunization but enhanced antibody responses after secondary immunization. The secondary immunoglobulin G (IgG) anti-PPS14 antibody response after immunization with PPS14-OVA was especially enhanced by complement depletion, was observed at doses as low as 0.2 mug of antigen, and was maximal when CVF was administered within 2 days of immunization. The avidity and opsonophagocytic functions of IgG anti-PPS14 antibodies were comparable in mice immunized with PPS14-OVA with or without complement depletion. Serum anti-PPS14 antibody concentrations were near normal, and the enhancing effects of CVF treatment on the secondary anti-PPS14 antibody response were also apparent in splenectomized mice immunized with PPS14-OVA. These results demonstrate that complement activation can have distinct effects on the primary and secondary antibody responses to a T-cell-independent type 2 antigen, either unmodified or conjugated to a T-cell-dependent protein carrier. These differences should be taken into consideration when using complement to modulate the immune response to vaccines. PMID:15618164

  12. Depletion of complement has distinct effects on the primary and secondary antibody responses to a conjugate of pneumococcal serotype 14 capsular polysaccharide and a T-cell-dependent protein carrier.

    PubMed

    Test, Samuel T; Mitsuyoshi, Joyce K; Hu, Yong

    2005-01-01

    Complement activation plays a critical role in the immune response to T-cell-dependent and T-cell-independent antigens. However, the effect of conjugation of T-cell-dependent protein carriers to T-cell-independent type 2 antigens on the requirement for complement in the humoral immune response to such antigens remains unknown. We studied the role of complement activation on the antibody response of BALB/c mice immunized with the T-cell-independent type 2 antigen serotype 14 pneumococcal capsular polysaccharide (PPS14), either in unmodified form or conjugated to ovalbumin (OVA). In mice immunized with either PPS14 or PPS14-OVA, depletion of endogenous complement at the time of primary immunization by treatment with cobra venom factor (CVF) diminished serum anti-PPS14 concentrations after primary immunization but enhanced antibody responses after secondary immunization. The secondary immunoglobulin G (IgG) anti-PPS14 antibody response after immunization with PPS14-OVA was especially enhanced by complement depletion, was observed at doses as low as 0.2 mug of antigen, and was maximal when CVF was administered within 2 days of immunization. The avidity and opsonophagocytic functions of IgG anti-PPS14 antibodies were comparable in mice immunized with PPS14-OVA with or without complement depletion. Serum anti-PPS14 antibody concentrations were near normal, and the enhancing effects of CVF treatment on the secondary anti-PPS14 antibody response were also apparent in splenectomized mice immunized with PPS14-OVA. These results demonstrate that complement activation can have distinct effects on the primary and secondary antibody responses to a T-cell-independent type 2 antigen, either unmodified or conjugated to a T-cell-dependent protein carrier. These differences should be taken into consideration when using complement to modulate the immune response to vaccines.

  13. Computer simulation and experimental study of the polysaccharide-polysaccharide interaction in the bacteria Azospirillum brasilense Sp245

    NASA Astrophysics Data System (ADS)

    Arefeva, Oksana A.; Kuznetsov, Pavel E.; Tolmachev, Sergey A.; Kupadze, Machammad S.; Khlebtsov, Boris N.; Rogacheva, Svetlana M.

    2003-09-01

    We have studied the conformational properties and molecular dynamics of polysaccharides by using molecular modeling methods. Theoretical and experimental results of polysaccharide-polysaccharide interactions are described.

  14. Chemical studies on the polysaccharides of Salicornia brachiata.

    PubMed

    Sanandiya, Naresh D; Siddhanta, A K

    2014-11-01

    A group of 12 polysaccharide extracts were prepared from the tips, stem and roots of an Indian halophyte Salicornia brachiata Roxb. obtained by sequential extractions with cold water (CW), hot water (HW), aqueous ammonium oxalate (OX) and aqueous sodium hydroxide (ALK) solutions. Monosaccharide composition analysis revealed that all the polysaccharide extract samples consisted primarily of rhamnose, arabinose, mannose, galactose, glucose, whereas ribose and xylose were present only in some of the extracts. All the extracts exhibited low apparent viscosity (1.47-2.02 cP) and sulphate and contained no prominent toxic metal ions. Fucose was detected only in OX extract of the roots. These polysaccharides were found to be heterogeneous and highly branched (glycoside linkage analysis, size-exclusion chromatography, (13)C-NMR, FT-IR, circular dichroism and optical rotation data). Physico-chemical analyses of these polysaccharides including uronic acid, sulphate and protein contents were also carried out. This constitutes the first report on the profiling of Salicornia polysaccharides.

  15. Biochemical And Genetic Modification Of Polysaccharides

    NASA Technical Reports Server (NTRS)

    Kern, Roger G.; Petersen, Gene R.; Richards, Gil F.

    1993-01-01

    Bacteriophages producing endopolysaccharase-type enzymes used to produce, isolate, and purify high yields of modified polysaccharides from polysaccharides produced by, and incorporated into capsules of, certain bacteria. Bacteriophages used in conversion of native polysaccharide materials into polymers of nearly uniform high molecular weight or, alternatively, into highly pure oligosaccharides. Also used in genetic selection of families of polysaccharides structurally related to native polysaccharide materials, but having altered properties. Resulting new polysaccharides and oligosaccharides prove useful in variety of products, including pharmaceutical chemicals, coating materials, biologically active carbohydrates, and drag-reducing additives for fluids.

  16. Lycium barbarum polysaccharides promotes in vivo proliferation of adult rat retinal progenitor cells

    PubMed Central

    Wang, Hua; Lau, Benson Wui-Man; Wang, Ning-li; Wang, Si-ying; Lu, Qing-jun; Chang, Raymond Chuen-Chung; So, Kwok-fai

    2015-01-01

    Lycium barbarum is a widely used Chinese herbal medicine prescription for protection of optic nerve. However, it remains unclear regarding the effects of Lycium barbarum polysaccharides, the main component of Lycium barbarum, on in vivo proliferation of adult ciliary body cells. In this study, adult rats were intragastrically administered low- and high-dose Lycium barbarum polysaccharides (1 and 10 mg/kg) for 35 days and those intragastrically administered phosphate buffered saline served as controls. The number of Ki-67-positive cells in rat ciliary body in the Lycium barbarum polysaccharides groups, in particular low-dose Lycium barbarum polysaccharides group, was significantly greater than that in the phosphate buffered saline group. Ki-67-positive rat ciliary body cells expressed nestin but they did not express glial fibrillary acidic protein. These findings suggest that Lycium barbarum polysaccharides can promote the proliferation of adult rat retinal progenitor cells and the proliferated cells present with neuronal phenotype. PMID:26889185

  17. Polysaccharides from Polygonatum Inhibit the Proliferation of Prostate Cancer-Associated Fibroblasts.

    PubMed

    Han, Shu-Yu; Hu, Ming-Hua; Qi, Guan-Yun; Ma, Chao-Xiong; Wang, Yuan-Yuan; Ma, Fang-Li; Tao, Ning; Qin, Zhi-Hai

    2016-01-01

    Inhibition of cancer-associated broblasts (CAFs) may improve the efficacy of cancer therapy. Polysaccharide extracted from polygonatum can selectively inhibit the growth of prostate-CAFs (<.001) without inhibiting the growth of normal broblasts (NAFs). Polysaccharides from polygonatum stimulate autophagy of prostate-CAFs. 3-methyl-adenine(3-MA) is an autophagy inhibitor. 3-MA was added to prostate-CAFs with polysaccharide from polygonatum to determine whether autophagy plays an important role in the restrained effect. Finally, polysaccharide from polygonatum treatment significantly increased the activation of Beclin-1 and LC3, key autophagy proteins. Polysaccharides from polygonatum stimulate autophagy of prostate-CAFs and inhibits prostate-CAF growth, indicating that a novel anti-cancer strategy involves inhibiting the growth of prostate- CAFs. PMID:27644624

  18. Split of chiral degeneracy in mechanical and structural properties of oligopeptide-polysaccharide biomaterials.

    PubMed

    Taraban, Marc B; Hyland, Laura L; Yu, Y Bruce

    2013-09-01

    Enantiomeric biomaterials which are mirror images of each other are characterized by chiral degeneracy--identical structural characteristics and bulk material properties. The addition of another chiral component, D-polysaccharide, has been shown to split such degeneracy and result in two distinct biomaterials. Dynamic oscillatory rheometry and small-angle X-ray scattering demonstrate that the natural biochirality combination of L-peptides and D-polysaccharides assembles faster, has higher elastic moduli (G'), and is structurally more beneficial as opposed to the alternative D-peptide and D-polysaccharide combination. Chemical modifications of the OH-groups in α-D-glucose units in D-polysaccharides weaken such splitting of chiral degeneracy. These findings form a basis to design novel biomaterials and provide additional insight on why proteins and polysaccharides have oppoiste chirality in the biological world. PMID:23879188

  19. Purification of aloe polysaccharides by using aqueous two-phase extraction with desalination.

    PubMed

    Xing, Jian-Min; Li, Fen-Fang

    2009-01-01

    A PEG6000/(NH4)2SO4 aqueous two-phase system was chosen as the separation system. The effects of NaCl, Gu(SCN)2 and pH on the partitioning of aloe polysaccharide and bovine serum albumin (BSA) were studied. The result shows that the recovery of aloe polysaccharide was decreased with the increase in pH and the recovery of protein was not changed; NaCl and Gu(SCN)2 can change the partitioning behaviour of the BSA. The separation of BSA and aloe polysaccharides was also carried out. Then, the aloe polysaccharides were obtained by using ultrafiltration membrane. The composition of the polysaccharides was analysed by gas chromatography. The results show that mannose is mainly monosaccharide, and it only contains a few glucose.

  20. Antiviral Potential of Algae Polysaccharides Isolated from Marine Sources: A Review

    PubMed Central

    Ahmadi, Azin; Zorofchian Moghadamtousi, Soheil; Abubakar, Sazaly; Zandi, Keivan

    2015-01-01

    From food to fertilizer, algal derived products are largely employed in assorted industries, including agricultural, biomedical, food, and pharmaceutical industries. Among different chemical compositions isolated from algae, polysaccharides are the most well-established compounds, which were subjected to a variety of studies due to extensive bioactivities. Over the past few decades, the promising results for antiviral potential of algae-derived polysaccharides have advocated them as inordinate candidates for pharmaceutical research. Numerous studies have isolated various algal polysaccharides possessing antiviral activities, including carrageenan, alginate, fucan, laminaran, and naviculan. In addition, different mechanisms of action have been reported for these polysaccharides, such as inhibiting the binding or internalization of virus into the host cells or suppressing DNA replication and protein synthesis. This review strives for compiling previous antiviral studies of algae-derived polysaccharides and their mechanism of action towards their development as natural antiviral agents for future investigations. PMID:26484353

  1. Chemical features of Ganoderma polysaccharides with antioxidant, antitumor and antimicrobial activities.

    PubMed

    Ferreira, Isabel C F R; Heleno, Sandrina A; Reis, Filipa S; Stojkovic, Dejan; Queiroz, Maria João R P; Vasconcelos, M Helena; Sokovic, Marina

    2015-06-01

    Ganoderma genus comprises one of the most commonly studied species worldwide, Ganoderma lucidum. However, other Ganoderma species have been also reported as important sources of bioactive compounds. Polysaccharides are important contributors to the medicinal properties reported for Ganoderma species, as demonstrated by the numerous publications, including reviews, on this matter. Yet, what are the chemical features of Ganoderma polysaccharides that have bioactivity? In the present manuscript, the chemical features of Ganoderma polysaccharides with reported antioxidant, antitumor and antimicrobial activities (the most studied worldwide) are analyzed in detail. The composition of sugars (homo- versus hetero-glucans and other polysaccharides), type of glycosidic linkages, branching patterns, and linkage to proteins are discussed. Methods for extraction, isolation and identification are evaluated and, finally, the bioactivity of polysaccharidic extracts and purified compounds are discussed. The integration of data allows deduction of structure-activity relationships and gives clues to the chemical aspects involved in Ganoderma bioactivity.

  2. Polysaccharides purified from wild Cordyceps activate FGF2/FGFR1c signaling

    NASA Astrophysics Data System (ADS)

    Zeng, Yangyang; Han, Zhangrun; Yu, Guangli; Hao, Jiejie; Zhang, Lijuan

    2015-02-01

    Land animals as well as all organisms in ocean synthesize sulfated polysaccharides. Fungi split from animals about 1.5 billion years ago. As fungi make the evolutionary journey from ocean to land, the biggest changes in their living environment may be a sharp decrease in salt concentration. It is established that sulfated polysaccharides interact with hundreds of signaling molecules and facilitate many signaling transduction pathways, including fibroblast growth factor (FGF) and FGF receptor signaling pathway. The disappearance of sulfated polysaccharides in fungi and plants on land might indicate that polysaccharides without sulfation might be sufficient in facilitating protein ligand/receptor interactions in low salinity land. Recently, it was reported that plants on land start to synthesize sulfated polysaccharides in high salt environment, suggesting that fungi might be able to do the same when exposed in such environment. Interestingly, Cordyceps, a fungus habituating inside caterpillar body, is the most valued traditional Chinese Medicine. One of the important pharmaceutical active ingredients in Cordyceps is polysaccharides. Therefore, we hypothesize that the salty environment inside caterpillar body might allow the fungi to synthesize sulfated polysaccharides. To test the hypothesis, we isolated polysaccharides from both lava and sporophore of wild Cordyceps and also from Cordyceps militaris cultured without or with added salts. We then measured the polysaccharide activity using a FGF2/FGFR1c signaling-dependent BaF3 cell proliferation assay and found that polysaccharides isolated from wild Cordyceps activated FGF2/FGFR signaling, indicating that the polysaccharides synthesized by wild Cordyceps are indeed different from those by the cultured mycelium.

  3. Whey protein/polysaccharide-stabilized emulsions: Effect of polymer type and pH on release and topical delivery of salicylic acid.

    PubMed

    Combrinck, Johann; Otto, Anja; du Plessis, Jeanetta

    2014-06-01

    Emulsions are widely used as topical formulations in the pharmaceutical and cosmetic industries. They are thermodynamically unstable and require emulsifiers for stabilization. Studies have indicated that emulsifiers could affect topical delivery of actives, and this study was therefore designed to investigate the effects of different polymers, applied as emulsifiers, as well as the effects of pH on the release and topical delivery of the active. O/w emulsions were prepared by the layer-by-layer technique, with whey protein forming the first layer around the oil droplets, while either chitosan or carrageenan was subsequently adsorbed to the protein at the interface. Additionally, the emulsions were prepared at three different pH values to introduce different charges to the polymers. The active ingredient, salicylic acid, was incorporated into the oil phase of the emulsions. Physical characterization of the resulting formulations, i.e., droplet size, zeta potential, stability, and turbidity in the water phase, was performed. Release studies were conducted, after which skin absorption studies were performed on the five most stable emulsions, by using Franz type diffusion cells and utilizing human, abdominal skin membranes. It was found that an increase in emulsion droplet charge could negatively affect the release of salicylic acid from these formulations. Contrary, positively charged emulsion droplets were found to enhance dermal and transdermal delivery of salicylic acid from emulsions. It was hypothesized that electrostatic complex formation between the emulsifier and salicylic acid could affect its release, whereas electrostatic interaction between the emulsion droplets and skin could influence dermal/transdermal delivery of the active.

  4. Recognition of bacterial capsular polysaccharides and lipopolysaccharides by the macrophage mannose receptor.

    PubMed

    Zamze, Susanne; Martinez-Pomares, Luisa; Jones, Hannah; Taylor, Philip R; Stillion, Richard J; Gordon, Siamon; Wong, Simon Y C

    2002-11-01

    The in vitro binding of the macrophage mannose receptor to a range of different bacterial polysaccharides was investigated. The receptor was shown to bind to purified capsular polysaccharides from Streptococcus pneumoniae and to the lipopolysaccharides, but not capsular polysaccharides, from Klebsiella pneumoniae. Binding was Ca(2+)-dependent and inhibitable with d-mannose. A fusion protein of the mannose receptor containing carbohydrate recognition domains 4-7 and a full-length soluble form of the mannose receptor containing all domains external to the transmembrane region both displayed very similar binding specificities toward bacterial polysaccharides, suggesting that domains 4-7 are sufficient for recognition of these structures. Surprisingly, no direct correlation could be made between polysaccharide structure and binding to the mannose receptor, suggesting that polysaccharide conformation may play an important role in recognition. The full-length soluble form of the mannose receptor was able to bind simultaneously both polysaccharide via the carbohydrate recognition domains and sulfated oligosaccharide via the cysteine-rich domain. The possible involvement of the mannose receptor, either cell surface or soluble, in the innate and adaptive immune responses to bacterial polysaccharides is discussed.

  5. Recognition of bacterial capsular polysaccharides and lipopolysaccharides by the macrophage mannose receptor.

    PubMed

    Zamze, Susanne; Martinez-Pomares, Luisa; Jones, Hannah; Taylor, Philip R; Stillion, Richard J; Gordon, Siamon; Wong, Simon Y C

    2002-11-01

    The in vitro binding of the macrophage mannose receptor to a range of different bacterial polysaccharides was investigated. The receptor was shown to bind to purified capsular polysaccharides from Streptococcus pneumoniae and to the lipopolysaccharides, but not capsular polysaccharides, from Klebsiella pneumoniae. Binding was Ca(2+)-dependent and inhibitable with d-mannose. A fusion protein of the mannose receptor containing carbohydrate recognition domains 4-7 and a full-length soluble form of the mannose receptor containing all domains external to the transmembrane region both displayed very similar binding specificities toward bacterial polysaccharides, suggesting that domains 4-7 are sufficient for recognition of these structures. Surprisingly, no direct correlation could be made between polysaccharide structure and binding to the mannose receptor, suggesting that polysaccharide conformation may play an important role in recognition. The full-length soluble form of the mannose receptor was able to bind simultaneously both polysaccharide via the carbohydrate recognition domains and sulfated oligosaccharide via the cysteine-rich domain. The possible involvement of the mannose receptor, either cell surface or soluble, in the innate and adaptive immune responses to bacterial polysaccharides is discussed. PMID:12196537

  6. Structural characterization and biomedical properties of sulfated polysaccharide from the gladius of Sepioteuthis lessoniana (Lesson, 1831).

    PubMed

    Seedevi, Palaniappan; Moovendhan, Meivelu; Vairamani, Shanmugam; Shanmugam, Annian

    2016-04-01

    Sulfated polysaccharide was extracted from the internal shell (gladius) of Sepioteuthis lessoniana. The sulfated polysaccharide contained 61.3% of carbohydrate, 0.8% of protein, 28.2% of ash and 1.33% of moisture respectively. The elemental composition was analyzed using CHNS/O analyzer. The molecular weight of sulfated polysaccharide determined through PAGE was found to be as 66 kDa. Monosaccharides analysis revealed that sulfated polysaccharide was composed of rhamnose, galactose, xylose and glucose. The structural features of sulfated polysaccharide were analyzed by FT-IR and NMR spectroscopy. Further the sulfated polysaccharide was evaluated for its antibacterial activity against selected human clinical pathogens, namely Staphylococcus aureus, Klebsiella pneumoniae, Salmonella typhi, Vibrio cholerae, Klebsiella oxytoca, Escherichia coli, Salmonella paratyphi, Proteus mirabilis, Vibrio parahaemolyticus and Streptococcus pyogenes using agar well diffusion method. The polysaccharide has showed good antibacterial activity and MIC and MBC have also been evaluated. The anticancer activity was tested against HeLa cell line by MTT assay. The Cytotoxic Concentration (CC50) was observed as 700 μg/ml and the maximum anticancer activity of 62.89% was recorded at 200 μg/ml; whereas, the lowest of 9.87% was observed at 25 μg/ml. In conclusion, the sulfated polysaccharide is an alternate, non-toxic and cheap source of substance that showed good antibacterial and anticancer acitivity. PMID:26724686

  7. Cell wall polysaccharides are mislocalized to the Vacuole in echidna mutants.

    PubMed

    McFarlane, Heather E; Watanabe, Yoichiro; Gendre, Delphine; Carruthers, Kimberley; Levesque-Tremblay, Gabriel; Haughn, George W; Bhalerao, Rishikesh P; Samuels, Lacey

    2013-11-01

    During cell wall biosynthesis, the Golgi apparatus is the platform for cell wall matrix biosynthesis and the site of packaging, of both matrix polysaccharides and proteins, into secretory vesicles with the correct targeting information. The objective of this study was to dissect the post-Golgi trafficking of cell wall polysaccharides using echidna as a vesicle traffic mutant of Arabidopsis thaliana and the pectin-secreting cells of the seed coat as a model system. ECHIDNA encodes a trans-Golgi network (TGN)-localized protein, which was previously shown to be required for proper structure and function of the secretory pathway. In echidna mutants, some cell wall matrix polysaccharides accumulate inside cells, rather than being secreted to the apoplast. In this study, live cell imaging of fluorescent protein markers as well as transmission electron microscopy (TEM)/immunoTEM of cryofixed seed coat cells were used to examine the consequences of TGN disorganization in echidna mutants under conditions of high polysaccharide production and secretion. While in wild-type seed coat cells, pectin is secreted to the apical surface, in echidna, polysaccharides accumulate in post-Golgi vesicles, the central lytic vacuole and endoplasmic reticulum-derived bodies. In contrast, proteins were partially mistargeted to internal multilamellar membranes in echidna. These results suggest that while secretion of both cell wall polysaccharides and proteins at the TGN requires ECHIDNA, different vesicle trafficking components may mediate downstream events in their secretion from the TGN. PMID:24058145

  8. Cadherin Domains in the Polysaccharide-Degrading Marine Bacterium Saccharophagus degradans 2-40 Are Carbohydrate-Binding Modules▿

    PubMed Central

    Fraiberg, Milana; Borovok, Ilya; Bayer, Edward A.; Weiner, Ronald M.; Lamed, Raphael

    2011-01-01

    The complex polysaccharide-degrading marine bacterium Saccharophagus degradans strain 2-40 produces putative proteins that contain numerous cadherin and cadherin-like domains involved in intercellular contact interactions. The current study reveals that both domain types exhibit reversible calcium-dependent binding to different complex polysaccharides which serve as growth substrates for the bacterium. PMID:21036994

  9. Rheologically interesting polysaccharides from yeasts

    NASA Technical Reports Server (NTRS)

    Petersen, G. R.; Nelson, G. A.; Cathey, C. A.; Fuller, G. G.

    1989-01-01

    We have examined the relationships between primary, secondary, and tertiary structures of polysaccharides exhibiting the rheological property of friction (drag) reduction in turbulent flows. We found an example of an exopolysaccharide from the yeast Cryptococcus laurentii that possessed high molecular weight but exhibited lower than expected drag reducing activity. Earlier correlations by Hoyt showing that beta 1 --> 3, beta 2 --> 4, and alpha 1 --> 3 linkages in polysaccharides favored drag reduction were expanded to include correlations to secondary structure. The effect of sidechains in a series of gellan gums was shown to be related to sidechain length and position. Disruption of secondary structure in drag reducing polysaccharides reduced drag reducing activity for some but not all exopolysaccharides. The polymer from C. laurentii was shown to be more stable than xanthan gum and other exopolysaccharides under the most vigorous of denaturing conditions. We also showed a direct relationship between extensional viscosity measurements and the drag reducing coefficient for four exopolysaccharides.

  10. Actinobacillus actinomycetemcomitans Y4 capsular-polysaccharide-like polysaccharide promotes osteoclast-like cell formation by interleukin-1 alpha production in mouse marrow cultures.

    PubMed Central

    Nishihara, T; Ueda, N; Amano, K; Ishihara, Y; Hayakawa, H; Kuroyanagi, T; Ohsaki, Y; Nagata, K; Noguchi, T

    1995-01-01

    The mechanism of osteoclast-like cell formation induced by periodontopathic bacterium Actinobacillus actinomycetemcomitans Y4 (serotype b) capsular-polysaccharide-like polysaccharide (capsular-like polysaccharide) was examined in a mouse bone marrow culture system. When mouse bone marrow cells were cultured with A. actinomycetemcomitans Y4 capsular-like polysaccharide for 9 days, many multinucleated cells were formed. The multinucleated cells showed several characteristics of osteoclasts, including tartrate-resistant acid phosphatase (TRACP) and the ability to resorb the calcified dentine. In this study, we examined the effects of antisera to interleukins on the formation of osteoclast-like cells induced by A. actinomycetemcomitans Y4 capsular-like polysaccharide. Monospecific anti-mouse recombinant interleukin-1 alpha (rIL-1 alpha) serum completely inhibited the formation of osteoclast-like cells in the presence of A. actinomycetemcomitans Y4 capsular-like polysaccharide. However, anti-mouse rIL-1 beta and anti-mouse rIL-6 sera showed no effect on osteoclast-like cell formation. IL-1 receptor antagonist significantly inhibited the osteoclast-like cell formation mediated by A. actinomycetemcomitans Y4 capsular-like polysaccharide in mouse marrow cultures. The bioactive IL-1 was detected in the culture media of mouse bone marrow cells stimulated with A. actinomycetemcomitans Y4 capsular-like polysaccharide. These results indicate that IL-1 alpha is involved in the mechanism of the formation of osteoclast-like cells induced by A. actinomycetemcomitans Y4 capsular-like polysaccharide. We sought to determine whether osteoclast-like cell formation induced by A. actinomycetemcomitans Y4 capsular-like polysaccharide could be modulated by the protein kinase inhibitors H8 and HA1004. The formation of osteoclast-like cells was suppressed by H8 and HA1004. These findings suggest that the signals by protein kinases may regulate osteoclast-like cell formation induced by A

  11. Nanoengineering of vaccines using natural polysaccharides.

    PubMed

    Cordeiro, Ana Sara; Alonso, María José; de la Fuente, María

    2015-11-01

    Currently, there are over 70 licensed vaccines, which prevent the pathogenesis of around 30 viruses and bacteria. Nevertheless, there are still important challenges in this area, which include the development of more active, non-invasive, and thermo-resistant vaccines. Important biotechnological advances have led to safer subunit antigens, such as proteins, peptides, and nucleic acids. However, their limited immunogenicity has demanded potent adjuvants that can strengthen the immune response. Particulate nanocarriers hold a high potential as adjuvants in vaccination. Due to their pathogen-like size and structure, they can enhance immune responses by mimicking the natural infection process. Additionally, they can be tailored for non-invasive mucosal administration (needle-free vaccination), and control the delivery of the associated antigens to a specific location and for prolonged times, opening room for single-dose vaccination. Moreover, they allow co-association of immunostimulatory molecules to improve the overall adjuvant capacity. The natural and ubiquitous character of polysaccharides, together with their intrinsic immunomodulating properties, their biocompatibility, and biodegradability, justify their interest in the engineering of nanovaccines. In this review, we aim to provide a state-of-the-art overview regarding the application of nanotechnology in vaccine delivery, with a focus on the most recent advances in the development and application of polysaccharide-based antigen nanocarriers.

  12. Hyperbranched acidic polysaccharide from green tea.

    PubMed

    Yang, Liqun; Fu, Shanshan; Zhu, Xiane; Zhang, Li-Ming; Yang, Yanrui; Yang, Xiaomin; Liu, Hui

    2010-12-13

    An acidic tea polysaccharide (ALTPS), isolated from green tea ( Camellia sinensis ), was characterized as a hyperbranched glycoprotein containing the acidic heteropolysaccharide chains and the protein residues from the results of UV-vis, FTIR, one- and two-dimensional NMR, GC, GC-MS, and amino acid analyses. Solution properties of ALTPS were investigated by static and dynamic light scattering analyses and viscometry. The results indicated that the viscosity behavior of ALTPS exhibited a typical polyelectrolyte effect in distilled water, which may be avoided by adding salts. The low intrinsic viscosity of ALTPS in the solutions (8-15 mL/g) is attributed to its hyperbranched structure. By application of the polymer solution theory, it was revealed that ALTPS was present in a sphere-like conformation in the solutions as a result of the hyperbranched structure. The TEM image further confirmed that ALTPS existed in a spherical conformation in aqueous NaCl solution. Glucose was absorbed by ALTPS, which may be one of blood glucose lowering mechanisms of tea polysaccharides.

  13. Panels of chemically-modified heparin polysaccharides and natural heparan sulfate saccharides both exhibit differences in binding to Slit and Robo, as well as variation between protein binding and cellular activity† †Electronic supplementary information (ESI) available: NMR chemical shift characterisation of modified heparins, protein sequence alignment methodology and data, protein binding and activity assay dose-response curves. See DOI: 10.1039/c6mb00432f Click here for additional data file.

    PubMed Central

    Ahmed, Yassir A.; Yates, Edwin A.; Moss, Diana J.; Loeven, Markus A.; Hussain, Sadaf-Ahmahni; Hohenester, Erhard; Turnbull, Jeremy E.

    2016-01-01

    Heparin/heparan sulfate (HS) glycosaminoglycans are required for Slit–Robo cellular responses. Evidence exists for interactions between each combination of Slit, Robo and heparin/HS and for formation of a ternary complex. Heparin/HS are complex mixtures displaying extensive structural diversity. The relevance of this diversity has been studied to a limited extent using a few select chemically-modified heparins as models of HS diversity. Here we extend these studies by parallel screening of structurally diverse panels of eight chemically-modified heparin polysaccharides and numerous natural HS oligosaccharide chromatographic fractions for binding to both Drosophila Slit and Robo N-terminal domains and for activation of a chick retina axon response to the Slit fragment. Both the polysaccharides and oligosaccharide fractions displayed variability in binding and cellular activity that could not be attributed solely to increasing sulfation, extending evidence for the importance of structural diversity to natural HS as well as model modified heparins. They also displayed differences in their interactions with Slit compared to Robo, with Robo preferring compounds with higher sulfation. Furthermore, the patterns of cellular activity across compounds were different to those for binding to each protein, suggesting that biological outcomes are selectively determined in a subtle manner that does not simply reflect the sum of the separate interactions of heparin/HS with Slit and Robo. PMID:27502551

  14. Polysaccharide-Modified Synthetic Polymeric Biomaterials

    PubMed Central

    Baldwin, Aaron D.; Kiick, Kristi L.

    2010-01-01

    This review presents an overview of polysaccharide-conjugated synthetic polymers and their use in tissue-engineered scaffolds and drug-delivery applications. This topic will be divided into four categories: (1) polymeric materials modified with non-mammalian polysaccharides such as alginate, chitin, and dextran; (2) polymers modified with mammalian polysaccharides such as hyaluronan, chondroitin sulfate, and heparin; (3) multi-polysaccharide-derivatized polymer conjugate systems; and (4) polymers containing polysaccharide-mimetic molecules. Each section will discuss relevant conjugation techniques, analysis, and the impact of these materials as micelles, particles, or hydrogels used in in-vitro and in-vivo biomaterial applications. PMID:20091875

  15. Polysaccharide-modified synthetic polymeric biomaterials.

    PubMed

    Baldwin, Aaron D; Kiick, Kristi L

    2010-01-01

    This review presents an overview of polysaccharide-conjugated synthetic polymers and their use in tissue-engineered scaffolds and drug-delivery applications. This topic will be divided into four categories: (1) polymeric materials modified with non-mammalian polysaccharides such as alginate, chitin, and dextran; (2) polymers modified with mammalian polysaccharides such as hyaluronan, chondroitin sulfate, and heparin; (3) multi-polysaccharide-derivatized polymer conjugate systems; and (4) polymers containing polysaccharide-mimetic molecules. Each section will discuss relevant conjugation techniques, analysis, and the impact of these materials as micelles, particles, or hydrogels used in in-vitro and in-vivo biomaterial applications. PMID:20091875

  16. Polysaccharides templates for assembly of nanosilver.

    PubMed

    Emam, Hossam E; Ahmed, Hanan B

    2016-01-01

    Polysaccharides are particularly attractive in biomedical applications due to its biodegradability and biocompatibility. In addition to its ecofriendly effects and easy processing into different hydrogel shapes, made polysaccharides used on a large scale as suitable media for preparation of silver nanoparticles (AgNPs). In spite of, most of polysaccharides are water insoluble, but it has shown to be quite efficient capping agents and/or nanoreactor matrices for production of AgNPs. Several methods have been developed to get the benefit of multi-functionality for polysaccharides' macromolecules in preparation of AgNPs. Therefore, recently, preparation of nanosilver using different polysaccharides have been the focus of an exponentially increasing number of works devoted to develop nanocomposites by blending AgNPs with different polysaccharides matrices. The current review represents a wide survey for the published studies which interested in using of polysaccharides in nanosilver preparations.

  17. O-Acetylation of Plant Cell Wall Polysaccharides

    PubMed Central

    Gille, Sascha; Pauly, Markus

    2011-01-01

    Plant cell walls are composed of structurally diverse polymers, many of which are O-acetylated. How plants O-acetylate wall polymers and what its function is remained elusive until recently, when two protein families were identified in the model plant Arabidopsis that are involved in the O-acetylation of wall polysaccharides – the reduced wall acetylation (RWA) and the trichome birefringence-like (TBL) proteins. This review discusses the role of these two protein families in polysaccharide O-acetylation and outlines the differences and similarities of polymer acetylation mechanisms in plants, fungi, bacteria, and mammals. Members of the TBL protein family had been shown to impact pathogen resistance, freezing tolerance, and cellulose biosynthesis. The connection of TBLs to polysaccharide O-acetylation thus gives crucial leads into the biological function of wall polymer O-acetylation. From a biotechnological point understanding the O-acetylation mechanism is important as acetyl-substituents inhibit the enzymatic degradation of wall polymers and released acetate can be a potent inhibitor in microbial fermentations, thus impacting the economic viability of, e.g., lignocellulosic based biofuel production. PMID:22639638

  18. Functional Exploration of the Polysaccharide Lyase Family PL6

    PubMed Central

    Mathieu, Sophie; Henrissat, Bernard; Labre, Flavien; Skjåk-Bræk, Gudmund; Helbert, William

    2016-01-01

    Alginate, the main cell-wall polysaccharide of brown algae, is composed of two residues: mannuronic acid (M-residues) and, its C5-epimer, guluronic acid (G-residues). Alginate lyases define a class of enzymes that cleave the glycosidic bond of alginate by β-elimination. They are classified according to their ability to recognize the distribution of M- and G-residues and are named M-, G- or MG-lyases. In the CAZy database, alginate lyases have been grouped by sequence similarity into seven distinct polysaccharide lyase families. The polysaccharide lyase family PL6 is subdivided into three subfamilies. Subfamily PL6_1 includes three biochemically characterized enzymes (two alginate lyases and one dermatan sulfatase lyase). No characterized enzymes have been described in the two other subfamilies (PL6_2 and PL6_3). To improve the prediction of polysaccharide-lyase activity in the PL6 family, we re-examined the classification of the PL6 family and biochemically characterized a set of enzymes reflecting the diversity of the protein sequences. Our results show that subfamily PL6_1 includes two dermatan sulfates lyases and several alginate lyases that have various substrate specificities and modes of action. In contrast, subfamilies PL6_2 and PL6_3 were found to contain only endo-poly-MG-lyases. PMID:27438604

  19. The pretreatment effects on the antioxidant activity of jujube polysaccharides.

    PubMed

    Qu, Chenling; Yu, Songcheng; Jin, Huali; Wang, Jinshui; Luo, Li

    2013-10-01

    Pretreatment is vital to keep the bioactivities of polysaccharides. In this paper, the effects of hot water, ultrasonic and microwave extraction, as well as the effects of protein and pigment removal steps, on the antioxidant activity of water soluble polysaccharides in jujube (WSPJ) were studied. Hydroxyl free radical (OH) scavenging activity was adopted to determine the antioxidant activity of WSPJ. The results showed that OH scavenging activity of WSPJ extracted by ultrasonic wave was higher than that extracted by hot water and by microwave. Furthermore, power parameter in both ultrasonic and microwave extraction affected the OH scavenging activity dramatically. On the other hand, Sevag reagent was better than trichloroacetic acid (TCA), TCA with 1-butanol (TCA-B) and hydrochloric acid for protein removal, and H2O2 was better than active carbon for pigment removal to keep the antioxidant activity of WSPJ.

  20. Antioxidant and anticoagulant activity of sulfated polysaccharide from Gracilaria debilis (Forsskal).

    PubMed

    Sudharsan, Sadhasivam; Subhapradha, Namasivayam; Seedevi, Palaniappan; Shanmugam, Vairamani; Madeswaran, Perumal; Shanmugam, Annaian; Srinivasan, Alagiri

    2015-11-01

    Sulfated polysaccharide was isolated from Gracilaria debilis and purified through gel chromatography and their molecular weight was determined through AGE and PAGE. The total sugars in the crude, fractionated and purified polysaccharide were estimated as 52.65%, 59.70% and 67.60%, respectively. The ash and moisture content of crude and purified polysaccharide was found to be 14.2% and 23.5% and the polysaccharide was free from protein contamination. The sulfate and uronic acid contents in the crude, fractionated and purified were estimated as 14.08%, 15.33% and 16.01% and 10.12%, 13.56%, 16.70%. The elemental composition including carbon (crude - 23.12%, purified - 21.05%), hydrogen (crude - 3.4%, purified - 4.13%) and nitrogen (crude - 1.22%, purified - 0.56%) were also analyzed. The anticoagulant activity of the sulfated polysaccharide through APTT and PT was estimated at 14.11 and 8.23IU/mg. The purified polysaccharide with the molecular mass of 20kDa showed highest antioxidant activity (38.57%, 43.48% and 38.88%) in all the assays tested such as DPPH hydroxyl radical, superoxide radical, hydroxyl radical scavenging activities and the structural property was analyzed through FT-IR and (1)H NMR spectrum. The results together suggest that the isolated low molecular weight sulfated polysaccharide will demonstrate as a enormously available alternative natural source of antioxidant for industrial uses.

  1. Controls of Polysaccharide Chemistry on the Kinetics and Thermodynamics of Heterogeneous Calcium Carbonate Nucleation

    NASA Astrophysics Data System (ADS)

    Giuffre, A. J.; Han, N.; Dove, P. M.

    2011-12-01

    Polysaccharide fibrils control the orientation of calcium carbonate (CaCO3) biominerals. Good examples are found in the multilayered extracellular mucilaginous sheath of green algae and cyanobacteria and in specialized vesicles inside coccolithophorids. More complex organisms such as arthropods and mollusks form biomineralized exoskeletons and shells that consist of insoluble polysaccharides and soluble acid-rich proteins. In these structures, CaCO3 mineral orientation occurs along fibers of the polysaccharide chitin. This raises the question of whether polysaccharide chemistry has specific roles in directing biomineralization. The last three decades of research show that acidic proteins influence CaCO3 polymorph selection, crystallographic orientation, and nucleation and growth rates but little is known about the function of polysaccharides. In fact, polysaccharides are long considered an inert component of organic frameworks. In this experimental investigation, we test the hypothesis that polysaccharides have chemistry-specific influences on calcification by measuring the kinetics of calcite nucleation onto three types of polysaccharide films under controlled solution compositions. Characterized polysaccharides of simple repeating monomer sequences were chosen as model compounds to represent the major carbohydrates seen in microbial and calcifying environments: 1) alginic acid with carboxyl groups, 2) hyaluronic acid with alternating carboxyl and acetylamine groups, and 3) chitosan with amine and acetylamine groups. Biosubstrates were prepared by electrodeposition of these compounds as thin gel-like films onto gold-coated silicon wafers. Using a flow-through cell, heterogeneous nucleation rates of calcite were measured for a suite of supersaturation conditions. These rate data were compared to similar measurements for carboxyl- and hydroxyl-terminated self-assembled monolayers. Calcite nucleation rates onto the three polysaccharides vary by a factor of 400x

  2. Cations modulate polysaccharide structure to determine FGF-FGFR signaling: a comparison of signaling and inhibitory polysaccharide interactions with FGF-1 in solution.

    PubMed

    Guimond, Scott E; Rudd, Timothy R; Skidmore, Mark A; Ori, Alessandro; Gaudesi, Davide; Cosentino, Cesare; Guerrini, Marco; Edge, Ruth; Collison, David; McInnes, Eric; Torri, Giangiacomo; Turnbull, Jeremy E; Fernig, David G; Yates, Edwin A

    2009-06-01

    For heparan sulfate (HS) to bind and regulate the activity of proteins, the polysaccharide must present an appropriate sequence and adopt a suitable conformation. The conformations of heparin derivatives, as models of HS, are altered via a change in the associated cations, and this can drastically modify their FGF signaling activities. Here, we report that changing the cations associated with an N-acetyl-enriched heparin polysaccharide, from sodium to copper(II), converted it from supporting signaling through the fibroblast growth factor receptor (FGF-1-FGFR1c) tyrosine kinase signaling system to being inhibitory in a cell-based BaF3 assay. Nuclear magnetic resonance and synchrotron radiation circular dichroism (SRCD) spectroscopy demonstrated that the polysaccharide conformation differed in the presence of sodium or copper(II) cations. Electron paramagnetic resonance confirmed the environment of the copper(II) ion on the N-acetyl-enriched polysaccharide was distinct from that previously observed with intact heparin, which supported signaling. Secondary structures in solution complexes of polysaccharides with FGF-1 (which either supported signaling through FGFR1c or were inhibitory) were determined by SRCD. This allowed direct comparison of the two FGF-1-polysaccharide complexes in solution, containing identical molecular components and differing only in their cation content. Subtle structural differences were revealed, including a reduction in the level of disordered structure in the inhibitory complex. PMID:19400583

  3. Structurally altered capsular polysaccharides produced by mutant bacteria

    NASA Technical Reports Server (NTRS)

    Kern, Roger G. (Inventor); Petersen, Gene R. (Inventor); Richards, Gil F. (Inventor)

    1995-01-01

    Structurally altered capsular polysaccharides are produced by mutant bacteria. These polysaccharides are isolated by selecting a wild type bacterial strain and a phage producing degradative enzymes that have substrate specificity for the capsular polysaccharides produced by the wild type bacteria. Phage-resistant mutants producing capsular polysaccharides are selected and the structurally altered capsular polysaccharide is isolated therefrom.

  4. Differential regulation of polysaccharide-specific antibody responses to isolated polysaccharides, conjugate vaccines, and intact Gram-positive versus Gram-negative extracellular bacteria.

    PubMed

    Snapper, Clifford M

    2016-06-24

    Bacterial capsular polysaccharides are major virulence factors and are key targets in a number of licensed anti-bacterial vaccines. Their major characteristics are their large molecular weight and expression of repeating antigenic epitopes that mediate multivalent B cell receptor cross-linking. In addition, since the majority of these antigens cannot associate with MHC-II they fail to recruit CD4+ T cell help and are referred to as T cell-independent antigens. In this review I will discuss a series of studies from my laboratory that have underscored the importance of understanding polysaccharide-specific antibody responses within the context in which the PS is expressed (i.e. in isolation, as a component of conjugate vaccines, and expressed naturally by intact bacteria). We have shown that multivalent B cell receptor crosslinking, as mediated by polysaccharides, uniquely determines the qualitative response of the B cell to subsequent stimuli, but by itself is insufficient to induce antibody secretion or class switching. For these latter events to occur, second signals must act in concert with primary signals derived from the B cell receptor. The co-expression of polysaccharide and protein within intact bacteria promotes recruitment of CD4+ T cell help for the associated PS-specific IgG response, in contrast to isolated polysaccharides. Further, the particulate nature of extracellular bacteria confers properties to the polysaccharide-specific IgG response that makes it distinct immunologically from soluble conjugate vaccines. Finally, the underlying biochemical and/or structural differences that distinguish Gram-positive and Gram-negative bacteria appear to play critical roles in differentially regulating the associated polysaccharide-specific IgG responses to these groups of pathogens. These studies have a number of implications for the understanding and future design of polysaccharide-based vaccines.

  5. Differential regulation of polysaccharide-specific antibody responses to isolated polysaccharides, conjugate vaccines, and intact Gram-positive versus Gram-negative extracellular bacteria.

    PubMed

    Snapper, Clifford M

    2016-06-24

    Bacterial capsular polysaccharides are major virulence factors and are key targets in a number of licensed anti-bacterial vaccines. Their major characteristics are their large molecular weight and expression of repeating antigenic epitopes that mediate multivalent B cell receptor cross-linking. In addition, since the majority of these antigens cannot associate with MHC-II they fail to recruit CD4+ T cell help and are referred to as T cell-independent antigens. In this review I will discuss a series of studies from my laboratory that have underscored the importance of understanding polysaccharide-specific antibody responses within the context in which the PS is expressed (i.e. in isolation, as a component of conjugate vaccines, and expressed naturally by intact bacteria). We have shown that multivalent B cell receptor crosslinking, as mediated by polysaccharides, uniquely determines the qualitative response of the B cell to subsequent stimuli, but by itself is insufficient to induce antibody secretion or class switching. For these latter events to occur, second signals must act in concert with primary signals derived from the B cell receptor. The co-expression of polysaccharide and protein within intact bacteria promotes recruitment of CD4+ T cell help for the associated PS-specific IgG response, in contrast to isolated polysaccharides. Further, the particulate nature of extracellular bacteria confers properties to the polysaccharide-specific IgG response that makes it distinct immunologically from soluble conjugate vaccines. Finally, the underlying biochemical and/or structural differences that distinguish Gram-positive and Gram-negative bacteria appear to play critical roles in differentially regulating the associated polysaccharide-specific IgG responses to these groups of pathogens. These studies have a number of implications for the understanding and future design of polysaccharide-based vaccines. PMID:27133879

  6. Sucrose release from polysaccharide gels.

    PubMed

    Nishinari, Katsuyoshi; Fang, Yapeng

    2016-05-18

    Sucrose release from polysaccharide gels has been studied extensively because it is expected to be useful in understanding flavour release from solid foods and to find a new processing method which produces more palatable and healthier foods. We provide an overview of the release of sucrose and other sugars from gels of agar and related polysaccharides. The addition of sucrose to agar solutions leads to the increase in transparency of the resulting gels and the decrease in syneresis, which is attributed to the decrease in mesh size in gels. The syneresis occurring in the quiescent condition and fluid release induced by compression is discussed. The relationship between the sugar release and the structural, rheological and thermal properties of gels is also discussed. Finally, the future research direction is proposed.

  7. Cellulose degradation by polysaccharide monooxygenases.

    PubMed

    Beeson, William T; Vu, Van V; Span, Elise A; Phillips, Christopher M; Marletta, Michael A

    2015-01-01

    Polysaccharide monooxygenases (PMOs), also known as lytic PMOs (LPMOs), enhance the depolymerization of recalcitrant polysaccharides by hydrolytic enzymes and are found in the majority of cellulolytic fungi and actinomycete bacteria. For more than a decade, PMOs were incorrectly annotated as family 61 glycoside hydrolases (GH61s) or family 33 carbohydrate-binding modules (CBM33s). PMOs have an unusual surface-exposed active site with a tightly bound Cu(II) ion that catalyzes the regioselective hydroxylation of crystalline cellulose, leading to glycosidic bond cleavage. The genomes of some cellulolytic fungi contain more than 20 genes encoding cellulose-active PMOs, suggesting a diversity of biological activities. PMOs show great promise in reducing the cost of conversion of lignocellulosic biomass to fermentable sugars; however, many questions remain about their reaction mechanism and biological function. This review addresses, in depth, the structural and mechanistic aspects of oxidative depolymerization of cellulose by PMOs and considers their biological function and phylogenetic diversity.

  8. Sucrose release from polysaccharide gels.

    PubMed

    Nishinari, Katsuyoshi; Fang, Yapeng

    2016-05-18

    Sucrose release from polysaccharide gels has been studied extensively because it is expected to be useful in understanding flavour release from solid foods and to find a new processing method which produces more palatable and healthier foods. We provide an overview of the release of sucrose and other sugars from gels of agar and related polysaccharides. The addition of sucrose to agar solutions leads to the increase in transparency of the resulting gels and the decrease in syneresis, which is attributed to the decrease in mesh size in gels. The syneresis occurring in the quiescent condition and fluid release induced by compression is discussed. The relationship between the sugar release and the structural, rheological and thermal properties of gels is also discussed. Finally, the future research direction is proposed. PMID:26952168

  9. Cellulose degradation by polysaccharide monooxygenases.

    PubMed

    Beeson, William T; Vu, Van V; Span, Elise A; Phillips, Christopher M; Marletta, Michael A

    2015-01-01

    Polysaccharide monooxygenases (PMOs), also known as lytic PMOs (LPMOs), enhance the depolymerization of recalcitrant polysaccharides by hydrolytic enzymes and are found in the majority of cellulolytic fungi and actinomycete bacteria. For more than a decade, PMOs were incorrectly annotated as family 61 glycoside hydrolases (GH61s) or family 33 carbohydrate-binding modules (CBM33s). PMOs have an unusual surface-exposed active site with a tightly bound Cu(II) ion that catalyzes the regioselective hydroxylation of crystalline cellulose, leading to glycosidic bond cleavage. The genomes of some cellulolytic fungi contain more than 20 genes encoding cellulose-active PMOs, suggesting a diversity of biological activities. PMOs show great promise in reducing the cost of conversion of lignocellulosic biomass to fermentable sugars; however, many questions remain about their reaction mechanism and biological function. This review addresses, in depth, the structural and mechanistic aspects of oxidative depolymerization of cellulose by PMOs and considers their biological function and phylogenetic diversity. PMID:25784051

  10. Structural characterization and DPPH· radical scavenging activity of a polysaccharide from Guara fruits.

    PubMed

    Hua, Dehong; Zhang, Dezhi; Huang, Bing; Yi, Pan; Yan, Chunyan

    2014-03-15

    The crude polysaccharides were extracted from fruits of Psidium guajava Linn. by hot water. After removal of proteins, isolation and purification by DEAE-52 Cellulose chromatography and Sephadex G-75 gel filtration, a polysaccharide (GP70-2) was obtained and structurally characterized. GP70-2 has a relative molecular weight of 74 kDa and was composed of D-galactose and L-arabinose in the ratio of 1:1, with a specific optical rotation of [a]D(25) = +101°. Structural characterization of this novel polysaccharide was carried out using infrared spectroscopy, methylation analyses, and NMR studies ((1)H, (13)C, (1)H-(1)H-COSY, HMQC, and HMBC). Based on the above data, the following structure was assigned to the repeated core unit of GP70-2: [Formula: see text]. This polysaccharide showed a concentration dependent DPPH· radical scavenging activity.

  11. Formation of water-soluble soybean polysaccharides from spent flakes by hydrogen peroxide treatment.

    PubMed

    Pierce, Brian C; Wichmann, Jesper; Tran, Tam H; Cheetamun, Roshan; Bacic, Antony; Meyer, Anne S

    2016-06-25

    In this paper we propose a novel chemical process for the generation of water-soluble polysaccharides from soy spent flake, a by-product of the soy food industry. This process entails treatment of spent flake with hydrogen peroxide at an elevated temperature, resulting in the release of more than 70% of the original insoluble material as high molar mass soluble polysaccharides. A design of experiment was used to quantify the effects of pH, reaction time, and hydrogen peroxide concentration on the reaction yield, average molar mass, and free monosaccharides generated. The resulting product is low in protein, fat, and minerals and contains predominantly water-soluble polysaccharides of high molar mass, including arabinan, type I arabinogalactan, homogalacturonan, xyloglucan, rhamnogalacturonan, and (glucurono)arabinoxylan. This treatment provides a straightforward approach for generation of soluble soy polysaccharides and opens a new range of opportunities for this abundant and underutilized material in future research and industrial applications. PMID:27083842

  12. Rapeseed polysaccharides as prebiotics on growth and acidifying activity of probiotics in vitro.

    PubMed

    Wang, Xiao; Huang, Meiying; Yang, Fan; Sun, Hanju; Zhou, Xianxuan; Guo, Ying; Wang, Xiaoli; Zhang, Manli

    2015-07-10

    In vitro digestibility, prebiotic activity and chemical composition of polysaccharides from rapeseed were deliberately studied in this paper. After preliminary treatments, two fractions of polysaccharides (RP1 and RP2) were obtained after purification by DEAE-cellulose and Sephadex G-100. Their primary structural feature and molecule weights were characterized. Furthermore, their digestibility was also evaluated by artificial gastric juice and α-amylase. Finally, their proliferative effect on bifidobacteria and lactobacilli and acid production of the resulting probiotics in vitro were investigated. The results showed that RP1 and RP2 were homogeneously protein-bound polysaccharides with molecular weights of 28.51 and 6.55 kDa, respectively. They were resistant to hydrolysis by artificial gastric juice and α-amylase. Moreover, they could also significantly stimulate the tested probiotics to proliferate and produce organic acids. These findings clearly suggest the polysaccharides from rapeseed are potential to be exploited as novel prebiotics.

  13. Apoptosis-induced anti-tumor effect of Curcuma kwangsiensis polysaccharides against human nasopharyngeal carcinoma cells.

    PubMed

    Zeng, Jianhong; Dai, Ping; Ren, Lingyun; Song, Bo; Chen, Xu; Wang, Xiaohua; Wang, Jianhong; Zhang, Tiecheng; Zhu, Weiguo

    2012-08-01

    This study aimed to investigate the effect of Curcuma kwangsiensis polysaccharides on the viability of human nasopharyngeal carcinoma cell line CNE-2 cells, and explore the possible mechanisms. CNE-2 cells were treated with various concentrations of C. kwangsiensis polysaccharides, then the proliferation, apoptosis and the protein expression of apoptosis-related regulators p53 and Bcl-2 were assessed. The results demonstrated that C. kwangsiensis polysaccharides can significantly inhibit the proliferation of CNE-2 cells, which was possibly through the induction of apoptosis mediated by attenuating Bcl-2 expression and promoting p53 expression. The present study therefore indicates that C. kwangsiensis polysaccharides could be developed into potential drugs for nasopharyngeal carcinoma treatment.

  14. Marine Polysaccharides in Pharmaceutical Applications: An Overview

    PubMed Central

    Laurienzo, Paola

    2010-01-01

    The enormous variety of polysaccharides that can be extracted from marine plants and animal organisms or produced by marine bacteria means that the field of marine polysaccharides is constantly evolving. Recent advances in biological techniques allow high levels of polysaccharides of interest to be produced in vitro. Biotechnology is a powerful tool to obtain polysaccharides from a variety of micro-organisms, by controlling the growth conditions in a bioreactor while tailoring the production of biologically active compounds. Following an overview of the current knowledge on marine polysaccharides, with special attention to potential pharmaceutical applications and to more recent progress on the discovering of new polysaccharides with biological appealing characteristics, this review will focus on possible strategies for chemical or physical modification aimed to tailor the final properties of interest. PMID:20948899

  15. Polysaccharide-based nanocomposites and their applications

    PubMed Central

    Zheng, Yingying; Monty, Jonathan; Linhardt, Robert J.

    2014-01-01

    Polysaccharide nanocomposites have become increasingly important materials over the past decade. Polysaccharides offer a green alternative to synthetic polymers in the preparation of soft nanomaterials. They have also been used in composites with hard nanomaterials, such as metal nanoparticles and carbon-based nanomaterials. This mini review describes methods for polysaccharide nanocomposite preparation and reviews the various types and diverse applications for these novel materials. PMID:25498200

  16. Advances on Bioactive Polysaccharides from Medicinal Plants.

    PubMed

    Xie, Jian-Hua; Jin, Ming-Liang; Morris, Gordon A; Zha, Xue-Qiang; Chen, Han-Qing; Yi, Yang; Li, Jing-En; Wang, Zhi-Jun; Gao, Jie; Nie, Shao-Ping; Shang, Peng; Xie, Ming-Yong

    2016-07-29

    In recent decades, the polysaccharides from the medicinal plants have attracted a lot of attention due to their significant bioactivities, such as anti-tumor activity, antioxidant activity, anticoagulant activity, antidiabetic activity, radioprotection effect, anti-viral activity, hypolipidemic and immunomodulatory activities, which make them suitable for medicinal applications. Previous studies have also shown that medicinal plant polysaccharides are non-toxic and show no side effects. Based on these encouraging observations, most researches have been focusing on the isolation and identification of polysaccharides, as well as their bioactivities. A large number of bioactive polysaccharides with different structural features and biological effects from medicinal plants have been purified and characterized. This review provides a comprehensive summary of the most recent developments in physiochemical, structural features and biological activities of bioactive polysaccharides from a number of important medicinal plants, such as polysaccharides from Astragalus membranaceus, Dendrobium plants, Bupleurum, Cactus fruits, Acanthopanax senticosus, Angelica sinensis (Oliv.) Diels, Aloe barbadensis Miller, and Dimocarpus longan Lour. Moreover, the paper has also been focused on the applications of bioactive polysaccharides for medicinal applications. Recent studies have provided evidence that polysaccharides from medicinal plants can play a vital role in bioactivities. The contents and data will serve as a useful reference material for further investigation, production, and application of these polysaccharides in functional foods and therapeutic agents.

  17. Advances on Bioactive Polysaccharides from Medicinal Plants.

    PubMed

    Xie, Jian-Hua; Jin, Ming-Liang; Morris, Gordon A; Zha, Xue-Qiang; Chen, Han-Qing; Yi, Yang; Li, Jing-En; Wang, Zhi-Jun; Gao, Jie; Nie, Shao-Ping; Shang, Peng; Xie, Ming-Yong

    2016-07-29

    In recent decades, the polysaccharides from the medicinal plants have attracted a lot of attention due to their significant bioactivities, such as anti-tumor activity, antioxidant activity, anticoagulant activity, antidiabetic activity, radioprotection effect, anti-viral activity, hypolipidemic and immunomodulatory activities, which make them suitable for medicinal applications. Previous studies have also shown that medicinal plant polysaccharides are non-toxic and show no side effects. Based on these encouraging observations, most researches have been focusing on the isolation and identification of polysaccharides, as well as their bioactivities. A large number of bioactive polysaccharides with different structural features and biological effects from medicinal plants have been purified and characterized. This review provides a comprehensive summary of the most recent developments in physiochemical, structural features and biological activities of bioactive polysaccharides from a number of important medicinal plants, such as polysaccharides from Astragalus membranaceus, Dendrobium plants, Bupleurum, Cactus fruits, Acanthopanax senticosus, Angelica sinensis (Oliv.) Diels, Aloe barbadensis Miller, and Dimocarpus longan Lour. Moreover, the paper has also been focused on the applications of bioactive polysaccharides for medicinal applications. Recent studies have provided evidence that polysaccharides from medicinal plants can play a vital role in bioactivities. The contents and data will serve as a useful reference material for further investigation, production, and application of these polysaccharides in functional foods and therapeutic agents. PMID:26463231

  18. Recombinant expression of Streptococcus pneumoniae capsular polysaccharides in Escherichia coli

    PubMed Central

    Kay, Emily J.; Yates, Laura E.; Terra, Vanessa S.; Cuccui, Jon; Wren, Brendan W.

    2016-01-01

    Currently, Streptococcus pneumoniae is responsible for over 14 million cases of pneumonia worldwide annually, and over 1 million deaths, the majority of them children. The major determinant for pathogenesis is a polysaccharide capsule that is variable and is used to distinguish strains based on their serotype. The capsule forms the basis of the pneumococcal polysaccharide vaccine (PPV23) that contains purified capsular polysaccharide from 23 serotypes, and the pneumococcal conjugate vaccine (PCV13), containing 13 common serotypes conjugated to CRM197 (mutant diphtheria toxin). Purified capsule from S. pneumoniae is required for pneumococcal conjugate vaccine production, and costs can be prohibitively high, limiting accessibility of the vaccine in low-income countries. In this study, we demonstrate the recombinant expression of the capsule-encoding locus from four different serotypes of S. pneumoniae within Escherichia coli. Furthermore, we attempt to identify the minimum set of genes necessary to reliably and efficiently express these capsules heterologously. These E. coli strains could be used to produce a supply of S. pneumoniae serotype-specific capsules without the need to culture pathogenic bacteria. Additionally, these strains could be applied to synthetic glycobiological applications: recombinant vaccine production using E. coli outer membrane vesicles or coupling to proteins using protein glycan coupling technology. PMID:27110302

  19. Polysaccharides from Chinese tea: recent advance on bioactivity and function.

    PubMed

    Cao, Hui

    2013-11-01

    Tea (Camellia sinensis) has a long history of medicinal use in Asian countries such as China, Japan, India and Thailand as ancient as 500,000 years ago. Tea is globally one of the most popular and lowest cost beverages, next only to water. Tea leaves are popularly consumed with unfermented (green tea), semi-fermented (oolong tea), and fermented (black and puerh) forms. The chemical composition of tea mainly includes polyphenols (TPP), proteins, polysaccharides (TPS), chlorophyll, and alkaloids. Great advances have been made in chemical and bioactive studies of catechins and polyphenols from tea in recent decades. However, polysaccharides from tea materials have received much less consideration than TPP. Recently, TPS from tea leaves and flowers have attracted great interest. The number of relevant publications has increased rapidly in recent years. Herein, the bioactivities and function aspects of TPS from Chinese tea were reviewed. PMID:23994784

  20. Incorporation of bacterial extracellular polysaccharide by black fly larvae (Simuliidae)

    USGS Publications Warehouse

    Couch, C.A.; Meyer, J.L.; Hall, R.O.

    1996-01-01

    Black fly larvae (Simulium) assimilated, with high efficiency (80-90%), bacterial extracellular polysaccharide (EPS) extracted from laboratory cultures of a pseudomonad isolated from the Ogeechee River. Incorporation was traced using 13C-labelled EPS offered to larvae as a coating on a mixture of 1-??m latex beads and kaolin particles. These EPS-coated particles were used to simulate natural particles, both living and dead. Solubility, protein, and nitrogen content of the EPS suggested it was a slime rather than a capsular polysaccharide. Glycosyl composition of the EPS was glucose and galactose in ?? and ?? linkages, with pyruvate, succinate, and possibly malonate constituent groups. To evaluate the incorporation of C derived from protein associated with the EPS matrix, feeding experiments were conducted using EPS with and without proteins extracted. Black fly larvae incorporated 7.2 ??g EPS C larva-1 d-1 from EPS that did not have proteins extracted, and 19.5 ??g EPS C larva-1 d-1 from EPS with proteins extracted. Carbon in protein that is typically associated with EPS was not solely or selectively incorporated. EPS incorporation rates are similar to rates of cellular bacterial carbon incorporation previously estimated for Ogeechee River black fly larvae. If EPS is generally available as a food resource, the importance of bacteria in detrital food webs may be underestimated by studies that examine only the consumption of bacterial cells.

  1. Serological, chemical, and structural analyses of the Escherichia coli cross-reactive capsular polysaccharides K13, K20, and K23.

    PubMed Central

    Vann, W F; Soderstrom, T; Egan, W; Tsui, F P; Schneerson, R; Orskov, I; Orskov, F

    1983-01-01

    The Escherichia coli K13, K20, and K23 capsular polysaccharide antigens are serologically related. All of these polysaccharides contain ribose and 2-keto-3-deoxyoctonate in equimolar quantities. The K13 and K20 polysaccharides are partially O-acetylated. A comparison of these polysaccharides after O-deacetylation, by nuclear magnetic resonance and permethylation analysis, showed that these polysaccharides contained the disaccharide repeat unit leads to)-beta-ribofuranosyl-(1 leads to 7)-beta-2-keto-3-deoxyoctonate. They differed in the presence and location of an acetyl moiety. The K13 polysaccharide was O-acetylated at C-4 of the 2-keto-3-deoxyoctonate. The K20 antigen was O-acetylated at C-5 of the ribose moiety. The K23 polymer was nonacetylated. The cross-reactivity of these antigens was demonstrated by tandem-crossed immunoelectrophoresis. Antibodies to K23 could be completely absorbed from OK K23 serum by K13, K20, and K23 antigenic extracts. The K13 and K20 antibodies could be completely absorbed from their respective antisera only by homologous antigenic extracts. Monoclonal antibodies were prepared against a protein conjugate of the K13 polysaccharide. Analyses of the reactions of these antibodies with the three polysaccharides suggest that the K13 polysaccharide has at least three antigenic sites, one of which is common to the K13, K20, and K23 polysaccharides. PMID:6187684

  2. Immunologically active polysaccharides of Arnica montana cell cultures.

    PubMed

    Puhlmann, J; Zenk, M H; Wagner, H

    1991-01-01

    From the nutrition medium of Arnica montana cell cultures two homogeneous polysaccharides, an acidic arabino-3,6-galactan-protein with mean Mr of 100,000 and a neutral fucogalactoxyloglucan with mean Mr of 22,500 have been isolated by DEAE-Sepharose CL-6B and Sephacryl S-400 column chromatography. Their structures were elucidated mainly by methylation analysis, partial acidic and enzymatic hydrolysis and 13C NMR spectroscopy. The fucogalactoxyloglucan shows a pronounced enhancement of phagocytosis in vivo. The arabino-3,6-galactan-protein displays a strong anticomplementary effect and stimulates macrophages to excrete the tumour necrosis factor (TNF alpha).

  3. Starch-degrading polysaccharide monooxygenases.

    PubMed

    Vu, Van V; Marletta, Michael A

    2016-07-01

    Polysaccharide degradation by hydrolytic enzymes glycoside hydrolases (GHs) is well known. More recently, polysaccharide monooxygenases (PMOs, also known as lytic PMOs or LPMOs) were found to oxidatively degrade various polysaccharides via a copper-dependent hydroxylation. PMOs were previously thought to be either GHs or carbohydrate binding modules (CBMs), and have been re-classified in carbohydrate active enzymes (CAZY) database as auxiliary activity (AA) families. These enzymes include cellulose-active fungal PMOs (AA9, formerly GH61), chitin- and cellulose-active bacterial PMOs (AA10, formerly CBM33), and chitin-active fungal PMOs (AA11). These PMOs significantly boost the activity of GHs under industrially relevant conditions, and thus have great potential in the biomass-based biofuel industry. PMOs that act on starch are the latest PMOs discovered (AA13), which has expanded our perspectives in PMOs studies and starch degradation. Starch-active PMOs have many common structural features and biochemical properties of the PMO superfamily, yet differ from other PMO families in several important aspects. These differences likely correlate, at least in part, to the differences in primary and higher order structures of starch and cellulose, and chitin. In this review we will discuss the discovery, structural features, biochemical and biophysical properties, and possible biological functions of starch-active PMOs, as well as their potential application in the biofuel, food, and other starch-based industries. Important questions regarding various aspects of starch-active PMOs and possible economical driving force for their future studies will also be highlighted. PMID:27170366

  4. Solution NMR spectroscopy of food polysaccharides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many polysaccharides are allowed for direct food use, where they serve a number of useful functions. In addition to possibly being a source of calories, a food polysaccharide may be a dietary fiber, bulking agent, crystallization inhibitor, thickener, encapsulant, gelling agent, foam and emulsion s...

  5. Bioactive polysaccharides and gut microbiome (abstract)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many polysaccharides have shown the ability to reduce plasma cholesterol or postprandial glycemia. Viscosity in the small intestine seems to be required to slow glucose uptake. Cereal mixed linkage beta-glucans, psyllium, glucomannans, and other polysaccharides also seem to require higher molecula...

  6. Applications of Important Polysaccharides in Drug Delivery.

    PubMed

    Huang, Gangliang; Mei, Xinya; Xiao, Feng; Chen, Xin; Tang, Qilin; Peng, Daquan

    2015-01-01

    Polysaccharide is a kind of biological material, which has good biocompatibility, non-toxicity, and non-immunogenicity. So, the polysaccharide has widely been applied in drug delivery system. The applications of chitosan, hyaluronic acid and dextran in drug delivery have been summarized herein. PMID:25578889

  7. Characterization, antioxidant and antitumor activities of polysaccharides from purple sweet potato.

    PubMed

    Wu, Qiongying; Qu, Hongsen; Jia, Junqiang; Kuang, Cong; Wen, Yan; Yan, Hui; Gui, Zhongzheng

    2015-11-01

    Three polysaccharides, PSPP1-1, PSPP2-1 and PSPP3-1, were isolated from purple sweet potato. The three polysaccharides belonged to β-type polysaccharides and contained low proportions of proteins and uronic acids. PSPP1-1 and PSPP3-1 with molecular weights of 33.3 and 75.3 kDa, respectively, were composed of rhamnose, xylose, glucose and galactose, whereas PSPP2-1 with molecular weight of 17.8 kDa was composed of rhamnose and galactose. The three polysaccharides possessed in vitro antioxidant (scavenging DPPH radicals, chelating ferrous ions and reducing power) and antitumor activities (against SGC7901 and SW620 cells) in a dose-dependent manner. Among the three polysaccharides, PSPP2-1 exhibited the strongest reducing power, scavenging activity on DPPH radicals and chelating capability on ferrous ions. PSPP1-1 showed the strongest inhibitory activities on the growth of SGC7901 and SW620 cells. In addition, flow cytometry results showed that PSPP1-1 could induce apoptosis in SGC7901 and SW620 cells. These results suggest that polysaccharides from purple sweet potato are potential natural antioxidant and antitumor agents that can be used as drugs or functional food ingredients. PMID:26256321

  8. Characterization of pectic polysaccharides extracted from apple pomace by hot-compressed water.

    PubMed

    Wang, Xin; Lü, Xin

    2014-02-15

    Response surface methodology (RSM) was used to optimize the extraction of pectic polysaccharides from apple pomace by hot-compressed water, by which the optimum levels of the parameters were obtained as follows: extraction temperature 140 °C, extraction time 5 min, S:W ratio 1:14. Compared with commercial pectin, the Mw, galacturonic acid content, DM and protein of the extracted pectic polysaccharides were lower while ash content and neutral sugars were higher. The endothermic transition temperature and fusion heat of the extracted pectic polysaccharides was lower than commercial one according to DSC analysis. For its rheological properties, it was found that the viscosity of the extracted pectic polysaccharides solution was slightly lower than commercial pectin at lower shear rate region while it decreased sharply when the shear rate increased. Besides, both G' and G" moduli of the extracted pectic polysaccharides were lower than the commercial pectin's possibly because of weaker polymer chain interaction, which was also reflected in gel textural properties. However, the extracted pectic polysaccharides showed higher in vitro antioxidant capability and inhibitory effect on HT-29 colon adenocarcinoma cells than commercial pectin.

  9. Corn pollen polysaccharides: composition of radiation-resistant nutrients and bioactivity

    NASA Astrophysics Data System (ADS)

    Lu, Weihong; Wenxin, Gao; Sun, Yeqing

    Corn pollen contains significant levels of free amino acids and protein, which greatly contribute to the biological function of corn pollen. However, to date there is no report in either China or abroad on research regarding the specific radiation-resistant composition in corn pollen includ-ing pollen polysaccharide. Reports on corn pollen polysaccharide have been mostly focused on immunological competence and anti-tumor functions. This study emphasized the optimization of the technical conditions for the extraction of corn pollen polysaccharide and the analysis of the corn pollen polysaccharide's structure. On that basis, we have developed in vitro experi-ments with corn pollen polysaccharide and report on its antioxidant functional activity. Our innovation lies in defining the specific composition of the radiation-resistant nutrients and active compounds as well as identifying the structure of the active compounds. We have successfully separated the active radiation-resistant functional factors, which are of great significance for astronauts and other special groups. Our results lay the groundwork for further research and development of corn pollen polysaccharide and ingredient technology.

  10. Effects of extraction methods on the antioxidant activities of polysaccharides from Agaricus blazei Murrill.

    PubMed

    Jia, Shaoyi; Li, Feng; Liu, Yong; Ren, Haitao; Gong, Guili; Wang, Yanyan; Wu, Songhai

    2013-11-01

    Five polysaccharides were obtained from Agaricus blazei Murrill (ABM) through different extraction methods including hot water extraction, single enzyme extraction (pectinase, cellulase or papain) and compound enzymes extraction (cellulase:pectinase:papain). Their characteristics such as the polysaccharide yield, polysaccharide content, protein content, infrared spectra were determined, and antioxidant activities were investigated on the basis of hydroxyl radical, DPPH free radical, ABTS free radical and reducing power. The results showed that five extracts exhibited antioxidant activities in a concentration-dependent manner. Compared with other methods, the compound enzymes extraction method was found to present the highest polysaccharides yield (17.44%). Moreover, compound enzymes extracts exhibited the strongest reducing power and highest scavenging rates on hydroxyl radicals, DPPH radicals and ABTS radicals. On the contrary, hot water extraction method had the lowest polysaccharides yield of 11.95%, whose extracts also exhibited the lowest antioxidant activities. Overall, the available data obtained in vitro models suggested that ABM extracts were natural antioxidants and compound enzymes extraction was an appropriate, mild and effective extracting method for obtaining the polysaccharide extracts from Agaricus blazei Murrill (ABM).

  11. Isolation and antiviral activity of water-soluble Cynomorium songaricum Rupr. polysaccharides.

    PubMed

    Tuvaanjav, Suvdmaa; Shuqin, Han; Komata, Masashi; Ma, Chunjie; Kanamoto, Taisei; Nakashima, Hideki; Yoshida, Takashi

    2016-01-01

    The plant, Cynomorium songaricum Rupr., is used as a traditional medicine in China and Mongolia. In the present study, two new water-soluble polysaccharides isolated from C. songaricum Rupr. were purified by successive Sephadex G-75 and G-50 column chromatographies and then characterized by high resolution NMR and IR spectroscopies. The molecular weights of two polysaccharides were determined by an aqueous GPC to be [Formula: see text] = 3.7 × 10(4) and 1.0 × 10(4), respectively. In addition, it was found that the polysaccharide with the larger molecular weight was an acidic polysaccharide. It was found that the iodine-starch reaction of both isolated polysaccharides was negative and the methylation analysis gave 2, 4, 6-tri-O-methyl alditol acetate as a main product. NMR and IR measurements and sugar analysis revealed that both polysaccharides had a (1 → 3)-α-d-glucopyranosidic main chain with a small number of branches. After sulfation, the sulfated C. songaricum Rupr. polysaccharides were found to have a potent inhibitory effect on HIV infection of MT-4 cells at a 50% effective concentration of 0.3-0.4 μg/ml, a concentration that has almost the same high activity as standard dextran and curdlan sulfates, EC50 = 0.35 and 0.14 μg/ml, respectively. The 50% cytotoxic concentration was low, CC50>1000 μg/ml. In addition, the interaction between the sulfated polysaccharides and poly-l-lysine as a model protein compound was investigated by a surface plasmon resonance to reveal the anti-HIV mechanism. PMID:26838028

  12. Thermo-mechanical and hydrophilic properties of polysaccharide/gluten-based bioplastics.

    PubMed

    Zárate-Ramírez, L S; Romero, A; Bengoechea, C; Partal, P; Guerrero, A

    2014-11-01

    The influence of adding different polysaccharides (locust bean gum, LBG; methyl cellulose, MC; and carboxymethyl cellulose, CMC) to gluten-based biodegradable polymeric materials was assessed in this work. Gluten/polysaccharide/plasticiser bioplastics were prepared at different polysaccharide concentrations (0-4.5%) and pH values by mixing in a two-blade counter-rotating batch mixer (at 25 °C under adiabatic conditions) and thermomoulding at 9MPa and 130 °C. Bioplastic probes were evaluated through dynamic mechanical thermal analysis, tensile strength and water absorption capacity tests. Results pointed out that a moderate enhancement of the network structure may be achieved by adding polysaccharide at a pH close to the protein isoelectric point (pH 6), which also conferred a further thermosetting capacity to the system. Moreover, the addition of MC and CMC was found to significantly enhance material elongation properties. However, the presence of charges induced by pH leaded to a higher incompatibility between the polysaccharide and protein domains forming the composite. The pH value played a relevant role in the material water absorption, which significantly increased under acidic or basic conditions (particularly at pH 3). PMID:25129712

  13. Thermo-mechanical and hydrophilic properties of polysaccharide/gluten-based bioplastics.

    PubMed

    Zárate-Ramírez, L S; Romero, A; Bengoechea, C; Partal, P; Guerrero, A

    2014-11-01

    The influence of adding different polysaccharides (locust bean gum, LBG; methyl cellulose, MC; and carboxymethyl cellulose, CMC) to gluten-based biodegradable polymeric materials was assessed in this work. Gluten/polysaccharide/plasticiser bioplastics were prepared at different polysaccharide concentrations (0-4.5%) and pH values by mixing in a two-blade counter-rotating batch mixer (at 25 °C under adiabatic conditions) and thermomoulding at 9MPa and 130 °C. Bioplastic probes were evaluated through dynamic mechanical thermal analysis, tensile strength and water absorption capacity tests. Results pointed out that a moderate enhancement of the network structure may be achieved by adding polysaccharide at a pH close to the protein isoelectric point (pH 6), which also conferred a further thermosetting capacity to the system. Moreover, the addition of MC and CMC was found to significantly enhance material elongation properties. However, the presence of charges induced by pH leaded to a higher incompatibility between the polysaccharide and protein domains forming the composite. The pH value played a relevant role in the material water absorption, which significantly increased under acidic or basic conditions (particularly at pH 3).

  14. Enzymatic method for improving the injectability of polysaccharides

    DOEpatents

    Griffith, William L.; Compere, Alicia L.; Holleman, James W.

    1982-01-01

    A method for enhancing the ability of polysaccharides in aqueous solution to flow through a porous medium comprises contacting the polysaccharides with an endoenzyme capable of hydrolyzing at least one of the linkages of the sugar units of the polysaccharides and maintaining the polysaccharides in contact with the enzyme under hydrolysis conditions for a time sufficient to decrease the tendency of the polysaccharides to plug the porous medium yet insufficient to decrease the viscosity of the aqueous polysaccharides by more than 25%. The partially hydrolyzed polysaccharides are useful as thickening agents for flooding water used to recover oil from oil-containing subterranean formations.

  15. Extracellular polysaccharide production by thraustochytrid protists.

    PubMed

    Jain, Ruchi; Raghukumar, Seshagiri; Tharanathan, R; Bhosle, N B

    2005-01-01

    Four strains of marine stramenopilan protists, the thraustochytrids, were studied for their ability to produce extracellular polysaccharides (EPSs). Observations by light and scanning electron microscopy revealed the production of a matrix of EPS around groups of cells in stationary cultures. EPS in shake culture filtrates ranged from 0.3 to 1.1 g/L. EPS production, which was studied in greater detail in 2 isolates, SC-1 and CW1, increased with age of cultures, reaching a peak in the stationary phase. Anion exchange chromatography yielded a single fraction of the EPS of both species. The EPS contained 39% to 53% sugars, besides proteins, lipids, uronic acids, and sulfates. Molecular weight of the EPS produced by SC-1 was approximately 94 kDa, and that of CW1, 320 kDa. Glucose formed the major component in the EPS of both isolates-galactose, mannose, and arabinose being the other components. Cultures of both isolates survived air-drying up to a period of 96 hours, suggesting a role for EPS in preventing desiccation of cells.

  16. Niches of two polysaccharide-degrading Polaribacter isolates from the North Sea during a spring diatom bloom

    PubMed Central

    Xing, Peng; Hahnke, Richard L; Unfried, Frank; Markert, Stephanie; Huang, Sixing; Barbeyron, Tristan; Harder, Jens; Becher, Dörte; Schweder, Thomas; Glöckner, Frank Oliver; Amann, Rudolf I; Teeling, Hanno

    2015-01-01

    Members of the flavobacterial genus Polaribacter thrive in response to North Sea spring phytoplankton blooms. We analyzed two respective Polaribacter species by whole genome sequencing, comparative genomics, substrate tests and proteomics. Both can degrade algal polysaccharides but occupy distinct niches. The liquid culture isolate Polaribacter sp. strain Hel1_33_49 has a 3.0-Mbp genome with an overall peptidase:CAZyme ratio of 1.37, four putative polysaccharide utilization loci (PULs) and features proteorhodopsin, whereas the agar plate isolate Polaribacter sp. strain Hel1_85 has a 3.9-Mbp genome with an even peptidase:CAZyme ratio, eight PULs, a mannitol dehydrogenase for decomposing algal mannitol-capped polysaccharides but no proteorhodopsin. Unlike other sequenced Polaribacter species, both isolates have larger sulfatase-rich PULs, supporting earlier assumptions that Polaribacter take part in the decomposition of sulfated polysaccharides. Both strains grow on algal laminarin and the sulfated polysaccharide chondroitin sulfate. For strain Hel1_33_49, we identified by proteomics (i) a laminarin-induced PUL, (ii) chondroitin sulfate-induced CAZymes and (iii) a chondroitin-induced operon that likely enables chondroitin sulfate recognition. These and other data suggest that strain Hel1_33_49 is a planktonic flavobacterium feeding on proteins and a small subset of algal polysaccharides, while the more versatile strain Hel1_85 can decompose a broader spectrum of polysaccharides and likely associates with algae. PMID:25478683

  17. Trapped translocation intermediates establish the route for export of capsular polysaccharides across Escherichia coli outer membranes.

    PubMed

    Nickerson, Nicholas N; Mainprize, Iain L; Hampton, Lauren; Jones, Michelle L; Naismith, James H; Whitfield, Chris

    2014-06-01

    The outer membrane (OM) of Gram-negative bacteria is designed to exclude potentially harmful molecules. This property presents a challenge for bacteria that must secrete proteins and large glycoconjugates to grow, divide, and persist. Proteins involved in trafficking such molecules have been identified, but their precise roles are often unresolved due to the difficulty in capturing "snapshots" during the export pathway. Wza is the prototype for the large family of OM polysaccharide export proteins. In Escherichia coli, Wza is essential for the assembly of a capsule, a protective surface coat composed of long-chain polysaccharides. Wza creates an octameric α-helical channel spanning the OM, but the bulk of the protein exists as a large periplasmic structure enclosing an extensive lumen. Residues within the lumen of Wza were targeted for site-specific incorporation of the UV photo-cross-linkable unnatural amino acid p-benzoyl-L-phenylalanine. Using this in vivo photo-cross-linking strategy, we were able to trap polysaccharide translocation intermediates within the lumen of Wza, providing the first unequivocal evidence to our knowledge that nascent capsular polysaccharide chains exit the cell through the Wza portal.

  18. Polysaccharides and bacterial plugging. [Leuconostoc bacteria

    SciTech Connect

    Fogler, H.S.

    1992-01-01

    The objectives of this research are to elucidate and model bacterial transport in porous media, to determine the importance of polysaccharides bridging as a retentive mechanism, and to identify key parameters that influence porous media plugging. This project has been subdivided into three tasks: Task 1 is the determination of the growth kinetics of the Leuconostoc bacteria and how they are affected by (1) the nutrient feed, and (2) surface effects; Task 2 will quantify the importance of polysaccharide production as a cell retention mechanism; and Task 3 is the elucidation of the rate of polysaccharide production and the combined effect that polysaccharide production and cell growth has upon plugging. Batch growth experiments have been conducted to determine the rate of cell growth, polymer production and nutrient consumption. The data from these experiments are currently being reduced to aid the development of an overall growth model.

  19. Quantitative Structure Activity Relationship Models for the Antioxidant Activity of Polysaccharides

    PubMed Central

    Nie, Kaiying; Wang, Zhaojing

    2016-01-01

    In this study, quantitative structure activity relationship (QSAR) models for the antioxidant activity of polysaccharides were developed with 50% effective concentration (EC50) as the dependent variable. To establish optimum QSAR models, multiple linear regressions (MLR), support vector machines (SVM) and artificial neural networks (ANN) were used, and 11 molecular descriptors were selected. The optimum QSAR model for predicting EC50 of DPPH-scavenging activity consisted of four major descriptors. MLR model gave EC50 = 0.033Ara-0.041GalA-0.03GlcA-0.025PC+0.484, and MLR fitted the training set with R = 0.807. ANN model gave the improvement of training set (R = 0.96, RMSE = 0.018) and test set (R = 0.933, RMSE = 0.055) which indicated that it was more accurately than SVM and MLR models for predicting the DPPH-scavenging activity of polysaccharides. 67 compounds were used for predicting EC50 of the hydroxyl radicals scavenging activity of polysaccharides. MLR model gave EC50 = 0.12PC+0.083Fuc+0.013Rha-0.02UA+0.372. A comparison of results from models indicated that ANN model (R = 0.944, RMSE = 0.119) was also the best one for predicting the hydroxyl radicals scavenging activity of polysaccharides. MLR and ANN models showed that Ara and GalA appeared critical in determining EC50 of DPPH-scavenging activity, and Fuc, Rha, uronic acid and protein content had a great effect on the hydroxyl radicals scavenging activity of polysaccharides. The antioxidant activity of polysaccharide usually was high in MW range of 4000–100000, and the antioxidant activity could be affected simultaneously by other polysaccharide properties, such as uronic acid and Ara. PMID:27685320

  20. A proteomic analysis of mushroom polysaccharide-treated HepG2 cells.

    PubMed

    Chai, Yangyang; Wang, Guibin; Fan, Lili; Zhao, Min

    2016-01-01

    The anti-tumor properties of fungal polysaccharides have gained significant recognition in Asia and tropical America. In this study, the differential expression of proteins in normal HepG2 cells and those treated with polysaccharides that had been isolated from Phellinus linteus (PL), Ganoderma lucidum (GL) and Auricularia auricula (AA) was investigated. Using two-dimensional electrophoresis (2DE), a total of 104 protein spots were determined to be overexpressed in these cells compared with noncancerous regions. A total of 59 differentially expressed proteins were identified through MALDI-TOF-MS. In addition, 400 biological processes (BP), 133 cell components (CC) and 146 molecular functions (MF) were enriched by Gene Ontology (GO) analysis, and 78 KEGG pathways were enriched by pathway enrichment. Protein-Protein Interaction (PPI) analysis demonstrated the interaction networks affected by polysaccharides in HepG2 cells. Then, DJ-1 and 14-3-3 were identified as the key proteins in the networks, and the expression of the mRNA and proteins were evaluated using Real-time quantitative PCR (qRT-PCR) and Western blotting (WB), respectively. The results were in agreement with the 2DE. These results provided information on significant proteins of hepatocellular carcinoma (HCC) and form an important basis for the future development of valuable medicinal mushroom resources. PMID:27020667

  1. A proteomic analysis of mushroom polysaccharide-treated HepG2 cells

    PubMed Central

    Chai, Yangyang; Wang, Guibin; Fan, Lili; Zhao, Min

    2016-01-01

    The anti-tumor properties of fungal polysaccharides have gained significant recognition in Asia and tropical America. In this study, the differential expression of proteins in normal HepG2 cells and those treated with polysaccharides that had been isolated from Phellinus linteus (PL), Ganoderma lucidum (GL) and Auricularia auricula (AA) was investigated. Using two-dimensional electrophoresis (2DE), a total of 104 protein spots were determined to be overexpressed in these cells compared with noncancerous regions. A total of 59 differentially expressed proteins were identified through MALDI-TOF-MS. In addition, 400 biological processes (BP), 133 cell components (CC) and 146 molecular functions (MF) were enriched by Gene Ontology (GO) analysis, and 78 KEGG pathways were enriched by pathway enrichment. Protein-Protein Interaction (PPI) analysis demonstrated the interaction networks affected by polysaccharides in HepG2 cells. Then, DJ-1 and 14-3-3 were identified as the key proteins in the networks, and the expression of the mRNA and proteins were evaluated using Real-time quantitative PCR (qRT-PCR) and Western blotting (WB), respectively. The results were in agreement with the 2DE. These results provided information on significant proteins of hepatocellular carcinoma (HCC) and form an important basis for the future development of valuable medicinal mushroom resources. PMID:27020667

  2. Polysaccharides and virulence of Burkholderia pseudomallei.

    PubMed

    Sarkar-Tyson, M; Thwaite, J E; Harding, S V; Smither, S J; Oyston, P C F; Atkins, T P; Titball, R W

    2007-08-01

    Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease of humans and animals. Gene clusters which encode capsular polysaccharide (type I O-PS) and LPS (type II O-PS), both of which play roles in virulence, have previously been identified. Here, the identification of two further putative clusters, type III O-PS and type IV O-PS, is reported. Mice challenged with type III O-PS or type IV O-PS mutants showed increased mean times to death (7.8 and 11.6 days) compared to those challenged with wild-type B. pseudomallei (3 days). To investigate the possible roles of polysaccharides in protection, mice were immunized with killed cells of wild-type B. pseudomallei or killed cells of B. pseudomallei with mutations in the O antigen, capsular polysaccharide, type III O-PS or type IV O-PS gene clusters. Immunization with all polysaccharide mutant strains resulted in delayed time to death compared to the naïve controls, following challenge with wild-type B. pseudomallei strain K96243. However, immunization with killed polysaccharide mutant strains conferred different degrees of protection, demonstrating the immunological importance of the polysaccharide clusters on the surface of B. pseudomallei.

  3. Monoclonal antibodies against plant cell wall polysaccharides

    SciTech Connect

    Hahn, M.G.; Bucheli, E.; Darvill, A.; Albersheim, P. )

    1989-04-01

    Monoclonal antibodies (McAbs) are useful tools to probe the structure of plant cell wall polysaccharides and to localize these polysaccharides in plant cells and tissues. Murine McAbs were generated against the pectic polysaccharide, rhamnogalacturonan I (RG-I), isolated from suspension-cultured sycamore cells. The McAbs that were obtained were grouped into three classes based upon their reactivities with a variety of plant polysaccharides and membrane glycoproteins. Eleven McAbs (Class I) recognize epitope(s) that appear to be immunodominant and are found in RG-I from sycamore and maize, citrus pectin, polygalacturonic acid, and membrane glycoproteins from suspension-cultured cells of sycamore, maize, tobacco, parsley, and soybean. A second group of five McAbs (Class II) recognize epitope(s) present in sycamore RG-I, but do not bind to any of the other polysaccharides or glycoproteins recognized by Class I. Lastly, one McAb (Class III) reacts with sycamore RG-I, sycamore and tamarind xyloglucan, and sycamore and rice glucuronoarabinoxylan, but does not bind to maize RG-I, polygalacturonic acid or the plant membrane glycoproteins recognized by Class I. McAbs in Classes II and III are likely to be useful in studies of the structure, biosynthesis and localization of plant cell wall polysaccharides.

  4. Structural Characterization of Arabidopsis Leaf Arabinogalactan Polysaccharides1[W

    PubMed Central

    Tryfona, Theodora; Liang, Hui-Chung; Kotake, Toshihisa; Tsumuraya, Yoichi; Stephens, Elaine; Dupree, Paul

    2012-01-01

    Proteins decorated with arabinogalactan (AG) have important roles in cell wall structure and plant development, yet the structure and biosynthesis of this polysaccharide are poorly understood. To facilitate the analysis of biosynthetic mutants, water-extractable arabinogalactan proteins (AGPs) were isolated from the leaves of Arabidopsis (Arabidopsis thaliana) plants and the structure of the AG carbohydrate component was studied. Enzymes able to hydrolyze specifically AG were utilized to release AG oligosaccharides. The released oligosaccharides were characterized by high-energy matrix-assisted laser desorption ionization-collision-induced dissociation mass spectrometry and polysaccharide analysis by carbohydrate gel electrophoresis. The Arabidopsis AG is composed of a β-(1→3)-galactan backbone with β-(1→6)-d-galactan side chains. The β-(1→6)-galactan side chains vary in length from one to over 20 galactosyl residues, and they are partly substituted with single α-(1→3)-l-arabinofuranosyl residues. Additionally, a substantial proportion of the β-(1→6)-galactan side chain oligosaccharides are substituted at the nonreducing termini with single 4-O-methyl-glucuronosyl residues via β-(1→6)-linkages. The β-(1→6)-galactan side chains are occasionally substituted with α-l-fucosyl. In the fucose-deficient murus1 mutant, AGPs lack these fucose modifications. This work demonstrates that Arabidopsis mutants in AGP structure can be identified and characterized. The detailed structural elucidation of the AG polysaccharides from the leaves of Arabidopsis is essential for insights into the structure-function relationships of these molecules and will assist studies on their biosynthesis. PMID:22891237

  5. Comparison of the preliminary characterizations and antioxidant properties of polysaccharides obtained from Phellinus baumii growth on different culture substrates.

    PubMed

    Zhang, Zuo-fa; Lv, Guo-ying; Song, Ting-ting; Jin, Qun-li; Huang, Jian-bo; Fan, Lei-fa; Cai, Wei-ming

    2015-11-01

    Three polysaccharides (PPB-MB, PPB-MW and PPB-MM) were obtained from the fruiting body of Phellinus baumii growth on different culture substrates (mulberry branches, mixed wood sawdust and an equal combination of the two materials) and their chemical composition was investigated. PPB-MM contained the highest contents of neutral sugar (66.59%) and uronic acid (23.38%), followed by PPB-MW and PPB-MB, with PPB-MW having the highest protein content. The three polysaccharides were all composed of six kinds of monosaccharides, namely fucose, mannose, glactose, xylose, arabinose and glucose. The antioxidant activities of the three polysaccharides were determined using lipid peroxidant inhibition, ABTS radical scavenging, and Fe(2+)-chelating assay. Results showed that PPB-MM exhibited the highest antioxidant properties in all the assays. As a result, an equal combination of mulberry branches and mixed wood sawdust serves as a good culture substrate for producing such antioxidant polysaccharides.

  6. Synthesis and characterization of Escherichia coli O18 O-polysaccharide conjugate vaccines.

    PubMed Central

    Cryz, S J; Cross, A S; Sadoff, J C; Fürer, E

    1990-01-01

    Nontoxic, serologically reactive O polysaccharide was derived from Escherichia coli O18 lipopolysaccharide by acid hydrolysis, extraction with organic solvents, and gel filtration chromatography. Oxidized O polysaccharide was covalently coupled to either Pseudomonas aeruginosa toxin A or cholera toxin by using adipic acid dihydrazide as a spacer molecule in the presence of carbodiimide. The resulting conjugates were composed of approximately equal amounts of O polysaccharide and protein and were nontoxic and nonpyrogenic. Both conjugates engendered an immunoglobulin G antibody response in rabbits that recognized native O18 lipopolysaccharide. Such antibody was able to promote the uptake and killing of an E. coli O18 strain bearing the K1 capsule by human polymorphonuclear leukocytes. Immunoglobulin G isolated from the sera of rabbits immunized with either conjugate afforded protection against an E. coli O18 challenge when passively transferred to mice. PMID:2105272

  7. Cellulase-assisted extraction of polysaccharides from Cucurbita moschata and their antibacterial activity.

    PubMed

    Qian, Zhi-Gang

    2014-01-30

    In this study, cellulase-assisted extraction of water soluble polysaccharides from pumpkin (Cucurbita moschata) and their antibacterial activity were investigated. The polysaccharides yield was monitored during the extraction process. The optimum extraction conditions were determined as follows: time, 40 min; temperature, 55°C; pH, 4.5; and cellulase amount, 4,000 U/g. The extracts were centrifuged, filtered, proteins removed by Sevag method, concentrated to approximately 15% (w/v), precipitated with 5 volumes of absolute ethanol, freeze-dried, and pulverized to yield a water soluble powder of pumpkin polysaccharides (PP). The sugar content of the product was 68.3%, and the yield was 17.34% (w/w), respectively. The PP had high antibacterial activity against Bacillus subtilis, Staphylococcus aureus, and Escherichia coli at the concentration of 100 mg/mL.

  8. Development and application of a suite of polysaccharide-degrading enzymes for analyzing plant cell walls

    PubMed Central

    Bauer, Stefan; Vasu, Prasanna; Persson, Staffan; Mort, Andrew J.; Somerville, Chris R.

    2006-01-01

    To facilitate analysis of plant cell wall polysaccharide structure and composition, we cloned 74 genes encoding polysaccharide-degrading enzymes from Aspergillus nidulans, Aspergillus fumigatus, and Neurospora crassa and expressed the genes as secreted proteins with C-terminal Myc and 6× His tags. Most of the recombinant enzymes were active in enzyme assays, and optima for pH and temperature were established. A subset of the enzymes was used to fragment polysaccharides from the irregular xylem 9 (irx9) mutant of Arabidopsis. The analysis revealed a decrease in the abundance of xylan in the mutant, indicating that the IRX9 gene, which encodes a putative family 43 glycosyltransferase, is required for xylan synthesis. PMID:16844780

  9. A biotechnological perspective on the application of iron oxide magnetic colloids modified with polysaccharides.

    PubMed

    Dias, A M G C; Hussain, A; Marcos, A S; Roque, A C A

    2011-01-01

    Iron oxide magnetic nanoparticles (MNPs) alone are suitable for a broad spectrum of applications, but the low stability and heterogeneous size distribution in aqueous medium represent major setbacks. These setbacks can however be reduced or diminished through the coating of MNPs with various polymers, especially biopolymers such as polysaccharides. Polysaccharides are biocompatible, non-toxic and renewable; in addition, they possess chemical groups that permit further functionalization of the MNPs. Multifunctional entities can be created through decoration with specific molecules e.g. proteins, peptides, drugs, antibodies, biomimetic ligands, transfection agents, cells, and other ligands. This development opens a whole range of applications for iron oxide nanoparticles. In this review the properties of magnetic structures composed of MNPs and several polysaccharides (Agarose, Alginate, Carrageenan, Chitosan, Dextran, Heparin, Gum Arabic, Pullulan and Starch) will be discussed, in view of their recent and future biomedical and biotechnological applications.

  10. Antioxidative mechanism of Lycium barbarum polysaccharides promotes repair and regeneration following cavernous nerve injury

    PubMed Central

    Zhao, Zhan-kui; Yu, Hong-lian; Liu, Bo; Wang, Hui; Luo, Qiong; Ding, Xie-gang

    2016-01-01

    Polysaccharides extracted from Lycium barbarum exhibit antioxidant properties. We hypothesized that these polysaccharides resist oxidative stress-induced neuronal damage following cavernous nerve injury. In this study, rat models were intragastrically administered Lycium barbarum polysaccharides for 2 weeks at 1, 7, and 14 days after cavernous nerve injury. Serum superoxide dismutase and glutathione peroxidase activities significantly increased at 1 and 2 weeks post-injury. Serum malondialdehyde levels decreased at 2 and 4 weeks. At 12 weeks, peak intracavernous pressure, the number of myelinated axons and nicotinamide adenine dinucleotide phosphate-diaphorase-positive nerve fibers, levels of phospho-endothelial nitric oxide synthase protein and 3-nitrotyrosine were higher in rats administered at 1 day post-injury compared with rats administered at 7 and 14 days post-injury. These findings suggest that application of Lycium barbarum polysaccharides following cavernous nerve crush injury effectively promotes nerve regeneration and erectile functional recovery. This neuroregenerative effect was most effective in rats orally administered Lycium barbarum polysaccharides at 1 day after cavernous nerve crush injury. PMID:27651780

  11. Antioxidative mechanism of Lycium barbarum polysaccharides promotes repair and regeneration following cavernous nerve injury.

    PubMed

    Zhao, Zhan-Kui; Yu, Hong-Lian; Liu, Bo; Wang, Hui; Luo, Qiong; Ding, Xie-Gang

    2016-08-01

    Polysaccharides extracted from Lycium barbarum exhibit antioxidant properties. We hypothesized that these polysaccharides resist oxidative stress-induced neuronal damage following cavernous nerve injury. In this study, rat models were intragastrically administered Lycium barbarum polysaccharides for 2 weeks at 1, 7, and 14 days after cavernous nerve injury. Serum superoxide dismutase and glutathione peroxidase activities significantly increased at 1 and 2 weeks post-injury. Serum malondialdehyde levels decreased at 2 and 4 weeks. At 12 weeks, peak intracavernous pressure, the number of myelinated axons and nicotinamide adenine dinucleotide phosphate-diaphorase-positive nerve fibers, levels of phospho-endothelial nitric oxide synthase protein and 3-nitrotyrosine were higher in rats administered at 1 day post-injury compared with rats administered at 7 and 14 days post-injury. These findings suggest that application of Lycium barbarum polysaccharides following cavernous nerve crush injury effectively promotes nerve regeneration and erectile functional recovery. This neuroregenerative effect was most effective in rats orally administered Lycium barbarum polysaccharides at 1 day after cavernous nerve crush injury. PMID:27651780

  12. Polysaccharides from the green seaweed Codium decorticatum. Structure and cell wall distribution.

    PubMed

    Fernández, Paula Virginia; Raffo, María Paula; Alberghina, Josefina; Ciancia, Marina

    2015-03-01

    The cell wall polysaccharides from Codium decorticatum and their assembly were studied and these results were compared with those obtained previously for this genus. The water soluble polysaccharides are: (i) Pyruvylated and sulfated 3- and 6-linked β-D-galactans with sulfate mainly on C-4 and also on C-6. Pyruvate ketals are linked to O-3 and O-4 of terminal β-D-galactose or O-4 and O-6 of 3-linked β-D-galactose. (ii) Sulfated 3-linked β-L-arabinans substituted on C-2 or C-2 and C-4 predominantly with sulfate, but also with single stubs of arabinose, and (iii) 4-linked β-D-mannans with a low degree of sulfation on C-2. The whole polysaccharide system comprises 6.9% of sulfated polysaccharides and 32.9% of fibrillar polysaccharides, mostly insoluble mannans. By in situ localization it was possible to detect two similar fibrillar layers separated by a zone rich in charged polymers. Besides, arabinogalactan proteins co-localized with the fibrillar components.

  13. Anti-fatigue activity of a novel polysaccharide conjugates from Ziyang green tea.

    PubMed

    Chi, Aiping; Li, Hong; Kang, Chenzhe; Guo, Huanhuan; Wang, Yimin; Guo, Fei; Tang, Liang

    2015-09-01

    The aim of this study was to investigate the anti-fatigue activity of polysaccharides from Ziyang green tea. Polysaccharides were isolated from Ziyang green tea and its physicochemical properties were analyzed. Meanwhile, a 4-week weight-loaded swimming test of mice was established and polysaccharides were orally administrated during exercise. The biochemical parameters related to fatigue were determined, such as exhaustive time, blood urea nitrogen (BUN), blood lactate acid (Bla) levels and lactic dehydrogenase (LDH) activity in serum, Superoxide dismutase (SOD), Glutathione peroxidase (GSH-Px) activities, Malondialdehyde (MDA) and glycogen levels in skeletal muscle. The results demonstrated that polysaccharide from Ziyang green tea was a selenium-polysaccharide-protein conjugate (Se-TP), and Se-TP administration significantly prolonged exhaustive time and increased glycogen level and GSH-Px activity in muscle, in addition, markedly decreased BUN, Bla levels and LDH activity in serum and MDA level in muscle. In conclusion, Se-TP treatment can significantly improve exercise-induced fatigue and decrease the oxidative stress induced by the exhaustive exercise.

  14. Antioxidative mechanism of Lycium barbarum polysaccharides promotes repair and regeneration following cavernous nerve injury

    PubMed Central

    Zhao, Zhan-kui; Yu, Hong-lian; Liu, Bo; Wang, Hui; Luo, Qiong; Ding, Xie-gang

    2016-01-01

    Polysaccharides extracted from Lycium barbarum exhibit antioxidant properties. We hypothesized that these polysaccharides resist oxidative stress-induced neuronal damage following cavernous nerve injury. In this study, rat models were intragastrically administered Lycium barbarum polysaccharides for 2 weeks at 1, 7, and 14 days after cavernous nerve injury. Serum superoxide dismutase and glutathione peroxidase activities significantly increased at 1 and 2 weeks post-injury. Serum malondialdehyde levels decreased at 2 and 4 weeks. At 12 weeks, peak intracavernous pressure, the number of myelinated axons and nicotinamide adenine dinucleotide phosphate-diaphorase-positive nerve fibers, levels of phospho-endothelial nitric oxide synthase protein and 3-nitrotyrosine were higher in rats administered at 1 day post-injury compared with rats administered at 7 and 14 days post-injury. These findings suggest that application of Lycium barbarum polysaccharides following cavernous nerve crush injury effectively promotes nerve regeneration and erectile functional recovery. This neuroregenerative effect was most effective in rats orally administered Lycium barbarum polysaccharides at 1 day after cavernous nerve crush injury.

  15. In Vitro Antioxidant and Anti-Proliferation Activities of Polysaccharides from Various Extracts of Different Mushrooms

    PubMed Central

    Li, Xiaoyu; Wang, Zhenyu; Wang, Lu; Walid, Elfalleh; Zhang, Hua

    2012-01-01

    Polysaccharides were extracted from eight kinds of Chinese mushrooms using three solvents and were evaluated for their total carbohydrate, polyphenolic and protein contents, and antioxidant and anti-proliferation activities. The results suggested that all the polysaccharides had significant antioxidant capacities (EC50 ranged from 1.70 ± 0.42 to 65.98 ± 1.74 μM TE/g crude polysaccharide inhibition of ABTS+, EC50 ranged from 5.06 ± 0.12 to 127.38 ± 1.58 mg VCE/g CP scavenging of OH· and EC50 ranged from 0.70 ± 0.04 to 33.54 ± 0.49 mg VCE/g CP inhibition of lipid peroxidation) (TE: trolox equivalent; VCE: VC equivalent; CP: crude polysaccharide). The acid extracts of Russula vinosa Lindblad had the highest ABTS+ scavenging activity. Aqueous extracts of Dictyophora indusiata and Hohenbuehelia serotina possessed, respectively, the highest OH· scavenging capacity and ability to inhibit lipid peroxidation. Mushroom extracts also inhibited proliferation of HeLa and HepG2 cells in a dose-dependent manner. These results indicate that the mushroom polysaccharides might be potential antioxidant resources. PMID:22754332

  16. Sulfated polysaccharides from Loligo vulgaris skin: potential biological activities and partial purification.

    PubMed

    Abdelmalek, Baha Eddine; Sila, Assaâd; Krichen, Fatma; Karoud, Wafa; Martinez-Alvarez, Oscar; Ellouz-Chaabouni, Semia; Ayadi, Mohamed Ali; Bougatef, Ali

    2015-01-01

    The characteristics, biological properties, and purification of sulfated polysaccharides extracted from squid (Loligo vulgaris) skin were investigated. Their chemical and physical characteristics were determined using X-ray diffraction and infrared spectroscopic analysis. Sulfated polysaccharides from squid skin (SPSS) contained 85.06% sugar, 2.54% protein, 1.87% ash, 8.07% sulfate, and 1.72% uronic acid. The antioxidant properties of SPSS were investigated based on DPPH radical-scavenging capacity (IC50 = 19.42 mg mL(-1)), hydrogen peroxide-scavenging activity (IC50 = 0.91 mg mL(-1)), and β-carotene bleaching inhibition (IC50 = 2.79 mg mL(-1)) assays. ACE-inhibitory activity of SPSS was also investigated (IC50 = 0.14 mg mL(-1)). Further antimicrobial activity assays indicated that SPSS exhibited marked inhibitory activity against the bacterial and fungal strains tested. Those polysaccharides did not display hemolytic activity towards bovine erythrocytes. Fractionation by DEAE-cellulose column chromatography showed three major absorbance peaks. Results of this study suggest that sulfated polysaccharides from squid skin are attractive sources of polysaccharides and promising candidates for future application as dietary ingredients.

  17. Chemical structure, conjugation, and cross-reactivity of Bacillus pumilus Sh18 cell wall polysaccharide.

    PubMed

    Kubler-Kielb, Joanna; Coxon, Bruce; Schneerson, Rachel

    2004-10-01

    Bacillus pumilus strain Sh18 cell wall polysaccharide (CWP), cross-reactive with the capsular polysaccharide of Haemophilus influenzae type b, was purified and its chemical structure was elucidated using fast atom bombardment mass spectrometry, nuclear magnetic resonance techniques, and sugar-specific degradation procedures. Two major structures, 1,5-poly(ribitol phosphate) and 1,3-poly(glycerol phosphate), with the latter partially substituted by 2-acetamido-2-deoxy-alpha-galactopyranose (13%) and 2-acetamido-2-deoxy-alpha-glucopyranose (6%) on position O-2, were found. A minor component was established to be a polymer of -->3-O-(2-acetamido-2-deoxy-beta-glucopyranosyl)-1-->4-ribitol-1-OPO3-->. The ratios of the three components were 56, 34, and 10 mol%, respectively. The Sh18 CWP was covalently bound to carrier proteins, and the immunogenicity of the resulting conjugates was evaluated in mice. Two methods of conjugation were compared: (i) binding of 1-cyano-4-dimethylaminopyridinium tetrafluoroborate-activated hydroxyl groups of the CWP to adipic acid dihydrazide (ADH)-derivatized protein, and (ii) binding of the carbodiimide-activated terminal phosphate group of the CWP to ADH-derivatized protein. The conjugate-induced antibodies reacted in an enzyme-linked immunosorbent assay with the homologous polysaccharide and with a number of other bacterial polysaccharides containing ribitol and glycerol phosphates, including H. influenzae types a and b and strains of Staphylococcus aureus and Staphylococcus epidermidis. PMID:15466043

  18. Viscoelastic properties of levan polysaccharides

    NASA Astrophysics Data System (ADS)

    Noll, Kenneth; Rende, Deniz; Ozisik, Rahmi; Toksoy-Oner, Ebru

    2014-03-01

    Levan is a naturally occurring polysaccharide that is composed of β-D-fructofuranose units with β(2-6) linkages between fructose rings. It is synthesized by the action of a secreted levansucrase (EC 2.4.1.10) that converts sucrose into the levan externally (exopolysaccharide). Levan is a homopolysaccharide that is non-toxic, water soluble,, and has anti-tumor activity and low immunological response. Therefore, levan presents great potential to be used as a novel functional biopolymer in foods, feeds, cosmetics, pharmaceutical and chemical industries. Despite these favorable properties, levan has a moderately low mechanical properties and poor film forming capability. In the current study, the agglomeration behavior of levan in water and in saline solutions was investigated at 298 and 310 K by dynamic light scattering and transmission electron microscopy (TEM). The viscoelastic properties of neat and oxidized levan films were studied via nanoindentation experiments in the quasi-static and dynamic modes The material is partially based upon work supported by NSF under Grant Nos. 1200270 and 1003574, and TUBITAK 111M232.

  19. New Insights into Regulation of Proteome and Polysaccharide in Cell Wall of Elsholtzia splendens in Response to Copper Stress

    PubMed Central

    Liu, Tingting; Shen, Chaofeng; Wang, Yi; Huang, Canke; Shi, Jiyan

    2014-01-01

    Background and Aims Copper (Cu) is an essential micronutrient for plants. However, excess amounts of Cu are toxic and result in a wide range of harmful effects on the physiological and biochemical processes of plants. Cell wall has a crucial role in plant defense response to toxic metals. To date, the process of cell wall response to Cu and the detoxification mechanism have not been well documented at the proteomic level. Methods An recently developed 6-plex Tandem Mass Tag was used for relative and absolute quantitation methods to achieve a comprehensive understanding of Cu tolerance/detoxification molecular mechanisms in the cell wall. LC–MS/MS approach was performed to analyze the Cu-responsive cell wall proteins and polysaccharides. Key Results The majority of the 22 up-regulated proteins were involved in the antioxidant defense pathway, cell wall polysaccharide remodeling, and cell metabolism process. Changes in polysaccharide amount, composition, and distribution could offer more binding sites for Cu ions. The 33 down-regulated proteins were involved in the signal pathway, energy, and protein synthesis. Conclusions Based on the abundant changes in proteins and polysaccharides, and their putative functions, a possible protein interaction network can provide new insights into Cu stress response in root cell wall. Cu can facilitate further functional research on target proteins associated with metal response in the cell wall. PMID:25340800

  20. Immobilized phosphorylase for synthesis of polysaccharides from glucose

    NASA Technical Reports Server (NTRS)

    Marshall, D. L.

    1972-01-01

    Continuous processes for enzymatic production of carbohydrates from glucose are discussed. Key reactant in process is identified as phosphorylase which catalyzes reversible formation or degradation of polysaccharide. Chemical compounds and reactions to synthesize polysaccharides are analyzed.

  1. Immunomodulatory effects of a polysaccharide from Tamarindus indica.

    PubMed

    Sreelekha, T T; Vijayakumar, T; Ankanthil, R; Vijayan, K K; Nair, M K

    1993-04-01

    A polysaccharide isolated and purified from Tamarindus indica shows immunomodulatory activities such as phagocytic enhancement, leukocyte migration inhibition and inhibition of cell proliferation. These properties suggest that this polysaccharide from T. indica may have some biological applications.

  2. Neisseria meningitidis serogroup A capsular polysaccharide acetyltransferase, methods and compositions

    DOEpatents

    Stephens, David S.; Gudlavalleti, Seshu K.; Tzeng, Yih-Ling; Datta, Anup K.; Carlson, Russell W.

    2011-02-08

    Provided are methods for recombinant production of an O-acetyltransferase and methods for acetylating capsular polysaccharides, especially those of a Serogroup A Neisseria meningitidis using the recombinant O-acetyltransferase, and immunogenic compositions comprising the acetylated capsular polysaccharide.

  3. Ice nucleation activity of polysaccharides

    NASA Astrophysics Data System (ADS)

    Bichler, Magdalena; Felgitsch, Laura; Haeusler, Thomas; Seidl-Seiboth, Verena; Grothe, Hinrich

    2015-04-01

    Heterogeneous ice nucleation is an important process in the atmosphere. It shows direct impact on our climate by triggering ice cloud formation and therefore it has much influence on the radiation balance of our planet (Lohmann et al. 2002; Mishchenko et al. 1996). The process itself is not completely understood so far and many questions remain open. Different substances have been found to exhibit ice nucleation activity (INA). Due to their vast differences in chemistry and morphology it is difficult to predict what substance will make good ice nuclei and which will not. Hence simple model substances must be found and be tested regarding INA. Our work aims at gaining to a deeper understanding of heterogeneous ice nucleation. We intend to find some reference standards with defined chemistry, which may explain the mechanisms of heterogeneous ice nucleation. A particular focus lies on biological carbohydrates in regards to their INA. Biological carbohydrates are widely distributed in all kingdoms of life. Mostly they are specific for certain organisms and have well defined purposes, e.g. structural polysaccharides like chitin (in fungi and insects) and pectin (in plants), which has also water-binding properties. Since they are widely distributed throughout our biosphere and mostly safe to use for nutrition purposes, they are well studied and easily accessible, rendering them ideal candidates as proxies. In our experiments we examined various carbohydrates, like the already mentioned chitin and pectin, as well as their chemical modifications. Lohmann U.; A Glaciation Indirect Aerosol Effect Caused by Soot Aerosols; J. Geoph. Res.; Vol. 24 No.4; pp 11-1 - 11-4; 2002 Mishchenko M.I., Rossow W.B., Macke A., Lacis A. A.; Sensitivity of Cirrus Cloud Albedo, Bidirectional Reflectance and Optical Thickness Retrieval Accuracy to Ice Particle Shape, J. Geoph. Res.; Vol. 101, No D12; pp. 16,973 - 16,985; 1996

  4. Mucosal immunogenicity of polysaccharides conjugated to a peptide or multiple-antigen peptide containing T- and B-cell epitopes.

    PubMed Central

    Lett, E; Klopfenstein, C; Klein, J P; Schöller, M; Wachsmann, D

    1995-01-01

    In this study we investigated the mucosal and systemic responses to two T-cell-independent polysaccharides, a serogroup f polysaccharide (formed of rhamnose glucose polymers [RGPs]) from Streptococcus mutans OMZ 175 and a mannan from Saccharomyces cerevisiae, covalently conjugated either to a linear peptide (peptide 3) or to a multiple-antigen peptide (MAP), both derived from S. mutans protein SR, an adhesin of the I/II protein antigen family of oral streptococci. Peptide 3 and MAP, which contained at least one B- and one T-cell epitope, were tested as carriers for the polysaccharides and as protective immunogens. Intragastric intubation of rats with the conjugates (RGPs-peptide 3, RGPs-MAP, mannan-peptide 3, and mannan-MAP) associated with liposomes produced salivary immunoglobulin A (IgA) antibodies which reacted with RGPs or mannan, peptide 3 or MAP, protein SR, and S. mutans or S. cerevisiae cells. Administration of conjugate boosters to the animals showed that both carriers conjugated to the polysaccharides were able to induce, in immunized animals, a salivary antipolysaccharide IgA memory. In contrast, animals primed and challenged with unconjugated polysaccharide showed no anamnestic response. Rats orally immunized with the conjugates also developed systemic primary antipolysaccharide and antipeptide IgM antibody responses which were characterized by a switch from IgM to IgG during the course of the secondary response. Data presented here demonstrated that both peptide 3 and the MAP construct can act as good carriers for orally administered polysaccharides. Unexpectedly, the use of a MAP did not further improve the immunogenicity of polysaccharides at the mucosal level; nevertheless, such a construct should be of great interest in overcoming the problem of genetic restriction induced by linear peptides. PMID:7790080

  5. Purification, characterization and biological activities in vitro of polysaccharides extracted from tea seeds.

    PubMed

    Wang, Yuanfeng; Liu, Yangyang; Mao, Fangfang; Liu, Yiran; Wei, Xinlin

    2013-11-01

    Three polysaccharide fractions (NTSPS, ATSPS1-1 and ATSPS2) were isolated and purified from the water extract of tea (Camellia sinensis) seeds with yields of 32.74%, 25.22%, and 11.09%, respectively. The average molecular weight of NTSPS, ATSPS1-1 and ATSPS2 was determined by HPGPC system, with an Mw of 4.588, 500 and 100 kDa, respectively. Monosaccharide analysis by ion chromatography revealed they differed from each other in monosaccharide kinds and molar ratio. Furthermore, ATSPS1-1 and ATSPS2 were heteropolysaccharide bounded with protein (2.83% and 2.12%) and 17 general amino acids were detected in them on an ion-chromatographic analyzer. The IR spectrum of ATSPS1-1 and ATSPS2 revealed the typical characteristics of polysaccharides and protein. In addition, the antitumor and immunological activity of the three polysaccharide fractions in vitro were also evaluated. It was found inhibition activity of NTSPS, ATSPS1-1 and ATSPS2 on K562 cells increased with increasing sample concentration and the inhibition ratios of them at 400 μg/mL were beyond 30.13 ± 3.54%, 36.61 ± 2.75% and 32.33 ± 2.53%, respectively. They also exhibited strong ability in promoting proliferation of mice splenic lymphocyte. Results of these studies indicated the three purified polysaccharides had a potential application as natural antitumor drugs.

  6. Isolation and prebiotic activity of water-soluble polysaccharides fractions from the bamboo shoots (Phyllostachys praecox).

    PubMed

    He, Shudong; Wang, Xin; Zhang, Yi; Wang, Jing; Sun, Hanju; Wang, Junhui; Cao, Xiaodong; Ye, Yongkang

    2016-10-20

    The water-soluble polysaccharides from bamboo shoots (Phyllostachys praecox) (WBP) were isolated, and the characterizations as well as prebiotic activities were investigated. The yield of WBP was 7.58±0.31% under optimal hot-water extraction conditions. Two fractions, i.e., WBP-1 and WBP-2 with molecular weight of 83.50kDa and 80.08kDa, respectively, were purified by chromatography. Both the polysaccharides fractions were identified as heteropolysaccharides-protein complexes composed of 15 kinds of common amino acids in protein part and rhamnose, arabinose, xylose, mannose, glucose and galactose in different molar ratios in polysaccharide part. The existence of α- and β-glycosidic linkages between the sugar units was confirmed by FTIR and NMR spectra. Compared with the blank control and the reference of FOS, WBP-1 and WBP-2 significantly increased the numbers of Bifidobacterium adolescentis and Bifidobacterium bifidum (P<0.05), which contributed to the production of organic acids, suggesting that the polysaccharides have potential prebiotic properties. PMID:27474570

  7. Recalcitrant polysaccharide degradation by novel oxidative biocatalysts.

    PubMed

    Dimarogona, Maria; Topakas, Evangelos; Christakopoulos, Paul

    2013-10-01

    The classical hydrolytic mechanism for the degradation of plant polysaccharides by saprophytic microorganisms has been reconsidered after the recent landmark discovery of a new class of oxidases termed lytic polysaccharide monooxygenases (LPMOs). LPMOs are of increased biotechnological interest due to their implication in lignocellulosic biomass decomposition for the production of biofuels and high-value chemicals. They act on recalcitrant polysaccharides by a combination of hydrolytic and oxidative function, generating oxidized and non-oxidized chain ends. They are copper-dependent and require molecular oxygen and an external electron donor for their proper function. In this review, we present the recent findings concerning the mechanism of action of these oxidative enzymes and identify issues and questions to be addressed in the future. PMID:23995228

  8. Learning from microbial strategies for polysaccharide degradation.

    PubMed

    Hemsworth, Glyn R; Déjean, Guillaume; Davies, Gideon J; Brumer, Harry

    2016-02-01

    Complex carbohydrates are ubiquitous in all kingdoms of life. As major components of the plant cell wall they constitute both a rich renewable carbon source for biotechnological transformation into fuels, chemicals and materials, and also form an important energy source as part of a healthy human diet. In both contexts, there has been significant, sustained interest in understanding how microbes transform these substrates. Classical perspectives of microbial polysaccharide degradation are currently being augmented by recent advances in the discovery of lytic polysaccharide monooxygenases (LPMOs) and polysaccharide utilization loci (PULs). Fundamental discoveries in carbohydrate enzymology are both advancing biological understanding, as well as informing applications in industrial biomass conversion and modulation of the human gut microbiota to mediate health benefits.

  9. Regulation of Polysaccharide Utilization Contributes to the Persistence of Group A Streptococcus in the Oropharynx▿

    PubMed Central

    Shelburne, Samuel A.; Okorafor, Nnaja; Sitkiewicz, Izabela; Sumby, Paul; Keith, David; Patel, Payal; Austin, Celest; Graviss, Edward A.; Musser, James M.

    2007-01-01

    Group A Streptococcus (GAS) genes that encode proteins putatively involved in polysaccharide utilization show growth phase-dependent expression in human saliva. We sought to determine whether the putative polysaccharide transcriptional regulator MalR influences the expression of such genes and whether MalR helps GAS infect the oropharynx. Analysis of 32 strains of 17 distinct M protein serotypes revealed that MalR is highly conserved across GAS strains. malR transcripts were detectable in patients with GAS pharyngitis, and the levels increased significantly during growth in human saliva compared to the levels during growth in glucose-containing or nutrient-rich media. To determine if MalR influenced the expression of polysaccharide utilization genes, we compared the transcript levels of eight genes encoding putative polysaccharide utilization proteins in the parental serotype M1 strain MGAS5005 and its ΔmalR isogenic mutant derivative. The transcript levels of all eight genes were significantly increased in the ΔmalR strain compared to the parental strain, especially during growth in human saliva. Following experimental infection, the ΔmalR strain persistently colonized the oropharynx in significantly fewer mice than the parental strain colonized, and the numbers of ΔmalR strain CFU recovered were significantly lower than the numbers of the parental strain CFU recovered. These data led us to conclude that MalR influences the expression of genes putatively involved in polysaccharide utilization and that MalR contributes to the persistence of GAS in the oropharynx. PMID:17403878

  10. 2-Fluoro-L-Fucose Is a Metabolically Incorporated Inhibitor of Plant Cell Wall Polysaccharide Fucosylation.

    PubMed

    Villalobos, Jose A; Yi, Bo R; Wallace, Ian S

    2015-01-01

    The monosaccharide L-fucose (L-Fuc) is a common component of plant cell wall polysaccharides and other plant glycans, including the hemicellulose xyloglucan, pectic rhamnogalacturonan-I (RG-I) and rhamnogalacturonan-II (RG-II), arabinogalactan proteins, and N-linked glycans. Mutations compromising the biosynthesis of many plant cell wall polysaccharides are lethal, and as a result, small molecule inhibitors of plant cell wall polysaccharide biosynthesis have been developed because these molecules can be applied at defined concentrations and developmental stages. In this study, we characterize novel small molecule inhibitors of plant fucosylation. 2-fluoro-L-fucose (2F-Fuc) analogs caused severe growth phenotypes when applied to Arabidopsis seedlings, including reduced root growth and altered root morphology. These phenotypic defects were dependent upon the L-Fuc salvage pathway enzyme L-Fucose Kinase/ GDP-L-Fucose Pyrophosphorylase (FKGP), suggesting that 2F-Fuc is metabolically converted to the sugar nucleotide GDP-2F-Fuc, which serves as the active inhibitory molecule. The L-Fuc content of cell wall matrix polysaccharides was reduced in plants treated with 2F-Fuc, suggesting that this molecule inhibits the incorporation of L-Fuc into these polysaccharides. Additionally, phenotypic defects induced by 2F-Fuc treatment could be partially relieved by the exogenous application of boric acid, suggesting that 2F-Fuc inhibits RG-II biosynthesis. Overall, the results presented here suggest that 2F-Fuc is a metabolically incorporated inhibitor of plant cellular fucosylation events, and potentially suggest that other 2-fluorinated monosaccharides could serve as useful chemical probes for the inhibition of cell wall polysaccharide biosynthesis. PMID:26414071

  11. 2-Fluoro-L-Fucose Is a Metabolically Incorporated Inhibitor of Plant Cell Wall Polysaccharide Fucosylation

    PubMed Central

    Wallace, Ian S.

    2015-01-01

    The monosaccharide L-fucose (L-Fuc) is a common component of plant cell wall polysaccharides and other plant glycans, including the hemicellulose xyloglucan, pectic rhamnogalacturonan-I (RG-I) and rhamnogalacturonan-II (RG-II), arabinogalactan proteins, and N-linked glycans. Mutations compromising the biosynthesis of many plant cell wall polysaccharides are lethal, and as a result, small molecule inhibitors of plant cell wall polysaccharide biosynthesis have been developed because these molecules can be applied at defined concentrations and developmental stages. In this study, we characterize novel small molecule inhibitors of plant fucosylation. 2-fluoro-L-fucose (2F-Fuc) analogs caused severe growth phenotypes when applied to Arabidopsis seedlings, including reduced root growth and altered root morphology. These phenotypic defects were dependent upon the L-Fuc salvage pathway enzyme L-Fucose Kinase/ GDP-L-Fucose Pyrophosphorylase (FKGP), suggesting that 2F-Fuc is metabolically converted to the sugar nucleotide GDP-2F-Fuc, which serves as the active inhibitory molecule. The L-Fuc content of cell wall matrix polysaccharides was reduced in plants treated with 2F-Fuc, suggesting that this molecule inhibits the incorporation of L-Fuc into these polysaccharides. Additionally, phenotypic defects induced by 2F-Fuc treatment could be partially relieved by the exogenous application of boric acid, suggesting that 2F-Fuc inhibits RG-II biosynthesis. Overall, the results presented here suggest that 2F-Fuc is a metabolically incorporated inhibitor of plant cellular fucosylation events, and potentially suggest that other 2-fluorinated monosaccharides could serve as useful chemical probes for the inhibition of cell wall polysaccharide biosynthesis. PMID:26414071

  12. 2-Fluoro-L-Fucose Is a Metabolically Incorporated Inhibitor of Plant Cell Wall Polysaccharide Fucosylation.

    PubMed

    Villalobos, Jose A; Yi, Bo R; Wallace, Ian S

    2015-01-01

    The monosaccharide L-fucose (L-Fuc) is a common component of plant cell wall polysaccharides and other plant glycans, including the hemicellulose xyloglucan, pectic rhamnogalacturonan-I (RG-I) and rhamnogalacturonan-II (RG-II), arabinogalactan proteins, and N-linked glycans. Mutations compromising the biosynthesis of many plant cell wall polysaccharides are lethal, and as a result, small molecule inhibitors of plant cell wall polysaccharide biosynthesis have been developed because these molecules can be applied at defined concentrations and developmental stages. In this study, we characterize novel small molecule inhibitors of plant fucosylation. 2-fluoro-L-fucose (2F-Fuc) analogs caused severe growth phenotypes when applied to Arabidopsis seedlings, including reduced root growth and altered root morphology. These phenotypic defects were dependent upon the L-Fuc salvage pathway enzyme L-Fucose Kinase/ GDP-L-Fucose Pyrophosphorylase (FKGP), suggesting that 2F-Fuc is metabolically converted to the sugar nucleotide GDP-2F-Fuc, which serves as the active inhibitory molecule. The L-Fuc content of cell wall matrix polysaccharides was reduced in plants treated with 2F-Fuc, suggesting that this molecule inhibits the incorporation of L-Fuc into these polysaccharides. Additionally, phenotypic defects induced by 2F-Fuc treatment could be partially relieved by the exogenous application of boric acid, suggesting that 2F-Fuc inhibits RG-II biosynthesis. Overall, the results presented here suggest that 2F-Fuc is a metabolically incorporated inhibitor of plant cellular fucosylation events, and potentially suggest that other 2-fluorinated monosaccharides could serve as useful chemical probes for the inhibition of cell wall polysaccharide biosynthesis.

  13. BBB-Permeable, Neuroprotective, and Neurotrophic Polysaccharide, Midi-GAGR

    PubMed Central

    Makani, Vishruti; Jang, Yong-gil; Christopher, Kevin; Judy, Wesley; Eckstein, Jacob; Hensley, Kenneth; Chiaia, Nicolas; Kim, Dong-Shik; Park, Joshua

    2016-01-01

    An enormous amount of efforts have been poured to find an effective therapeutic agent for the treatment of neurodegenerative diseases including Alzheimer’s disease (AD). Among those, neurotrophic peptides that regenerate neuronal structures and increase neuron survival show a promise in slowing neurodegeneration. However, the short plasma half-life and poor blood-brain-barrier (BBB)-permeability of neurotrophic peptides limit their in vivo efficacy. Thus, an alternative neurotrophic agent that has longer plasma half-life and better BBB-permeability has been sought for. Based on the recent findings of neuroprotective polysaccharides, we searched for a BBB-permeable neuroprotective polysaccharide among natural polysaccharides that are approved for human use. Then, we discovered midi-GAGR, a BBB-permeable, long plasma half-life, strong neuroprotective and neurotrophic polysaccharide. Midi-GAGR is a 4.7kD cleavage product of low acyl gellan gum that is approved by FDA for human use. Midi-GAGR protected rodent cortical neurons not only from the pathological concentrations of co-/post-treated free reactive radicals and Aβ42 peptide but also from activated microglial cells. Moreover, midi-GAGR showed a good neurotrophic effect; it enhanced neurite outgrowth and increased phosphorylated cAMP-responsive element binding protein (pCREB) in the nuclei of primary cortical neurons. Furthermore, intra-nasally administered midi-GAGR penetrated the BBB and exerted its neurotrophic effect inside the brain for 24 h after one-time administration. Midi-GAGR appears to activate fibroblast growth factor receptor 1 (FGFR1) and its downstream neurotrophic signaling pathway for neuroprotection and CREB activation. Additionally, 14-day intranasal administration of midi-GAGR not only increased neuronal activity markers but also decreased hyperphosphorylated tau, a precursor of neurofibrillary tangle, in the brains of the AD mouse model, 3xTg-AD. Taken together, midi-GAGR with good BBB

  14. The structure and function of glycoproteins synthesized during slime-polysaccharide production by membranes of the root-cap cells of maize (Zea mays).

    PubMed Central

    Green, J R; Northcote, D H

    1978-01-01

    The synthesis of the maize root slime polysaccharides was investigated by using [1-3H]-fucose as a marker for slime production. Three fractions were separated by centrifugation in a CsCl density gradient. Two of these were glycoproteins and occurred within the membranes of the cells of the root tip; the third was the slime polysaccharides. Radioactive pulse-chase experiments showed that the glycoproteins were precursors of the slime polysaccharides, and the carbohydrate portion of the glycoproteins had a similar composition to that of the free slime. The linkage between the protein and the carbohydrate of one of the glycoproteins was shown to be a xylose-threonine bond. It is postulated that the slime polysaccharides are synthesized and transported on proteins within the membrane system of the root tip. PMID:646802

  15. Polysaccharide-based strategies for heart tissue engineering.

    PubMed

    Silva, Amanda K A; Juenet, Maya; Meddahi-Pellé, Anne; Letourneur, Didier

    2015-02-13

    Polysaccharides are abundant biomolecules in nature presenting important roles in a wide variety of living systems processes. Considering the structural and biological functions of polysaccharides, their properties have raised interest for tissue engineering. Herein, we described the latest advances in cardiac tissue engineering mediated by polysaccharides. We reviewed the data already obtained in vitro and in vivo in this field with several types of polysaccharides. Cardiac injection, intramyocardial in situ polymerization strategies, and scaffold-based approaches involving polysaccharides for heart tissue engineering are thus discussed.

  16. Fermentation and addition of enzymes to a diet based on high-moisture corn, rapeseed cake, and peas improve digestibility of nonstarch polysaccharides, crude protein, and phosphorus in pigs.

    PubMed

    Jakobsen, G V; Jensen, B B; Bach Knudsen, K E; Canibe, N

    2015-05-01

    Fluctuating prices of cereals have led to an interest in alternative ingredients for feed. The aim of this study was to evaluate the effect of fermentation and the addition of nonstarch polysaccharide (NSP)-degrading enzymes on the ileal and total tract digestibility of nutrients of a diet based on locally grown crops. Four diets were fed including a nonfermented liquid standard grower diet (Control) and 3 experimental diets based on high-moisture corn, rapeseed cake, and peas fed as nonfermented liquid feed (nFLF), fermented liquid feed (FLF), or FLF supplemented with an enzyme mixture of β-glucanase + xylanase + pectinase (FLF+Enz). The FLF was prepared by mixing feed and water (1:2.5, wt/wt) and, once daily, replacing 50% of the mixture with an equal amount of fresh feed and water. The diets were fed to 8 ileal cannulated barrows in a double Latin square design. Ileal digesta and feces were collected after an adaption period of 10 d. Results showed microbiologically good-quality fermented diets. The levels of Enterobacteriaceae were 5.1 to 5.4 log cfu/g in FLF and FLF+Enz vs. 6.3 log cfu/g in nFLF in the ileum and 5.1 to 5.2 log cfu/g in FLF and FLF+Enz vs. 6.3 log cfu/g in nFLF in the feces. Apparent total tract digestibility (ATTD) of CP was increased by fermentation (73.2% in FLF vs. 69.0% in nFLF; P = 0.033), and digestibility of P showed a tendency (P = 0.073) toward an increase. Addition of the enzyme mixture resulted in a pronounced reduction of dietary NSP compared with FLF (12.8% total NSP in FLF+Enz vs. 15.9% total NSP in FLF; P< 0.001), which also led to increased apparent ileal digestibility (AID) of total and insoluble NSP (total NSP, 31.1% in FLF+Enz vs. 13.6% in FLF; P = 0.002). The Control did not, in general, show higher digestibility values than the experimental diet. However, in the cases were it did, fermentation and enzyme addition brought the digestibility to the level of the Control. In conclusion, fermentation increased the ATTD of CP

  17. Anticorrosive Microbial Polysaccharides: Structure-Function Relationships

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Water-soluble microbial polysaccharides are often implicated in biofilm formation and are believed to mediate cell-cell aggregation and adhesion to surfaces. Generally, biofilm formation is considered harmful or undesirable, as it leads to increased drag, plugging of pores, dimished heat transfer, ...

  18. Antitumor activity of methylan polysaccharide derivatives.

    PubMed

    Ramachandran, Priyadharshini; Jeya, Marimuthu; Moon, Hee-Jung; Lee, Kyoung-Mi; Kim, In-Won; Kim, Jung-Hoe; Lee, Jung-Kul

    2010-07-01

    Methylan polysaccharide derivatives were prepared by dialkylaminoalkylation and reductive amination followed by quaternization. Their antitumor activity was investigated and a relationship between structure and activity is suggested. For quaternized DEAE-methylan at only 75 mug ml(-1), tumor cell proliferation was suppressed by 58-84% in three cell lines tested in the order Colo < Hela < HepG2. PMID:20349111

  19. Fucoidans — sulfated polysaccharides of brown algae

    NASA Astrophysics Data System (ADS)

    Usov, Anatolii I.; Bilan, M. I.

    2009-08-01

    The methods of isolation of fucoidans and determination of their chemical structures are reviewed. The fucoidans represent sulfated polysaccharides of brown algae, the composition of which varies from simple fucan sulfates to complex heteropolysaccharides. The currently known structures of such biopolymers are presented. A variety of the biological activities of fucoidans is briefly summarised.

  20. Plant-Polysaccharide-Degrading Enzymes from Basidiomycetes

    PubMed Central

    Rytioja, Johanna; Hildén, Kristiina; Yuzon, Jennifer; Hatakka, Annele; de Vries, Ronald P.

    2014-01-01

    SUMMARY Basidiomycete fungi subsist on various types of plant material in diverse environments, from living and dead trees and forest litter to crops and grasses and to decaying plant matter in soils. Due to the variation in their natural carbon sources, basidiomycetes have highly varied plant-polysaccharide-degrading capabilities. This topic is not as well studied for basidiomycetes as for ascomycete fungi, which are the main sources of knowledge on fungal plant polysaccharide degradation. Research on plant-biomass-decaying fungi has focused on isolating enzymes for current and future applications, such as for the production of fuels, the food industry, and waste treatment. More recently, genomic studies of basidiomycete fungi have provided a profound view of the plant-biomass-degrading potential of wood-rotting, litter-decomposing, plant-pathogenic, and ectomycorrhizal (ECM) basidiomycetes. This review summarizes the current knowledge on plant polysaccharide depolymerization by basidiomycete species from diverse habitats. In addition, these data are compared to those for the most broadly studied ascomycete genus, Aspergillus, to provide insight into specific features of basidiomycetes with respect to plant polysaccharide degradation. PMID:25428937

  1. Antitumor activity of methylan polysaccharide derivatives.

    PubMed

    Ramachandran, Priyadharshini; Jeya, Marimuthu; Moon, Hee-Jung; Lee, Kyoung-Mi; Kim, In-Won; Kim, Jung-Hoe; Lee, Jung-Kul

    2010-07-01

    Methylan polysaccharide derivatives were prepared by dialkylaminoalkylation and reductive amination followed by quaternization. Their antitumor activity was investigated and a relationship between structure and activity is suggested. For quaternized DEAE-methylan at only 75 mug ml(-1), tumor cell proliferation was suppressed by 58-84% in three cell lines tested in the order Colo < Hela < HepG2.

  2. Aldehyde-containing urea-absorbing polysaccharides

    NASA Technical Reports Server (NTRS)

    Mueller, W. A.; Hsu, G. C.; Marsh, H. E., Jr. (Inventor)

    1977-01-01

    A novel aldehyde containing polymer (ACP) is prepared by reaction of a polysaccharide with periodate to introduce aldehyde groups onto the C2 - C3 carbon atoms. By introduction of ether and ester groups onto the pendant primary hydroxyl solubility characteristics are modified. The ACP is utilized to absorb nitrogen bases such as urea in vitro or in vivo.

  3. Purification and characterization of Staphylococcus aureus type 8 capsular polysaccharide.

    PubMed Central

    Fournier, J M; Vann, W F; Karakawa, W W

    1984-01-01

    Clinical isolates of Staphylococcus aureus have been previously classified into eight types on the basis of their capsular polysaccharide. The high prevalence of the type 8 capsular polysaccharide among bacteremic isolates suggests the importance of this capsular antigen in staphylococcal disease. The capsular polysaccharide was purified from extracts of three clinical isolates of S. aureus type 8 of different geographic and temperal origin by ion-exchange chromatography and gel filtration. Gas chromatography, gas chromatography-mass spectrometry, and 13C-nuclear magnetic resonance showed that the type 8 capsular polysaccharide is composed of O-acetyl groups, N-acetylfucosamine, and an aminouronic acid similar to N-acetylgalactosaminouronic acid. The purified polysaccharide reacted only with type 8 antiserum in double diffusion experiments. Our analysis shows that the type 8 polysaccharide is both chemically and serologically distinct from teichoic acid and previously characterized polysaccharides of S. aureus. Images PMID:6429051

  4. Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro.

    PubMed

    Chen, Chun; You, Li-Jun; Abbasi, Arshad Mehmood; Fu, Xiong; Liu, Rui Hai

    2015-10-01

    Single-factor experiment and Box-Behnken design (BBD) were applied to optimize the ultrasound-assisted extraction of mulberry fruits polysaccharides (MFP). Under optimum conditions: ratio of water to raw material 40.25, extraction temperature 69°C, ultrasonic power 190W and extraction time 75 min, the MFP yield was 3.13% (±0.07%), in accordance to the predicted value of 3.04%. The mulberry fruits polysaccharides fractions was obtained by deproteinization (MFP-1), followed by decolorization and deionization (MFP-2). Carbohydrate content in MFP, MFP-1 and MFP-2 was 58.61% (±1.47%), 69.98% (±0.91%), 81.18% (±1.29%), as well as proteins was estimated 16.50% (±0.86%), 1.57% (±0.63%), 1.02% (±0.18%), respectively. The FT-IR indicated that MFP, MFP-1 and MFP-2 were acidic polysaccharides. The MFP-1 exhibited the strongest antioxidant activity, while MFP-2 showed the strongest hyperglycemic activity in vitro. This may be caused by their different compositions and physical properties in the different mulberry fruit polysaccharides fractions.

  5. Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro.

    PubMed

    Chen, Chun; You, Li-Jun; Abbasi, Arshad Mehmood; Fu, Xiong; Liu, Rui Hai

    2015-10-01

    Single-factor experiment and Box-Behnken design (BBD) were applied to optimize the ultrasound-assisted extraction of mulberry fruits polysaccharides (MFP). Under optimum conditions: ratio of water to raw material 40.25, extraction temperature 69°C, ultrasonic power 190W and extraction time 75 min, the MFP yield was 3.13% (±0.07%), in accordance to the predicted value of 3.04%. The mulberry fruits polysaccharides fractions was obtained by deproteinization (MFP-1), followed by decolorization and deionization (MFP-2). Carbohydrate content in MFP, MFP-1 and MFP-2 was 58.61% (±1.47%), 69.98% (±0.91%), 81.18% (±1.29%), as well as proteins was estimated 16.50% (±0.86%), 1.57% (±0.63%), 1.02% (±0.18%), respectively. The FT-IR indicated that MFP, MFP-1 and MFP-2 were acidic polysaccharides. The MFP-1 exhibited the strongest antioxidant activity, while MFP-2 showed the strongest hyperglycemic activity in vitro. This may be caused by their different compositions and physical properties in the different mulberry fruit polysaccharides fractions. PMID:26076608

  6. PelA Deacetylase Activity Is Required for Pel Polysaccharide Synthesis in Pseudomonas aeruginosa

    PubMed Central

    Colvin, Kelly M.; Alnabelseya, Noor; Baker, Perrin; Whitney, John C.

    2013-01-01

    The Pel polysaccharide serves as an intercellular adhesin for the formation and maintenance of biofilms in the opportunistic pathogen Pseudomonas aeruginosa. Pel biosynthesis requires the products of a seven-gene operon, pelA-pelG, all of which are necessary for Pel-dependent biofilm formation and Pel-related phenotypes. One of the genes, pelA, encodes a protein with a predicted polysaccharide deacetylase domain. In this work, the role of the putative deacetylase domain in Pel production was examined. We first established that purified recombinant PelA hydrolyzed the pseudosubstrate p-nitrophenyl acetate in vitro, and site-specific mutations of predicted deacetylase active-site residues reduced activity greater than 10-fold. Additionally, these mutants were deficient in Pel-dependent biofilm formation and wrinkly colony morphology in vivo. Subcellular fractionation experiments demonstrate that PelA localizes to both the membrane and periplasmic fractions. Finally, antiserum against the Pel polysaccharide was generated, and PelA deacetylase mutants do not produce Pel-reactive material. Taken together, these results suggest that the deacetylase activity of PelA is important for the production of the Pel polysaccharide. PMID:23504011

  7. Alfalfa Root Nodule Invasion Efficiency Is Dependent on Sinorhizobium meliloti Polysaccharides

    PubMed Central

    Pellock, Brett J.; Cheng, Hai-Ping; Walker, Graham C.

    2000-01-01

    The soil bacterium Sinorhizobium meliloti is capable of entering into a nitrogen-fixing symbiosis with Medicago sativa (alfalfa). Particular low-molecular-weight forms of certain polysaccharides produced by S. meliloti are crucial for establishing this symbiosis. Alfalfa nodule invasion by S. meliloti can be mediated by any one of three symbiotically important polysaccharides: succinoglycan, EPS II, or K antigen (also referred to as KPS). Using green fluorescent protein-labeled S. meliloti cells, we have shown that there are significant differences in the details and efficiencies of nodule invasion mediated by these polysaccharides. Succinoglycan is highly efficient in mediating both infection thread initiation and extension. However, EPS II is significantly less efficient than succinoglycan at mediating both invasion steps, and K antigen is significantly less efficient than succinoglycan at mediating infection thread extension. In the case of EPS II-mediated symbioses, the reduction in invasion efficiency results in stunted host plant growth relative to plants inoculated with succinoglycan or K-antigen-producing strains. Additionally, EPS II- and K-antigen-mediated infection threads are 8 to 10 times more likely to have aberrant morphologies than those mediated by succinoglycan. These data have important implications for understanding how S. meliloti polysaccharides are functioning in the plant-bacterium interaction, and models are discussed. PMID:10894742

  8. Characterization and free radical scavenging activity of rapeseed meal polysaccharides WPS-1 and APS-2.

    PubMed

    Zhu, Jianfei; Wu, Moucheng

    2009-02-11

    Two major polysaccharide fractions, WPS-1 and APS-2, were isolated from water-soluble and alkali-soluble extracts of Huaza No. 4 rapeseed meal with a stepwise procedure of D3520 macroporous adsorption resin column chromatography, ethanol precipitation, and DE-52 cellulose column chromatography. Physicochemical properties of the polysaccharides were determined by chemical methods, high -performance liquid chromatography (HPLC), gel permeation chromatography (GPC), and Fourier transform infrared spectrometry (FT-IR). The chemiluminescence (CL) method was used to investigate the free radical scavenging activity of the polysaccharide fractions. The polysaccharides were primarily polymers of arabinose, galactose, and glucose, associated with protein portions consisting of 13 different amino acids. The average molecular masses of WPS-1 and APS-2 were 7.20 x 10(5) and 1.61 x 10(5) Da, respectively. Compared with APS-2, WPS-1 was more effective at scavenging superoxide radical (O(2)(*-)) and hydroxyl radical (HO(*)), but less effective at scavenging hydrogen peroxide (H(2)O(2)). In decreasing order, the free radical scavenging activity of WPS-1 and APS-2 toward reactive oxygen species (ROS) was H(2)O(2) > HO(*) > O(2)(*-).

  9. Structural and immunological feature of rhamnogalacturonan I-rich polysaccharide from Korean persimmon vinegar.

    PubMed

    Kim, Hoon; Hong, Hee-Do; Suh, Hyung-Joo; Shin, Kwang-Soon

    2016-08-01

    The crude polysaccharide (KPV-0) isolated from Korean persimmon vinegar was fractionated using gel filtration chromatography to enhance the immunostimulatory activity and to identify the structural features of active fraction. Among three fractions, KPV-I obtained in a void volume, demonstrated the potent production of macrophage-stimulating mediators, including tumor necrosis factor-α, interleukin (IL)-6, IL-12, and nitric oxide. KPV-I showed a combined single peak with high molecular weight of 55,000Da by high performance size exclusion chromatography. Component sugar analysis revealed that KPV-I contained mainly of arabinose, mannose, galactose, rhamnose and galacturonic acid. Single radial gel diffusion assay using β-glucosyl Yariv reagent showed that KPV-I contained arabinogalactan protein with 13.7%. Methylation analysis indicated that KPV-I contained 21 kinds of neutral glycosidic linkages, which seemed to be composed three kinds of polysaccharide; that is a rhamnogalacturonan-I (65-70%) derived from persimmon as a raw material, a mannan (20-25%) derived from fermentation-associated microorganisms, and a linear glucans (less than 10%). In conclusion, polysaccharide isolated from persimmon vinegar could augment the macrophage stimulation, and a large amounts of RG-I polysaccharide derived from persimmon is likely a crucial role in expression of the activity in persimmon vinegar. PMID:27131731

  10. Structural and immunological feature of rhamnogalacturonan I-rich polysaccharide from Korean persimmon vinegar.

    PubMed

    Kim, Hoon; Hong, Hee-Do; Suh, Hyung-Joo; Shin, Kwang-Soon

    2016-08-01

    The crude polysaccharide (KPV-0) isolated from Korean persimmon vinegar was fractionated using gel filtration chromatography to enhance the immunostimulatory activity and to identify the structural features of active fraction. Among three fractions, KPV-I obtained in a void volume, demonstrated the potent production of macrophage-stimulating mediators, including tumor necrosis factor-α, interleukin (IL)-6, IL-12, and nitric oxide. KPV-I showed a combined single peak with high molecular weight of 55,000Da by high performance size exclusion chromatography. Component sugar analysis revealed that KPV-I contained mainly of arabinose, mannose, galactose, rhamnose and galacturonic acid. Single radial gel diffusion assay using β-glucosyl Yariv reagent showed that KPV-I contained arabinogalactan protein with 13.7%. Methylation analysis indicated that KPV-I contained 21 kinds of neutral glycosidic linkages, which seemed to be composed three kinds of polysaccharide; that is a rhamnogalacturonan-I (65-70%) derived from persimmon as a raw material, a mannan (20-25%) derived from fermentation-associated microorganisms, and a linear glucans (less than 10%). In conclusion, polysaccharide isolated from persimmon vinegar could augment the macrophage stimulation, and a large amounts of RG-I polysaccharide derived from persimmon is likely a crucial role in expression of the activity in persimmon vinegar.

  11. A polysaccharide from Dendrobium huoshanense prevents hepatic inflammatory response caused by carbon tetrachloride

    PubMed Central

    Tian, Chang-Cheng; Zha, Xue-Qiang; Luo, Jian-Ping

    2015-01-01

    Dendrobium huoshanense is a precious herbal medicine in China, which exhibits a variety of restorative and therapeutic effects. This study aimed at investigating the hepatoprotective effects of a polysaccharide (DHP1A) isolated from D. huoshanense via water extraction, diethylaminoethyl (DEAE) cellulose anion exchange and size exclusion chromatography. The animal experiment indicated that the oral administration of DHP1A obviously reduced the levels of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and 8-hydroxy-2′-deoxyguanosine in the serum of mice treated with carbon tetrachloride (CCl4), suggesting the hepatoprotective potential of this polysaccharide. Moreover, DHP1A decreased the expressions of tumor necrosis factor-α, interleukin-1β, monocyte chemoattractant protein-1, macrophage inflammatory protein-2, CD68 and phosphorylated IκBα (p-IκBα) in the CCl4-treated mice. These results revealed that the hepatoprotective effect of DHP1A was partly attributed to its anti-inflammatory action. PMID:26019626

  12. Extraction and characterization of the auricularia auricular polysaccharide

    NASA Astrophysics Data System (ADS)

    Zhang, Q. T.

    2016-07-01

    To study a new protein drugs carrier, the Auricularia auricular polysaccharide (AAP) was extracted and purified from Auricularia auricular, and then characterized by the micrOTOF-Q mass spectrometer, UV/Vis spectrophotometer, moisture analyzer and SEM. The results showed that the AAP sample was water- soluble and white flocculence, its molecular weight were 20506.9 Da∼⃒63923.7 Da, and the yield, moisture, and total sugar contents of the AAP were 4.5%, 6.2% and 90.12%(w/w), respectively. The results of the SEM revealed that the AAP dried by vacuum were spherical particles with a smooth surface, and the AAP freeze-dried had continuous porous sheet shape with the loose structure.

  13. Adhesion of marine fouling organisms on hydrophilic and amphiphilic polysaccharides.

    PubMed

    Bauer, Stella; Arpa-Sancet, Maria Pilar; Finlay, John A; Callow, Maureen E; Callow, James A; Rosenhahn, Axel

    2013-03-26

    Polysaccharides are a promising material for nonfouling surfaces because their chemical composition makes them highly hydrophilic and able to form water-storing hydrogels. Here we investigated the nonfouling properties of hyaluronic acid (HA) and chondroitin sulfate (CS) against marine fouling organisms. Additionally, the free carboxyl groups of HA and CS were postmodified with the hydrophobic trifluoroethylamine (TFEA) to block free carboxyl groups and render the surfaces amphiphilic. All coatings were tested with respect to their protein resistance and against settlement and adhesion of different marine fouling species. Both the settlement and adhesion strength of a marine bacterium (Cobetia marina), zoospores of the seaweed Ulva linza, and cells of a diatom (Navicula incerta) were reduced compared to glass control surfaces. In most cases, TFEA capping increased or maintained the performance of the HA coatings, whereas for the very well performing CS coatings the antifouling performance was reduced after capping. PMID:23425225

  14. Immunochemical characterization of cross-reactivity of pneumococcal group 9 capsular polysaccharide types 9N, 9A, 9L, and 9V.

    PubMed Central

    Szu, S; Lee, C J; Carlo, D; Henrichsen, J

    1981-01-01

    The chemical composition and immunochemical characterization of the four cross-reactive pneumococcal capsular polysaccharides within group 9 (types 9N, 9A, 9L, and 9V) were investigated. Their serological reactions were studied by using unabsorbed antisera prepared by immunizing rabbits with pneumococci of each of the four group 9 capsular polysaccharide types. Type 9A antiserum showed the most extensive cross-reactions with the four group 9 polysaccharides. Absorption with type 9N, 9L, or 9V polysaccharide removed 63, 96, or 87%, respectively, of the heterologous antibodies from the type 9A antiserum. All four of the group 9 polysaccharides contained glucose, N-acetylmannosamine, and glucuronic acid. In addition, types 9N and 9L had N-acetylglucosamine, and types 9A, 9L, and 9V contained galactose. Reduction of the uronic acid residues of the type 9 polysaccharides removed most of their homologous and much of their heterologous reactivities, indicating an important role for the uronic acid component in their antigenicity. The four group 9 polysaccharide preparations had comparable molecular sizes and only traces of protein and nucleic acid. Further studies to evaluate the most protective type among the group 9 strains to be included in the current pneumococcal vaccine are discussed. Images PMID:7216451

  15. Polysaccharide intercellular adhesin in biofilm: structural and regulatory aspects

    PubMed Central

    Arciola, Carla Renata; Campoccia, Davide; Ravaioli, Stefano; Montanaro, Lucio

    2015-01-01

    Staphylococcus aureus and Staphylococcus epidermidis are the leading etiologic agents of implant-related infections. Biofilm formation is the main pathogenetic mechanism leading to the chronicity and irreducibility of infections. The extracellular polymeric substances of staphylococcal biofilms are the polysaccharide intercellular adhesin (PIA), extracellular-DNA, proteins, and amyloid fibrils. PIA is a poly-β(1-6)-N-acetylglucosamine (PNAG), partially deacetylated, positively charged, whose synthesis is mediated by the icaADBC locus. DNA sequences homologous to ica locus are present in many coagulase-negative staphylococcal species, among which S. lugdunensis, however, produces a biofilm prevalently consisting of proteins. The product of icaA is an N-acetylglucosaminyltransferase that synthetizes PIA oligomers from UDP-N-acetylglucosamine. The product of icaD gives optimal efficiency to IcaA. The product of icaC is involved in the externalization of the nascent polysaccharide. The product of icaB is an N-deacetylase responsible for the partial deacetylation of PIA. The expression of ica locus is affected by environmental conditions. In S. aureus and S. epidermidis ica-independent alternative mechanisms of biofilm production have been described. S. epidermidis and S. aureus undergo to a phase variation for the biofilm production that has been ascribed, in turn, to the transposition of an insertion sequence in the icaC gene or to the expansion/contraction of a tandem repeat naturally harbored within icaC. A role is played by the quorum sensing system, which negatively regulates biofilm formation, favoring the dispersal phase that disseminates bacteria to new infection sites. Interfering with the QS system is a much debated strategy to combat biofilm-related infections. In the search of vaccines against staphylococcal infections deacetylated PNAG retained on the surface of S. aureus favors opsonophagocytosis and is a potential candidate for immune-protection. PMID

  16. Hyaluronan: A Simple Polysaccharide with Diverse Biological Functions

    PubMed Central

    Dicker, Kevin T.; Gurski, Lisa A.; Pradhan-Bhatt, Swati; Witt, Robert L.; Farach-Carson, Mary C.; Jia, Xinqiao

    2014-01-01

    Hyaluronan (HA) is a linear polysaccharide with disaccharide repeats of D-glucuronic acid and N-acetyl-D-glucosamine. It is evolutionary conserved and abundantly expressed in the extracellular matrix (ECM), on the cell surface and even inside cells. Being a simple polysaccharide, HA exhibits an astonishing array of biological functions. HA interacts with various proteins or proteoglycans to organize the ECM and to maintain tissue homeostasis. The unique physical and mechanical properties of HA contribute to the maintenance of tissue hydration, the mediation of solute diffusion through the extracellular space and the lubrication of certain tissues. The diverse biological functions of HA are manifested through its complex interactions with matrix components and resident cells. Binding of HA with cell surface receptors activates various signaling pathways that regulate cell function, tissue development, inflammation, wound healing and tumor progression and metastasis. Taking advantage of the inherent biocompatibility and biodegradability of HA, as well as its susceptibility to chemical modification, researchers have developed various HA-based biomaterials and tissue constructs with promising and broad clinical potential. In this article, we illustrate the properties of HA from a matrix biology perspective by first introducing principles underlying the biosynthesis and biodegradation of HA, as well as the interactions of HA with various proteins and proteoglycans. We next highlight the roles of HA in physiological and pathological states, including morphogenesis, wound healing and tumor metastasis. A deeper understanding of the mechanisms underlying the roles of HA in various physiological processes can provide new insights and tools for the engineering of complex tissues and tissue models. PMID:24361428

  17. Polysaccharide intercellular adhesin in biofilm: structural and regulatory aspects.

    PubMed

    Arciola, Carla Renata; Campoccia, Davide; Ravaioli, Stefano; Montanaro, Lucio

    2015-01-01

    Staphylococcus aureus and Staphylococcus epidermidis are the leading etiologic agents of implant-related infections. Biofilm formation is the main pathogenetic mechanism leading to the chronicity and irreducibility of infections. The extracellular polymeric substances of staphylococcal biofilms are the polysaccharide intercellular adhesin (PIA), extracellular-DNA, proteins, and amyloid fibrils. PIA is a poly-β(1-6)-N-acetylglucosamine (PNAG), partially deacetylated, positively charged, whose synthesis is mediated by the icaADBC locus. DNA sequences homologous to ica locus are present in many coagulase-negative staphylococcal species, among which S. lugdunensis, however, produces a biofilm prevalently consisting of proteins. The product of icaA is an N-acetylglucosaminyltransferase that synthetizes PIA oligomers from UDP-N-acetylglucosamine. The product of icaD gives optimal efficiency to IcaA. The product of icaC is involved in the externalization of the nascent polysaccharide. The product of icaB is an N-deacetylase responsible for the partial deacetylation of PIA. The expression of ica locus is affected by environmental conditions. In S. aureus and S. epidermidis ica-independent alternative mechanisms of biofilm production have been described. S. epidermidis and S. aureus undergo to a phase variation for the biofilm production that has been ascribed, in turn, to the transposition of an insertion sequence in the icaC gene or to the expansion/contraction of a tandem repeat naturally harbored within icaC. A role is played by the quorum sensing system, which negatively regulates biofilm formation, favoring the dispersal phase that disseminates bacteria to new infection sites. Interfering with the QS system is a much debated strategy to combat biofilm-related infections. In the search of vaccines against staphylococcal infections deacetylated PNAG retained on the surface of S. aureus favors opsonophagocytosis and is a potential candidate for immune-protection. PMID

  18. Molecular cloning and expression of the O4 polysaccharide gene cluster from Escherichia coli.

    PubMed

    Haraguchi, G E; Hull, R A; Krallmann-Wenzel, U; Hull, S I

    1989-02-01

    The Escherichia coli O4 serotype is common among isolates from urinary tract infections. The genes responsible for the biosynthesis of the O4 polysaccharide in a human uropathogenic E. coli were cloned and expressed in E. coli K-12. The recombinant plasmid pGH60, which conferred the O4 phenotype, encoded eight proteins with apparent molecular weights of 39, 36.5, 35, 32.8, 26, 24, 20.7 and 13 kDa.

  19. α-Amylase-assisted extraction of polysaccharides from Panax ginseng.

    PubMed

    Sun, Lin; Wu, Di; Ning, Xin; Yang, Guang; Lin, Ziheng; Tian, Meihong; Zhou, Yifa

    2015-04-01

    In this paper, α-amylase-assisted extraction was used to isolate the polysaccharide that remained in hot water-extracted ginseng. The yield of the polysaccharide was 9.0%, almost equal to that of the hot water-extracted polysaccharide. Using anion exchange and gel permeation chromatography, the polysaccharide was fractionated into a neutral polysaccharide fraction and six pectic fractions. The neutral fraction accounted for 76% of the polysaccharide and contained both amylopectin and amylose. The pectic polysaccharide fractions were identified to be arabinogalactan, type-I rhamnogalacturonan and homogalacturonan-type pectin by high-performance liquid chromatography, Fourier transform-infrared and nuclear magnetic resonance analysis. Structural and lymphocyte proliferation activity results showed that these polysaccharides were different from those extracted by hot water, indicating that ginseng contains complex polysaccharides with diverse structures, which results in its diverse pharmacological activities. The α-amylase-assisted extraction is a novel method for preparing ginseng polysaccharides and could be applied toward the further study and exploration of ginseng. These findings provide technical and theoretical support for ginseng pharmacology.

  20. The Glycobiome of the Rumen Bacterium Butyrivibrio proteoclasticus B316T Highlights Adaptation to a Polysaccharide-Rich Environment

    PubMed Central

    Kelly, William J.; Leahy, Sinead C.; Altermann, Eric; Yeoman, Carl J.; Dunne, Jonathan C.; Kong, Zhanhao; Pacheco, Diana M.; Li, Dong; Noel, Samantha J.; Moon, Christina D.; Cookson, Adrian L.; Attwood, Graeme T.

    2010-01-01

    Determining the role of rumen microbes and their enzymes in plant polysaccharide breakdown is fundamental to understanding digestion and maximising productivity in ruminant animals. Butyrivibrio proteoclasticus B316T is a Gram-positive, butyrate-forming rumen bacterium with a key role in plant polysaccharide degradation. The 4.4Mb genome consists of 4 replicons; a chromosome, a chromid and two megaplasmids. The chromid is the smallest reported for all bacteria, and the first identified from the phylum Firmicutes. B316 devotes a large proportion of its genome to the breakdown and reassembly of complex polysaccharides and has a highly developed glycobiome when compared to other sequenced bacteria. The secretion of a range of polysaccharide-degrading enzymes which initiate the breakdown of pectin, starch and xylan, a subtilisin family protease active against plant proteins, and diverse intracellular enzymes to break down oligosaccharides constitute the degradative capability of this organism. A prominent feature of the genome is the presence of multiple gene clusters predicted to be involved in polysaccharide biosynthesis. Metabolic reconstruction reveals the absence of an identifiable gene for enolase, a conserved enzyme of the glycolytic pathway. To our knowledge this is the first report of an organism lacking an enolase. Our analysis of the B316 genome shows how one organism can contribute to the multi-organism complex that rapidly breaks down plant material in the rumen. It can be concluded that B316, and similar organisms with broad polysaccharide-degrading capability, are well suited to being early colonizers and degraders of plant polysaccharides in the rumen environment. PMID:20689770

  1. A polysaccharide-degrading marine bacterium Flammeovirga sp. MY04 and its extracellular agarase system

    NASA Astrophysics Data System (ADS)

    Han, Wenjun; Gu, Jingyan; Yan, Qiujie; Li, Jungang; Wu, Zhihong; Gu, Qianqun; Li, Yuezhong

    2012-09-01

    Bacteria of the genus Flammeovirga can digest complex polysaccharides (CPs), but no details have been reported regarding the CP depolymerases of these bacteria. MY04, an agarolytic marine bacterium isolated from coastal sediments, has been identified as a new member of the genus Flammeovirga. The MY04 strain is able to utilize multiple CPs as a sole carbon source and grows well on agarose, mannan, or xylan. This strain produces high concentrations of extracellular proteins (490 mg L-1 ± 18.2 mg L-1 liquid culture) that exhibit efficient and extensive degradation activities on various polysaccharides, especially agarose. These proteins have an activity of 310 U mg-1 ± 9.6 U mg-1 proteins. The extracellular agarase system (EAS) in the crude extracellular enzymes contains at least four agarose depolymerases, which are with molecular masses of approximately 30-70 kDa. The EAS is stable at a wide range of pH values (6.0-11.0), temperatures (0-50°C), and sodium chloride (NaCl) concentrations (0-0.9 mol L-1). Two major degradation products generated from agarose by the EAS are identified to be neoagarotetraose and neoagarohexaose, suggesting that β-agarases are the major constituents of the MY04 EAS. These results suggest that the Flammeovirga strain MY04 and its polysaccharide-degradation system hold great promise in industrial applications.

  2. Effects of selenizing angelica polysaccharide and selenizing garlic polysaccharide on immune function of murine peritoneal macrophage.

    PubMed

    Gao, Zhenzhen; Liu, Kuanhui; Tian, Weijun; Wang, Hongchao; Liu, Zhenguang; Li, Youying; Li, Entao; Liu, Cui; Li, Xiuping; Hou, Ranran; Yue, Chanjuan; Wang, Deyun; Hu, Yuanliang

    2015-07-01

    The effects of two selenizing polysaccharides (sCAP2 and sGPS6) on immune function of murine peritoneal macrophages taking two non-selenizing polysaccharides (CAP and GPS) and modifier Na2SeO3 as control. In vitro test, the changes of selenizing polysaccharides, non-selenizing polysaccharides and Na2SeO3 on murine macrophages function were evaluated by phagocytosis and nitric oxide (NO) secretion tests. In vivo test, the mice were injected respectively with 0.2, 0.4 and 0.6 mg of sCAP2, sGPS6, CAP and GPS, or Na2SeO3 80 μg or normal saline 0.4 mL. The peritoneal macrophages were collected and cultured to determine the contents of TNF-α, IL-6 and IL-10 in supernatants by enzyme-linked immunosorbent assay. The results showed that sCAP2 and sGPS6 could significantly promote the phagocytosis and secretion of NO and three cytokines of macrophages in comparison with CAP and GPS. sCAP2 possessed the strongest activity. This indicates that selenylation modification can further improve the immune-enhancing activity of polysaccharide, and sCAP2 could be as a new immunopotentiator.

  3. Influence of extraction parameters on physico-chemical characters and antioxidant activity of water soluble polysaccharides from Macrocybe gigantea (Massee) Pegler & Lodge.

    PubMed

    Khatua, Somanjana; Acharya, Krishnendu

    2016-04-01

    Polysaccharides from mushrooms are potentially active pharmaceutical ingredients and their action is dependent upon conformation, composition, size etc. In this context, three water soluble crude polysaccharide rich fractions viz. hot water extracted polysaccharide (HWP), cold alkaline extracted polysaccharide (CAP) and hot alkaline extracted polysaccharide (HAP) have been isolated using varying extraction parameters from Macrocybe gigantea, a well-known edible mushroom collected from Gangetic plain of West Bengal and authenticated by DNA barcoding of nrDNA ITS region. Physico-chemical investigation revealed that the fractions were mainly composed of β-configuration in pyranose form of sugars conjugated with small amount of protein. Further analysis presented that polysaccharides were composed of same monosaccharide even in similar order of ratio (D-glucose > D-galactose > D-mannose > D-xylose). However, D-glucose as well as β-glucan were found to be in the highest amount in CAP. The helical structure was determined by Congo red assay which indicated that polysaccharides were in aggregate forms except HWP which consisted of tertiary structure. These diverse structural features may have imparted effect on free radical scavenging activity of polysaccharides where HWP was the most active in all assays. HWP was proved to be a good scavenger of free radicals, strong chelator of ferrous ion and had high reducing power. Thus it can be inferred that HWP may foster further studies for searching active compound which might be used as ingredients of functional foods, nutraceuticals and pharmaceuticals. Moreover, to the best of our knowledge this is the first report on chemical composition and antioxidant activity of different crude polysaccharides from M. gigantea. PMID:27413214

  4. A serogroup A meningococcal polysaccharide vaccine

    PubMed Central

    Erwa, H. H.; Haseeb, M. A.; Idris, A. A.; Lapeyssonnie, L.; Sanborn, W. R.; Sippel, J. E.

    1973-01-01

    Vaccination against cerebrospinal meningitis (CSM) has regained interest with the use of capsular polysaccharides (or polyosides) of the meningococcus as specific immunizing agents. These compounds proved to be effective in the USA against meningitis caused by Neisseria meningitidis serotype C. This study considers whether the polysaccharides of the serotype A meningococcus, which is prevalent in the African CSM belt, could be protective in epidemic conditions. Taking advantage of the usual seasonal peak of CSM cases, controlled field trials were undertaken in the Sudan early in 1973. 21 640 persons were vaccinated, half of them with a meningococcal polyoside A vaccine and the other half with tetanus toxoid as a placebo. In the former group there were no cases of meningitis, whereas in the latter 10 cases were reported, of which 7 were confirmed by laboratory tests. These studies indicate that the meningococcal polyoside A vaccine is efficient in epidemic conditions and could be used to control outbreaks of meningococcal meningitis. PMID:4211056

  5. Microbial extracellular polysaccharides and plagioclase dissolution

    NASA Astrophysics Data System (ADS)

    Welch, S. A.; Barker, W. W.; Banfield, J. F.

    1999-05-01

    Bytownite feldspar was dissolved in batch reactors in solutions of starch (glucose polymer), gum xanthan (glucose, mannose, glucuronic acid), pectin (poly-galacturonic acid), and four alginates (mannuronic and guluronic acid) with a range of molecular weights (low, medium, high and uncharacterized) to evaluate the effect of extracellular microbial polymers on mineral dissolution rates. Solutions were analyzed for dissolved Si and Al as an indicator of feldspar dissolution. At neutral pH, feldspar dissolution was inhibited by five of the acid polysaccharides, gum xanthan, pectin, alginate low, alginate medium, alginate high, compared to an organic-free control. An uncharacterized alginate substantially enhanced both Si and Al release from the feldspar. Starch, a neutral polysaccharide, had no apparent effect. Under mildly acidic conditions, initial pH ≈ 4, all of the polymers enhanced feldspar dissolution compared to the inorganic controls. Si release from feldspar in starch solution exceeded the control by a factor of three. Pectin and gum xanthan increased feldspar dissolution by a factor of 10, and the alginates enhanced feldspar dissolution by a factor of 50 to 100. Si and Al concentrations increased with time, even though solutions were supersaturated with respect to several possible secondary phases. Under acidic conditions, initial pH ≈ 3, below the pK a of the carboxylic acid groups, dissolution rates increased, but the relative increase due to the polysaccharides is lower, approximately a factor of two to ten. Microbial extracellular polymers play a complex role in mineral weathering. Polymers appear to inhibit dissolution under some conditions, possibly by irreversibly binding to the mineral surfaces. The extracellular polysaccharides can also enhance dissolution by providing protons and complexing with ions in solution.

  6. Golgi-Mediated Synthesis and Secretion of Matrix Polysaccharides of the Primary Cell Wall of Higher Plants

    PubMed Central

    Driouich, Azeddine; Follet-Gueye, Marie-Laure; Bernard, Sophie; Kousar, Sumaira; Chevalier, Laurence; Vicré-Gibouin, Maïté; Lerouxel, Olivier

    2012-01-01

    The Golgi apparatus of eukaryotic cells is known for its central role in the processing, sorting, and transport of proteins to intra- and extra-cellular compartments. In plants, it has the additional task of assembling and exporting the non-cellulosic polysaccharides of the cell wall matrix including pectin and hemicelluloses, which are important for plant development and protection. In this review, we focus on the biosynthesis of complex polysaccharides of the primary cell wall of eudicotyledonous plants. We present and discuss the compartmental organization of the Golgi stacks with regards to complex polysaccharide assembly and secretion using immuno-electron microscopy and specific antibodies recognizing various sugar epitopes. We also discuss the significance of the recently identified Golgi-localized glycosyltransferases responsible for the biosynthesis of xyloglucan (XyG) and pectin. PMID:22639665

  7. A sulfated polysaccharide, fucans, isolated from brown algae Sargassum vulgare with anticoagulant, antithrombotic, antioxidant and anti-inflammatory effects.

    PubMed

    Dore, Celina Maria P Guerra; das C Faustino Alves, Monique Gabriela; Will, Luiza Sheyla E Pofírio; Costa, Thiago G; Sabry, Diego A; de Souza Rêgo, Leonardo Augusto R; Accardo, Camila M; Rocha, Hugo Alexandre O; Filgueira, Luciana Guimarães A; Leite, Edda Lisboa

    2013-01-01

    Fucan (SV1) sulfated polysaccharides from the brown algae Sargassum vulgare were extracted, fractionated in acetone and examined with respect to chemical composition, anticoagulant, anti-inflammatory, antithrombotic effects and cellular proliferation. These polysaccharides contain low levels of protein, high level of carbohydrate and sulfate. Monosaccharides analysis revealed that SV1 was composed of fucose, galactose, xylose, glucuronic acid and mannose. SV1 polysaccharide prolonged activated partial thromboplastin time (aPTT) and exhibited high antithrombotic action in vivo, with a concentration ten times higher than heparin activity. PSV1, a purified form in gel filtration showed very low biological activities. SV1 stimulated the enzymatic activity of FXa. Its action on DPPH radical scavenging activity was 22%. This polymer has no cytotoxic action (hemolytic) on ABO and Rh blood types in different erythrocyte groups. It displays strong anti-inflammatory action at all concentrations tested in the carrageenan-induced paw edema model, demonstrated by reduced edema and cellular infiltration. PMID:23044157

  8. Isolation and rheological properties of tamarind seed polysaccharide from tamarind kernel powder using protease enzyme and high-intensity ultrasound.

    PubMed

    Poommarinvarakul, Sukhum; Tattiyakul, Jirarat; Muangnapoh, Chirakarn

    2010-06-01

    The effectiveness of using protease and combinations of protease and high-intensity ultrasound for high-purity, high-yield tamarind seed polysaccharide (TSP) production was investigated. Tamarind kernel powder (TKP) suspension was treated with protease alone at 0.16, 0.48, and 0.80 units/mL and with protease-ultrasound combinations over 3 different orders of sequence (before, simultaneous with, and after protease digestion) using combinations of 0.48 units/mL protease and high-intensity ultrasound at 25% and 50% amplitude for 15 and 30 min. The long protease digestion time could produce high-purity isolated TSP, but the polysaccharide yields were lower. The polysaccharide purity and yield were highly improved, even at a shorter protease digestion time, when the protease treatment was combined with high-intensity ultrasound. The increased amplitude level and sonication time decreased the average molecular weight of the polysaccharide. The rheological properties of the TKP and the isolated TSP, from nondestructive oscillatory measurements, demonstrated that the latter present a viscoelastic solution. The decreasing of protein content resulted in better elasticity of the solution. The power law model could be used to fit the down curve between shear rate and shear stress data. The consistency coefficient (K) increased while the flow behavior index decreased with the increased purity of the polysaccharide as a result of increasing increased digestion time, enzyme concentration, sonication power, and sonication time.

  9. Perspective on the Use of Sulfated Polysaccharides from Marine Organisms as a Source of New Antithrombotic Drugs

    PubMed Central

    Mourão, Paulo A. S.

    2015-01-01

    Thromboembolic diseases are increasing worldwide and always require anticoagulant therapy. We still need safer and more secure antithrombotic drugs than those presently available. Sulfated polysaccharides from marine organisms may constitute a new source for the development of such drugs. Investigation of these compounds usually attempts to reproduce the therapeutic effects of heparin. However, we may need to follow different routes, focusing particularly in the following aspects: (1) defining precisely the specific structures required for interaction of these sulfated polysaccharides with proteins of the coagulation system; (2) looking for alternative mechanisms of action, distinct from those of heparin; (3) identifying side effects (mostly pro-coagulant action and hypotension rather than bleeding) and preparing derivatives that retain the desired antithrombotic action but are devoid of side effects; (4) considering that sulfated polysaccharides with low anticoagulant action on in vitro assays may display potent effects on animal models of experimental thrombosis; and finally (5) investigating the antithrombotic effect of these sulfated polysaccharides after oral administration or preparing derivatives that may achieve this effect. If these aspects are successfully addressed, sulfated polysaccharides from marine organisms may conquer the frontier of antithrombotic therapy and open new avenues for treatment or prevention of thromboembolic diseases. PMID:25955754

  10. Polysaccharide Nanosystems for Future Progress in Cardiovascular Pathologies

    PubMed Central

    Silva, Amanda Karine Andriola; Letourneur, Didier; Chauvierre, Cédric

    2014-01-01

    Natural polysaccharides have received a lot of attention in the biomedical field. Indeed, sources of polysaccharides, extracted or produced from plants, bacteria, fungi or algae, are diverse and renewable. Moreover, recent progresses in polysaccharide chemistry and nanotechnologies allow elaborating new dedicated nanosystems. Polysaccharide-based nanosystems may be designed for interacting in several biological processes. In particular, the atherothrombotic pathology is highly concerned by polysaccharide-mediated recognition. Atherothrombotic diseases, regardless of the anatomical localization, remain the main causes of morbidity and mortality in the industrialized world. This review intends to provide an overview on polysaccharide-based nanosystems as drug delivery systems and targeted contrast agents for molecular imaging with an emphasis on the treatment and imaging of cardiovascular pathologies. PMID:24723980

  11. Iron oxyhydroxide mineralization on microbial extracellular polysaccharides

    NASA Astrophysics Data System (ADS)

    Chan, Clara S.; Fakra, Sirine C.; Edwards, David C.; Emerson, David; Banfield, Jillian F.

    2009-07-01

    Iron biominerals can form in neutral pH microaerophilic environments where microbes both catalyze iron oxidation and create polymers that localize mineral precipitation. In order to classify the microbial polymers that influence FeOOH mineralogy, we studied the organic and mineral components of biominerals using scanning transmission X-ray microscopy (STXM), micro X-ray fluorescence (μXRF) microscopy, and high-resolution transmission electron microscopy (HRTEM). We focused on iron microbial mat samples from a creek and abandoned mine; these samples are dominated by iron oxyhydroxide-coated structures with sheath, stalk, and filament morphologies. In addition, we characterized the mineralized products of an iron-oxidizing, stalk-forming bacterial culture isolated from the mine. In both natural and cultured samples, microbial polymers were found to be acidic polysaccharides with carboxyl functional groups, strongly spatially correlated with iron oxyhydroxide distribution patterns. Organic fibrils collect FeOOH and control its recrystallization, in some cases resulting in oriented crystals with high aspect ratios. The impact of polymers is particularly pronounced as the materials age. Synthesis experiments designed to mimic the biomineralization processes show that the polysaccharide carboxyl groups bind dissolved iron strongly but release it as mineralization proceeds. Our results suggest that carboxyl groups of acidic polysaccharides are produced by different microorganisms to create a wide range of iron oxyhydroxide biomineral structures. The intimate and potentially long-term association controls the crystal growth, phase, and reactivity of iron oxyhydroxide nanoparticles in natural systems.

  12. Marine Origin Polysaccharides in Drug Delivery Systems

    PubMed Central

    Cardoso, Matias J.; Costa, Rui R.; Mano, João F.

    2016-01-01

    Oceans are a vast source of natural substances. In them, we find various compounds with wide biotechnological and biomedical applicabilities. The exploitation of the sea as a renewable source of biocompounds can have a positive impact on the development of new systems and devices for biomedical applications. Marine polysaccharides are among the most abundant materials in the seas, which contributes to a decrease of the extraction costs, besides their solubility behavior in aqueous solvents and extraction media, and their interaction with other biocompounds. Polysaccharides such as alginate, carrageenan and fucoidan can be extracted from algae, whereas chitosan and hyaluronan can be obtained from animal sources. Most marine polysaccharides have important biological properties such as biocompatibility, biodegradability, and anti-inflammatory activity, as well as adhesive and antimicrobial actions. Moreover, they can be modified in order to allow processing them into various shapes and sizes and may exhibit response dependence to external stimuli, such as pH and temperature. Due to these properties, these biomaterials have been studied as raw material for the construction of carrier devices for drugs, including particles, capsules and hydrogels. The devices are designed to achieve a controlled release of therapeutic agents in an attempt to fight against serious diseases, and to be used in advanced therapies, such as gene delivery or regenerative medicine. PMID:26861358

  13. Nanofiltration of polysaccharides from Agaricus subrufescens.

    PubMed

    Camelini, C M; Rezzadori, K; Benedetti, S; Proner, M C; Fogaça, L; Azambuja, A A; Giachini, A J; Rossi, M J; Petrus, J C C

    2013-12-01

    A simplified submerged airlift cultivation was established for the production of biomass from Agaricus subrufescens. In this work, soluble polysaccharides extracted from fungal mycelium, fruiting bodies, and the residual culture media were concentrated by nanofiltration. Total and high molar mass polysaccharides and soluble solids were determined in the concentrate for the three extracts. Additionally, the permeate flow, the influences of temperature and pressure, and the resistance to the permeate flow during filtration were also evaluated. Ayield of 5.5 g/L of biomass with 35%glucose conversion was obtained when 0.5 g/L of initial inoculum was employed. Average specific speed of growth was 0.4/day, with biomass productivity of about 0.76 g/(L day). Nanofiltration has yielded polysaccharide increases of 85, 82, and 92% in the extracts from fruiting bodies, mycelium, and liquid media, respectively. A reduction in the permeate flow was observed during filtration, and it was compensated by higher pressures and temperatures. The higher resistance to the permeate flux was caused by polarization due to concentration (polarized gel layer), reaching values of 88% for the culture media. Maximal resistance caused by the membrane reached values of 40% for the extract from the fruiting bodies. On the other hand, resistance caused by fouling was responsible for less than 3.5%. In conclusion, nanofiltration is efficient to concentrate these functional compounds extracted from A. subrufescens and can, therefore, be applied in different biotechnological areas.

  14. Iron oxyhydroxide mineralization on microbial extracellular polysaccharides

    SciTech Connect

    Chan, Clara S.; Fakra, Sirine C.; Edwards, David C.; Emerson, David; Banfield, Jillian F.

    2010-06-22

    Iron biominerals can form in neutral pH microaerophilic environments where microbes both catalyze iron oxidation and create polymers that localize mineral precipitation. In order to classify the microbial polymers that influence FeOOH mineralogy, we studied the organic and mineral components of biominerals using scanning transmission X-ray microscopy (STXM), micro X-ray fluorescence ({mu}XRF) microscopy, and high-resolution transmission electron microscopy (HRTEM). We focused on iron microbial mat samples from a creek and abandoned mine; these samples are dominated by iron oxyhydroxide-coated structures with sheath, stalk, and filament morphologies. In addition, we characterized the mineralized products of an iron-oxidizing, stalk-forming bacterial culture isolated from the mine. In both natural and cultured samples, microbial polymers were found to be acidic polysaccharides with carboxyl functional groups, strongly spatially correlated with iron oxyhydroxide distribution patterns. Organic fibrils collect FeOOH and control its recrystallization, in some cases resulting in oriented crystals with high aspect ratios. The impact of polymers is particularly pronounced as the materials age. Synthesis experiments designed to mimic the biomineralization processes show that the polysaccharide carboxyl groups bind dissolved iron strongly but release it as mineralization proceeds. Our results suggest that carboxyl groups of acidic polysaccharides are produced by different microorganisms to create a wide range of iron oxyhydroxide biomineral structures. The intimate and potentially long-term association controls the crystal growth, phase, and reactivity of iron oxyhydroxide nanoparticles in natural systems.

  15. Immunoregulatory activities of polysaccharides from mung bean.

    PubMed

    Yao, Yang; Zhu, Yingying; Ren, Guixing

    2016-03-30

    Ultrasonic treatment was performed on water-extractable polysaccharides from the seed of mung beans. Purified by anion-exchange and gel filtration chromatography, MWP-1' and MWP-2' were obtained. Average molecular weights (Mws) of MWP-1' and MWP-2' were 68.4 kDa, and 52.4 kDa, respectively. Monosaccharides components analysis indicated that MWP-1' was composed of Rha, Ara, Man and Gal in a molar percent of 0.4:2.6:5.3:0.7. MWP-2' was composed of Ara, Man, Gal and Glc in a molar percent of 0.5:1.4:2.1:0.4. In vitro study showed that both polysaccharides samples were able to stimulate the production of secretory molecules (NO, TNF-α and IL-6) of RAW264.7 murine macrophages in a dosage dependent manner. MWP-2' seemed to be the most potent and induced significantly higher the NO production. These findings suggest that the ultrasonic treatment polysaccharides isolated in our study have immune potentiation effects on macrophages.

  16. Immunochemical characterization of Brucella lipopolysaccharides and polysaccharides.

    PubMed Central

    Moreno, E; Speth, S L; Jones, L M; Berman, D T

    1981-01-01

    Purified lipopolysaccharide (LPS) extracted with phenol-water from smooth Brucella abortus was hydrolyzed with 1% acetic acid at 100 degrees C. The degraded polysaccharide (AH) released gave reactions of identity with the native polysaccharide hapten (NH) in phenol-water- or trichloroacetic acid-extracted endotoxin preparations of B. abortus and with the polysaccharide (poly B) extracted by trichloroacetic acid from rough B. melitensis strain B115. Poly B was present in the soluble cytoplasmic fraction but not in the membrane fraction, of disrupted B115 cells. It could not be extracted from three rough mutants of B. abortus or from B canis or B. ovis cells. Both AH and NH shared determinants present on smooth LPS and missing from poly B. Sugars found in purified LPS, NH, and AH included mannose, glucose, quinovosamine, glucosamine, and 2-keto-3-deoxyoctonate. Poly B contained only a trace amount of quinovosamine and no 2-keto-3-deoxyoctonate detectable by the thiobarbiturate assay. Sera from some rabbits immunized with pure smooth LPS and some, but not all, cows infected with field strains of B. abortus recognized the determinants missing from poly B. A subclass-specific enzyme-linked immunoassay showed that most of the antibody in sera from infected cows which binds to smooth LPS and to NH is of the immunoglobulin G1 subclass. Images PMID:6163716

  17. Evaluation of Phosphorylated Psyllium Seed Polysaccharide as a Release Retardant.

    PubMed

    Rao, Monica R P; Warrier, Deepa U; Rao, Shivani H

    2015-01-01

    The aim of the present study was to modify psyllium seed polysaccharide and evaluate the modified polysaccharide as release retardant in tablets employing ciprofloxacin hydrochloride as model drug. Studies on polysaccharide from psyllium husk has been reported but no work has been reported on characterization and modification of the polysaccharide present in the psyllium (Plantago ovata) seed and the use of the modified polysaccharide as a release retardant in tablets. In this study, the seed gum was modified using sodium trimetaphosphate as crosslinking agent. Sustained release matrix tablets of ciprofloxacin hydrochloride were prepared by wet granulation using various drug-polymer ratios. The polymers investigated were psyllium polysaccharide, phosphorylated psyllium polysaccharide and widely used release retardant hydroxypropyl methylcellulose K100M. The tablets were evaluated for hardness, friability, drug content, swelling profile and in vitro dissolution studies. The matrix tablets containing 1:3 proportion of drug-phosphorylated psyllium polysaccharide was found to have higher hardness as compared to tablets containing 1:1 and 1:2 proportions. The results of swelling behavior in water showed that the tablets containing 1:3 drug:phosphorylated psyllium polysaccharide ratio had swelling comparable to that of tablets containing 1:3 drug:hydroxypropyl methylcellulose ratio. The in vitro dissolution studies shows that the dissolution rate was retarded from 98.41 to 37.6% in 6 h with increase in concentration of phosphorylated psyllium polysaccharide from 100 to 300 mg. Formulations containing psyllium polysaccharide showed complete drug release in 8 h whereas those formulated with phosphorylated psyllium polysaccharide exhibited extended drug release over the 12 h period. Drug release kinetic studies revealed that drug release followed Korsmeyer-Peppas model. PMID:26798177

  18. Evaluation of Phosphorylated Psyllium Seed Polysaccharide as a Release Retardant

    PubMed Central

    Rao, Monica R. P.; Warrier, Deepa U.; Rao, Shivani H.

    2015-01-01

    The aim of the present study was to modify psyllium seed polysaccharide and evaluate the modified polysaccharide as release retardant in tablets employing ciprofloxacin hydrochloride as model drug. Studies on polysaccharide from psyllium husk has been reported but no work has been reported on characterization and modification of the polysaccharide present in the psyllium (Plantago ovata) seed and the use of the modified polysaccharide as a release retardant in tablets. In this study, the seed gum was modified using sodium trimetaphosphate as crosslinking agent. Sustained release matrix tablets of ciprofloxacin hydrochloride were prepared by wet granulation using various drug-polymer ratios. The polymers investigated were psyllium polysaccharide, phosphorylated psyllium polysaccharide and widely used release retardant hydroxypropyl methylcellulose K100M. The tablets were evaluated for hardness, friability, drug content, swelling profile and in vitro dissolution studies. The matrix tablets containing 1:3 proportion of drug-phosphorylated psyllium polysaccharide was found to have higher hardness as compared to tablets containing 1:1 and 1:2 proportions. The results of swelling behavior in water showed that the tablets containing 1:3 drug:phosphorylated psyllium polysaccharide ratio had swelling comparable to that of tablets containing 1:3 drug:hydroxypropyl methylcellulose ratio. The in vitro dissolution studies shows that the dissolution rate was retarded from 98.41 to 37.6% in 6 h with increase in concentration of phosphorylated psyllium polysaccharide from 100 to 300 mg. Formulations containing psyllium polysaccharide showed complete drug release in 8 h whereas those formulated with phosphorylated psyllium polysaccharide exhibited extended drug release over the 12 h period. Drug release kinetic studies revealed that drug release followed Korsmeyer-Peppas model. PMID:26798177

  19. Antibacterial and antiviral study of dialdehyde polysaccharides

    NASA Astrophysics Data System (ADS)

    Song, Le

    Concerns for microbial contamination and infection to the general population, especially the spread of drug-resistant microorganisms, have greatly increased. Polymeric biocides have been found to be a feasible strategy to inactivate drug-resistant bacteria. However, current polymeric biocide systems involve multi-step chemical reactions and they are not cost-effective. Desirable antimicrobial systems need to be designed to be environmentally friendly, broad-spectrum effective against microorganisms, flexible for various delivery methods and economically affordable. We demonstrated that dialdehyde polysaccharides (including dialdehyde starch and dialdehdye cellulose) were broad-spectrum polymeric biocides against gram-positive/negative bacteria, bacteriophages and human virus. These polymers can be easily converted from starch and cellulose through one-step periodate oxidation. Destructions of microorganism by dialdehyde polysaccharides have been achieved in aqueous suspension or by solid surface contact. The dialdehdye functions of dialdehdye polysaccharides were found to be the dominant action against microorganism. The reactivity of the dialdehyde functionality was found to be pH-dependent as well as related to the dispersion of dialdehyde polysaccharides. Degradation of dialdehyde starch during cooking was confirmed. Degradation of dialdehyde starch was more liable in alkaline condition. Carboxylic acid and conjugated aldehyde functionalities were the two main degradation products, confirmed from the spectroscopic studies. The pH effect on the polysaccharide structure and the corresponding antimicrobial activity was very complicated. No decisive conclusions could be obtained from this study. Liner inactivation kinetics was found for dialdehyde starch aqueous suspension against bacteria. This linear inactivation kinetics was derived from the pseudo-first chemical reaction between the dialdehyde starch and the bacteria. The established inactivation kinetics was

  20. Fungal Polysaccharides: Biological Activity Beyond the Usual Structural Properties

    PubMed Central

    Rodrigues, Marcio L.; Nimrichter, Leonardo; Cordero, Radames J. B.; Casadevall, Arturo

    2011-01-01

    Studies on structure and function of polysaccharides in biological systems classically involve sequence and compositional analyses, anomeric configuration, type of glycosidic linkage, and presence of substituents. Recent studies, however, indicates that other structural parameters, so far little explored, can directly influence the biological activity of microbial polysaccharides. Among these parameters, we highlight the molecular dimensions of Cryptococcus neoformans polysaccharides, which appear to be inversely correlated with their immunobiological activity. These recent observations raise new concepts about the structure and function of polysaccharides, which stimulates the design of new experimental approaches and suggests previously unknown applications. PMID:21886639

  1. Polysaccharide based Copolymers as Supramolecular Systems in Biomedical Applications.

    PubMed

    Célia Monteiro de Paula, Regina; Andrade Feitosa, Judith Pessoa; Beserra Paula, Haroldo César

    2015-01-01

    Polysaccharides are natural polymers, obtained from a large variety of sources ranging from fungi to more complex organisms such as birds and whales. Their use for pharmaceutical and biomedical applications has been the subject of numerous researches by the world´s academia. Polysaccharide chemical/physical modifications leading to graft copolymers are discussed in this review, focusing on those nanosystems that are potential candidates for drug delivery applications. Therefore, this review focuses on the biomedical application of polysaccharide based copolymers, particularly as nanocarriers. Copolymer of polysaccharides such as alginate, cellulose, chitosan, dextran, guar, hyaluronic acid, pullulan and starch as drug delivery nanocarriers will be discussed. PMID:26424388

  2. Burkholderia pseudomallei Capsular Polysaccharide Conjugates Provide Protection against Acute Melioidosis

    PubMed Central

    Burtnick, Mary N.; Stokes, Margaret G. M.; Whelan, Adam O.; Williamson, E. Diane; Atkins, Timothy P.; Prior, Joann L.; Brett, Paul J.

    2014-01-01

    Burkholderia pseudomallei, the etiologic agent of melioidosis, is a CDC tier 1 select agent that causes severe disease in both humans and animals. Diagnosis and treatment of melioidosis can be challenging, and in the absence of optimal chemotherapeutic intervention, acute disease is frequently fatal. Melioidosis is an emerging infectious disease for which there are currently no licensed vaccines. Due to the potential malicious use of B. pseudomallei as well as its impact on public health in regions where the disease is endemic, there is significant interest in developing vaccines for immunization against this disease. In the present study, type A O-polysaccharide (OPS) and manno-heptose capsular polysaccharide (CPS) antigens were isolated from nonpathogenic, select-agent-excluded strains of B. pseudomallei and covalently linked to carrier proteins. By using these conjugates (OPS2B1 and CPS2B1, respectively), it was shown that although high-titer IgG responses against the OPS or CPS component of the glycoconjugates could be raised in BALB/c mice, only those animals immunized with CPS2B1 were protected against intraperitoneal challenge with B. pseudomallei. Extending upon these studies, it was also demonstrated that when the mice were immunized with a combination of CPS2B1 and recombinant B. pseudomallei LolC, rather than with CPS2B1 or LolC individually, they exhibited higher survival rates when challenged with a lethal dose of B. pseudomallei. Collectively, these results suggest that CPS-based glycoconjugates are promising candidates for the development of subunit vaccines for immunization against melioidosis. PMID:24866807

  3. Structural characterization and immunomodulatory effect of a polysaccharide HCP-2 from Houttuynia cordata.

    PubMed

    Cheng, Bao-Hui; Chan, Judy Yuet-Wa; Chan, Ben Chung-Lap; Lin, Huang-Quan; Han, Xiao-Qiang; Zhou, Xuelin; Wan, David Chi-Cheong; Wang, Yi-Fen; Leung, Ping-Chung; Fung, Kwok-Pui; Lau, Clara Bik-San

    2014-03-15

    Immunomodulation of natural polysaccharides has been the hot topic of research in recent years. In order to explore the immunomodulatory effect of Houttuynia cordata Thunb., the water extract was studied and a polysaccharide HCP-2 with molecular weight of 60,000 Da was isolated by chromatography using DEAE Sepharose CL-6B and Sephacryl S-500 [corrected] HR columns. The structure characterization of HCP-2 was performed by Fourier transform infrared spectroscopy (FTIR), acidic hydrolysis, PMP derivation, HPLC analysis and nuclear magnetic resonance spectra (NMR). HCP-2 was elucidated as a pectic polysaccharide with a linear chain of 1,4-linked α-D-galacturonic acid residues in which part of the 6-carboxyl groups were methyl esterified and part of 2-hydroxyl groups were acetylated. The bioactivity assays showed that HCP-2 could increase the secretions of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), macrophage inhibitory protein-1α (MIP-1α), macrophage inhibitory protein-1β (MIP-1β), and RANTES (regulated on activation, normal T cell expressed and secreted) in human peripheral blood mononuclear cells (PBMCs), which play critical roles in the innate immune system and shape the adaptive immunity. Our results implied that HCP-2 could be an immune enhancer. PMID:24528726

  4. Isolation, preliminary characterization and hepatoprotective activity of polysaccharides from Tamarindus indica L.

    PubMed

    Samal, Predeep Kumar; Dangi, Jawahar Singh

    2014-02-15

    Polysaccharide was isolated from Tamarindus indica L. (TIP) and was characterized in terms of moisture and ash content, pH, water holding capacity, particle size, tapped density, bulk density, carr's index, Hausners ratio, angle of repose, content of glucose, uronic acid and sulfate. Morphological, spectral (UV-vis, FTIR) and DSC thermal analysis reveals polysaccharide nature of the isolated starch. DPPH radical scavenging activity of TIP shows RSA comparable to that of silymarin. Hepatoprotective potential of TIP in terms of biochemical parameters, SGOT, SGPT, ALP and BRN were significantly increased (P<0.05) and reduction of serum Total protein in the group of rats given thioacetamide (100mg/kg s.c.). Histopathology reveals that TIP under antagonize the effect of thioacetamide by acting, either as membrane stabilizer, thereby preventing the distortion of the cellular ionic environment associated with thioacetamide intoxication, or by preventing interaction of thioacetamide with the transcriptional machinery of the cells.

  5. The core and O-polysaccharide structure of the Caulobacter crescentus lipopolysaccharide.

    PubMed

    Jones, Michael D; Vinogradov, Evgeny; Nomellini, John F; Smit, John

    2015-01-30

    Here we describe the analysis of the structure of the lipopolysaccharide (LPS) from Caulobacter crescentus strain JS1025, a derivative of C. crescentus CB15 NA1000 with an engineered amber mutation in rsaA, leading to the loss of the protein S-layer and gene CCNA_00471 encoding a putative GDP-L-fucose synthase. LPS was isolated using an aqueous membrane disruption method. Polysaccharide and core oligosaccharide were produced by mild acid hydrolysis and analyzed by nuclear magnetic resonance spectroscopy and chemical methods. Spectra revealed the presence of two polysaccharides, one of them, a rhamnan, could be removed using periodate oxidation. Another polymer, built from 4-amino-4-deoxy-D-rhamnose (perosamine), mannose, and 3-O-methyl-glucose, should be the O-chain of the LPS according to genetic data. The attribution of the rhamnan as a part of LPS or a separate polymer was not possible.

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

    PubMed

    Yu, Zeyuan; Liu, Lu; Xu, Yaqin; Wang, Libo; Teng, Xin; Li, Xingguo; Dai, Jing

    2015-11-01

    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 4013 Da, 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.

  7. Structural Characterization of the Extracellular Polysaccharide from Vibrio cholerae O1 El-Tor

    PubMed Central

    Yildiz, Fitnat; Fong, Jiunn; Sadovskaya, Irina; Grard, Thierry; Vinogradov, Evgeny

    2014-01-01

    The ability to form biofilms is important for environmental survival, transmission, and infectivity of Vibrio cholerae, the causative agent of cholera in humans. To form biofilms, V. cholerae produces an extracellular matrix composed of proteins, nucleic acids and a glycoconjugate, termed Vibrio exopolysaccharide (VPS). Here, we present the data on isolation and characterization of the polysaccharide part of the VPS (VPS-PS), which has the following structure: where α-D-Glc is partially (∼20%) replaced with α-D-GlcNAc. α-GulNAcAGly is an amide between 2-acetamido-2-deoxy-α-guluronic acid and glycine. Apparently, the polysaccharide is bound to a yet unidentified component, which gives it high viscosity and completely suppresses any NMR signals belonging to the sugar chains of the VPS. The only reliable method to remove this component at present is a treatment of the whole glycoconjugate with concentrated hydrochloric acid. PMID:24520310

  8. Expression and characterization of a lytic polysaccharide monooxygenase from Bacillus thuringiensis.

    PubMed

    Zhang, Huiyan; Zhao, Yong; Cao, Hailong; Mou, Guangqing; Yin, Heng

    2015-08-01

    Lytic polysaccharide monooxygenases (LPMOs) are recently discovered oxidative enzymes that are capable of oxidative cleavage of recalcitrant polysaccharides such as chitin or cellulose. Despite the importance of LPMOs in biomass conversion and the large number of lpmo genes in microorganisms, only a few LPMOs have been well studied, and further characterization of these proteins is thus of interest. In this study, a chitin-active AA10 family LPMO from Bacillus thuringiensis subsp. kurstaki, BtLPMO10A, is described. This enzyme generates even-numbered oxidized oligosaccharides as the dominated products from crystalline chitin, however, interestingly, when colloidal chitin is used as the substrate, a ladder of oxidized oligosaccharides is observed. These results provide new insights into the action mode of LPMOs that may be affected by the substrates. PMID:25936286

  9. Adsorbents for apheresis prepared from polysaccharides of algae that threaten ecosystem services.

    PubMed

    Kanno, Kenichi; Tanigawa, Tetsuya; Fujita, Yoshihiro; Ohnaka, Naoto; Kubo, Yuichiro; Yoshida, Kaito; Kojima, Kota; Nakata, Koshiro; Shimohara, Masafumi

    2014-08-01

    In this work, we investigated whether materials isolated from algae that threaten ecosystems can be used for human benefit. We converted acidic polysaccharides (ulvan) from the alga Ulva pertusa into soft hydrogel materials. In addition to ulvan, the hydrogels also contained alginate in a polyion complex with chitosan. Cross-linking the hydrogel with glutaraldehyde reduced polysaccharide elution from the polyion complex gel. We also found that both ulvan-chitosan and alginate-chitosan gels were able to remove urea and heavy metals from aqueous solution. This is clinically significant, since during apheresis, toxic compounds such as urea have to be removed from the bloodstream of patients. Importantly, albumin was not removed by the hydrogels, implying that this vital protein can be returned to the bloodstream following dialysis. PMID:25146760

  10. Extraction, chemical analysis of Angelica sinensis polysaccharides and antioxidant activity of the polysaccharides in ischemia-reperfusion rats.

    PubMed

    Zhang, Song; He, Ben; Ge, Junbo; Li, Huibin; Luo, Xiuying; Zhang, Hui; Li, Yuhui; Zhai, Changlin; Liu, Pingang; Liu, Xin; Fei, Xuetao

    2010-11-01

    Angelica sinensis polysaccharides were analyzed using high performance liquid chromatography (HPLC) and Fourier Transform Infrared (FT-IR). The major sugar of the polysaccharide was saccharose (18.55%); and the sugar constituted about 83% of the monomer content. Glucose and fructose were found as minor components of the polysaccharides. The FT-IR spectra of A. sinensis polysaccharides are used for determination of their structural features. The FT-IR spectrum of A. sinensis polysaccharides showed bands at 1641 cm(-1), 1415 cm(-1), 1050 cm(-1) and 926 cm(-1) characteristic for the carboxylic group. Absorptions at 2920-2930 cm(-1) are attributed to asymmetrical stretching vibration of CH(2)-group. Medium stretch observed in the range 1650-1400 cm(-1) is assigned to C-C stretching of polysaccharides. Cardioprotective effects of A. sinensis polysaccharides were evaluated by using myocardial ischemia/reperfusion (IR) rats. A. sinensis polysaccharides treatment significantly reduced myocardial infarction size, enhanced CT-1 and antioxidant enzymes activity, downregulated caspase-12 mRNA expression in rats. The study strongly suggests the cardioprotective activity of A. sinensis polysaccharides in limiting ischemia-reperfusion induced myocardial injury. PMID:20691723

  11. Immunomodulatory dietary polysaccharides: a systematic review of the literature

    PubMed Central

    2010-01-01

    Background A large body of literature suggests that certain polysaccharides affect immune system function. Much of this literature, however, consists of in vitro studies or studies in which polysaccharides were injected. Their immunologic effects following oral administration is less clear. The purpose of this systematic review was to consolidate and evaluate the available data regarding the specific immunologic effects of dietary polysaccharides. Methods Studies were identified by conducting PubMed and Google Scholar electronic searches and through reviews of polysaccharide article bibliographies. Only articles published in English were included in this review. Two researchers reviewed data on study design, control, sample size, results, and nature of outcome measures. Subsequent searches were conducted to gather information about polysaccharide safety, structure and composition, and disposition. Results We found 62 publications reporting statistically significant effects of orally ingested glucans, pectins, heteroglycans, glucomannans, fucoidans, galactomannans, arabinogalactans and mixed polysaccharide products in rodents. Fifteen controlled human studies reported that oral glucans, arabinogalactans, heteroglycans, and fucoidans exerted significant effects. Although some studies investigated anti-inflammatory effects, most studies investigated the ability of oral polysaccharides to stimulate the immune system. These studies, as well as safety and toxicity studies, suggest that these polysaccharide products appear to be largely well-tolerated. Conclusions Taken as a whole, the oral polysaccharide literature is highly heterogenous and is not sufficient to support broad product structure/function generalizations. Numerous dietary polysaccharides, particularly glucans, appear to elicit diverse immunomodulatory effects in numerous animal tissues, including the blood, GI tract and spleen. Glucan extracts from the Trametes versicolor mushroom improved survival and

  12. Proteins.

    ERIC Educational Resources Information Center

    Doolittle, Russell F.

    1985-01-01

    Examines proteins which give rise to structure and, by virtue of selective binding to other molecules, make genes. Binding sites, amino acids, protein evolution, and molecular paleontology are discussed. Work with encoding segments of deoxyribonucleic acid (exons) and noncoding stretches (introns) provides new information for hypotheses. (DH)

  13. Protein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Proteins are the major structural and functional components of all cells in the body. They are macromolecules that comprise 1 or more chains of amino acids that vary in their sequence and length and are folded into specific 3-dimensional structures. The sizes and conformations of proteins, therefor...

  14. Methods of saccharification of polysaccharides in plants

    DOEpatents

    Howard, John; Fake, Gina

    2014-04-29

    Saccharification of polysaccharides of plants is provided, where release of fermentable sugars from cellulose is obtained by adding plant tissue composition. Production of glucose is obtained without the need to add additional .beta.-glucosidase. Adding plant tissue composition to a process using a cellulose degrading composition to degrade cellulose results in an increase in the production of fermentable sugars compared to a process in which plant tissue composition is not added. Using plant tissue composition in a process using a cellulose degrading enzyme composition to degrade cellulose results in decrease in the amount of cellulose degrading enzyme composition or exogenously applied cellulase required to produce fermentable sugars.

  15. In vitro antioxidant activity of polysaccharide from Gardenia jasminoides ellis

    USGS Publications Warehouse

    Fan, Y.; Ge, Z.; Luo, A.

    2011-01-01

    A water-soluble polysaccharide, GP, was isolated from Gardenia jasminoides Ellis through hot water extraction followed by ethanol precipitation. The in vitro free radicals scavenging tests exhibited that GP has significant scavenging abilities especially for ABTS, DPPH, and hydroxyl radicals, which suggests that the polysaccharide GP is a novel antioxidant. ?? 2011 Academic Journals.

  16. Structural modification of polysaccharides: A biochemical-genetic approach

    NASA Technical Reports Server (NTRS)

    Kern, Roger G.; Petersen, Gene R.

    1991-01-01

    Polysaccharides have a wide range of industrial and biomedical applications. An industry trend is underway towards the increased use of bacteria to produce polysaccharides. Long term goals of this work are the adaptation and enhancement of saccharide properties for electronic and optic applications. In this report we illustrate the application of enzyme-bearing bacteriophage on strains of the enteric bacterium Klebsiella pneumoniae, which produces a polysaccharide with the relatively rare rheological property of drag-reduction. This has resulted in the production of new polysaccharides with enhanced rheological properties. Our laboratory is developing techniques for processing and structurally modifying bacterial polysaccharides and oligosaccharides which comprise their basic polymeric repeat units. Our research has focused on bacteriophage which produce specific polysaccharide degrading enzymes. This has lead to the development of enzymes generated by bacteriophage as tools for polysaccharide modification and purification. These enzymes were used to efficiently convert the native material to uniform-sized high molecular weight polymers, or alternatively into high-purity oligosaccharides. Enzyme-bearing bacteriophage also serve as genetic selection tools for bacteria that produce new families of polysaccharides with modified structures.

  17. Effect of Nitrogen on Polysaccharide Production in a Porphyridium sp.

    PubMed

    Arad, S M; Friedman, O D; Rotem, A

    1988-10-01

    Porphyridium cultures grown on either nitrate or ammonium as the nitrogen source showed similar patterns of growth and cell wall polysaccharide production. The effect of nitrogen on growth and cell wall polysaccharide production was studied by applying three regimens of supply: batch mode, in which nitrate was supplied at the beginning of the experiment and became depleted at day 6; continual mode, in which nitrate was added daily; and deficient mode, in which the cells were cultured in a nitrate-free medium. Growth was similar in the batch- and continual-mode cultures, whereas it was totally inhibited in the deficient-mode culture. Polysaccharide content (per volume) was highest in the batch-mode culture and lowest in the deficient-mode culture. However, polysaccharide production per cell was similar in the continual- and deficient-mode cultures, the highest value being found in the batch-mode culture. In addition to its effect on polysaccharide content, nitrogen affected the polysaccharide distribution between soluble and bound polysaccharides. In the deficientmode culture, most of the cell wall polysaccharide was dissolved in the medium.

  18. Inhibitory Effects of Angelica Polysaccharide on Activation of Mast Cells

    PubMed Central

    Mao, Wei-An; Sun, Yuan-Yuan; Mao, Jing-Yi; Wang, Li; Zhang, Jian; Zhou, Jie; Rahman, Khalid; Ye, Ying

    2016-01-01

    This study was designed to investigate the inhibitory effects of Angelica polysaccharide (AP) on activation of mast cells and its possible molecular mechanism. In our study, we determined the proinflammatory cytokines and allergic mediators in anti-DNP IgE stimulated RBL-2H3 cells and found that AP (50, 100, and 200 μg/mL) significantly decreased the release of histamine, β-hexosaminidase, leukotrienes C4 (LTC4), IL-1, IL-4, TNF-α, IL-6, and human monocyte chemotactic protein-1 (MCP-1/CCL2) (p < 0.05). In addition, Ca2+ entry was inhibited by treatment with AP. AP also downregulated the protein expressions of p-Fyn, p-Akt, p-P38, IL-4, TNF-α, and NF-κB p65 in both Fyn gene upregulated and normal RBL-2H3 cells (p < 0.05). Collectively, our results showed that AP could inhibit the activation of mast cells via suppressing the releases of proinflammatory cytokines allergic mediators, Gab2/PI3-K/Akt and Fyn/Syk pathways. PMID:27200102

  19. The role of ascorbate peroxidase, guaiacol peroxidase, and polysaccharides in cassava (Manihot esculenta Crantz) roots under postharvest physiological deterioration.

    PubMed

    Uarrota, Virgílio Gavicho; Moresco, Rodolfo; Schmidt, Eder Carlos; Bouzon, Zenilda Laurita; Nunes, Eduardo da Costa; Neubert, Enilto de Oliveira; Peruch, Luiz Augusto Martins; Rocha, Miguel; Maraschin, Marcelo

    2016-04-15

    This study aimed to investigate the role of ascorbate peroxidase (APX), guaiacol peroxidase (GPX), polysaccharides, and protein contents associated with the early events of postharvest physiological deterioration (PPD) in cassava roots. Increases in APX and GPX activity, as well as total protein contents occurred from 3 to 5 days of storage and were correlated with the delay of PPD. Cassava samples stained with Periodic Acid-Schiff (PAS) highlighted the presence of starch and cellulose. Degradation of starch granules during PPD was also detected. Slight metachromatic reaction with toluidine blue is indicative of increasing of acidic polysaccharides and may play an important role in PPD delay. Principal component analysis (PCA) classified samples according to their levels of enzymatic activity based on the decision tree model which showed GPX and total protein amounts to be correlated with PPD. The Oriental (ORI) cultivar was more susceptible to PPD.

  20. Quantification and characterization of cell wall polysaccharides released by non-Saccharomyces yeast strains during alcoholic fermentation.

    PubMed

    Giovani, Giovanna; Rosi, Iolanda; Bertuccioli, Mario

    2012-11-15

    In order to improve knowledge about the oenological characteristics of non-Saccharomyces yeast strains, and to reconsider their contribution to wine quality, we studied the release of polysaccharides by 13 non-Saccharomyces strains of different species (three wine yeasts, six grape yeasts, and three spoilage yeasts) during alcoholic fermentation in synthetic must. Three Saccharomyces cerevisiae strains were included for comparison. All of the non-Saccharomyces strains released polysaccharides into fermentation medium; the amount released depended on the yeast species, the number of cells formed and their physiological conditions. Normalizing the quantity of macromolecules released to the cell biomass revealed that most non-Saccharomyces strains produced a greater quantity of polysaccharides compared to S. cerevisiae strains after 7 and 14days of fermentation. This capacity was particularly expressed in the studied wine spoilage yeasts (Saccharomycodes ludwigii, Zygosaccharomyces bailii, and Brettanomyces bruxellensis). Chemical characterization of exocellular polysaccharides produced by non-Saccharomyces yeasts revealed them to essentially be mannoproteins with high mannose contents, ranging from 93% for S'codes. ludwigii to 73-74% for Pichia anomala and Starmerella bombicola. Protein contents varied from 9% for P. anomala to 29% for Z. bailii. These compositions were very similar to those of the S. cerevisiae strains, and to the chemical composition of the cell wall mannoproteins of different yeast species. The presence of galactose, in addition to mannose and glucose, in the exocellular polysaccharides released by Schizosaccharomyces pombe, confirmed the parietal nature of the polysaccharides released by non-Saccharomyces yeasts; only this species has a galactomannan located in the outer layer of the cell wall.

  1. The first application of a new polysaccharide from Acanthophyllum bracteatum for the health improvement of Atlantic salmon exposed to mercury chloride.

    PubMed

    Jahanbin, Kambiz; Hedayati, Aliakbar; Moini, Sohrab; Gohari, Ahmad Reza; Emam-Djomeh, Zahra; Esposito, Antonio; Bagheri, Tahere

    2012-05-01

    Polysaccharides are biopolymers that play key roles as therapeutic agents. The current study examined the efficiency of a polysaccharide newly derived from a plant resource for detection of sub-acute mercury (Hg) exposure on blood characteristics of Atlantic salmon. The polysaccharide was extracted from the roots of Acanthophyllum bracteatum, and the Atlantic salmon were divided into three groups and placed in fiberglass tanks: one with no Hg as a control, one filled with 60 µg l(-1) of Hg solution, and one with 60 µg l(-1) Hg solution plus 1% weight/weight (w/w) of polysaccharide, which had been added 24 hours before the addition of Hg solution. Serum hematological and biochemical parameters were analysed. Results of the study showed that the blood factors were significantly adjusted in the group that had been exposed to the polysaccharide and Hg compared to the group that had been exposed only to Hg. The polysaccharide had an adjustment effect on lymphocytes, granulocytes, red blood cells (RBCs), Hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), protein, chlorine (Cl), sodium (Na), and glucose. It did not have a significant effect on hematocrit (Ht), mean corpuscular hemoglobin concentration (MCHC), potassium (K), and monocytes. Finally, the results confirmed that the new polysaccharide has useful effects on the host, possibly acting as a probiotic by selectively motivating the activity of some useful bacteria, which can improve the host health. This is the first report on application of this polysaccharide for the health improvement of Atlantic salmon exposed to mercury chloride. PMID:21911428

  2. Methods for degrading or converting plant cell wall polysaccharides

    DOEpatents

    Berka, Randy; Cherry, Joel

    2008-08-19

    The present invention relates to methods for converting plant cell wall polysaccharides into one or more products, comprising: treating the plant cell wall polysaccharides with an effective amount of a spent whole fermentation broth of a recombinant microorganism, wherein the recombinant microorganism expresses one or more heterologous genes encoding enzymes which degrade or convert the plant cell wall polysaccharides into the one or more products. The present invention also relates to methods for producing an organic substance, comprising: (a) saccharifying plant cell wall polysaccharides with an effective amount of a spent whole fermentation broth of a recombinant microorganism, wherein the recombinant microorganism expresses one or more heterologous genes encoding enzymes which degrade or convert the plant cell wall polysaccharides into saccharified material; (b) fermenting the saccharified material of step (a) with one or more fermenting microoganisms; and (c) recovering the organic substance from the fermentation.

  3. The Antiviral Activities and Mechanisms of Marine Polysaccharides: An Overview

    PubMed Central

    Wang, Wei; Wang, Shi-Xin; Guan, Hua-Shi

    2012-01-01

    Recently, the studies on the antiviral activities of marine natural products, especially marine polysaccharides, are attracting more and more attention all over the world. Marine-derived polysaccharides and their lower molecular weight oligosaccharide derivatives have been shown to possess a variety of antiviral activities. This paper will review the recent progress in research on the antiviral activities and the mechanisms of these polysaccharides obtained from marine organisms. In particular, it will provide an update on the antiviral actions of the sulfated polysaccharides derived from marine algae including carrageenans, alginates, and fucans, relating to their structure features and the structure–activity relationships. In addition, the recent findings on the different mechanisms of antiviral actions of marine polysaccharides and their potential for therapeutic application will also be summarized in detail. PMID:23235364

  4. Characterization of polysaccharides from Ganoderma spp. using saccharide mapping.

    PubMed

    Wu, Ding-Tao; Xie, Jing; Hu, De-Jun; Zhao, Jing; Li, Shao-Ping

    2013-09-12

    Polysaccharides from Ganoderma spp. and their adulterants were firstly investigated and compared using saccharide mapping, enzymatic (endo-1,3-β-D-glucanase and pectinase) digestion followed by polysaccharide analysis using carbohydrate gel electrophoresis analysis. The results showed that both 1,3-β-D-glucosidic and 1,4-α-D-galactosiduronic linkages were existed in Lingzhi (Ganoderma lucidum and Ganoderma sinense), and the similarity of polysaccharides from G. lucidum and G. sinense was high, which may contribute to rational use of Lingzhi. Different species of Ganoderma and their adulterants can be differentiated based on the saccharide mapping, which is helpful to well understand the structural characters of polysaccharides from different species of Ganoderma and to improve the quality control of polysaccharides in Lingzhi.

  5. [Antigenic polysaccharides of bacteria. 22. Structure of the O-specific polysaccharide chain of Proteus hauseri lipopolysaccharide].

    PubMed

    Vinogradov, E V; Shashkov, A S; Knirel', Iu A; Kochetkov, N K; Kholodkova, E V

    1987-05-01

    The following structure of the repeating unit of the Proteus hauseri O-specific polysaccharide was established on the basis of monosaccharide composition and 13C NMR data of the polysaccharide and products of its Smith degradation and partial cleavage with hydrogen fluoride: (Formula: see text).

  6. Structural characterization of polysaccharides from bamboo

    NASA Astrophysics Data System (ADS)

    Kamil, Ruzaimah Nik Mohamad; Yusuf, Nur'aini Raman; Yunus, Normawati M.; Yusup, Suzana

    2014-10-01

    The alkaline and water soluble polysaccharides were isolate by sequential extractions with distilled water, 60% ethanol containing 1%, 5% and 8% NaOH. The samples were prepared at 60 °C for 3 h from local bamboo. The functional group of the sample were examined using FTIR analysis. The most precipitate obtained is from using 60% ethanol containing 8% NaOH with yield of 2.6%. The former 3 residues isolated by sequential extractions with distilled water, 60% ethanol containing 1% and 5% NaOH are barely visible after filtering with cellulose filter paper. The FTIR result showed that the water-soluble polysaccharides consisted mainly of OH group, CH group, CO indicates the carbohydrate and sugar chain. The sample weight loss was slightly decreased with increasing of temperature.

  7. Organized polysaccharide fibers as stable drug carriers.

    PubMed

    Janaswamy, Srinivas; Gill, Kristin L; Campanella, Osvaldo H; Pinal, Rodolfo

    2013-04-15

    Many challenges arise during the development of new drug carrier systems, and paramount among them are safety, solubility and controlled release requirements. Although synthetic polymers are effective, the possibility of side effects imposes restrictions on their acceptable use and dose limits. Thus, a new drug carrier system that is safe to handle and free from side effects is very much in need and food grade polysaccharides stand tall as worthy alternatives. Herein, we demonstrate for the first time the feasibility of sodium iota-carrageenan fibers and their distinctive water pockets to embed and release a wide variety of drug molecules. Structural analysis has revealed the existence of crystalline network in the fibers even after encapsulating the drug molecules, and iota-carrageenan maintains its characteristic and reproducible double helical structure suggesting that the composites thus produced are reminiscent of cocrystals. The melting properties of iota-carrageenan:drug complexes are distinctly different from those of either drug or iota-carrageenan fiber. The encapsulated drugs are released in a sustained manner from the fiber matrix. Overall, our research provides an elegant opportunity for developing effective drug carriers with stable network toward enhancing and/or controlling bioavailability and extending shelf-life of drug molecules using GRAS excipients, food polysaccharides, that are inexpensive and non-toxic.

  8. Organized polysaccharide fibers as stable drug carriers

    PubMed Central

    Janaswamy, Srinivas; Gill, Kristin L.; Campanella, Osvaldo H.; Pinal, Rodolfo

    2013-01-01

    Many challenges arise during the development of new drug carrier systems, and paramount among them are safety, solubility and controlled release requirements. Although synthetic polymers are effective, the possibility of side effects imposes restrictions on their acceptable use and dose limits. Thus, a new drug carrier system that is safe to handle and free from side effects is very much in need and food grade polysaccharides stand tall as worthy alternatives. Herein, we demonstrate for the first time the feasibility of sodium iota-carrageenan fibers and their distinctive water pockets to embed and release a wide variety of drug molecules. Structural analysis has revealed the existence of crystalline network in the fibers even after encapsulating the drug molecules, and iota-carrageenan maintains its characteristic and reproducible double helical structure suggesting that the composites thus produced are reminiscent of cocrystals. The melting properties of iota-carrageenan:drug complexes are distinctly different from those of either drug or iota-carrageenan fiber. The encapsulated drugs are released in a sustained manner from the fiber matrix. Overall, our research provides an elegant opportunity for developing effective drug carriers with stable network toward enhancing and/or controlling bioavailability and extending shelf-life of drug molecules using GRAS excipients, food polysaccharides, that are inexpensive and non–toxic. PMID:23544530

  9. Coordinate regulation of glycan degradation and polysaccharide capsule biosynthesis by a prominent human gut symbiont.

    PubMed

    Martens, Eric C; Roth, Robyn; Heuser, John E; Gordon, Jeffrey I

    2009-07-01

    Bacteria in the distal human gut have evolved diverse abilities to metabolize complex glycans, including the capacity to degrade these compounds as nutrients and to assemble their component sugars into new polymers such as extracellular capsules. The human gut bacterium Bacteroides thetaiotaomicron is well endowed with the ability to metabolize both host- and diet-derived glycans. Its genome contains 88 different polysaccharide utilization loci (PULs) for complex glycan catabolism and eight different gene clusters for capsular polysaccharide biosynthesis. Here, we investigate one of the prominent mechanisms by which this gut symbiont regulates many PULs involved in host mucin O-glycan degradation; namely, transcriptional regulation via the concerted interactions of cell-envelope-localized TonB-dependent transporters, extra-cytoplasmic function sigma factors and anti-sigma factors, which participate together in a regulatory pathway termed trans-envelope signaling. Unexpectedly, we found that several different trans-envelope signaling switches involved in PUL-mediated O-glycan degradation also modulate capsular polysaccharide synthesis. A novel regulatory pathway, which is dependent on expression of O-glycan-targeting outer membrane proteins, governs this coordinated regulation of glycan catabolism and capsule synthesis. This latter finding provides a new link in the dynamic interplay between complex glycan metabolism, microbial physiology, and host responses that occurs during colonization of the gut.

  10. Effect of ultrasonic treatment on the physicochemical properties and antioxidant activities of polysaccharide from Cyclocarya paliurus.

    PubMed

    Tang, Wei; Lin, Lihua; Xie, Jianhua; Wang, Zhijun; Wang, Hui; Dong, Yujia; Shen, Mingyue; Xie, Mingyong

    2016-10-20

    The effects of ultrasonic treatment on the physicochemical properties and antioxidant activities of the polysaccharide from Cyclocarya paliurus leaves were investigated. The physicochemical properties were detected by high-performance gel permeation chromatography (HPGPC), high-performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) and fourier transform infrared spectrometer (FT-IR) as well. The antioxidant activities were evaluated by the assay of DPPH, hydroxyl free radical, β-carotene-linoleic acid, respectfully. Results showed that no considerable change was observed on the structure between the polysaccharide of Cyclocarya paliurus (CP) and the ultrasonic polysaccharide of Cyclocarya paliurus (UCP) by FT-IR, and no apparent differences were found on the content of sugar, uronic acid and protein after the ultrasonic treatment, but difference appeared in the ratio of monosaccharide composition of CP (1.0:4.6:3.5:4.5) and UCP (1.0:5.9:3.9:4.4), and the antioxidant activities of CP were promoted after ultrasonic treatment. PMID:27474571

  11. Chemical compositions and bioactivities of crude polysaccharides from tea leaves beyond their useful date.

    PubMed

    Xiao, Jianbo; Huo, Jianglei; Jiang, Huixian; Yang, Fan

    2011-12-01

    The chemical compositions and bioactivities of crude tea polysaccharides (TPS) from the out-of-date tea leaves (beyond their useful date), namely Xihu Longjing (XTPS), Anxi Tieguanyin (TTPS), Chawentianxia (CTPS) and Huizhoulvcha (HTPS), in market were investigated. These TPS showed similar neutral sugar content and different distribution of molecular weight (1-800 kD). These crude TPS were mainly composed of rhamnose, arabinose, galactose, glucose, xylose, mannose, and galacturonic acid. IR spectra confirmed that these crude TPS were composed of polysaccharide, protein and uronic acids. These TPS showed similar DPPH scavenging activity and exhibited lower DPPH scavenging activities than Vc within 25-200 μg/mL. However, these TPS with higher concentrations (200-400 μg/mL) showed similar DPPH scavenging activity with Vc. HTPS exhibited significant higher superoxide anion scavenging activity than others TPS and gallic acid. XTPS showed significant higher inhibitory effects on α-glucosidase and α-amylase with inhibitory percentages of 64.35% and 82.24% than others TPS. TTPS, XTPS, and HTPS exhibited similar inhibition ability on α-d-glucosidase and α-amylase. The overdue tea leaves can be a resource of tea polysaccharides as function food.

  12. Purification, characterization and antioxidant activities of polysaccharides from thinned-young apple.

    PubMed

    Dou, Jiao; Meng, Yonghong; Liu, Lei; Li, Jie; Ren, Daoyuan; Guo, Yurong

    2015-01-01

    Purification, characterization and antioxidant activities of polysaccharides from thinned-young apple (TYA) were investigated in the study. A water-soluble polysaccharide (TYAP) was obtained with hot water extraction, which was further purified by chromatography of Cellulose DEAE-52 and Sephadex G-150 to get three purified fractions of TYAP-1, TYAP-2 and TYAP-3. HPLC analysis showed that the three fractions were mainly composed of galactose and arabinose with the average molecular weights of 115 kDa, 479 kDa and 403 kDa, respectively and the monosaccharide compositions of TYAP-2 and TYAP-3 were more complicated than that of TYAP-1. Moreover, TYAP fractions of three had no absorptions at 260 nm and 280 nm in the UV spectrum, indicating the absence of protein and nucleic acid. Their structure properties were further confirmed by FT-IR. In vitro antioxidant activities of three fractions were characterized by DPPH(•), HO(•), O2(•-) and ferric-reducing antioxidant power systems. The results indicated that TYAP-1, TYAP-2 and TYAP-3 possessed significant antioxidant effects in a concentration-dependant manner in the tested concentration range of 0.25-5.0mg/mL. Among the fractions tested, TYAP-3 showed significantly higher antioxidant activity than that of TYAP-1 and TYAP-2. All of these findings provide a scientific basis for the further use of polysaccharides from thinned-young apple. PMID:25109456

  13. Antioxidant and anti-aging activities of polysaccharides from Calocybe indica var. APK2.

    PubMed

    Govindan, Sudha; Johnson, Elizabeth Elcy Rani; Christopher, Jabapramila; Shanmugam, Jayasakthi; Thirumalairaj, Vinothkumar; Gopalan, Jayanthi

    2016-06-01

    The crude polysaccharides were extracted from the fruiting bodies of Calocybe indica (CIP). The antioxidant activities of CIP were evaluated both in vitro and in vivo. Chemical characteristics of the polysaccharides were investigated. In in vitro antioxidant assay, CIP showed noticeable 2,2-diphenyl-1-picryl-hydrazyl (DPPH), hydroxyl radical scavenging activities, reducing power and lipid peroxidation inhibition. Chemical analysis showed the presence of carbohydrate, protein and the FTIR spectra revealed the presence of general characteristic absorption peak of the polysaccharides. For in vivo antioxidant activity, two different doses of CIP were orally administrated over a period of 6 weeks in a d-galactose (d-gal) induced aged mice model. Significantly lowered activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), levels of glutathione (GSH) and elevated malondialdehyde (MDA) levels were observed in brain and serum of d-galactose induced rats, when compared to control rats. Administration of CIP significantly raised the activities of SOD, CAT, GPx, levels of GSH and lowered the levels of MDA in mice brain and serum in a dose-dependent manner. The results suggested that CIP had potent antioxidant activity and could minimize the occurrence of age-associated disorders associated with involvement of free radicals. PMID:27174669

  14. Influences of Ulva fasciata polysaccharide on the rheology and stabilization of cinnamaldehyde emulsions.

    PubMed

    Shao, Ping; Shao, Jiamei; Jiang, Yike; Sun, Peilong

    2016-01-01

    Emulsifying properties of water soluble polysaccharides from Ulva fasciata (UFP) were evaluated in cinnamaldehyde/water emulsions in terms of droplet size distribution, rheological properties, visual phase separation, and zeta-potential. The cinnamaldehyde/water (10%, wt/wt) emulsions were formulated and stabilized by different concentrations of UFP (0.1-4%, wt/wt). The obtained emulsions showed monomodal droplet size distributions with average droplet size (D[3,2]) below 1.0μm, when 3% (wt/wt) UFP was added as the emulsifying agent under a homogenization pressure of 75MPa. The rheological properties and zeta-potential of the emulsions appeared to be dependent on the UFP concentration. Furthermore, the UFP exhibited better emulsifying and stabilizing properties in the investigated system when compared to other commercial polysaccharides of gum Arabic and gum Ghatti. The results also suggested that the emulsifying and stabilizing mechanism of the UFP may not only be ascribed to its surface-active protein moiety, but also to the hydrophobicity of the polysaccharide itself. These findings provided a theoretical basis for potential utilization of UFP as a novel hydrocolloid emulsifying agent. PMID:26453847

  15. Bacterial glycobiology: rhamnose-containing cell wall polysaccharides in Gram-positive bacteria

    PubMed Central

    Mistou, Michel-Yves; Sutcliffe, Iain C.; van Sorge, Nina M.

    2016-01-01

    The composition of the Gram-positive cell wall is typically described as containing peptidoglycan, proteins and essential secondary cell wall structures called teichoic acids, which comprise approximately half of the cell wall mass. The cell walls of many species within the genera Streptococcus, Enterococcus and Lactococcus contain large amounts of the sugar rhamnose, which is incorporated in cell wall-anchored polysaccharides (CWP) that possibly function as homologues of well-studied wall teichoic acids (WTA). The presence and chemical structure of many rhamnose-containing cell wall polysaccharides (RhaCWP) has sometimes been known for decades. In contrast to WTA, insight into the biosynthesis and functional role of RhaCWP has been lacking. Recent studies in human streptococcal and enterococcal pathogens have highlighted critical roles for these complex polysaccharides in bacterial cell wall architecture and pathogenesis. In this review, we provide an overview of the RhaCWP with regards to their biosynthesis, genetics and biological function in species most relevant to human health. We also briefly discuss how increased knowledge in this field can provide interesting leads for new therapeutic compounds and improve biotechnological applications. PMID:26975195

  16. Purification, characterization and antitumor activity of polysaccharides extracted from Phellinus igniarius mycelia.

    PubMed

    Li, Shi-Chao; Yang, Xiao-Ming; Ma, Hai-Le; Yan, Jing-Kun; Guo, Dan-Zhao

    2015-11-20

    Water-soluble intracellular polysaccharides (IPS) were extracted from cultured mycelia of Phellinus igniarius. The IPS were purified by ethanol fractional precipitation, ion-exchange and size exclusion chromatography in that order. Homogeneous polysaccharide IPSW-1, IPSW-2, IPSW-3, and IPSW-4 were obtained, which molecular characteristics were examined using multiangle laser-light scattering and refractive index detector system. The average molecular weights of them were 34.1, 17.7, 15.1, 21.7kDa, respectively. GC analysis indicated that IPSW-1, IPSW-2 and IPSW-3 all only contained glucose, while IPSW-4 was composed of rhamnose, xylose, mannose, glucose and galactose in a molar ratio of 1.29:1.21:1:43.86:1.86. UV and IR analysis suggested they belonged to α-type of the pyran group and didn't contain protein. These homogeneous polysaccharides could inhibit the growth of SW480 and HepG2 cells to a certain extent in a dose-dependent manner. So they could be beneficial for the further development of a natural carcinoma preventive agent and functional food. PMID:26344250

  17. Bacterial glycobiology: rhamnose-containing cell wall polysaccharides in Gram-positive bacteria.

    PubMed

    Mistou, Michel-Yves; Sutcliffe, Iain C; van Sorge, Nina M

    2016-07-01

    The composition of the Gram-positive cell wall is typically described as containing peptidoglycan, proteins and essential secondary cell wall structures called teichoic acids, which comprise approximately half of the cell wall mass. The cell walls of many species within the genera Streptococcus, Enterococcus and Lactococcus contain large amounts of the sugar rhamnose, which is incorporated in cell wall-anchored polysaccharides (CWP) that possibly function as homologues of well-studied wall teichoic acids (WTA). The presence and chemical structure of many rhamnose-containing cell wall polysaccharides (RhaCWP) has sometimes been known for decades. In contrast to WTA, insight into the biosynthesis and functional role of RhaCWP has been lacking. Recent studies in human streptococcal and enterococcal pathogens have highlighted critical roles for these complex polysaccharides in bacterial cell wall architecture and pathogenesis. In this review, we provide an overview of the RhaCWP with regards to their biosynthesis, genetics and biological function in species most relevant to human health. We also briefly discuss how increased knowledge in this field can provide interesting leads for new therapeutic compounds and improve biotechnological applications.

  18. Purification and Evaluation of Polysaccharide Intercellular Adhesion (PIA) Antigen from Staphylococcus epidermidis.

    PubMed

    Mirzaei, Bahman; Moosavi, Seyed Fazlollah; Babaei, Ryhane; Siadat, Seyed Davar; Vaziri, Farzam; Shahrooei, Mohammad

    2016-11-01

    The polysaccharide intercellular adhesin (PIA) confers major functional effects in biofilm formation, which bears an important role in the pathogenicity of Staphylococcus epidermidis. Following the identification of biofilm-forming strains by biochemical and molecular methods, isogenic strain was prepared and in vitro biofilm formation assay was performed consequently. By parallel analysis of both the PIA-positive and PIA-negative strains using size exclusion chromatography by Fast protein liquid chromatography (FPLC) method, the respective PIA was purified. Recovered PIA was examined using colorimetric and hemagglutination assays. Finally, the recovered PIA was analyzed using Fourier-transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy methods. By the parallel purification process and comparison of the obtained graphs from the FPLC detector, fractions near the void volume were determined as PIA. The colorimetric and hemagglutination assays were applied and the content of carbohydrates (hexose = 620 µg/ml, hexosamine = 5700 µg/ml and ketoses = 170 µg/ml) and hemagglutination titer (1:128) in recovered polysaccharide were determined. This study shows that PIA has a significant role in the biofilm formation in S. epidermidis strains. The recovered polysaccharide and its molecular weight were analyzed within the near void volume of the utilized column.

  19. Purification and Evaluation of Polysaccharide Intercellular Adhesion (PIA) Antigen from Staphylococcus epidermidis.

    PubMed

    Mirzaei, Bahman; Moosavi, Seyed Fazlollah; Babaei, Ryhane; Siadat, Seyed Davar; Vaziri, Farzam; Shahrooei, Mohammad

    2016-11-01

    The polysaccharide intercellular adhesin (PIA) confers major functional effects in biofilm formation, which bears an important role in the pathogenicity of Staphylococcus epidermidis. Following the identification of biofilm-forming strains by biochemical and molecular methods, isogenic strain was prepared and in vitro biofilm formation assay was performed consequently. By parallel analysis of both the PIA-positive and PIA-negative strains using size exclusion chromatography by Fast protein liquid chromatography (FPLC) method, the respective PIA was purified. Recovered PIA was examined using colorimetric and hemagglutination assays. Finally, the recovered PIA was analyzed using Fourier-transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy methods. By the parallel purification process and comparison of the obtained graphs from the FPLC detector, fractions near the void volume were determined as PIA. The colorimetric and hemagglutination assays were applied and the content of carbohydrates (hexose = 620 µg/ml, hexosamine = 5700 µg/ml and ketoses = 170 µg/ml) and hemagglutination titer (1:128) in recovered polysaccharide were determined. This study shows that PIA has a significant role in the biofilm formation in S. epidermidis strains. The recovered polysaccharide and its molecular weight were analyzed within the near void volume of the utilized column. PMID:27460584

  20. Cell wall carbohydrates from fruit pulp of Argania spinosa: structural analysis of pectin and xyloglucan polysaccharides.

    PubMed

    Aboughe-Angone, Sophie; Nguema-Ona, Eric; Ghosh, Partha; Lerouge, Patrice; Ishii, Tadashi; Ray, Bimalendu; Driouich, Azeddine

    2008-01-14

    Isolated cell walls of Argania spinosa fruit pulp were fractionated into their polysaccharide constituents and the resulting fractions were analysed for monosaccharide composition and chemical structure. The data reveal the presence of homogalacturonan, rhamnogalacturonan-I (RG-I) and rhamnogalacturonan-II (RG-II) in the pectic fraction. RG-I is abundant and contains high amounts of Ara and Gal, indicative of an important branching in this polysaccharide. RG-II is less abundant than RG-I and exists as a dimer. Structural characterisation of xyloglucan using enzymatic hydrolysis, gas chromatography, MALDI-TOF-MS and methylation analysis shows that XXGG, XXXG, XXLG and XLLG are the major subunit oligosaccharides in the ratio of 0.6:1:1.2:1.6. This finding demonstrates that the major neutral hemicellulosic polysaccharide is a galacto-xyloglucan. In addition, Argania fruit xyloglucan has no XUFG, a novel xyloglucan motif recently discovered in Argania leaf cell walls. Finally, the isolation and analysis of arabinogalactan-proteins showed that Argania fruit pulp is rich in these proteoglycans. PMID:18005949

  1. Preparation and immunogenicity-evaluation of typhoid O-specific polysaccharides bio-conjugate vaccines.

    PubMed

    Zhehui, Peng; Chao, Pan; Peng, Sun; Erling, Feng; Jun, Wu; Li, Zhu; Qingzhong, Peng; Hengliang, Wang

    2015-05-01

    Typhoid fever caused by Salmonella Typhi is still a major public health problem in developing countries. In this study, we constructed a genetically modified Salmonella Typhi strain expressing O-specific polysaccharides (OPS) antigen conjugated to a carrier, recombinant Pseudomonas aeruginosa exotoxin A(rEPA N29). The conjugates (OPS-rEPA N29) were further purified and evaluated for their immunogenicity. The results of ELISA showed that the conjugates evoked higher titers of IgG than OPS, suggesting that rEPAN29 increased immunogenicity of OPS significantly as a carrier. Moreover, three injections with 3-week interval evoked slightly higher titers of IgG than three injections with 2-week interval. However, injection of excess conjugates could not evoke higher titers of IgG against lipid polysaccharide (LPS). In summary, our study provides a new strategy for preparing polysaccharides-protein conjugate vaccines as well as similar bio-conjugate vaccines of other Gram-negative pathogens.

  2. Up regulation of annexin A2 on murine H22 hepatocarcinoma cells induced by cartilage polysaccharide.

    PubMed

    Liu, Anjun; Liu, Dongyue; Zhao, Sisi; Zheng, Jie; Cao, Dongxu; Zhang, Hong

    2011-10-01

    In this study, we investigated the antitumor effect of a tumor vaccine prepared from H22 hepatocarcinoma cells induced by cartilage polysaccharide. We found out there were specific antigens which combined with antigen-specific antibodies from immune murine serum. Results of western blot analysis showed that about 36 kDa make specific antibodies appeared specific antibodies in antiserum of immune mice, whereas the best immune effects became visible at the induction time of 48 h. Analyses of 2-dimensional electrophoresis identified the specific antigen was annexin A2, which was a glycosylated protein that contained a glycosylation site, closely related to oncogenesis, cancer development, invasion and metastasis. Proteomics indicated that both quantity and conformation of annexin A2 were changed after induced by cartilage polysaccharide. Lastly, we found there was a major increase of annexin A2 mRNA on H22 cells induced by cartilage polysaccharide. In summary, our data suggested that annexin A2, a specific antigen played a key role in antitumor immune response and activating the immune system. It would be a potential type of tumor vaccine which provided new ideas for tumor immunoprophylaxis. PMID:21111695

  3. Purification, characterization and antitumor activity of polysaccharides extracted from Phellinus igniarius mycelia.

    PubMed

    Li, Shi-Chao; Yang, Xiao-Ming; Ma, Hai-Le; Yan, Jing-Kun; Guo, Dan-Zhao

    2015-11-20

    Water-soluble intracellular polysaccharides (IPS) were extracted from cultured mycelia of Phellinus igniarius. The IPS were purified by ethanol fractional precipitation, ion-exchange and size exclusion chromatography in that order. Homogeneous polysaccharide IPSW-1, IPSW-2, IPSW-3, and IPSW-4 were obtained, which molecular characteristics were examined using multiangle laser-light scattering and refractive index detector system. The average molecular weights of them were 34.1, 17.7, 15.1, 21.7kDa, respectively. GC analysis indicated that IPSW-1, IPSW-2 and IPSW-3 all only contained glucose, while IPSW-4 was composed of rhamnose, xylose, mannose, glucose and galactose in a molar ratio of 1.29:1.21:1:43.86:1.86. UV and IR analysis suggested they belonged to α-type of the pyran group and didn't contain protein. These homogeneous polysaccharides could inhibit the growth of SW480 and HepG2 cells to a certain extent in a dose-dependent manner. So they could be beneficial for the further development of a natural carcinoma preventive agent and functional food.

  4. Cell Wall Polysaccharide Synthases Are Located in Detergent-Resistant Membrane Microdomains in Oomycetes ▿ †

    PubMed Central

    Briolay, Anne; Bouzenzana, Jamel; Guichardant, Michel; Deshayes, Christian; Sindt, Nicolas; Bessueille, Laurence; Bulone, Vincent

    2009-01-01

    The pathways responsible for cell wall polysaccharide biosynthesis are vital in eukaryotic microorganisms. The corresponding synthases are potential targets of inhibitors such as fungicides. Despite their fundamental and economical importance, most polysaccharide synthases are not well characterized, and their molecular mechanisms are poorly understood. With the example of Saprolegnia monoica as a model organism, we show that chitin and (1→3)-β-d-glucan synthases are located in detergent-resistant membrane microdomains (DRMs) in oomycetes, a phylum that comprises some of the most devastating microorganisms in the agriculture and aquaculture industries. Interestingly, no cellulose synthase activity was detected in the DRMs. The purified DRMs exhibited similar biochemical features as lipid rafts from animal, plant, and yeast cells, although they contained some species-specific lipids. This report sheds light on the lipid environment of the (1→3)-β-d-glucan and chitin synthases, as well as on the sterol biosynthetic pathways in oomycetes. The results presented here are consistent with a function of lipid rafts in cell polarization and as platforms for sorting specific sets of proteins targeted to the plasma membrane, such as carbohydrate synthases. The involvement of DRMs in the biosynthesis of major cell wall polysaccharides in eukaryotic microorganisms suggests a function of lipid rafts in hyphal morphogenesis and tip growth. PMID:19201970

  5. Comparison of physicochemical properties and immunomodulatory activity of polysaccharides from fresh and dried litchi pulp.

    PubMed

    Huang, Fei; Zhang, Ruifen; Yi, Yang; Tang, Xiaojun; Zhang, Mingwei; Su, Dongxiao; Deng, Yuanyuan; Wei, Zhencheng

    2014-03-31

    Drying is commonly used for preservation and processing of litchi. However, its polysaccharide structure may be altered by the drying process, resulting in biological activity changes. Polysaccharides from fresh and dried litchi pulp (denoted as LPF and LPD, respectively) were isolated, investigated by GC-MS, GPC and UV/IR spectrum analysis and their antitumor and immunomodulatory activities were evaluated in vitro. LPD, the molecular weight of which was lower than that of LPF, contained more protein, uronic acid, arabinose, galactose and xylose. Compared with LPF, LPD exhibited a higher inhibitory effect on the proliferation of HepG2, Hela and A549 cells from 50-750 μg/mL. LPD was also a better stimulator of spleen lymphocyte proliferation, NK cells cytotoxicity and macrophage phagocytosis from 50-400 μg/mL. In summary, drying could change the physicochemical properties and enhance the bioactivity of polysaccharides from litchi pulp. This finding is supported by the fact that dried litchi pulps are used in Traditional Chinese Medicine.

  6. Phase separation behavior of egg yolk suspensions after anionic polysaccharides addition.

    PubMed

    Navidghasemizad, Sahar; Temelli, Feral; Wu, Jianping

    2015-03-01

    The objectives of this study were to understand the interactions between three anionic polysaccharides (gum arabic, xanthan gum and ι-carrageenan) and egg yolk at pH 3, 5, 6, 8, 10 and possible phase separation behavior. Zeta potential of egg yolk was not affected by gum arabic addition while it became more negative at pH 5 after xanthan gum and ι-carrageenan addition. The particle size of ι-carrageenan yolk suspension was considerably higher than the other polysaccharide yolk suspensions at pH below 6 but was dramatically decreased at alkaline pH. Most polysaccharide yolk suspensions formed either a biphasic or a monophasic system, whereas three distinct phases were observed for xanthan gum yolk suspension at pH 6. Protein profile analysis of the lipid-rich cream phase obtained from xanthan gum added yolk showed similarities to apoproteins from low density lipoproteins (LDL) of egg yolk. Microscopy analysis indicated the co-presence of xanthan gum and LDL in the creamy phase, within a network formed by xanthan gum. It was suggested that electrostatic and hydrophobic interactions between the egg yolk and xanthan gum as well as xanthan gum's rheological properties could be responsible for the unique phase separation observed in the study. The findings of this study can form the basis for future studies to develop a new method to separate LDL from egg yolk.

  7. Proteins

    NASA Astrophysics Data System (ADS)

    Regnier, Fred E.; Gooding, Karen M.

    Because of the complexity of cellular material and body fluids, it is seldom possible to analyze a natural product directly. Qualitative and quantitative analyses must often be preceded by some purification step that separates the molecular species being examined from interfering materials. In the case of proteins, column liquid chromatography has been used extensively for these fractionations. With the advent of gel permeation, cation exchange, anion exchange, hydrophobic, and affinity chromatography, it became possible to resolve proteins through their fundamental properties of size, charge, hydrophobicity, and biological affinity. The chromatographic separations used in the early isolation and characterization of many proteins later became analytical tools in their routine analysis. Unfortunately, these inherently simple and versatile column chromatographic techniques introduced in the 50s and 60s have a severe limitation in routine analysis-separation time. It is common to encounter 1-24 h separation times with the classical gel-type supports.

  8. Role of sulphated polysaccharides from Sargassum Wightii in Cyclosporine A-induced oxidative liver injury in rats

    PubMed Central

    Josephine, Anthony; Nithya, Kalaiselvam; Amudha, Ganapathy; Veena, Coothan Kandaswamy; Preetha, Sreenivasan P; Varalakshmi, Palaninathan

    2008-01-01

    Background Seaweeds or marine algae have long been made up a key part of the Asian diet, and as an antioxidant, sulphated polysaccharides have piqued the interest of many researchers as one of the ocean's greatest treasures. The present investigation suggests the therapeutic potential of sulphated polysaccharides from marine brown algae "Sargassum wightii" in Cyclosporine A (CsA)- induced liver injury. CsA is a potent immunosuppressive agent used in the field of organ transplantations and various autoimmune disorders. However, hepatotoxicity due to CsA remains to be one of the major clinical challenges. Methods The effect of sulphated polysaccharides on CsA-induced hepatotoxicity was studied in adult male albino rats of Wistar strain, and the animals were randomized into four groups with six rats in each. Group I served as vehicle control. Group II rats were given CsA at a dosage of 25 mg/kg body weight, orally for 21 days. Group III rats were given sulphated polysaccharides at a dosage of 5 mg/kg body weight, subcutaneously for 21 days. Group IV rats were given sulphated polysaccharides simultaneously along with CsA, as mentioned in Group II for 21 days. Results CsA provoked hepatotoxicity was evident from the decreased activities of hepatic marker enzymes. A significant rise in the level of oxidants, along with a striking decline in both the enzymic and non-enzymic antioxidants, marks the severity of oxidative stress in CsA-induced rats. This in turn led to enhanced levels of lipid peroxidation, 8-hydroxy-2-deoxy guanosine and protein carbonyls, along with a decrease in ATPase activities and alterations in lipid profile. Histopathological changes also strongly support the above aberrations. However, concomitant treatment with sulphated polysaccharides restored the above deformities to near control and prevented the morphological alterations significantly. Conclusion Thus, the present study highlights that sulphated polysaccharides can act therapeutically against

  9. Antiangiogenic activity and direct antitumor effect from a sulfated polysaccharide isolated from seaweed.

    PubMed

    Guerra Dore, Celina Maria P; Faustino Alves, Monique Gabriela C; Santos, Nednaldo D; Cruz, Ana Katarina M; Câmara, Rafael Barros G; Castro, Allisson Jonathan G; Guimarães Alves, Luciana; Nader, Helena B; Leite, Edda Lisboa

    2013-07-01

    Angiogenesis is a dynamic proliferation and differentiation process. It requires endothelial proliferation, migration and tube formation. In this context, endothelial cells are a preferred target for several studies and therapies. Anionic polysaccharides (SV1 and PSV1) from brown seaweed Sargassum vulgare were fractionated (SV1), purified (PSV1) and displayed with high total sugars, sulfate content and very low level of protein. The antiangiogenic efficacy of polysaccharides was examined in vivo in the chick chorioallantoic membrane (CAM) model by using fertilized eggs. Decreases in the density of the capillaries were assessed and scored. The results showed that SV1 and PSV1 have an inhibitory effect on angiogenesis. These results were also confirmed by the inhibition of tubulogenesis in rabbit aorta endothelial cell (RAEC) in matrigel. These compounds were assessed in an apoptosis assay (Annexin V-FITC/PI) and cell viability by MTT assay of RAEC. These polysaccharides did not affect the viability and did not have apoptotic or necrotic action. RAEC cell when incubated with SV1and PSV1 showed inhibition of VEGF secretion, observed when compounds were incubated at 25, 50 and 100 μg/μL. The VEGF secretion with the RAEC cell line for 24 h was more effective for PSV1 at 50 μg/μL (71.4%) than for SV1 at 100 μg/μL (75.9%). SV1 and PSV1 had an antiproliferative action (47%) against tumor cell line HeLa. Our results indicate that these sulfated polysaccharides have antiangiogenic and antitumor actions. PMID:23507505

  10. Immunogenicity of polysaccharides conjugated to peptides containing T- and B-cell epitopes.

    PubMed Central

    Lett, E; Gangloff, S; Zimmermann, M; Wachsmann, D; Klein, J P

    1994-01-01

    To develop a general model of polysaccharide-peptide vaccine, we have investigated the efficiency of linear peptides derived from protein SR, and adhesin of the I/II protein antigen family of oral streptococci, to act as carriers for two T cell-independent polysaccharides: serogroup f polysaccharide from Streptococcus mutans OMZ 175 (poly f) and Saccharomyces cerevisiae mannan. Peptide 3 (YEKEPTPPTRTPDQ) and peptide 6 (TPEDPTDPTDPQDPSS), accessible on the native SR protein as demonstrated by their reactivity in enzyme-linked immunosorbent assays with rat antisera raised against protein SR, correspond to immunodominant regions of SR. Peptide 3 contains at least one B- and one T-cell epitope, as demonstrated by its ability to induce peptide- and SR-specific antibody responses without any carrier and to stimulate the proliferation of rat lymph node cells primed either with free peptide or native SR, whereas peptide 6 contains only B-cell epitope(s). Peptide 3 was then covalently coupled though reductive amination to either poly f or mannan, and peptide 6 was coupled to poly f. Subcutaneous immunizations of rats with poly f-peptide 3 or mannan-peptide 3 conjugates produced a systemic immunoglobulin M (IgM) and IgG antibody response, and the elicited antibodies reacted with free poly f or mannan, peptide 3, protein SR, and S. mutans or S. cerevisiae whole cells. Rats immunized with poly f-peptide 6 did not develop any antipeptide or anti-SR response. Furthermore, a booster immunization of animals with poly f-peptide 3 or mannan-peptide 3 conjugates induced high titers of anti-peptide 3, anti-poly f, and antimannan antibodies, which occurred quickly. The response is anamnestic for the peptide and the polysaccharides and is characterized by an Ig switch from IgM to IgG. The data presented here confirm that the presence of B- and T-cell epitopes is necessary to induce an anamnestic antipeptide response and that a peptide containing relevant B- and T-cell epitopes can act

  11. The immunostimulating role of lichen polysaccharides: a review.

    PubMed

    Shrestha, Gajendra; St Clair, Larry L; O'Neill, Kim L

    2015-03-01

    The immune system has capacity to suppress the development or progression of various malignancies including cancer. Research on the immunomodulating properties of polysaccharides obtained from plants, microorganisms, marine organisms, and fungi is growing rapidly. Among the various potential sources, lichens, symbiotic systems involving a fungus and an alga and/or a cyanobacterium, show promise as a potential source of immunomodulating compounds. It is well known that lichens produce an abundance of structurally diverse polysaccharides. However, only a limited number of studies have explored the immunostimulating properties of lichen polysaccharides. Published studies have shown that some lichen polysaccharides enhance production of nitrous oxide (NO) by macrophages and also alter the production levels of various proinflammatory and antiinflammatory cytokines (IL-10, IL-12, IL-1β, TNF-α, and IFN-α/β) by macrophages and dendritic cells. Although there are only a limited number of studies examining the role of lichen polysaccharides, all results suggest that lichen polysaccharides can induce immunomodulatory responses in macrophages and dendritic cells. Thus, a detailed evaluation of immunomodulatory capacity of lichen polysaccharides could provide a unique opportunity for the discovery of novel therapeutic agents. PMID:25339289

  12. Antibiofilm activity of Actinobacillus pleuropneumoniae serotype 5 capsular polysaccharide.

    PubMed

    Karwacki, Michael T; Kadouri, Daniel E; Bendaoud, Meriem; Izano, Era A; Sampathkumar, Vandana; Inzana, Thomas J; Kaplan, Jeffrey B

    2013-01-01

    Cell-free extracts isolated from colony biofilms of Actinobacillus pleuropneumoniae serotype 5 were found to inhibit biofilm formation by Staphylococcus aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans, but not by A. pleuropneumoniae serotype 5 itself, in a 96-well microtiter plate assay. Physical and chemical analyses indicated that the antibiofilm activity in the extract was due to high-molecular-weight polysaccharide. Extracts isolated from a mutant strain deficient in the production of serotype 5 capsular polysaccharide did not exhibit antibiofilm activity. A plasmid harboring the serotype 5 capsule genes restored the antibiofilm activity in the mutant extract. Purified serotype 5 capsular polysaccharide also exhibited antibiofilm activity against S. aureus. A. pleuropneumoniae wild-type extracts did not inhibit S. aureus growth, but did inhibit S. aureus intercellular adhesion and binding of S. aureus cells to stainless steel surfaces. Furthermore, polystyrene surfaces coated with A. pleuropneumoniae wild-type extracts, but not with capsule-mutant extracts, resisted S. aureus biofilm formation. Our findings suggest that the A. pleuropneumoniae serotype 5 capsule inhibits cell-to-cell and cell-to-surface interactions of other bacteria. A. pleuropneumoniae serotype 5 capsular polysaccharide is one of a growing number of bacterial polysaccharides that exhibit broad-spectrum, nonbiocidal antibiofilm activity. Future studies on these antibiofilm polysaccharides may uncover novel functions for bacterial polysaccharides in nature, and may lead to the development of new classes of antibiofilm agents for industrial and clinical applications. PMID:23691104

  13. Antibiofilm Activity of Actinobacillus pleuropneumoniae Serotype 5 Capsular Polysaccharide

    PubMed Central

    Karwacki, Michael T.; Kadouri, Daniel E.; Bendaoud, Meriem; Izano, Era A.; Sampathkumar, Vandana; Inzana, Thomas J.; Kaplan, Jeffrey B.

    2013-01-01

    Cell-free extracts isolated from colony biofilms of Actinobacillus pleuropneumoniae serotype 5 were found to inhibit biofilm formation by Staphylococcus aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans, but not by A. pleuropneumoniae serotype 5 itself, in a 96-well microtiter plate assay. Physical and chemical analyses indicated that the antibiofilm activity in the extract was due to high-molecular-weight polysaccharide. Extracts isolated from a mutant strain deficient in the production of serotype 5 capsular polysaccharide did not exhibit antibiofilm activity. A plasmid harboring the serotype 5 capsule genes restored the antibiofilm activity in the mutant extract. Purified serotype 5 capsular polysaccharide also exhibited antibiofilm activity against S. aureus. A. pleuropneumoniae wild-type extracts did not inhibit S. aureus growth, but did inhibit S. aureus intercellular adhesion and binding of S. aureus cells to stainless steel surfaces. Furthermore, polystyrene surfaces coated with A. pleuropneumoniae wild-type extracts, but not with capsule-mutant extracts, resisted S. aureus biofilm formation. Our findings suggest that the A. pleuropneumoniae serotype 5 capsule inhibits cell-to-cell and cell-to-surface interactions of other bacteria. A. pleuropneumoniae serotype 5 capsular polysaccharide is one of a growing number of bacterial polysaccharides that exhibit broad-spectrum, nonbiocidal antibiofilm activity. Future studies on these antibiofilm polysaccharides may uncover novel functions for bacterial polysaccharides in nature, and may lead to the development of new classes of antibiofilm agents for industrial and clinical applications. PMID:23691104

  14. Cell-Wall Polysaccharides of Developing Flax Plants.

    PubMed Central

    Gorshkova, T. A.; Wyatt, S. E.; Salnikov, V. V.; Gibeaut, D. M.; Ibragimov, M. R.; Lozovaya, V. V.; Carpita, N. C.

    1996-01-01

    Flax (Linum usitatissimum L.) fibers originate from procambial cells of the protophloem and develop in cortical bundles that encircle the vascular cylinder. We determined the polysaccharide composition of the cell walls from various organs of the developing flax plant, from fiber-rich strips peeled from the stem, and from the xylem. Ammonium oxalate-soluble polysaccharides from all tissues contained 5-linked arabinans with low degrees of branching, rhamnogalacturonans, and polygalacturonic acid. The fiber-rich peels contained, in addition, substantial amounts of a buffer-soluble, 4-linked galactan branched at the 0-2 and 0-3 positions with nonreducing terminal-galactosyl units. The cross-linking glycans from all tissues were (fucogalacto)xyloglucan, typical of type-I cell walls, xylans containing (1->)-[beta]-D-xylosyl units branched exclusively at the xylosyl O-2 with t-(4-O-methyl)-glucosyluronic acid units, and (galacto)glucomannans. Tissues containing predominantly primary cell wall contained a larger proportion of xyloglucan. The xylem cells were composed of about 60% 4-xylans, 32% cellulose, and small amounts of pectin and the other cross-linking polysaccharides. The noncellulosic polysaccharides of flax exhibit an uncommonly low degree of branching compared to similar polysaccharides from other flowering plants. Although the relative abundance of the various noncellulosic polysaccharides varies widely among the different cell types, the linkage structure and degree of branching of several of the noncellulosic polysaccharides are invariant. PMID:12226214

  15. The immunostimulating role of lichen polysaccharides: a review.

    PubMed

    Shrestha, Gajendra; St Clair, Larry L; O'Neill, Kim L

    2015-03-01

    The immune system has capacity to suppress the development or progression of various malignancies including cancer. Research on the immunomodulating properties of polysaccharides obtained from plants, microorganisms, marine organisms, and fungi is growing rapidly. Among the various potential sources, lichens, symbiotic systems involving a fungus and an alga and/or a cyanobacterium, show promise as a potential source of immunomodulating compounds. It is well known that lichens produce an abundance of structurally diverse polysaccharides. However, only a limited number of studies have explored the immunostimulating properties of lichen polysaccharides. Published studies have shown that some lichen polysaccharides enhance production of nitrous oxide (NO) by macrophages and also alter the production levels of various proinflammatory and antiinflammatory cytokines (IL-10, IL-12, IL-1β, TNF-α, and IFN-α/β) by macrophages and dendritic cells. Although there are only a limited number of studies examining the role of lichen polysaccharides, all results suggest that lichen polysaccharides can induce immunomodulatory responses in macrophages and dendritic cells. Thus, a detailed evaluation of immunomodulatory capacity of lichen polysaccharides could provide a unique opportunity for the discovery of novel therapeutic agents.

  16. Optimization for ultrasonic-microwave synergistic extraction of polysaccharides from Cornus officinalis and characterization of polysaccharides.

    PubMed

    Yin, Xiulian; You, Qinghong; Jiang, Zhonghai; Zhou, Xinghai

    2016-02-01

    Ultrasonic-microwave synergistic extraction (UMSE) of polysaccharides from Cornus officinalis was optimized by response surface methodology (RSM). The effect of four different factors on the yield of C. officinalis polysaccharides (COP) was studied. RSM results showed that the optimal conditions were extraction time of 31.49823 min, microwave power of 99.39769 W, and water-to-raw material ratio of 28.16273. The COP yield was 11.38±0.31% using the modified optimal conditions, which was consistent with the value predicted by the model. The crude COP was purified by DEAE-Cellulose 52 chromatography and Sephadex G-100 chromatography. Five fractions, namely, crude COP, COP-1, COP-2, COP-3, and COP-4, were obtained. Monosaccharide composition analysis revealed that the COP was composed of glucose, arabinose, fucose, xylose, mannose, and rhamnose. Preliminary structural characterizations of COP were conducted by scanning electron microscopy and Fourier transform infrared spectroscopy.

  17. Enzymatic method for improving the injectability of polysaccharides. [US Patent Application

    DOEpatents

    Griffith, W.L.; Compere, A.L.; Holleman, J.W.

    A method for enhancing the ability of polysaccharides in aqueous solution to flow through a porous medium comprises contacting the polysaccharides with an endoenzyme capable of hydrolyzing at least one of the linkages of the sugar units of the polysaccharides and maintaining the polysaccharides in contact with the enzyme under hydrolysis conditions for a time sufficient to decrease the tendency of the polysaccharides to plug the porous medium yet insufficient to decrease the viscosity of the aqueous polysaccharides by more than 25%. The partially hydrolyzed polysaccharides are useful as thickening agents for flooding water used to recover oil from oil-containing subterranean formations.

  18. Properties of bread dough with added fiber polysaccharides and phenolic antioxidants: a review.

    PubMed

    Sivam, Anusooya S; Sun-Waterhouse, Dongxiao; Quek, SiewYoung; Perera, Conrad O

    2010-10-01

    During breadmaking, different ingredients are used to ensure the development of a continuous protein network that is essential for bread quality. Interests in incorporating bioactive ingredients such as dietary fiber (DF) and phenolic antioxidants into popular foods such as bread have grown rapidly, due to the increased consumer health awareness. The added bioactive ingredients may or may not promote the protein cross-links. Appropriate cross-links among wheat proteins, fiber polysaccharides, and phenolic antioxidants could be the most critical factor for bread dough enhanced with DF and phenolic antioxidants. Such cross-links may influence the structure and properties of a bread system during baking. This article presents a brief overview of our current knowledge of the fate of the key components (wheat proteins, fibers, and phenolic antioxidants) and how they might interact during bread dough development and baking.

  19. Properties of Bread Dough with Added Fiber Polysaccharides and Phenolic Antioxidants: A Review

    PubMed Central

    Sivam, Anusooya S; Sun-Waterhouse, Dongxiao; Quek, SiewYoung; Perera, Conrad O

    2010-01-01

    During breadmaking, different ingredients are used to ensure the development of a continuous protein network that is essential for bread quality. Interests in incorporating bioactive ingredients such as dietary fiber (DF) and phenolic antioxidants into popular foods such as bread have grown rapidly, due to the increased consumer health awareness. The added bioactive ingredients may or may not promote the protein cross-links. Appropriate cross-links among wheat proteins, fiber polysaccharides, and phenolic antioxidants could be the most critical factor for bread dough enhanced with DF and phenolic antioxidants. Such cross-links may influence the structure and properties of a bread system during baking. This article presents a brief overview of our current knowledge of the fate of the key components (wheat proteins, fibers, and phenolic antioxidants) and how they might interact during bread dough development and baking. PMID:21535512

  20. Chemical Structures and Bioactivities of Sulfated Polysaccharides from Marine Algae

    PubMed Central

    Jiao, Guangling; Yu, Guangli; Zhang, Junzeng; Ewart, H. Stephen

    2011-01-01

    Sulfated polysaccharides and their lower molecular weight oligosaccharide derivatives from marine macroalgae have been shown to possess a variety of biological activities. The present paper will review the recent progress in research on the structural chemistry and the bioactivities of these marine algal biomaterials. In particular, it will provide an update on the structural chemistry of the major sulfated polysaccharides synthesized by seaweeds including the galactans (e.g., agarans and carrageenans), ulvans, and fucans. It will then review the recent findings on the anticoagulant/antithrombotic, antiviral, immuno-inflammatory, antilipidemic and antioxidant activities of sulfated polysaccharides and their potential for therapeutic application. PMID:21566795

  1. Spectroscopic manifestation of stretching vibrations of glycosidic linkage in polysaccharides

    NASA Astrophysics Data System (ADS)

    Nikonenko, N. A.; Buslov, D. K.; Sushko, N. I.; Zhbankov, R. G.

    2005-10-01

    Manifestation of stretching vibrations of glycosidic linkage in the infrared spectra of polysaccharides (native, microcrystalline, mercerized celluloses, amylose, starches) has been studied using the regularized method of deconvolution. It has been shown that the glycosidic linkage formation in the polysaccharides is characterized by the appearance of new absorption bands in the 1175-1140 cm -1 range as compared to their corresponding monomers. In the 1000-920 cm -1 region differences between the infrared spectra of polysaccharides due to the changes in the glycosidic linkage configuration have been found.

  2. Polysaccharide Nanoparticles for Efficient siRNA Targeting in Cancer Cells by Supramolecular pKa Shift

    PubMed Central

    Zhang, Ying-Ming; Yang, Yang; Zhang, Yu-Hui; Liu, Yu

    2016-01-01

    Biomacromolecular pKa shifting is considered as one of the most ubiquitous processes in biochemical events, e.g., the enzyme-catalyzed reaction and protein conformational stabilization. In this paper, we report on the construction of biocompatible polysaccharide nanoparticle with targeting ability and lower toxicity by supramolecular pKa shift strategy. This was realized through a ternary assembly constructed by the dual host‒guest interactions of an adamantane-bis(diamine) conjugate (ADA) with cucurbit[6]uril (CB[6]) and a polysaccharide. The potential application of such biocompatible nanostructure was further implemented by the selective transportation of small interfering RNA (siRNA) in a controlled manner. It is demonstrated that the strong encapsulation of the ADA’s diammonium tail by CB[6] not only reduced the cytotoxicity of the nano-scaled vehicle but also dramatically enhanced cation density through an obvious positive macrocycle-induced pKa shift, which eventually facilitated the subsequent siRNA binding. With a targeted polysaccharide shell containing a cyclodextrin‒hyaluronic acid conjugate, macrocycle-incorporated siRNA polyplexes were specifically delivered into malignant human prostate PC-3 cells. The supramolecular polysaccharide nanoparticles, the formation of which was enabled and promoted by the complexation-assisted pKa shift, may be used as a versatile tool for controlled capture and release of biofunctional substrates. PMID:27363811

  3. Polysaccharide Nanoparticles for Efficient siRNA Targeting in Cancer Cells by Supramolecular pKa Shift

    NASA Astrophysics Data System (ADS)

    Zhang, Ying-Ming; Yang, Yang; Zhang, Yu-Hui; Liu, Yu

    2016-07-01

    Biomacromolecular pKa shifting is considered as one of the most ubiquitous processes in biochemical events, e.g., the enzyme-catalyzed reaction and protein conformational stabilization. In this paper, we report on the construction of biocompatible polysaccharide nanoparticle with targeting ability and lower toxicity by supramolecular pKa shift strategy. This was realized through a ternary assembly constructed by the dual host‒guest interactions of an adamantane-bis(diamine) conjugate (ADA) with cucurbit[6]uril (CB[6]) and a polysaccharide. The potential application of such biocompatible nanostructure was further implemented by the selective transportation of small interfering RNA (siRNA) in a controlled manner. It is demonstrated that the strong encapsulation of the ADA’s diammonium tail by CB[6] not only reduced the cytotoxicity of the nano-scaled vehicle but also dramatically enhanced cation density through an obvious positive macrocycle-induced pKa shift, which eventually facilitated the subsequent siRNA binding. With a targeted polysaccharide shell containing a cyclodextrin‒hyaluronic acid conjugate, macrocycle-incorporated siRNA polyplexes were specifically delivered into malignant human prostate PC-3 cells. The supramolecular polysaccharide nanoparticles, the formation of which was enabled and promoted by the complexation-assisted pKa shift, may be used as a versatile tool for controlled capture and release of biofunctional substrates.

  4. Influence of O polysaccharides on biofilm development and outer membrane vesicle biogenesis in Pseudomonas aeruginosa PAO1.

    PubMed

    Murphy, Kathleen; Park, Amber J; Hao, Youai; Brewer, Dyanne; Lam, Joseph S; Khursigara, Cezar M

    2014-04-01

    Pseudomonas aeruginosa is a common opportunistic human pathogen known for its ability to adapt to changes in its environment during the course of infection. These adaptations include changes in the expression of cell surface lipopolysaccharide (LPS), biofilm development, and the production of a protective extracellular exopolysaccharide matrix. Outer membrane vesicles (OMVs) have been identified as an important component of the extracellular matrix of P. aeruginosa biofilms and are thought to contribute to the development and fitness of these bacterial communities. The goal of this study was to examine the relationships between changes in the cell surface expression of LPS O polysaccharides, biofilm development, and OMV biogenesis in P. aeruginosa. We compared wild-type P. aeruginosa PAO1 with three chromosomal knockouts. These knockouts have deletions in the rmd, wbpM, and wbpL genes that produce changes in the expression of common polysaccharide antigen (CPA), O-specific antigen (OSA), or both. Our results demonstrate that changes in O polysaccharide expression do not significantly influence OMV production but do affect the size and protein content of OMVs derived from both CPA(-) and OSA(-) cells; these mutant cells also exhibited different physical properties from wild-type cells. We further examined biofilm growth of the mutants and determined that CPA(-) cells could not develop into robust biofilms and exhibit changes in cell morphology and biofilm matrix production. Together these results demonstrate the importance of O polysaccharide expression on P. aeruginosa OMV composition and highlight the significance of CPA expression in biofilm development. PMID:24464462

  5. Antibodies to Streptococcus pneumoniae Capsular Polysaccharide Enhance Pneumococcal Quorum Sensing

    PubMed Central

    Yano, Masahide; Gohil, Shruti; Coleman, J. Robert; Manix, Catherine; Pirofski, Liise-anne

    2011-01-01

    ABSTRACT The use of pneumococcal capsular polysaccharide (PPS)-based vaccines has resulted in a substantial reduction in invasive pneumococcal disease. However, much remains to be learned about vaccine-mediated immunity, as seven-valent PPS-protein conjugate vaccine use in children has been associated with nonvaccine serotype replacement and 23-valent vaccine use in adults has not prevented pneumococcal pneumonia. In this report, we demonstrate that certain PPS-specific monoclonal antibodies (MAbs) enhance the transformation frequency of two different Streptococcus pneumoniae serotypes. This phenomenon was mediated by PPS-specific MAbs that agglutinate but do not promote opsonic effector cell killing of the homologous serotype in vitro. Compared to the autoinducer, competence-stimulating peptide (CSP) alone, transcriptional profiling of pneumococcal gene expression after incubation with CSP and one such MAb to the PPS of serotype 3 revealed changes in the expression of competence (com)-related and bacteriocin-like peptide (blp) genes involved in pneumococcal quorum sensing. This MAb was also found to induce a nearly 2-fold increase in CSP2-mediated bacterial killing or fratricide. These observations reveal a novel, direct effect of PPS-binding MAbs on pneumococcal biology that has important implications for antibody immunity to pneumococcus in the pneumococcal vaccine era. Taken together, our data suggest heretofore unsuspected mechanisms by which PPS-specific antibodies could affect genetic exchange and bacterial viability in the absence of host cells. PMID:21917597

  6. Biosynthesis of non-cellulosic polysaccharides of plant cell walls.

    PubMed

    Dhugga, Kanwarpal S

    2012-02-01

    Enzymes that make the polymer backbones of plant cell wall polysaccharides have proven to be recalcitrant to biochemical purification. Availability of mutational genetics and genomic tools paved the way for rapid progress in identifying genes encoding various cell wall glycan synthases. Mutational genetics, the primary tool used in unraveling cellulose biosynthesis, was ineffective in assigning function to any of the hemicellulosic, polymerizing glycan synthases. A combination of comparative genomics and functional expression in a heterologous system allowed identification of various cellulose synthase-like (Csl) sequences as being involved in the formation of β-1,4-mannan, β-1,4-glucan, and mixed-linked glucan. A number of xylose-deficient mutants have led to a variety of genes, none of which thus far possesses the motifs known to be conserved among polymerizing β-glycan synthases. Except for xylan synthase, which appears to be an agglomerate of proteins just like cellulose synthase, Golgi glycan synthases already identified suggest that the catalytic polypeptide by itself is sufficient for enzyme activity, most likely as a homodimer. Several of the Csl genes remain to be assigned a function. The possibility of the involvement of various Csl genes in making more than one product remains.

  7. Purification, characterization and immunomodulatory effects of Plantago depressa polysaccharides.

    PubMed

    Zhao, Hong; Wang, Qiuhong; Sun, Yanping; Yang, Bingyou; Wang, Zhibin; Chai, Guifang; Guan, Yongzhou; Zhu, Weiguo; Shu, Zunpeng; Lei, Xia; Kuang, Haixue

    2014-11-01

    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 1000 kDa 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. PMID:25129717

  8. [Immunomodulatory and antitumor properties of polysaccharide peptide (PSP)].

    PubMed

    Piotrowski, Jakub; Jędrzejewski, Tomasz; Kozak, Wiesław

    2015-01-01

    Modern medicine successfully uses multiple immunomodulators of natural origin, that can affect biological reactions and support body's natural defense mechanisms including antitumor activities. Among them is a group of products derived from fungi, including schizophyllan, lentinan, polysaccharide Krestin (PSK), and polysaccharidepeptide (PSP). Present paper is focused on polysaccharidepeptide, which due to the negligible toxicity and numerous benefits for health, is increasingly used in China and Japan as an adjuvant in the treatment of cancer. PSP is a protein-polisaccharide complex with a molecular weight 100 kDa derived from Coriolus versicolor mushroom. The results of numerous studies and clinical trials confirm that it inhibits the growth of cancer cells in in vitro and in vivo settings as well as decreases cancer treatment-related adverse side effects such as fatigue, loss of appetite, nausea, vomiting, and pain. PSP is able to restore weakened immune response observed in patients with cancer during chemotherapy. Its anti-tumor effects seemed to be mediated through immunomodulatory regulation. PSP stimulates cells of the immune system, induces synthesis of cytokines such as interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α), eicosanoids including prostaglandin E2 (PGE2), histamine, reactive oxygen species and nitrogen mediators. There is a growing interest in understanding the mechanisms of PSP action. Because of its unique properties and safety, PSP may become a widely used therapeutic agent in the near future. PMID:25614677

  9. Degradation mechanism of polysaccharides on irradiated sugarcane bagasse

    NASA Astrophysics Data System (ADS)

    Ribeiro, M. A.; Oikawa, H.; Mori, M. N.; Napolitano, C. M.; Duarte, C. L.

    2013-03-01

    Sugarcane bagasse is composed of cellulose, hemicelluloses, lignin, and a minor amount of protein and inorganic materials. Cellulose consists of linear macromolecular chains of glucose, linked by β-1,4-glucosidic bonds between the number one and the number four carbon atoms of the adjacent glucose units. Hemicelluloses are heterogeneous polymers, unlike cellulose, and are usually composed of 50-200 monomer units of pentose such as xylose and arabinose. Lignin is a complex polymer of p-hydroxyphenylpropanoid units connected by CC and COC links. Radiation-induced reactions in the macromolecules of the cellulose materials are known to be initiated through fast distribution of the absorbed energy within the molecules to produce long- and short-lived radicals. The present study was carried out using sugarcane bagasse samples irradiated by a Radiation Dynamics electron beam accelerator with 1.5 MeV and 37 kW, with the objective to evaluate the cleavage of the polysaccharides and the by-products formed as a result of the absorbed dose. The electron beam processing in 30 kGy of absorbed dose changed the sugarcane bagasse structure and composition, causing some cellulose and hemicelluloses cleavage. These cleavages were partial, forming oligosaccharides and liberating the sugars glucose and arabinose. The main by-product was acetic acid, originated from the de-acetylating of hemicelluloses.

  10. Use of paramagnetic chelated metal derivatives of polysaccharides and spin-labeled polysaccharides as contrast agents in magnetic resonance imaging

    SciTech Connect

    Bligh, S.W.; Harding, C.T.; Sadler, P.J.; Bulman, R.A.; Bydder, G.M.; Pennock, J.M.; Kelly, J.D.; Latham, I.A.; Marriott, J.A. )

    1991-02-01

    Soluble and insoluble polysaccharides were derivatized with diethylenetriaminepentaacetic acid (DTPA) and/or spin-labeled with 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO). Polysaccharides derivatized with DTPA were prepared via cyanogen bromide activation, coupling to a diamine linker, and to DTPA anhydride. Spin-labeled polysaccharides were also prepared via cyanogen bromide activation. The extent of derivatization for dextran (18 kDa) was about 120 glucose units per DTPA, and for cellulose and starch about 15-30 units per DTPA. For spin-labeled polysaccharides, the average loading ranged from 1 nitroxide per 16 glucose units for starch to 181 for dextran (82 kDa). These derivatized paramagnetic polysaccharides were shown to be more effective relaxants than the small paramagnetic molecules alone. Both soluble and insoluble polysaccharide-linker-DTPA-Gd(3) complexes were effectively cleared from the body (rats) after oral administration. After intravenous administration, the biodistribution of dextran-linker-DTPA-Gd(3) complexes differed significantly from that of GdDTPA. Reduction of the nitroxide by ascorbic acid was retarded in the polysaccharide derivatives, particularly in starch derivatized with both nitroxide and linker-DTPA-Cu(2). These agents showed contrast enhancement in the gastrointestinal tract of rabbits.

  11. Evidence for bioadhesive effects of polysaccharides and polysaccharide-containing herbs in an ex vivo bioadhesion assay on buccal membranes.

    PubMed

    Schmidgall, J; Schnetz, E; Hensel, A

    2000-02-01

    Aqueous extracts of polysaccharide-containing plants are widely used in therapy for irritated mucus membranes in the pharynx region. In order to prove the existence of mucilaginous effects of polysaccharide hydrocolloids on epithelia an ex vivo system based on porcine buccal membranes was established. The tissue culture was stable and there was no indication of cytolytic processes during the 5 hour incubation period. This was confirmed through histological studies and the respective LDH values as toxicity marker. The test system was shown to discriminate the adhesive effects of different raw polysaccharides, obtained from a variety of medicinal plants. While polysaccharides from Altheae officinalis, Plantago lanceolata, Malva moschata, or Tilia cordata showed only moderate bioadhesion to epithelial tissue, strong adhesive processes were observed with polysaccharides from Fucus vesiculosus and Calendula officinalis. The adhesive effects were concentration-dependent. Histological studies of membranes, incubated with a fluorescence-labelled rhamnogalacturonan, indicated the presence of distinct polysaccharide layers on the apical membrane surface. With these results, adsorption effects of certain polysaccharides on mucus membranes were shown for the first time. Such effects suggest that this may account, at least in part, for the therapeutic effects of mucilage-containing plants in the treatment of irritated buccal membranes. PMID:10705734

  12. Lytic Polysaccharide Monooxygenases in Biomass Conversion.

    PubMed

    Hemsworth, Glyn R; Johnston, Esther M; Davies, Gideon J; Walton, Paul H

    2015-12-01

    The derivation of second-generation biofuels from non-edible biomass is viewed as crucial for establishing a sustainable bio-based economy for the future. The inertness of lignocellulosic biomass makes its breakdown for conversion into fuels and other compounds a challenge. Enzyme cocktails can be utilized in the bio-refinery for lignocellulose deconstruction but until recently their costs were regarded as high. Lytic polysaccharide monooxygenases (LPMOs) offer tremendous promise for further process improvements owing to their ability to boost the activity of biomass-degrading enzyme consortia. Combining data from multiple disciplines, progress has been made in understanding the biochemistry of LPMOs. We review the academic literature in this area and highlight some of the key questions that remain.

  13. Nutraceutical functionalities of polysaccharides from marine invertebrates.

    PubMed

    Choi, Byeong-Dae; Choi, Yeung Joon

    2012-01-01

    Many researchers are seeking functional materials from marine resources. These marine resources can be used as traditional food additives, and specifically, these are based on polysaccharides. To date, there is a big opportunity to develop new high-value added products with indispensable functional characteristics, which can be used in nutraceuticals either as additives or supplements. Also, a crossover in the pharmaceutical market may be established. Some glycosaminoglycans (GAGs) mimetic-type molecules are already being utilized in the field of nutrition as well as in the cosmetics industry. This chemical is used as a dietary supplement to maintain the structure and function of cartilages, for the relief of pain caused by osteoarthritic joints, and can also be used as an anti-inflammatory agent. Recently, in relation to the prevalence of mad cow disease and avian influenza, the production of GAGs from marine invertebrates offers new market opportunities as compared with that obtained from bovine or avian livestock. PMID:22361178

  14. Ganoderma lucidum polysaccharide exerts anti-tumor activity via MAPK pathways in HL-60 acute leukemia cells.

    PubMed

    Yang, Guohua; Yang, Lei; Zhuang, Yun; Qian, Xifeng; Shen, Yunfeng

    2016-01-01

    In this study, we investigated the anti-tumor activity both in vitro and in vivo of a polysaccharide obtained from Ganoderma lucidum on HL-60 acute myeloid leukemia cells, and focused on its targeting effect on mitogen-activated protein kinase (MAPK) pathways. It was found by the methods such as western blot and flow cytometry (FCM), that G. lucidum polysaccharide (GLP) blocked the extracellular signal-regulated kinase/MAPK signaling pathway, simultaneously activated p38 and JNK MAPK pathways, and therefore regulated their downstream genes and proteins, including p53, c-myc, c-fos, c-jun, Bcl-2, Bax, cleaved caspase-3 and cyclin D1. As a result, cycle arrest and apoptosis of HL-60 cells were induced. Therefore, GLP exerted anti-tumor activity via MAPK pathways in HL-60 acute leukemia cells.

  15. Campylobacter Polysaccharide Capsules: Virulence and Vaccines

    PubMed Central

    Guerry, Patricia; Poly, Frédéric; Riddle, Mark; Maue, Alexander C.; Chen, Yu-Han; Monteiro, Mario A.

    2012-01-01

    Campylobacter jejuni remains a major cause of bacterial diarrhea worldwide and is associated with numerous sequelae, including Guillain Barré Syndrome, inflammatory bowel disease, reactive arthritis, and irritable bowel syndrome. C. jejuni is unusual for an intestinal pathogen in its ability to coat its surface with a polysaccharide capsule (CPS). These capsular polysaccharides vary in sugar composition and linkage, especially those involving heptoses of unusual configuration and O-methyl phosphoramidate linkages. This structural diversity is consistent with CPS being the major serodeterminant of the Penner scheme, of which there are 47 C. jejuni serotypes. Both CPS expression and expression of modifications are subject to phase variation by slip strand mismatch repair. Although capsules are virulence factors for other pathogens, the role of CPS in C. jejuni disease has not been well defined beyond descriptive studies demonstrating a role in serum resistance and for diarrhea in a ferret model of disease. However, perhaps the most compelling evidence for a role in pathogenesis are data that CPS conjugate vaccines protect against diarrheal disease in non-human primates. A CPS conjugate vaccine approach against this pathogen is intriguing, but several questions need to be addressed, including the valency of CPS types required for an effective vaccine. There have been numerous studies of prevalence of CPS serotypes in the developed world, but few studies from developing countries where the disease incidence is higher. The complexity and cost of Penner serotyping has limited its usefulness, and a recently developed multiplex PCR method for determination of capsule type offers the potential of a more rapid and affordable method. Comparative studies have shown a strong correlation of the two methods and studies are beginning to ascertain CPS-type distribution worldwide, as well as examination of correlation of severity of illness with specific CPS types. PMID:22919599

  16. The extracellular polysaccharide of Porphyridium sp.: an NMR study of lithium-resistant oligosaccharidic fragments.

    PubMed

    Gloaguen, Vincent; Ruiz, Gaël; Morvan, Henri; Mouradi-Givernaud, Aziza; Maes, Emmanuel; Krausz, Pierre; Strecker, Gérard

    2004-01-01

    This study deals with the chemical characterization of an extracellular polysaccharide produced by the unicellular red alga Porphyridium sp. The sugar moiety of this polymer is composed of three neutral monosaccharides (Xyl, Glc, and Gal) and one uronic acid (GlcA). Proteins represent 5.5% of the dry weight of the polymer. Uronic degradation of this exopolysaccharide with lithium in ethylenediamine yielded two different oligosaccharides. The absolute configuration of the constitutive monosaccharides was chemically determined and revealed the presence of D-Xyl, D-Glc, D-, and L-Gal. The following oligosaccharide structures were established by NMR spectroscopy: [carbohydrate structure: see text].

  17. Matriglycan: a novel polysaccharide that links dystroglycan to the basement membrane

    PubMed Central

    Yoshida-Moriguchi, Takako; Campbell, Kevin P

    2015-01-01

    Associations between cells and the basement membrane are critical for a variety of biological events including cell proliferation, cell migration, cell differentiation and the maintenance of tissue integrity. Dystroglycan is a highly glycosylated basement membrane receptor, and is involved in physiological processes that maintain integrity of the skeletal muscle, as well as development and function of the central nervous system. Aberrant O-glycosylation of the α subunit of this protein, and a concomitant loss of dystroglycan's ability to function as a receptor for extracellular matrix (ECM) ligands that bear laminin globular (LG) domains, occurs in several congenital/limb-girdle muscular dystrophies (also referred to as dystroglycanopathies). Recent genetic studies revealed that mutations in DAG1 (which encodes dystroglycan) and at least 17 other genes disrupt the ECM receptor function of dystroglycan and cause disease. Here, we summarize recent advances in our understanding of the enzymatic functions of two of these disease genes: the like-glycosyltransferase (LARGE) and protein O-mannose kinase (POMK, previously referred to as SGK196). In addition, we discuss the structure of the glycan that directly binds the ECM ligands and the mechanisms by which this functional motif is linked to dystroglycan. In light of the fact that dystroglycan functions as a matrix receptor and the polysaccharide synthesized by LARGE is the binding motif for matrix proteins, we propose to name this novel polysaccharide structure matriglycan. PMID:25882296

  18. Using naturally occurring polysaccharides to align molecules with nonlinear optical activity

    NASA Technical Reports Server (NTRS)

    Prasthofer, Thomas

    1996-01-01

    The Biophysics and Advanced Materials Branch of the Microgravity Science and Applications Division at Marshall Space Flight Center has been investigating polymers with the potential for nonlinear optical (NLO) applications for a number of years. Some of the potential applications for NLO materials include optical communications, computing, and switching. To this point the branch's research has involved polydiacetylenes, phthalocyanins, and other synthetic polymers which have inherent NLO properties. The aim of the present research is to investigate the possibility of using naturally occurring polymers such as polysaccharides or proteins to trap and align small organic molecules with useful NLO properties. Ordering molecules with NLO properties enhances 3rd order nonlinear effects and is required for 2nd order nonlinear effects. Potential advantages of such a system are the flexibility to use different small molecules with varying chemical and optical properties, the stability and cost of the polymers, and the ability to form thin, optically transparent films. Since the quality of any polymer films depends on optimizing ordering and minimizing defects, this work is particularly well suited for microgravity experiments. Polysaccharide and protein polymers form microscopic crystallites which must align to form ordered arrays. The ordered association of crystallites is disrupted by gravity effects and NASA research on protein crystal growth has demonstrated that low gravity conditions can improve crystal quality.

  19. Antiobesity properties of mushroom polysaccharides – A Review

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mushrooms are widely consumed for their nutritional and health benefits. To stimulate broader interest in the reported health-promoting properties of bioactive mushroom polysaccharides, this presentation will survey the chemistry (isolation and structural characterization) and reported antiobesity ...

  20. Enzymatic synthesis of oligo- and polysaccharide fatty acid esters.

    PubMed

    van den Broek, Lambertus A M; Boeriu, Carmen G

    2013-03-01

    Amphiphilic oligo- and polysaccharides (e.g. polysaccharide alkyl or alkyl-aryl esters) form a new class of polymers with exceptional properties. They function as polymeric surfactants, whilst maintaining most of the properties of the starting polymeric material such as emulsifying, gelling, and film forming properties combined with partial water solubility or permeability. At present carbohydrate fatty acid esters are generally obtained by chemical methods using toxic solvents and organic and inorganic catalysts that leave residual traces in the final products. Enzymatic reactions offer an attractive alternative route for the synthesis of polysaccharide esters. In this review the state of the art of enzymatic synthesis of oligo- and polysaccharides fatty esters has been described.

  1. Synbiotic matrices derived from plant oligosaccharides and polysaccharides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A porous synbiotic matrix was prepared by lyophilization of alginate and pectin or fructan oligosaccharides and polysaccharides cross-linked with calcium. These synbiotic matrices were excellent physical structures to support the growth of Lactobacillus acidophilus (1426) and Lactobacillus reuteri (...

  2. Extraction and antioxidant activity of polysaccharides from Rana chensinensis skin.

    PubMed

    Wang, Zhanyong; Zhao, Yuanyuan; Su, Tingting

    2015-01-22

    The extraction process of polysaccharides from Rana chensinensis skin was optimized by using a Box-Behnken design. The optimum extraction conditions were as follows: extraction time, 4.96h; extraction temperature, 100°C; ratio of water to raw material, 60; and extraction frequency, 1. Under these conditions, the experimental polysaccharide yield was 2.03±0.14%, which agreed with the predicted yield. The purified polysaccharide RCSP II was successfully obtained by diethylaminoethanol-Sepharose and Sepharose CL-6B column chromatography. In vitro experiments showed that RCSP II exhibited a strong scavenging activity against superoxide anion and 1,1-diphenyl-2-picrylhydrazyl radicals but a weak scavenging activity against hydroxyl radicals. RCSP II also showed a strong reducing capacity. Thus, this polysaccharide can be used as a natural antioxidant in functional foods or medicines.

  3. Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup.

    PubMed

    Sun, Jiadong; Ma, Hang; Seeram, Navindra P; Rowley, David C

    2016-09-28

    Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides.

  4. Microwave-assisted extraction of polysaccharides from mulberry leaves.

    PubMed

    Thirugnanasambandham, K; Sivakumar, V; Maran, J Prakash

    2015-01-01

    In this study, microwave-assisted extraction (MAE) of polysaccharides from mulberry leaves was investigated using response surface methodology (RSM). The effects of three extraction factors on the yield of polysaccharides was examined. The results showed that optimum extraction conditions were determined as follows: weight of the sample of 20 g, microwave power of 170 W, extraction time of 10 min. Under these optimal extraction conditions, polysaccharide yield was found to be 9.41%. Three factors-three level Box-Behnken response surface design (BBD) coupled with RSM was used to model the extraction process. ANOVA was used to examine the statistical significance of the developed model. Extracted polysaccharide was analyzed using Fourier transform infrared (FT-IR) spectroscopy.

  5. Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup.

    PubMed

    Sun, Jiadong; Ma, Hang; Seeram, Navindra P; Rowley, David C

    2016-09-28

    Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. PMID:27612524

  6. Safety Assessment of Microbial Polysaccharide Gums as Used in Cosmetics.

    PubMed

    Fiume, Monice M; Heldreth, Bart; Bergfeld, Wilma F; Belsito, Donald V; Hill, Ronald A; Klaassen, Curtis D; Liebler, Daniel C; Marks, James G; Shank, Ronald C; Slaga, Thomas J; Snyder, Paul W; Andersen, F Alan

    2016-07-01

    The Cosmetic Ingredient Review Expert Panel assessed the safety of 34 microbial polysaccharide gums for use in cosmetics, finding that these ingredients are safe in cosmetic formulations in the present practices of use and concentration. The microbial polysaccharide gums named in this report have a variety of reported functions in cosmetics, including emulsion stabilizer, film former, binder, viscosity-increasing agent, and skin-conditioning agent. The Panel reviewed available animal and clinical data in making its determination of safety. PMID:27383198

  7. Protein carriers of conjugate vaccines

    PubMed Central

    Pichichero, Michael E

    2013-01-01

    The immunogenicity of polysaccharides as human vaccines was enhanced by coupling to protein carriers. Conjugation transformed the T cell-independent polysaccharide vaccines of the past to T cell-dependent antigenic vaccines that were much more immunogenic and launched a renaissance in vaccinology. This review discusses the conjugate vaccines for prevention of infections caused by Hemophilus influenzae type b, Streptococcus pneumoniae, and Neisseria meningitidis. Specifically, the characteristics of the proteins used in the construction of the vaccines including CRM, tetanus toxoid, diphtheria toxoid, Neisseria meningitidis outer membrane complex, and Hemophilus influenzae protein D are discussed. The studies that established differences among and key features of conjugate vaccines including immunologic memory induction, reduction of nasopharyngeal colonization and herd immunity, and antibody avidity and avidity maturation are presented. Studies of dose, schedule, response to boosters, of single protein carriers with single and multiple polysaccharides, of multiple protein carriers with multiple polysaccharides and conjugate vaccines administered concurrently with other vaccines are discussed along with undesirable consequences of conjugate vaccines. The clear benefits of conjugate vaccines in improving the protective responses of the immature immune systems of young infants and the senescent immune systems of the elderly have been made clear and opened the way to development of additional vaccines using this technology for future vaccine products. PMID:23955057

  8. Xylan polysaccharides fabricated into nanofibrous substrate for myocardial infarction.

    PubMed

    Venugopal, J; Rajeswari, R; Shayanti, M; Sridhar, R; Sundarrajan, S; Balamurugan, R; Ramakrishna, S

    2013-04-01

    Myocardial infarction, a main cause of heart failure, leads to loss of cardiac tissue impairment of left ventricular function. Repair of diseased myocardium with in vitro engineered cardiac muscle patch/injectable biopolymers with cells may become a viable option for myocardial infarction. We attempted to solve these problems by in vitro study by selecting a plant based polysaccharides beech wood Xylan for the normal functioning of infarcted myocardium. The present study fabricated Xylan based nanofibrous scaffolds cross-linked with glutaraldehyde (Glu) vapors for 24 h, 48 h and 1% Glu blended fibers for the culture of neonatal rat cardiac cells for myocardial infarction. These nanofibers were characterized by SEM, FT-IR, tensile testing and cell culture studies for the normal expression of cardiac proteins. The observed results showed that the Xylan/polyvinyl alcohol (PVA) 24h Glu vapor cross-linked nanofibers (427 nm) having mechanical strength of 2.43 MPa and Young modulus of 3.74 MPa are suitable for the culture of cardiac cells. Cardiac cells proliferation increased only by 11% in Xylan/PVA 24h Glu cross-linked nanofibers compared to control tissue culture plate (TCP). The normal cardiac cell morphology was observed in 24h cross-linked Xylan/PVA nanofibers but 48 h cross-linked fibers cell morphology was changed to flattened and elongated on the fibrous surfaces. Confocal analysis for cardiac expression proteins actinin, connexin 43 was observed normally in 24h Glu cross-linked nanofibers compared to all other nanofibrous scaffolds. The fabricated Xylan/PVA nanofibrous scaffold may have good potential for the normal functioning of infarcted myocardium.

  9. Composition of the Capsular and Extracellular Polysaccharides of Rhizobium japonicum

    PubMed Central

    Mort, Andrew J.; Bauer, Wolfgang D.

    1980-01-01

    The chemical compositions of the capsular and extracellular polysaccharides of two strains of Rhizobium japonicum (311b 138 and 110) have been determined and correlated as a function of culture age with the ability of the bacteria from which they were obtained to bind soybean seed lectin. Each of the polysaccharides contains approximately constant amounts of mannosyl, glucosyl, and galacturonosyl residues in a molar ratio of 1:2:1. In addition they contain variable amounts of galactosyl and 4-O-methyl galactosyl residues. The total of galactose plus 4-O-methyl galactose, however, is constant and equivalent to the amount of mannose, indicating that the 4-O-methyl galactose residues arise by methylation of galactose residues in the polysaccharides. In both strains the proportion of galactose to methyl galactose is considerably greater in the polysaccharide from bacteria which do bind lectin than in the polysaccharide from bacteria which do not bind lectin. In addition to the changes in polysaccharide composition, there is a reduction of about 50% in the percentage of cells which are encapsulated as the cultures mature from early to late log phase. Since only capsulated cells bind lectin, the combination of the change in capsular composition and loss of encapsulation is probably sufficient to account for the loss of lectin binding capacity during growth of cultures of Rhizobium japonicum 311b 138 and 110. Images PMID:16661379

  10. Mapping the polysaccharide degradation potential of Aspergillus niger

    PubMed Central

    2012-01-01

    Background The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential of a given fungus for polysaccharide degradation. Results Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list of 188 genes coding for carbohydrate-active enzymes from Aspergillus niger, thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono- and polysaccharide substrates has allowed elucidation of concerted gene expression from this organism. One such example is the identification of a full set of extracellular polysaccharide-acting genes for the degradation of oat spelt xylan. Conclusions The mapping of plant polysaccharide structures along with the corresponding enzymatic activities is a powerful framework for expression analysis of carbohydrate-active enzymes. Applying this network-based approach, we provide the first genome-scale characterization of all genes coding for carbohydrate-active enzymes identified in A. niger. PMID:22799883

  11. [Analysis of thickening polysaccharides by the improved diethyldithioacetal derivatization method].

    PubMed

    Akiyama, Takumi; Yamazaki, Takeshi; Tanamoto, Kenichi

    2011-01-01

    The identification test for thickening polysaccharides containing neutral saccharides and uronic acids was investigated by GC analysis of constituent monosaccharides. The reported method, in which monosaccharides were converted to diethyldithioacetal derivatives with ethanethiol followed by trimethylsilylation, was improved in terms of operability and reproducibility of GC/MS analysis. The suitability of the improved diethyldithioacetal derivatization method was determined for seven thickening polysaccharides, i.e., carob bean gum, guar gum, karaya gum, gum arabic, gum ghatti, tragacanth gum and peach gum. The samples were acid-hydrolyzed to form monosaccharides. The hydrolysates were derivatized and analyzed with GC/FID. Each sugar derivative was detected as a single peak and was well separated from others on the chromatograms. The amounts of constituent monosaccharides in thickening polysaccharides were successfully estimated. Seven polysaccharides were distinguished from each other on the basis of constituent monosaccharides. Further examination of the time period of hydrolysis of polysaccharides using peach gum showed that the optimal times were not the same for all monosaccharides. A longer time was needed to hydrolyze glucuronic acid than neutral saccharides. The findings suggest that hydrolysis time may sometimes affect the analytical results on composition of constituent monosaccharides in polysaccharides.

  12. Antitussive activity of polysaccharides isolated from the Malian medicinal plants.

    PubMed

    Sutovská, M; Franová, S; Priseznaková, L; Nosálová, G; Togola, A; Diallo, D; Paulsen, B S; Capek, P

    2009-04-01

    From the leaves of popular Malian medicinal plants Trichilia emetica (TE) and Opilia celtidifolia (OC), and fruits of Crossopteryx febrifuga (CF) water and water-ethanol soluble polysaccharide materials were isolated. The results of chemical analysis of the crude polysaccharides showed the dominance of the arabinogalactan ( approximately 54%) and the rhamnogalacturonan ( approximately 30%) in T. emetica leaves, the arabinogalactan ( approximately 60%), the rhamnogalacturonan ( approximately 14%) and the glucuronoxylan ( approximately 14%) in O. celtidifolia leaves, and pectic type of polysaccharides ( approximately 75%) with a lower content of the arabinogalactan ( approximately 17%) in C. febrifuga fruits. The plant polysaccharides showed various biological effects on the citric acid-induced cough reflex and reactivity of airways smooth muscle in vivo conditions. T. emetica and O. celtidifolia polysaccharides possessed significant cough-suppressive effect on chemically induced cough. Furthermore, values of specific airways resistance pointed on bronchodilatory property of polysaccharides isolated from O. celtidifolia. However, the crude extract from C. febrifuga in the same dose as T. emetica and O. celtidifolia did not influence the experimentally induced cough as well as reactivity of airways smooth muscle despite of the fact that the water-ethanol extract is recommended for cough therapy in Mali in the form of syrup.

  13. Antitussive activity of polysaccharides isolated from the Malian medicinal plants.

    PubMed

    Sutovská, M; Franová, S; Priseznaková, L; Nosálová, G; Togola, A; Diallo, D; Paulsen, B S; Capek, P

    2009-04-01

    From the leaves of popular Malian medicinal plants Trichilia emetica (TE) and Opilia celtidifolia (OC), and fruits of Crossopteryx febrifuga (CF) water and water-ethanol soluble polysaccharide materials were isolated. The results of chemical analysis of the crude polysaccharides showed the dominance of the arabinogalactan ( approximately 54%) and the rhamnogalacturonan ( approximately 30%) in T. emetica leaves, the arabinogalactan ( approximately 60%), the rhamnogalacturonan ( approximately 14%) and the glucuronoxylan ( approximately 14%) in O. celtidifolia leaves, and pectic type of polysaccharides ( approximately 75%) with a lower content of the arabinogalactan ( approximately 17%) in C. febrifuga fruits. The plant polysaccharides showed various biological effects on the citric acid-induced cough reflex and reactivity of airways smooth muscle in vivo conditions. T. emetica and O. celtidifolia polysaccharides possessed significant cough-suppressive effect on chemically induced cough. Furthermore, values of specific airways resistance pointed on bronchodilatory property of polysaccharides isolated from O. celtidifolia. However, the crude extract from C. febrifuga in the same dose as T. emetica and O. celtidifolia did not influence the experimentally induced cough as well as reactivity of airways smooth muscle despite of the fact that the water-ethanol extract is recommended for cough therapy in Mali in the form of syrup. PMID:19150368

  14. Regioselective sulfation of Artemisia sphaerocephala polysaccharide: Characterization of chemical structure.

    PubMed

    Wang, Junlong; Yang, Wen; Wang, Jiancheng; Wang, Xia; Wu, Fang; Yao, Jian; Zhang, Ji; Lei, Ziqiang

    2015-11-20

    The biological activities of sulfated polysaccharides are related to the substitution positions of functional groups. In this study, regioselective sulfation of Artemisia sphaerocephala polysaccharides (SRSASP) was prepared by using triphenylchloromethane (TrCl) as protecting precursor. FT-IR spectra and X-ray photoelectron spectroscopy (XPS) showed that SO(3-) group (S(6+), high binding energy of 168.7eV) was widely present in sulfated polysaccharides. (13)C NMR spectroscopy showed that C-2 and C-3 substitution was occurred but not fully sulfation. Meanwhile, C-6 substituted signals near 65ppm were not observed. The degree of substitution varied from 0.44 to 0.63 in SRSASP which could be attributed to the low reactivity at secondary hydroxyl. Monosaccharide composition result showed a decrease in the ratio of mannose/glucose, indicating the change of chemical composition in sulfated polysaccharides. In size-exclusion chromatograph analysis, a decrease in molecular weight and broadening of molecular weight distribution of sulfated polysaccharides was also observed. It could be attributed to the hydrolysis of polysaccharide in the sulfated reaction.

  15. Marine derived polysaccharides for biomedical applications: chemical modification approaches.

    PubMed

    d'Ayala, Giovanna Gomez; Malinconico, Mario; Laurienzo, Paola

    2008-01-01

    Polysaccharide-based biomaterials are an emerging class in several biomedical fields such as tissue regeneration, particularly for cartilage, drug delivery devices and gelentrapment systems for the immobilization of cells. Important properties of the polysaccharides include controllable biological activity, biodegradability, and their ability to form hydrogels. Most of the polysaccharides used derive from natural sources; particularly, alginate and chitin, two polysaccharides which have an extensive history of use in medicine, pharmacy and basic sciences, and can be easily extracted from marine plants (algae kelp) and crab shells, respectively. The recent rediscovery of poly-saccharidebased materials is also attributable to new synthetic routes for their chemical modification, with the aim of promoting new biological activities and/or to modify the final properties of the biomaterials for specific purposes. These synthetic strategies also involve the combination of polysaccharides with other polymers. A review of the more recent research in the field of chemical modification of alginate, chitin and its derivative chitosan is presented. Moreover, we report as case studies the results of our recent work concerning various different approaches and applications of polysaccharide-based biomaterials, such as the realization of novel composites based on calcium sulphate blended with alginate and with a chemically modified chitosan, the synthesis of novel alginate-poly(ethylene glycol) copolymers and the development of a family of materials based on alginate and acrylic polymers of potential interest as drug delivery systems. PMID:18830142

  16. Degradation of hemicellulosic and cellulosic polysaccharides in pickled green olives.

    PubMed

    Sánchez-Romero, C; Guillén, R; Heredia, A; Jiménez, A; Fernández-Bolaños, J

    1998-01-01

    Changes that take place in the hemicellulosic and cellulosic polysaccharide fractions of the cell wall of olives (Olea europaea pomiformis, Manzanilla variety) during "Spanish style" processing have been studied. A comparative study of the extraction of hemicellulosic polysaccharides with and without prior delignification showed that these compounds could be extracted without previous delignification of the cell wall material. The depectinated material was sequentially extracted with 1 M and 4 M potassium hydroxide. In the unprocessed fruit, the neutral polysaccharides of the 1 M potassium hydroxide-soluble fraction contained mainly xyloglucans with significant amounts of arabinans. In the 4 M potassium hydroxide-soluble fraction, xyloglucans were the most important polysaccharide. The apparent molecular weight of these polysaccharides was 40 to 250 kDa. In addition, hemicelluloses (xylans and xyloglucans), which it was not possible to isolate in the previous stages of fractionation, were also found to be closely linked to the cellulose fraction. The most important changes during processing were the decrease in the molecular weight of xyloglucans in the 4 M potassium hydroxide-soluble fraction and the substantial decrease in the cellulose fraction, which in quantitative terms was one of the largest decreases that took place in the components of the total cell wall polysaccharides. PMID:9708258

  17. Marine derived polysaccharides for biomedical applications: chemical modification approaches.

    PubMed

    d'Ayala, Giovanna Gomez; Malinconico, Mario; Laurienzo, Paola

    2008-09-03

    Polysaccharide-based biomaterials are an emerging class in several biomedical fields such as tissue regeneration, particularly for cartilage, drug delivery devices and gelentrapment systems for the immobilization of cells. Important properties of the polysaccharides include controllable biological activity, biodegradability, and their ability to form hydrogels. Most of the polysaccharides used derive from natural sources; particularly, alginate and chitin, two polysaccharides which have an extensive history of use in medicine, pharmacy and basic sciences, and can be easily extracted from marine plants (algae kelp) and crab shells, respectively. The recent rediscovery of poly-saccharidebased materials is also attributable to new synthetic routes for their chemical modification, with the aim of promoting new biological activities and/or to modify the final properties of the biomaterials for specific purposes. These synthetic strategies also involve the combination of polysaccharides with other polymers. A review of the more recent research in the field of chemical modification of alginate, chitin and its derivative chitosan is presented. Moreover, we report as case studies the results of our recent work concerning various different approaches and applications of polysaccharide-based biomaterials, such as the realization of novel composites based on calcium sulphate blended with alginate and with a chemically modified chitosan, the synthesis of novel alginate-poly(ethylene glycol) copolymers and the development of a family of materials based on alginate and acrylic polymers of potential interest as drug delivery systems.

  18. Oral versus postingestive origin of polysaccharide appetite in the rat.

    PubMed

    Sclafani, A; Nissenbaum, J W

    1987-01-01

    Previous studies have revealed that rats consume substantial amounts of polysaccharide solutions, even if the solutions are made bitter with the addition of sucrose octa acetate (SOA). The present experiment used the gastric sham-feeding preparation to determine if it is the orosensory or postingestive properties of polysaccharides that motivate rats to consume polysaccharide (Polycose) solutions. In Experiment 1, food deprived rats sham fed less of a 0.05% SOA + 32% Polycose solution than they did of a 32% glucose solution, but their SOA-Polycose intake was still considerable (44 ml/hr). The same rats refused to sham feed SOA-gum and SOA-sugar solutions that were similar to the SOA-Polycose solution in bitter taste, viscosity and free sugar content. In Experiment 2, rats sham fed as much of a 32% Polycose solution as they did of a 32% sucrose solution. Despite the gastric fistula, some of the ingested Polycose was absorbed as evidenced by an increase in the rats' blood glucose levels. The addition of acarbose, a drug that inhibits polysaccharide digestion, to the Polycose solution blocked the increase in blood glucose, but did not reduce the rats' sham feeding of the solution. These findings indicate that it is the orosensory (presumably taste) properties of polysaccharide solutions, not their postingestive effects, that initially attract rats to the solutions. The results question the assumption that polysaccharides are "tasteless" to animals.

  19. Correlation Between Chain Architecture and Hydration Water Structure in Polysaccharides

    NASA Astrophysics Data System (ADS)

    Grossutti, Michael; Dutcher, John

    The physical properties of confined water can differ dramatically from those of bulk water. Hydration water associated with polysaccharides provides a particularly important example of confined water, with differences in polysaccharide structure providing different spatially confined environments for water adsorption. We have used attenuated total reflection infrared (ATR-IR) spectroscopy to investigate the structure of hydration water in films of three different polysaccharides under controlled relative humidity (RH) conditions. We compare the results obtained for films of highly branched, monodisperse phytoglycogen nanoparticles to those obtained for two unbranched polysaccharides, hyaluronic acid (HA) and chitosan. We find similarities between water structuring in the two linear polysaccharides, and significant differences for phytoglycogen. In particular, the phytoglycogen nanoparticles exhibited high network water connectivity, and a large increase in the fraction of multimer water clusters with increasing RH, whereas the water structure for HA and chitosan was found to be insensitive to changes in RH. These measurements provide unique insight into the relationship between the chain architecture and hydration of polysaccharides.

  20. Correlation Between Chain Architecture and Hydration Water Structure in Polysaccharides.

    PubMed

    Grossutti, Michael; Dutcher, John R

    2016-03-14

    The physical properties of confined water can differ dramatically from those of bulk water. Hydration water associated with polysaccharides provides a particularly interesting example of confined water, because differences in polysaccharide structure provide different spatially confined environments for water sorption. We have used attenuated total reflection infrared (ATR-IR) spectroscopy to investigate the structure of hydration water in films of three different polysaccharides under controlled relative humidity (RH) conditions. We compare the results obtained for films of highly branched, dendrimer-like phytoglycogen nanoparticles to those obtained for two unbranched polysaccharides, hyaluronic acid (HA), and chitosan. We find similarities between the water structuring in the two linear polysaccharides and significant differences for phytoglycogen. In particular, the results suggest that the high degree of branching in phytoglycogen leads to a much more well-ordered water structure (low density, high connectivity network water), indicating the strong influence of chain architecture on the structuring of water. These measurements provide unique insight into the relationship between the structure and hydration of polysaccharides, which is important for understanding and exploiting these sustainable nanomaterials in a wide range of applications.

  1. Structure of plant cell walls: XIX. Isolation and characterization of wall polysaccharides from suspension-cultured Douglas fir cells

    SciTech Connect

    Thomas, J.R.; McNeil, M.; Darvill, A.G.; Albersheim, P.

    1987-03-01

    The partial purification and characterization of cell wall polysaccharides isolated from suspension-cultured Douglas fir (Pseudotsuga menziesii) cells are described. Extraction of isolated cell walls from 1.0 M LiCl solubilized pectic polysaccharides with glycosyl-linkage compositions similar to those of rhamnogalacturonans I and II, pectic polysaccharides isolated from walls of suspension-cultured sycamore cells. Treatment of LiCl-extracted Douglas fir walls with an endo-..cap alpha..-1,4-polygalacturonase released only small, additional amounts of pectic polysaccharide, which had a glycosyl-linkage composition similar to that of rhamnogalacturonan I. Xyloglucan oligosaccharides were released from the endo-..cap alpha..-1,4-polygalacturonase-treated walls by treatment with an endo-..beta..-1,4-glucanase. These oligosaccharides included hepta- and nonasaccharides similar or identical to those released from sycamore cell walls by the same enzyme, and structurally related octa- and decasaccharides similar to those isolated from various angiosperms. Finally, additional xyloglucan and small amounts of xylan were extracted from the endo-..beta..-1,4-glucanase-treated walls by 0.5 N NaOH. The xylan resembled that extracted by NaOH from dicot cell walls in that it contained 2,4- but not 3,4-linked xylosyl residues. In this study, a total of 15% of the cell wall was isolated as pectic material, 10% as xyloglucan, and less than 1% as xylan. The noncellulosic polysaccharides accounted for 25% of the cell walls, cellulose for 23%, protein for 34%, and ash for 5%, for a total of 88% of the cell wall.

  2. Molar mass, entanglement, and associations of the biofilm polysaccharide of Staphylococcus epidermidis.

    PubMed

    Ganesan, Mahesh; Stewart, Elizabeth J; Szafranski, Jacob; Satorius, Ashley E; Younger, John G; Solomon, Michael J

    2013-05-13

    Biofilms are microbial communities that are characterized by the presence of a viscoelastic extracellular polymeric substance (EPS). Studies have shown that polysaccharides, along with proteins and DNA, are a major constituent of the EPS and play a dominant role in mediating its microstructure and rheological properties. Here, we investigate the possibility of entanglements and associative complexes in solutions of extracellular polysaccharide intercellular adhesin (PIA) extracted from Staphylococcus epidermidis biofilms. We report that the weight average molar mass and radius of gyration of PIA isolates are 2.01×10(5)±1200 g/mol and 29.2±1.2 nm, respectively. The coil overlap concentration, c*, was thus determined to be (32±4)×10(-4) g/mL. Measurements of the in situ concentration of PIA (cPIA,biofilm) was found to be (10±2)×10(-4) g/mL.Thus, cPIA,biofilmpolysaccharide entanglements. Furthermore, the onset of self-association and protein complexation of PIA occurs at concentrations far lower than the native PIA concentration in biofilms. This finding therefore suggests a critical role for those two association mechanisms in mediating biofilm viscoelasticity. PMID:23540609

  3. Polysaccharide based nanogels in the drug delivery system: Application as the carrier of pharmaceutical agents.

    PubMed

    Debele, Tilahun Ayane; Mekuria, Shewaye Lakew; Tsai, Hsieh-Chih

    2016-11-01

    Polysaccharide-based nanoparticles have fascinated attention as a vesicle of different pharmaceutical agents due to their unique multi-functional groups in addition to their physicochemical properties, including biocompatibility and biodegradability. The existence of multi-functional groups on the polysaccharide backbone permits facile chemical or biochemical modification to synthesize polysaccharide based nanoparticles with miscellaneous structures. Polysaccharide-based nanogels have high water content, large surface area for multivalent bioconjugation, tunable size, and interior network for the incorporation of different pharmaceutical agents. These unique properties offer great potential for the utilization of polysaccharide-based nanogels in the drug delivery systems. Hence, this review describes chemistry of certain common polysaccharides, several methodologies used to synthesize polysaccharide nanoparticles and primarily focused on the polysaccharide (or polysaccharide derivative) based nanogels as the carrier of pharmaceutical agents. PMID:27524098

  4. Structural studies of the O-specific polysaccharide(s) from the lipopolysaccharide of Azospirillum brasilense type strain Sp7.

    PubMed

    Sigida, Elena N; Fedonenko, Yuliya P; Shashkov, Alexander S; Zdorovenko, Evelina L; Konnova, Svetlana A; Ignatov, Vladimir V; Knirel, Yuriy A

    2013-10-18

    Lipopolysaccharide was obtained by phenol-water extraction from dried bacterial cells of Azospirillum brasilense type strain Sp7. Mild acid hydrolysis of the lipopolysaccharide followed by GPC on Sephadex G-50 resulted in a polysaccharide mixture, which was studied by composition and methylation analyses, Smith degradation and (1)H and (13)C NMR spectroscopy. The following polysaccharide structures were established, where italics indicate a non-stoichiometric (∼40%) 2-O-methylation of l-rhamnose.

  5. Sugar compositions, α-glucosidase inhibitory and amylase inhibitory activities of polysaccharides from leaves and flowers of Camellia sinensis obtained by different extraction methods.

    PubMed

    Wang, Yuanfeng; Yang, Zhiwei; Wei, Xinlin

    2010-11-01

    The sugar compositions, α-glucosidase inhibitory and α-amylase inhibitory activities of polysaccharides from leaves and flowers of green tea (Camellia sinensis) obtained by hot water extraction (HWE), boiled water extraction (BWE) and enzymatic extraction (EE) were investigated. The yields, sugar contents and monosaccharide compositions of tea leaves polysaccharides (TLPS) and tea flower polysaccharides (TFPS) were all significantly affected by extraction methods. The contents of acidic polysaccharides (APS) extracted by BWE and EE were both much more than those by HWE. The yields of TLPS and TFPS were determined as EE>BWE>HWE. Enzyme and higher temperature could improve the contents of APS and yields of TLPS and TFPS. TLPS and TFPS were all mainly composed of Rha, Ara, Gal, Glu and GalA, very little molar contents of GluA, Xyl and Man. It seemed that enzyme extraction could be more conducive to increase the content of Ara, Gal and GaLA. The molecular weights of TFPS were larger than those of TLPS. The molecular weights of polysaccharides obtained by EE decreased. Proteins in tea leaves and tea flowers might be decomposed by EE by observing UV peaks and IR absorption. peaks. The α-glucosidase inhibitory and amylase inhibitory activities of TLPS and TFPS obtained by EE were lower than those by water extracted method. The inhibitory percentages of TLPS and TFPS against α-amylase were all lower than α-glucosidase for different extractions.

  6. The polysaccharide from Tamarindus indica (TS-polysaccharide) protects cultured corneal-derived cells (SIRC cells) from ultraviolet rays.

    PubMed

    Raimondi, L; Lodovici, M; Guglielmi, F; Banchelli, G; Ciuffi, M; Boldrini, E; Pirisino, R

    2003-03-01

    The aim of this work was to investigate the possible protective effect of a new viscosising agent, TS-polysaccharide, on corneal-derived cells (SIRC) exposed to ultraviolet-B rays. To verify this, SIRC cells were first exposed, in the absence or in the presence of TS-polysaccharide (1% w/v), for 9 s at the UV-B source and then post-incubated for 45 min at 37 degrees C. After this period the hydrogen peroxide (H(2)O(2)) accumulated in the medium and the concentration of 8-hydroxy-2'-deoxy-guanosine (8-OHdG) in cell DNA was measured. In addition, the amount of (3)H-methyl-thymidine incorporated in cellular DNA was evaluated after 18 h from irradiation. Our results show that cells exposed to UV-B rays accumulate H(2)O(2), and have higher levels of 8OHdG and a lower amount of (3)H-methyl-thymidine incorporated in DNA than control cells. In the presence of TS-polysaccharide, the H(2)O(2) and 8-OHdG accumulation, and the (3)H-methyl-thymidine incorporation were significantly reduced with respect to the values measured in cells exposed in the absence of the polysaccharide. We propose a protective role of the polysaccharide in reducing UV-B derived DNA damage to eye cells. This finding could be of some clinical importance when the polysaccharide is used as a delivery system for ophthalmic preparations.

  7. Hypoglycemic Effects of Crude Polysaccharide from Purslane

    PubMed Central

    Gong, Fayong; Li, Fenglin; Zhang, Lili; Li, Jing; Zhang, Zhong; Wang, Guangyao

    2009-01-01

    The effects of crude polysaccharide from Purslane (CPP) on body weight (bw), blood glucose, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-c), triglyceride (TG) and serum insulin levels were studied in diabetes mellitus mice. CPP treatment (200, 400 mg/kg bw) for 28 days resulted in a significant decrease in the concentrations of fasting blood glucose (FBG), TC and TG. Furthermore, CPP significantly increased the concentration of HDL-c, body weight and serum insulin level in the mice. In addition, according to acute toxicity studies and single cell gel electrophoresis analysis, CPP did not produce any physical or behavioral signs of toxicity. More significantly, our data demonstrated CPP exhibited the best effects at the dose of 400 mg/kg bw. The above results suggest that CPP can control blood glucose and modulate the metabolism of glucose and blood lipids in diabetes mellitus mice, so we conclude that CPP should be evaluated as a candidate for future studies on diabetes mellitus. PMID:19399226

  8. Immunomodulatory Polysaccharide from Chlorophytum borivilianum Roots

    PubMed Central

    Thakur, Mayank; Connellan, Paul; Deseo, Myrna A.; Morris, Carol; Dixit, Vinod K.

    2011-01-01

    Chlorophytum borivilianum Santapau & Fernandes (Liliaceae) is an ayurvedic Rasayana herb with immunostimulating properties. The polysaccharide fraction (CBP) derived from hot water extraction of C. borivilianum (CB), comprising of ∼31% inulin-type fructans and ∼25% acetylated mannans (of hot water-soluble extract), was evaluated for its effect on natural killer (NK) cell activity (in vitro). Human peripheral blood mononuclear cells (PBMCs), isolated from whole blood on a Ficoll-Hypaque density gradient, were tested in the presence or absence of varying concentrations of each C. borivilianum fraction for modulation of NK cell cytotoxic activity toward K562 cells. Preliminary cytotoxicity evaluation against P388 cells was performed to establish non-cytotoxic concentrations of the different fractions. Testing showed the observed significant stimulation of NK cell activity to be due to the CBP of C. borivilianum. Furthermore, in vivo evaluation carried out on Wistar strain albino rats for humoral response to sheep red blood cells (SRBCs) and immunoglobulin-level determination using enzyme-linked immunosorbent assay (ELISA), exhibited an effectiveness of C. borivilianum aqueous extract in improving immune function. Present results provide useful information for understanding the role of CBP in modulating immune function. PMID:21792363

  9. Molecular investigations of flaxseed mucilage polysaccharides.

    PubMed

    Roulard, Romain; Petit, Emmanuel; Mesnard, François; Rhazi, Larbi

    2016-05-01

    The molecular properties of flaxseed mucilage were determined using a multi-angle laser light scattering (MALLS) detector coupled on-line to size exclusion chromatography (SEC) and asymmetric flow field-flow fractionation (AF4). Water and salt solution were tested as mobile phases. The SEC-MALLS method gave partial information and enabled molecular characterization of disaggregated mucilage molecules. Regardless of the eluent used, the observed Mw ranged from about 1.6 × 10(6) to more than 10 × 10(6) g/mol for mucilage polysaccharides. The AF4-MALLS system enabled a complete analysis of mucilage carbohydrate aggregates in water, in which two populations were satisfactorily separated. The molecular weight distribution (MWD) of molecules ranged from 1.5 × 10(6) to more than 4 × 10(8) g/mol. Experiments showed that the conformational structure of mucilage molecules was strongly influenced by ionic strength. Mucilage carbohydrates exhibited a spherical and compact structure in NaCl solution while they displayed a random-coil conformation in water. PMID:26851358

  10. Molecular investigations of flaxseed mucilage polysaccharides.

    PubMed

    Roulard, Romain; Petit, Emmanuel; Mesnard, François; Rhazi, Larbi

    2016-05-01

    The molecular properties of flaxseed mucilage were determined using a multi-angle laser light scattering (MALLS) detector coupled on-line to size exclusion chromatography (SEC) and asymmetric flow field-flow fractionation (AF4). Water and salt solution were tested as mobile phases. The SEC-MALLS method gave partial information and enabled molecular characterization of disaggregated mucilage molecules. Regardless of the eluent used, the observed Mw ranged from about 1.6 × 10(6) to more than 10 × 10(6) g/mol for mucilage polysaccharides. The AF4-MALLS system enabled a complete analysis of mucilage carbohydrate aggregates in water, in which two populations were satisfactorily separated. The molecular weight distribution (MWD) of molecules ranged from 1.5 × 10(6) to more than 4 × 10(8) g/mol. Experiments showed that the conformational structure of mucilage molecules was strongly influenced by ionic strength. Mucilage carbohydrates exhibited a spherical and compact structure in NaCl solution while they displayed a random-coil conformation in water.

  11. Inhibition of inflammatory injure by polysaccharides from Bupleurum chinense through antagonizing P-selectin.

    PubMed

    Tong, Haibin; Tian, Dan; Li, Tianbao; Wang, Bo; Jiang, Guiquan; Sun, Xin

    2014-05-25

    P-selectin-mediated adhesion between endothelium and neutrophils is a crucial process leading to acute inflammatory injure. Thus, P-selectin has been considered as promising target for therapeutics of acute inflammatory-related diseases. In the present study, the water-soluble polysaccharides (BCPs) were isolated from Bupleurum chinense, and we evaluated their therapeutical effects on acute inflammatory injure and antagonistic function against P-selectin-mediated neutrophil adhesion. Our results showed that BCPs significantly impaired the leukocyte infiltration and relieve lung injury in LPS-induced acute pneumonia model. BCPs significantly blocked the binding of P-selectin to neutrophils and inhibited P-selectin-mediated neutrophils rolling along CHO-P cell monolayer. The result from in vitro protein binding assay showed a direct evidence indicating that BCPs-treatment significantly eliminated the interaction between rhP-Fc and its physiological ligand PSGL-1 at protein level. Together, these results provide a novel therapeutical strategy for amelioration of inflammation-related disease processes by polysaccharides from B. chinense.

  12. Polysaccharide isolated from Angelica sinensis inhibits hepcidin expression in rats with iron deficiency anemia.

    PubMed

    Liu, Jin-Yu; Zhang, Yu; You, Ru-Xu; Zeng, Fang; Guo, Dan; Wang, Kai-Ping

    2012-10-01

    A novel polysaccharide named Angelica sinensis polysaccharide (ASP) was obtained from the powdered and defatted roots of A. sinensis (Oliv.) Diels. The molecular weight of ASP was determined to be 78 kDa and was 95.0% sugars consisting of mostly arabinose, glucose, and galactose with a molar ratio of 1:5.68:3.91. A previous study indicated that ASP may increase plasma iron levels by suppressing the expression of hepcidin, a negative regulator of body iron metabolism, in the liver. The present study aims to clarify the inhibitory effect of ASP on hepcidin expression in rat models of iron deficiency anemia (IDA), and clarify the mechanisms involved. It was demonstrated that ASP significantly reduced hepcidin expression by inhibiting the expression of mothers against decapentaplegic protein 4 (SMAD4) in liver and stimulating the secretion of erythropoietin, which further downregulated hepcidin by repressing CCAAT/enhancer-binding protein α (C/EBPα) and the phosphorylation of signal transducer and activator of transcription 3/5. The results indicate that ASP can suppress the expression of hepcidin in rats with IDA, and may be useful for the treatment of IDA.

  13. Bacterially induced mineralization of calcium carbonate: the role of exopolysaccharides and capsular polysaccharides.

    PubMed

    Ercole, Claudia; Cacchio, Paola; Botta, Anna Lucia; Centi, Valeria; Lepidi, Aldo

    2007-02-01

    Bacterially induced carbonate mineralization has been proposed as a new method for the restoration of limestones in historic buildings and monuments. We describe here the formation of calcite crystals by extracellular polymeric substances isolated from Bacillus firmus and Bacillus sphaericus. We isolated bacterial outer structures (glycocalix and parietal polymers), such as exopolysaccharides (EPS) and capsular polysaccharides (CPS) and checked for their influence on calcite precipitation. CPS and EPS extracted from both B. firmus and B. sphaericus were able to mediate CaCO3 precipitation in vitro. X-ray microanalysis showed that in all cases the formed crystals were calcite. Scanning electron microscopy showed that the shape of the crystals depended on the fractions utilized. These results suggest the possibility that biochemical composition of CPS or EPS influences the resulting morphology of CaCO3. There were no precipitates in the blank samples. CPS and EPS comprised of proteins and glycoproteins. Positive alcian blue staining also reveals acidic polysaccharides in CPS and EPS fractions. Proteins with molecular masses of 25-40 kDa and 70 kDa in the CPS fraction were highly expressed in the presence of calcium oxalate. This high level of synthesis could be related to the binding of calcium ions and carbonate deposition.

  14. Polyphenolic-polysaccharide conjugates from plants of Rosaceae/Asteraceae family as potential radioprotectors.

    PubMed

    Zbikowska, Halina Malgorzata; Szejk, Magdalena; Saluk, Joanna; Pawlaczyk-Graja, Izabela; Gancarz, Roman; Olejnik, Alicja Klaudia

    2016-05-01

    Polyphenolic-polysaccharide macromolecular, water-soluble glycoconjugates, isolated from the selected medicinal plants of Rosaceae/Asteraceae family: from leaves of Fragaria vesca L., Rubus plicatus Whe. et N. E., and from flowering parts of Sanguisorba officinalis L., and Erigeron canadensis L., were investigated for their ability to protect proteins and lipids of human plasma against γ-radiation-induced oxidative damage. Treatment of plasma with plant conjugates (6, 30, 150 μg/ml) prior exposure to 100 Gy radiation resulted in a significant inhibition of lipid peroxidation, evaluated by TBARS levels; conjugates isolated from E. canadensis and R. plicatus and a reference flavonoid quercetin showed similar high potential (approx. 70% inhibition, at 6 μg/ml). The conjugates prevented radiation-induced oxidation of protein thiols and significantly improved plasma total antioxidant capacity, estimated with Ellman's reagent and ABTS(.+) assay, respectively. The results demonstrate by the first time a significant radioprotective capability of the polyphenolic-polysaccharide conjugates isolated from E. canadensis, R. plicatus, S. officinalis and to the less extent from F. vesca. The abilities of these substances to inhibit radiation-induced lipid peroxidation and thiol oxidation in plasma seems to be mediated, but not limited to ROS scavenging activity. PMID:26848834

  15. The Anti-Oxidant and Antitumor Properties of Plant Polysaccharides.

    PubMed

    Jiao, Rui; Liu, Yingxia; Gao, Hao; Xiao, Jia; So, Kwok Fai

    2016-01-01

    Oxidative stress has been increasingly recognized as a major contributing factor in a variety of human diseases, from inflammation to cancer. Although certain parts of signaling pathways are still under investigation, detailed molecular mechanisms for the induction of diseases have been elucidated, especially the link between excessive oxygen reactive species (ROS) damage and tumorigenesis. Emerging evidence suggests anti-oxidant therapy can play a key role in treating those diseases. Among potential drug resources, plant polysaccharides are natural anti-oxidant constituents important for human health because of their long history in ethnopharmacology, wide availability and few side effects upon consumption. Plant polysaccharides have been shown to possess anti-oxidant, anti-inflammation, cell viability promotion, immune-regulation and antitumor functions in a number of disease models, both in laboratory studies and in the clinic. In this paper, we reviewed the research progress of signaling pathways involved in the initiation and progression of oxidative stress- and cancer-related diseases in humans. The natural sources, structural properties and biological actions of several common plant polysaccharides, including Lycium barbarum, Ginseng, Zizyphus Jujuba, Astragalus lentiginosus, and Ginkgo biloba are discussed in detail, with emphasis on their signaling pathways. All of the mentioned common plant polysaccharides have great potential to treat oxidative stress and cancinogenic disorders in cell models, animal disease models and clinical cases. ROS-centered pathways (e.g. mitochondrial autophagy, MAPK and JNK) and transcription factor-related pathways (e.g. NF-[Formula: see text]B and HIF) are frequently utilized by these polysaccharides with or without the further involvement of inflammatory and death receptor pathways. Some of the polysaccharides may also influence tumorigenic pathways, such as Wnt and p53 to play their anti-tumor roles. In addition, current

  16. The S. aureus polysaccharide capsule and Efb-dependent fibrinogen shield act in concert to protect against phagocytosis

    PubMed Central

    Kuipers, Annemarie; Stapels, Daphne A. C.; Weerwind, Lleroy T.; Ko, Ya-Ping; Ruyken, Maartje; Lee, Jean C.; van Kessel, Kok P.M.; Rooijakkers, Suzan H. M.

    2016-01-01

    Staphylococcus aureus has developed many mechanisms to escape from human immune responses. In order to resist phagocytic clearance, S. aureus expresses a polysaccharide capsule, which effectively masks the bacterial surface and surface-associated proteins, such as opsonins, from recognition by phagocytic cells. Additionally, secretion of the Extracellular fibrinogen binding protein (Efb) potently blocks phagocytic uptake of the pathogen. Efb creates a fibrinogen shield surrounding the bacteria by simultaneously binding complement C3b and fibrinogen at the bacterial surface. By means of neutrophil phagocytosis assays with fluorescently labeled encapsulated serotype 5 (CP5) and serotype 8 (CP8) strains we now compare the immune-modulating function of these shielding mechanisms. Our data indicate that, in highly encapsulated S. aureus strains, the polysaccharide capsule is able to prevent phagocytic uptake at plasma concentrations <10%, but loses its protective ability at higher concentrations of plasma. Interestingly, Efb shows a strong inhibitory effect on both capsule-negative as well as encapsulated strains at all tested plasma concentrations. Furthermore our results suggest that both shielding mechanisms can exist simultaneously and collaborate to provide optimal protection against phagocytosis at a broad range of plasma concentrations. Since opsonizing antibodies will be shielded from recognition by either mechanism, incorporating both capsular polysaccharides and Efb in future vaccines could be of great importance. PMID:27112346

  17. Cytochemistry and immunolocalisation of polysaccharides and proteoglycans in the endosperm of green Arabica coffee beans.

    PubMed

    Sutherland, P W; Hallett, I C; MacRae, E; Fischer, M; Redgwell, R J

    2004-06-01

    The major noncellulosic polysaccharides and proteoglycans in the coffee bean (Coffea arabica) cell wall are (galacto)mannans and arabinogalactan proteins. Immunological and chemical probes demonstrated that the mannans and arabinogalactan proteins were located continuously across the width of the cell wall, but that the concentration of different structural epitopes within these polysaccharide types showed considerable spatial variation. For the mannans this was implied by the striated pattern demonstrated by fluctuation of the affinity between the mannan monoclonal antibody BGM C6 and (galacto)mannan. The arabinogalactan proteins labelled by the Yariv reagent and the arabinogalactan protein-specific antibody LM2 appeared to be located in all regions of the wall except the middle lamella, but showed some differences in intensity of labelling. However, the LM6 antibody, specific for (1-->5)-alpha-arabinan epitopes, was located only as a compact region adjacent to the cell lumen in the body of the endosperm; though, it did label throughout the wall of epidermal cells. This implied that either some of the more highly arabinosylated arabinogalactan proteins contained contiguous 5-arabinosyl residues or that a rhamnogalacturonan which contained 5-arabinosyl residues as side chains existed in the cell wall. In either case the polymers were very restricted in their distribution. A second category of pectin, a homogalacturonan detected by JIM7, was located only in the middle lamella region. The architecture of the wall, as revealed by resin etching, appeared to reflect the chemical heterogeneity, with three distinct physical zones identifiable in a cross section across a single wall.

  18. Adjuvanticity of compound astragalus polysaccharide and sulfated epimedium polysaccharide per os.

    PubMed

    Chen, Xingying; Chen, Xiaolan; Qiu, Shulei; Hu, Yuanliang; Wang, Deyun; Liu, Xu; Zhao, Xiaojuan; Liu, Cui; Chen, Xinghui

    2013-11-01

    On the basis of previous researches, compound astragalus polysaccharide (APS) and sulfated epimedium polysaccharide (sEPS) oral liquid (AEO) was prepared. Three hundred and twenty 14-day-old chickens were randomly assigned into eight groups and vaccinated with ND vaccine except for blank control (BC) group, repeated vaccination at 28 days old. At the same time of each vaccination, the chickens in three experimental groups were taken orally with AEO, respectively, at three doses, in two component control groups with APS and sEPS, once a day for three successive days; in injection control group were injected with AEI once, and in vaccination control (VC) and BC groups were not administrated. On days 7, 14, 21, 28 and 35 after the first vaccination, peripheral lymphocyte proliferation, the serum antibody titer, IFN-γ and IL-2 concentrations and on day 35 immune organ index were measured. The results showed that AEO at high and medium doses could significantly promote lymphocyte proliferation and development of immune organ, enhance antibody titer and IFN-γ and IL-2 concentration, which was stronger than actions of AEI and two components. The results confirmed that AEO possessed reliable immunoenhancement and could be exploited into an oral immunopotentiator.

  19. The directionality of processive enzymes acting on recalcitrant polysaccharides is reflected in the kinetic signatures of oligomer degradation.

    PubMed

    Hamre, Anne Grethe; Schaupp, Daniel; Eijsink, Vincent G H; Sørlie, Morten

    2015-07-01

    The enzymatic degradation of the closely related insoluble polysaccharides; cellulose (β(1-4)-linked glucose) by cellulases and chitin (β(1-4)-linked N-acetylglucosamine) by chitinases, is of large biological and economical importance. Processive enzymes with different inherent directionalities, i.e. attacking the polysaccharide chains from opposite ends, are crucial for the efficiency of this degradation process. While processive cellulases with complementary functions differ in structure and catalytic mechanism, processive chitinases belong to one single protein family with similar active site architectures. Using the unique model system of Serratia marcescens with two processive chitinases attacking opposite ends of the substrate, we here show that different directionalities of processivity are correlated to distinct differences in the kinetic signatures for hydrolysis of oligomeric tetra-N-acetyl chitotetraose.

  20. Mass preparation of oligosaccharides by the hydrolysis of chondroitin sulfate polysaccharides with a subcritical water microreaction system.

    PubMed

    Yamada, Shuhei; Matsushima, Keiichiro; Ura, Haruo; Miyamoto, Nobuyuki; Sugahara, Kazuyuki

    2013-04-19

    The biological functions of chondroitin sulfate (CS) are executed by the interaction of specific oligosaccharide sequences in the polysaccharide chain with effective proteins. Thus, CS oligosaccharides are expected to have pharmacological applications. Furthermore, the demand for CS in health food supplements and medication is growing. However, the absorbency of CS polysaccharides in the digestive system is very low. Since the activity of orally administered CS is expected to increase by depolymerization, industrial production of CS oligosaccharides is required. In this study, hydrolysis with subcritical and super-critical water was applied to the depolymerization of CS for the first time, and hydrolytic conditions for oligosaccharide production were examined. CS oligosaccharides principally containing an N-acetyl-D-galactosamine residue at their reducing ends were successfully obtained. No significant desulfation was found in CS oligosaccharides prepared under optimized conditions. The production of CS oligosaccharides by this method will have a strong influence on the CS-related materials market.

  1. Mass preparation of oligosaccharides by the hydrolysis of chondroitin sulfate polysaccharides with a subcritical water microreaction system.

    PubMed

    Yamada, Shuhei; Matsushima, Keiichiro; Ura, Haruo; Miyamoto, Nobuyuki; Sugahara, Kazuyuki

    2013-04-19

    The biological functions of chondroitin sulfate (CS) are executed by the interaction of specific oligosaccharide sequences in the polysaccharide chain with effective proteins. Thus, CS oligosaccharides are expected to have pharmacological applications. Furthermore, the demand for CS in health food supplements and medication is growing. However, the absorbency of CS polysaccharides in the digestive system is very low. Since the activity of orally administered CS is expected to increase by depolymerization, industrial production of CS oligosaccharides is required. In this study, hydrolysis with subcritical and super-critical water was applied to the depolymerization of CS for the first time, and hydrolytic conditions for oligosaccharide production were examined. CS oligosaccharides principally containing an N-acetyl-D-galactosamine residue at their reducing ends were successfully obtained. No significant desulfation was found in CS oligosaccharides prepared under optimized conditions. The production of CS oligosaccharides by this method will have a strong influence on the CS-related materials market. PMID:23454651

  2. A family of starch-active polysaccharide monooxygenases.

    PubMed

    Vu, Van V; Beeson, William T; Span, Elise A; Farquhar, Erik R; Marletta, Michael A

    2014-09-23

    The recently discovered fungal and bacterial polysaccharide monooxygenases (PMOs) are capable of oxidatively cleaving chitin, cellulose, and hemicelluloses that contain β(1→4) linkages between glucose or substituted glucose units. They are also known collectively as lytic PMOs, or LPMOs, and individually as AA9 (formerly GH61), AA10 (formerly CBM33), and AA11 enzymes. PMOs share several conserved features, including a monocopper center coordinated by a bidentate N-terminal histidine residue and another histidine ligand. A bioinformatic analysis using these conserved features suggested several potential new PMO families in the fungus Neurospora crassa that are likely to be active on novel substrates. Herein, we report on NCU08746 that contains a C-terminal starch-binding domain and an N-terminal domain of previously unknown function. Biochemical studies showed that NCU08746 requires copper, oxygen, and a source of electrons to oxidize the C1 position of glycosidic bonds in starch substrates, but not in cellulose or chitin. Starch contains α(1→4) and α(1→6) linkages and exhibits higher order structures compared with chitin and cellulose. Cellobiose dehydrogenase, the biological redox partner of cellulose-active PMOs, can serve as the electron donor for NCU08746. NCU08746 contains one copper atom per protein molecule, which is likely coordinated by two histidine ligands as shown by X-ray absorption spectroscopy and sequence analysis. Results indicate that NCU08746 and homologs are starch-active PMOs, supporting the existence of a PMO superfamily with a much broader range of substrates. Starch-active PMOs provide an expanded perspective on studies of starch metabolism and may have potential in the food and starch-based biofuel industries.

  3. Dynamics of water solutions of natural polysaccharides by fast field cycling nmr relaxometry

    NASA Astrophysics Data System (ADS)

    Prusova, Alena; Conte, Pellegrino; Kucerik, Jiri; de Pasquale, Claudio; Alonzo, Giuseppe

    2010-05-01

    Cryobiology studies the effect of low temperatures on living systems such as microorganisms and plants. In particular, plants growing in cold or frozen environments can survive such extreme conditions due to the cold hardening process. Hardening is a three step process during which, first, translocation of polysaccharides to the plant roots affects water structure in the cell-soil surface. For this reason, increase of cell-membrane permeability and resistance to temperatures from -5°C to -10°C is achieved. In a second step, chemical alteration of cell membrane arises and resistance to temperatures up to -20°C is obtained. The last hardening step consists in the vitrification of the plant tissues which allow plants to survive at temperatures as low as -50°C. Since polysaccharides play a very important role in the initial part of the cold hardening process, it is of paramount importance to study the effect of such natural biopolymers on water structure. Here, we present preliminary data obtained by fast field cycling NMR relaxometry on the effect of hyaluronan (an anionic, non-sulfated glycosaminoglycan) on water structure at different concentrations of the polysaccharide. Although hyaluronan is a polysaccharide found exceptionally in animal, human or bacterial bodies, in the present work it was used as a model "pilot" compound. In fact, it has an unique ability to hold water and it contains both polysaccharide and protein-like acetamido functionalities. For this reason, hyaluronan promotes the future research on other plant biopolymers such as, for instance, starch and other very specific proteins. Results revealed that different water-structure systems surround the molecule of hyaluronan in diluted and semidiluted systems. Namely, at the lowest hyaluronan concentration, three hydration shells can be recognized. The first hydration shell is made by bound water (BW) which is strongly fixed to the hyaluronan surface mainly through electrostatic interactions. A

  4. Extraction, purification and elicitor activities of polysaccharides from Chrysanthemum indicum.

    PubMed

    Du, Ningning; Tian, Wei; Zheng, Dongfang; Zhang, Xinyi; Qin, Pinyan

    2016-01-01

    Polysaccharides isolated from Chrysanthemum indicum were studied for their pathogen-derived resistance against Sclerotium rolfsii sacc in Atractylodis maceocephalae koidz. The total sugar content and monosaccharide analysis were determined by phenol-sulfuric acid method and gas chromatography, and infrared spectroscopy performed for simple structure information. The activities of CAT and POD as protective enzymes in A. maceocephalae leaves were evaluated. The purified polysaccharides exhibited strong CAT and POD activities in inoculated with S. rolfsii in A. macrocephala leaves, attained the maximum value 568.3 Ug(-1)min(-1) and 604.4 Ug(-1)min(-1)respectively. Whereas, when compared with the control plants, 20mg/ml purified polysaccharides exhibited the strongest CAT and POD activities. Notably, the treatments of A. macepcephalae seedlings with C. indicum polysaccharides (CIP) decreased disease index development caused by S. rolfsii. The disease index after 10 days was significantly reduced when the seedlings treated with 20mg/ml CIP, 4.41 compared to the control plants 32.00. Given together, these results indicated that purified polysaccharides derived from C. indicum may be useful as a natural inducer. PMID:26562553

  5. Masquerading microbial pathogens: Capsular polysaccharides mimic host-tissue molecules

    PubMed Central

    Cress, Brady F.; Englaender, Jacob A.; He, Wenqin; Kasper, Dennis; Linhardt, Robert J.; Koffas, Mattheos A. G.

    2014-01-01

    Summary Bacterial pathogens bearing capsular polysaccharides identical to mammalian glycans benefit from an additional level of protection from host immune response. The increasing prevalence of antibiotic resistant bacteria portends an impending post-antibiotic age, characterized by diminishing efficacy of common antibiotics and routine application of multifaceted, complementary therapeutic approaches to treat bacterial infections, particularly multidrug-resistant organisms. The first line of defense for most bacterial pathogens consists of a physical and immunological barrier known as the capsule, commonly composed of a viscous layer of carbohydrates that are covalently bound to the cell wall in Gram-positive bacteria or often to lipids of the outer membrane in many Gram-negative bacteria. Bacterial capsular polysaccharides are a diverse class of high molecular weight polysaccharides contributing to virulence of many human pathogens in the gut, respiratory tree, urinary tract, and other host tissues, by hiding cell-surface components that might otherwise elicit host immune response. This review highlights capsular polysaccharides that are structurally identical or similar to polysaccharides found in mammalian tissues, including polysialic acid and glycosaminoglycan capsules hyaluronan, heparosan, and chondroitin. Such non-immunogenic coatings render pathogens insensitive to certain immune responses, effectively increasing residence time in host tissues and enabling pathologically relevant population densities to be reached. Biosynthetic pathways and capsular involvement in immune system evasion are described providing a basis for potential therapies aimed at supplementing or replacing antibiotic treatment. PMID:24372337

  6. Aspergillus Enzymes Involved in Degradation of Plant Cell Wall Polysaccharides

    PubMed Central

    de Vries, Ronald P.; Visser, Jaap

    2001-01-01

    Degradation of plant cell wall polysaccharides is of major importance in the food and feed, beverage, textile, and paper and pulp industries, as well as in several other industrial production processes. Enzymatic degradation of these polymers has received attention for many years and is becoming a more and more attractive alternative to chemical and mechanical processes. Over the past 15 years, much progress has been made in elucidating the structural characteristics of these polysaccharides and in characterizing the enzymes involved in their degradation and the genes of biotechnologically relevant microorganisms encoding these enzymes. The members of the fungal genus Aspergillus are commonly used for the production of polysaccharide-degrading enzymes. This genus produces a wide spectrum of cell wall-degrading enzymes, allowing not only complete degradation of the polysaccharides but also tailored modifications by using specific enzymes purified from these fungi. This review summarizes our current knowledge of the cell wall polysaccharide-degrading enzymes from aspergilli and the genes by which they are encoded. PMID:11729262

  7. Crosslinked ionic polysaccharides for stimuli-sensitive drug delivery.

    PubMed

    Alvarez-Lorenzo, Carmen; Blanco-Fernandez, Barbara; Puga, Ana M; Concheiro, Angel

    2013-08-01

    Polysaccharides are gaining increasing attention as components of stimuli-responsive drug delivery systems, particularly since they can be obtained in a well characterized and reproducible way from the natural sources. Ionic polysaccharides can be readily crosslinked to render hydrogel networks sensitive to a variety of internal and external variables, and thus suitable for switching drug release on-off through diverse mechanisms. Hybrids, composites and grafted polymers can reinforce the responsiveness and widen the range of stimuli to which polysaccharide-based systems can respond. This review analyzes the state of the art of crosslinked ionic polysaccharides as components of delivery systems that can regulate drug release as a function of changes in pH, ion nature and concentration, electric and magnetic field intensity, light wavelength, temperature, redox potential, and certain molecules (enzymes, illness markers, and so on). Examples of specific applications are provided. The information compiled demonstrates that crosslinked networks of ionic polysaccharides are suitable building blocks for developing advanced externally activated and feed-back modulated drug delivery systems.

  8. Rapid bacterial degradation of polysaccharides in anoxic marine systems

    SciTech Connect

    Arnosti, C.; Repeta, D.J.; Blough, N.V. )

    1994-06-01

    Extracellular hydrolysis of organic macromolecules is often assumed to be the slow step in remineralization of organic matter. The authors tested this assumption by comparing the degradation of four polysaccharides (pullulan, laminarin, and two polysaccharides isolated from the marine cyanobacterium Synechococcus WH7335) to determine whether size, linkage position, or anomeric linkage affected rates or mechanisms of carbohydrate degradation by mixed cultures of anaerobic bacteria enriched from marine sediments. Gel permeation chromatography and nuclear magnetic resonance spectroscopy (NMR) were used to follow the extracellular conversion of high molecular weight polysaccharides to lower molecular weight polysaccharides and oligosaccharides which were subsequently remineralized. In all cases, substrate degradation was rapid. NMR spectra showed that preferential hydrolysis occurred at specific chemical linkages, and extracellular enzymatic hydrolysis of polysaccharides occurred far more rapidly than bacterial uptake and remineralization of the lower molecular weight oligosaccharides produced through enzymatic hydrolysis. Substrate size was not a significant determinant of remineralization rate: High molecular weight does not always correlate with slow degradation rate. The hypothesis that extracellular enzymatic hydrolysis is a slow step in the degradation of macromolecular organic matter in marine systems needs to be critically re-examined.

  9. Hot-compressed water extraction of polysaccharides from soy hulls.

    PubMed

    Liu, Hua-Min; Wang, Fei-Yun; Liu, Yu-Lan

    2016-07-01

    The polysaccharides of soy hulls were extracted by hot-compressed water at temperatures of 110 from 180°C and various treatment times (10-150min) in a batch system. It was determined that a moderate temperature and short time are suitable for the preparation of polysaccharides. The structure of xylan and the inter- and intra-chain hydrogen bonding of cellulose fibrils in the soy hulls were not significantly broken down. The polysaccharides obtained were primarily composed of α-L-arabinofuranosyl units, 4-O-methyl-glucuronic acid units and α-D-galactose units attached with substituted units. A sugar analysis indicated that arabinose was the major component, constituting 35.6-46.9% of the polysaccharide products extracted at 130°C, 140°C, and 150°C. This investigation contributes to the knowledge of the polysaccharides of soy by-products, which can reduce the environmental impact of waste from the food industries. PMID:26920272

  10. Phase I clinical trial of O-Acetylated pectin conjugate, a plant polysaccharide based typhoid vaccine

    PubMed Central

    Szu, Shousun C.; Lin, Kimi F-Y; Hunt, Steven; Chu, Chiayung; Thinh, Nguyen Duc

    2014-01-01

    Background Typhoid fever remains an important cause of morbidity and mortality in the developing countries. Vi capsular polysaccharide conjugate vaccine demonstrated safety and efficacy in young children in high endemic regions. A novel typhoid conjugate vaccine based on plant polysaccharide pectin was studied in a phase I trial. Methods Fruit pectin, having the same carbohydrate backbone structure as Vi, was purified from citrus peel and used as the polysaccharide source to prepare a semi-synthetic typhoid conjugate vaccine. Pectin was chemically O-acetylated (OAcPec) to antigenically resemble Vi and conjugated to carrier protein rEPA, a recombinant exoprotein A from Pseudomonas aeruginosa. 25 healthy volunteers, 18–45 years old, were injected once with OAcPec-rEPA. Safety and IgG antibodies reactive with Vi and pectin were analyzed. Results No vaccine associated serious adverse reaction was reported. Six weeks after the injection of OAcPec-rEPA, 64% of the volunteers elicited >4 fold rise of anti-Vi IgG. At 26 weeks the level declined, but the difference between the levels at 6 and 26 weeks are not statistically significant. There is a direct correlation between the level of anti-Vi IgG before and after the injection (R2 = 0.96). The anti-Vi IgG can be absorbed by Vi, but not by pectin. There was no corresponding increase of anti-pectin after the injection, indicating the antibody response to OAcPec-rEPA was specific to Vi. There is no Vi-rEPA data in US adults for comparison of immune responses. The OAcPec-rEPA elicited significantly less IgG anti-Vi in US adults than those by Vi-rEPA in Vietnamese adults. Conclusion The O-acetylated pectin conjugate, a plant based typhoid vaccine, is safe and immunogenic in adult volunteers. PMID:24657719

  11. Immunomodulatory and Hemagglutinating Activities of Acidic Polysaccharides Isolated from Combretum racemosum

    PubMed Central

    Schepetkin, Igor A.; Kouakou, Koffi; Yapi, Ahoua; Kirpotina, Liliya N.; Jutila, Mark A.; Quinn, Mark T.

    2013-01-01

    Extracts of leaves of different species of the genus Combretum have been used historically to treat a variety of medicinal problems. However, little is known about the active components conferring therapeutic properties to these extracts. In the present studies, we evaluated biochemical properties and immunomodulatory activity of polysaccharides isolated from the leaves of Combretum racemosum. Water-soluble polysaccharides from leaves of C. racemosum were extracted and fractionated by DEAE-cellulose and Diaion HP-20 to obtain a Diaion-bound fraction, designated Combretum polysaccharide-acidic bound or CP-AB, which was eluted with methanol, and an unbound fraction, designated as CP-AU. Molecular weight determination, sugar analysis, and other physical and chemical characterization of the fractions were performed. Fraction CP-AU (mol. weight 5.0 kDa) contained type II arabinogalactan and had potent immunomodulatory activity, inducing the production of interleukin (IL)-1β, -6, -10, and tumor necrosis factor-α (TNF-α) by human peripheral blood mononuclear cells (PBMC) and MonoMac-6 monocytic cells. Likewise, intraperitoneal administration of CP-AU increased in vivo serum levels of IL-6 and monocyte chemoattractant protein-1 (MCP-1) in mice. CP-AU-induced secretion of TNF-α in PBMC was prevented by Toll-like receptor 4 (TLR4) antagonist LPS-RS. Treatment with CP-AU induced phosphorylation of Akt2, Akt3, GSK-3β, HSP27, mTOR, and all p38 MAPK isoforms (α, β, δ, and γ), as well as stimulation of AP-1/NF-κB transcriptional activity. In addition, CP-AU effectively agglutinated erythrocytes from several species, including human, mouse, and rabbit. In contrast, fraction CP-AB was inactive in all biological tests, including cytokine production and hemagglutination. These data suggest that at least part of the beneficial therapeutic effects reported for the water extracts of leaves from C. racemosum are due to modulation of leukocyte functions. PMID:23380150

  12. [Comparison on polysaccharide content and PMP-HPLC fingerprints of polysaccharide in stems and leaves of Dendrobium officinale].

    PubMed

    Zhou, Gui-Fen; Pang, Min-Xia; Chen, Su-Hong; Lv, Gui-Yuan; Yan, Mei-Qiu

    2014-03-01

    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.

  13. Human monoclonal macroglobulins with specificity for Klebsiella K polysaccharides that contain 3,4-pyruvylated-D-galactose and 4,6- pyruvylated-D-galactose

    PubMed Central

    1980-01-01

    Two human IgM myeloma proteins, IgMWEA and IgMMAY, were found to react with agar and Klebsiella polysaccharides that contain pyruvylated D- galactose (DGal). Quantitative precipitin data and precipitin inhibition studies with methyl alpha- and beta-glycosides of 4,6- pyruvylated-D-galactose showed their combining sites to be different, although each was directed against the pyruvylated-D-Gal, one reacting most specifically with Klebsiella polysaccharides with terminal nonreducing beta-linked 2,4 pyruvylated-D-Gal, whereas the other reacted equally well with Klebsiella polysaccharides that contain 3,4 beta-linked and 4,6 alpha-linked terminal nonreducing pyruvylated-DGal. Inhibition studies showed that both sites are directed toward one of the two space isomers of 3,4- or 4,6-pyruvylated DGal, the form in which the methyl group of the pyruvate is equatorial, or endo, and its carboxyl group axial, or exo, to the plane of the acetal ring. Coprecipitation studies showed the combining site of IgMWEA to be located on an (Fab')2 fragment and not on the (Fc)5mu fragment. The monoclonal peak in the serum of IgMMAY was specifically precipitated by Klebsiella polysaccharide. Myeloma proteins with specificities of this type may occur with reasonable frequency in humans and may be a consequence of clonal expansion from inapparent infection, carrier states, or disease produced by various Klebsiella organisms. PMID:6158553

  14. [Effects of germanium on cell growth, polysaccharide production and cellular redox status in suspension cultures of protocorm-like bodies of Dendrobium huoshanense].

    PubMed

    Wei, Ming; Yang, Chaoying; Jiang, Shaotong

    2010-03-01

    To solve the problem of low growth rate and metabolism level in suspension cultures of protocorm-like bodies (PLBs) of Dendrobium huoshanense. The effects of germanium on PLB proliferation and accumulation of polysaccharides together with nutrient utilization were investigated and the contents of reducing sugars, soluble proteins, the activities of antioxidant enzymes and redox status of the cells of PLB were analyzed. The results indicated that the optimum concentration of germanium dioxide (4.0 mg/L) significantly enhanced the cell growth and accumulation of polysaccharides, greatly improved contents of reducing sugars and soluble proteins, increased the activities of superoxide dismutase (SOD) and catalase (CAT) but decreased the activity of peroxidase(POD). The cell dry weight and production of polysaccharides were 32.6 g/L and 3.78 g/L, respectively. The analysis of cellular redox status showed that the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) in cells and the activity of glutathione reductase were significantly increased by the addition of germanium dioxide. The suitable concentration of germanium dioxide was beneficial to the cell growth and the accumulation of polysaccharides.

  15. Polysaccharide-Based Membranes in Food Packaging Applications.

    PubMed

    Ferreira, Ana R V; Alves, Vítor D; Coelhoso, Isabel M

    2016-01-01

    Plastic packaging is essential nowadays. However, the huge environmental problem caused by landfill disposal of non-biodegradable polymers in the end of life has to be minimized and preferentially eliminated. The solution may rely on the use of biopolymers, in particular polysaccharides. These macromolecules with film-forming properties are able to produce attracting biodegradable materials, possibly applicable in food packaging. Despite all advantages of using polysaccharides obtained from different sources, some drawbacks, mostly related to their low resistance to water, mechanical performance and price, have hindered their wider use and commercialization. Nevertheless, with increasing attention and research on this field, it has been possible to trace some strategies to overcome the problems and recognize solutions. This review summarizes some of the most used polysaccharides in food packaging applications. PMID:27089372

  16. Marine Polysaccharides from Algae with Potential Biomedical Applications

    PubMed Central

    de Jesus Raposo, Maria Filomena; de Morais, Alcina Maria Bernardo; de Morais, Rui Manuel Santos Costa

    2015-01-01

    There is a current tendency towards bioactive natural products with applications in various industries, such as pharmaceutical, biomedical, cosmetics and food. This has put some emphasis in research on marine organisms, including macroalgae and microalgae, among others. Polysaccharides with marine origin constitute one type of these biochemical compounds that have already proved to have several important properties, such as anticoagulant and/or antithrombotic, immunomodulatory ability, antitumor and cancer preventive, antilipidaemic and hypoglycaemic, antibiotics and anti-inflammatory and antioxidant, making them promising bioactive products and biomaterials with a wide range of applications. Their properties are mainly due to their structure and physicochemical characteristics, which depend on the organism they are produced by. In the biomedical field, the polysaccharides from algae can be used in controlled drug delivery, wound management, and regenerative medicine. This review will focus on the biomedical applications of marine polysaccharides from algae. PMID:25988519

  17. [Discussion on polysaccharide determination methods in new Chinese drug research].

    PubMed

    Li, Ji-Ping

    2014-09-01

    According to existing problems in polysaccharide determination methods in new Chinese drug applications, the method suitability, chemical reference selection, components interference and method research were introduced. The author suggests that suitable determination method should be selected according to the structure and property of the polysaccharide, and validated. Some influent factors should be examined to assure the accuracy of the method, such as the stability and using amount of the visualizing reagent, visualizing time, maximum detection wavelength etc. Monosaccharide and other water soluble components should be removed from polysaccharide sample, and suitable reference substance and detection wavelength should be selected. It should pay attention to mutual interference of neutral and acidic saccharide, and use inhibitor to eliminate the interference. Because the slopes of the standard curves are different for different monosaccharide, it is proposed that the types and ratios of the monosaccharide in heteroglycan should be understood, and mixed reference substance solution in the ratio is prepared for determination. PMID:25522636

  18. Antioxidant activity of herbal polysaccharides and cough reflex.

    PubMed

    Nosalova, G; Jurecek, L; Hromadkova, Z; Kostalova, Z; Sadlonova, V

    2013-01-01

    The extraction of Fallopia sachalinensis leaves resulted in two fractions (FS-1 and FS-2). Chemical and spectral analyses of samples revealed the prevalence of pectic polysaccharides with high galacturonic acid, arabinose, galactose, and rhamnose content. Arabinogalactan with a higher content of phenolic prevailed in the FS-1, whereas rhamnogalacturonan predominated in the FS-2 fraction. Both polysaccharides showed significant antioxidant activity according to DPPH and FRAP assays. Evaluation of antitussive activity in healthy adult conscious guinea pigs after oral application of 50 and 75 mg/kg of the FS-2 polysaccharide extracts showed a significant suppression of cough reflex, without an influence on specific airway resistance. The suppression of cough was comparable with that of codeine.

  19. Release of polysaccharide by sonication of cells ( Porphyridium sp.)

    NASA Astrophysics Data System (ADS)

    Faerman, V.; Mukmenev, I.; Shreiber, I.

    2009-03-01

    The paper deals with experimental data concerning the interaction of acoustical waves with microbial cell (Porphyridium sp.). The aim of the present paper was to increase the amount of biopolymer released from the microorganisms biomass with the aid ultrasound irradiation without scission or a decrease in the molecular weight. The results indicated that the amount of polysaccharide (for example) released from the cell pellet could be enhanced by ultrasound, depending on the frequency and energy of the ultrasound. The sugar composition remain the same, but the apparent viscosity of polysaccharide aqueous solutions decreased, indicated that some changes in the molecular shape and size occurred. When ultrasound irradiation was applied in the presence of either CO2 or CO2 + H2, the apparent viscosity of polysaccharide aqueous solutions increased (versus usual ultrasound treatment).

  20. Polysaccharide-Based Membranes in Food Packaging Applications.

    PubMed

    Ferreira, Ana R V; Alves, Vítor D; Coelhoso, Isabel M

    2016-01-01

    Plastic packaging is essential nowadays. However, the huge environmental problem caused by landfill disposal of non-biodegradable polymers in the end of life has to be minimized and preferentially eliminated. The solution may rely on the use of biopolymers, in particular polysaccharides. These macromolecules with film-forming properties are able to produce attracting biodegradable materials, possibly applicable in food packaging. Despite all advantages of using polysaccharides obtained from different sources, some drawbacks, mostly related to their low resistance to water, mechanical performance and price, have hindered their wider use and commercialization. Nevertheless, with increasing attention and research on this field, it has been possible to trace some strategies to overcome the problems and recognize solutions. This review summarizes some of the most used polysaccharides in food packaging applications.

  1. Polysaccharide-Based Membranes in Food Packaging Applications

    PubMed Central

    Ferreira, Ana R. V.; Alves, Vítor D.; Coelhoso, Isabel M.

    2016-01-01

    Plastic packaging is essential nowadays. However, the huge environmental problem caused by landfill disposal of non-biodegradable polymers in the end of life has to be minimized and preferentially eliminated. The solution may rely on the use of biopolymers, in particular polysaccharides. These macromolecules with film-forming properties are able to produce attracting biodegradable materials, possibly applicable in food packaging. Despite all advantages of using polysaccharides obtained from different sources, some drawbacks, mostly related to their low resistance to water, mechanical performance and price, have hindered their wider use and commercialization. Nevertheless, with increasing attention and research on this field, it has been possible to trace some strategies to overcome the problems and recognize solutions. This review summarizes some of the most used polysaccharides in food packaging applications. PMID:27089372

  2. Bioactivity and Applications of Sulphated Polysaccharides from Marine Microalgae

    PubMed Central

    de Jesus Raposo, Maria Filomena; de Morais, Rui Manuel Santos Costa; de Morais, Alcina Maria Miranda Bernardo

    2013-01-01

    Marine microalgae have been used for a long time as food for humans, such as Arthrospira (formerly, Spirulina), and for animals in aquaculture. The biomass of these microalgae and the compounds they produce have been shown to possess several biological applications with numerous health benefits. The present review puts up-to-date the research on the biological activities and applications of polysaccharides, active biocompounds synthesized by marine unicellular algae, which are, most of the times, released into the surrounding medium (exo- or extracellular polysaccharides, EPS). It goes through the most studied activities of sulphated polysaccharides (sPS) or their derivatives, but also highlights lesser known applications as hypolipidaemic or hypoglycaemic, or as biolubricant agents and drag-reducers. Therefore, the great potentials of sPS from marine microalgae to be used as nutraceuticals, therapeutic agents, cosmetics, or in other areas, such as engineering, are approached in this review. PMID:23344113

  3. Preparation and characterization of mucilage polysaccharide for biomedical applications.

    PubMed

    Archana, G; Sabina, K; Babuskin, S; Radhakrishnan, K; Fayidh, Mohammed A; Babu, P Azhagu Saravana; Sivarajan, M; Sukumar, M

    2013-10-15

    In the present investigation, the polysaccharide/mucilage from waste of Abelmoscus esculentus by modification in hot extraction using two different solvents (Acetone, Methanol) were extracted, characterized and further compared with seaweed polysaccharide for their potential applications. The percentage yield, emulsifying capacity and swelling index of this mucilage were determined. The macro algae and okra waste, gave high % yield (22.2% and 8.6% respectively) and good emulsifying capacity (EC%=52.38% and 54.76% respectively) with acetone, compared to methanol (11.3% and 0.28%; EC%=50%) (PH=7) while swelling index was greater with methanol than acetone extracts respectively. The infrared (I.R.) spectrum of the samples was recorded to investigate the chemical structure of mucilage. Thermal analysis of the mucilage was done with TGA (Thermal Gravimetric Analyzer) and DSC (Differential Scanning Calorimeter) which showed both okra and algal polysaccharide were thermostable hydrogels. PMID:23987320

  4. [Study on oral absorption enhancers of astragalus polysaccharides].

    PubMed

    Chen, Xiao-Yun; Tan, Xiao-Bin; Sun, E; Liu, Dan; Jia, Xiao-Bin; Zhang, Zhen-Hai

    2014-04-01

    Astragalus polysaccharides was lounded to 4-(2-aminoethylphenol), followed by labeling the APS-Tyr with fluorescein-5-isothiocyanate (FITC) at the secondary amino group. The absorption enhancement effects of low molecular weight chitosan and protamine on astragalus polysaccharides were evaluated via Caco-2 cell culture model. The results show that the fluorecent labeling compound has good stability and high sensitivity. On the other hand low molecular weight chitosan and protamine also can promoted absorption of the astragalus polysaccharides without any cytotoxity, and the absorption increase was more significant with increasing the amount of low molecular weight chitosan and protamine. At the same time, the low molecular weight chitosan has slightly better effect. The transepithelial electric resistance (TEER) of Caco-2 cells show that absorption enhancers could improve its membrane transport permeability by opening tight junctions between cells and increasing the cell membrane fluidity.

  5. Reviews on Mechanisms of In Vitro Antioxidant Activity of Polysaccharides

    PubMed Central

    Wang, Junqiao; Hu, Shuzhen; Nie, Shaoping; Yu, Qiang; Xie, Mingyong

    2016-01-01

    It is widely acknowledged that the excessive reactive oxygen species (ROS) or reactive nitrogen species (RNS) induced oxidative stress will cause significant damage to cell structure and biomolecular function, directly or indirectly leading to a number of diseases. The overproduction of ROS/RNS will be balanced by nonenzymatic antioxidants and antioxidant enzymes. Polysaccharide or glycoconjugates derived from natural products are of considerable interest from the viewpoint of potent in vivo and in vitro antioxidant activities recently. Particularly, with regard to the in vitro antioxidant systems, polysaccharides are considered as effective free radical scavenger, reducing agent, and ferrous chelator in most of the reports. However, the underlying mechanisms of these antioxidant actions have not been illustrated systematically and sometimes controversial results appeared among various literatures. To address this issue, we summarized the latest discoveries and advancements in the study of antioxidative polysaccharides and gave a detailed description of the possible mechanisms. PMID:26682009

  6. Fine Structure of Extracellular Polysaccharide of Erwinia amylovora†

    PubMed Central

    Politis, D. J.; Goodman, R. N.

    1980-01-01

    Virulent E9 and avirulent E8 strains of Erwinia amylovora were shown by means of light, transmission, and scanning microscopy to be, respectively, encapsulated and unencapsulated. Difficulty was encountered in stabilizing the fibrillar-appearing capsular extracellular polysaccharide. We suggest that the ephemeral nature of extracellular polysaccharide is due to the collapse of its extended structure upon dehydration. This occurs when bacteria are prepared for either transmission or scanning electron microscopy. The electron micrographs support our previous biochemical and immunological studies contending that the capsule is composed of tightly bound and loosely held components. The preparation of bacteria in freeze-dried colonies has permitted us to observe and explain the fluidity of the encapsulated strain. We suggest that this fluidity is a reflection of the loosely held extracellular polysaccharide or slime. Images PMID:16345638

  7. [Insertional mutation in the AZOBR_p60120 gene is accompanied by defects in the synthesis of lipopolysaccharide and calcofluor-binding polysaccharides in the bacterium Azospirillum brasilense Sp245].

    PubMed

    Katsy, E I; Prilipov, A G

    2015-03-01

    In the bacterium Azospirillum brasilense Sp245, extracellular calcofluor-binding polysaccharides (Cal+ phenotype) and two types of lipopolysaccharides, LPSI and LPSII, were previously identified. These lipopolysaccharides share the same repeating O-polysaccharide unit but have different antigenic structures and different charges of their O-polysaccharides and/or core oligosaccharides. Several dozens of predicted genes involved in the biosynthesis of polysaccharides have been localized in the AZOBR_p6 plasmid of strain Sp245 (GenBank accession no. HE577333). In the present work, it was demonstrated that an artificial transposon Omegon-Km had inserted into the central region of the AZOBR_p60120 gene in the A. brasilense Sp245 LPSI- Cal- KM252 mutant. In A. brasilense strain Sp245, this plasmid gene encodes a putative glycosyltransferase containing conserved domains characteristic of the enzymes participating in the synthesis of O-polysaccharides and capsular polysaccharides (accession no. YP004987664). In mutant KM252, a respective predicted protein is expected to be completely inactivated. As a result of the analysis of the EcoRI fragment of the AZOBR_p6 plasmid, encompassing the AZOBR_p60120 gene and a number of other loci, novel data on the structure of AZOBR_p6 were obtained: an approximately 5-kb gap (GenBank accession no. KM189439) was closed in the nucleotide sequence of this plasmid.

  8. Degradation of pectic polysaccharides in pickled green olives.

    PubMed

    Sánchez-Romero, C; Guillén, R; Heredia, A; Jiménez, A; Fernández-Bolaños, J

    1998-01-01

    The changes that occur in the pectic fractions in the cell wall of olives of the Manzanilla variety (Olea europaea pomiformis) during processing (initial treatment at high pH and subsequent lactic fermentation) have been researched. After studying various conditions for fractionating the pectic polysaccharides, the most adequate were chosen, involving sequential extraction with water, imidazole-hydrochloric acid buffer, sodium carbonate, 1 M potassium hydroxide, and 4 M potassium hydroxide. In the unprocessed fruit, the fractions studied consist mainly of high-molecular-weight acidic polysaccharides (70 to 250 kDa): homogalacturonans, rhamnogalacturonans, and branched arabinans. These were found in different proportions depending on the extraction agent used. At the same time, significant amounts of relatively low-molecular-weight (10 to 10.5 kDa) neutral branched arabinans were found in the water-soluble fraction. As a result of the processing, changes occurred in the proportions of the different groups of polysaccharides in accordance with changes in their solubility characteristics. These changes were reflected in the processed fruit by (i) and increase in the neutral branched arabinans in the water-soluble fraction due to the increased presence of such polysaccharides originally found in the carbonate and 4 M KOH-soluble fractions; (ii) an increase in homogalacturonans and rhamnogalacturonans, without significant changes in molecular weights, in the imidazole-soluble fraction as a result of the increased presence of corresponding polysaccharides originally found in the carbonate-soluble and water-soluble fractions; (iii) a substantial increase in uronic acids in the 1 M potassium hydroxide-soluble fraction, preferentially as low-molecular-weight polysaccharides; and (iv) a solubilization of arabinans in the 4 M potassium hydroxide-soluble fraction. PMID:9708257

  9. Structural features of the exocellular polysaccharides of Mycobacterium tuberculosis.

    PubMed Central

    Lemassu, A; Daffé, M

    1994-01-01

    The cell envelope which surrounds pathogenic mycobacteria is postulated to be a defence barrier against phagocytic cells and its outermost constituents have a tendency to accumulate in the culture medium. The present work demonstrates that the exocellular material of Mycobacterium tuberculosis contains large amounts of polysaccharides with only traces, if any at all, of lipids. Three types of polysaccharides were purified by anion-exchange and gel-filtration chromatography; all were found to be neutral compounds devoid of acyl substituents. They consisted of D-glucan, D-arabino-D-mannan and D-mannan, which were eluted from gel-filtration columns in positions corresponding to molecular masses of 123, 13 and 4 kDa respectively. Their predominant structural features were determined by the characterization of the per-O-methyl derivatives of enzymic, acetolysis and Smith-degradation products and by 1H- and 13C-n.m.r. spectroscopy of the purified polysaccharides, using mono- and two-dimensional homonuclear chemical-shift correlated spectroscopy and two-dimensional heteronuclear (1H/13C) spectroscopy. The glucan which represented up to 90% of the polysaccharides was composed of repeating units of five or six-->4-alpha-D-Glcp-1--> residues and a -->4-alpha-D-Glcp substituted at position 6 with an alpha-D-Glcp, indicating a glycogen-like highly branched structure not related to the so-called polysaccharide-II previously identified in tuberculin. The arabinomannan consisted of a mannan segment composed of a -->6-alpha-D-Man-1--> core substituted at some positions 2 with an alpha-D-Manp. The arabinan termini of the arabinomannan were found to be extensively capped with mannosyl residues. The possibility that these polysaccharides contribute to the persistence of the tubercle bacillus in the macrophage by molecular mimicry is discussed. PMID:8297342

  10. Autologous albumin enhances the humoral immune response to capsular polysaccharide covalently coattached to bacteria-sized latex beads.

    PubMed

    Colino, Jesus; Duke, Leah; Snapper, Clifford M

    2014-05-01

    Abundant autologous proteins, like serum albumin, should be immunologically inert. However, individuals with no apparent predisposition to autoimmune disease can develop immune responses to autologous therapeutic proteins. Protein aggregation is a potential major trigger of these responses. Adsorption of proteins to particles provides macromolecular size and may generate structural changes in the protein, resembling aggregation. Using aldehyde/sulfate latex beads coated with murine serum albumin (MSA), we found that BALB/c mice mounted MSA-specific IgG responses that were dependent on CD4(+) T cells. IgGs were specific for MSA adsorbed to solid surfaces and noncross-reactive with human, bovine, or pig albumins. T cells induced in response to MSA augmented the primary and induced boosted secondary IgG and IgM responses specific for the T cell-independent antigen, capsular polysaccharide of Streptococcus pneumoniae type 14 (PPS14), when the latter was attached to the same bead. Similar to the anti-MSA IgG response, the boosted PPS14-specific IgG secondary response was CD4(+) T-cell dependent, displayed a typical carrier effect, and was enhanced by, but did not require, Toll-like receptor stimulation. These results provide a potential mechanism for the induction of responses to autoantigens unable to induce specific T-cell responses, and provide new insights into polysaccharide-specific immunity.

  11. A polysaccharide fraction of adlay seed (Coixlachryma-jobi L.) induces apoptosis in human non-small cell lung cancer A549 cells

    SciTech Connect

    Lu, Xiangyi; Liu, Wei; Wu, Junhua; Li, Mengxian; Wang, Juncheng; Wu, Jihui; Luo, Cheng

    2013-01-11

    Highlights: Black-Right-Pointing-Pointer A polysaccharide from adlay seed, its molecular mass, optical rotation and sugars was determined. Black-Right-Pointing-Pointer We demonstrated that a polysaccharide from adlay can induce apoptosis in cancer cells. Black-Right-Pointing-Pointer The polysaccharide inhibited the metabolism and proliferation of NSCLC A549 cells. Black-Right-Pointing-Pointer The polysaccharide may trigger apoptosis via the mitochondria-dependent pathway. -- Abstract: Different seed extracts from Coix lachryma-jobi (adlay seed) have been used for the treatment of various cancers in China, and clinical data support the use of these extracts for cancer therapy; however, their underlying molecular mechanisms have not been well defined. A polysaccharide fraction, designated as CP-1, was extracted from the C.lachryma-jobi L. var. using the ethanol subsiding method. CP-1 induced apoptosis in A549 cells in a dose-dependent manner, as determined by MTT assay. Apoptotic bodies were observed in the cells by scanning electronic microscopy. Apoptosis and DNA accumulation during S-phase of the cell cycle were determined by annexin V-FITC and PI staining, respectively, and measured by flow cytometry. CP-1 also extended the comet tail length on single cell gel electrophoresis, and disrupted the mitochondrial membrane potential. Further analysis by western blotting showed that the expression of caspase-3 and caspase-9 proteins was increased. Taken together, our results demonstrate that CP-1 is capable of inhibiting A549 cell proliferation and inducing apoptosis via a mechanism primarily involving the activation of the intrinsic mitochondrial pathway. The assay data suggest that in addition to its nutritional properties, CP-1 is a very promising candidate polysaccharide for the development of anti-cancer medicines.

  12. Chitosan: A Promising Marine Polysaccharide for Biomedical Research

    PubMed Central

    Periayah, Mercy Halleluyah; Halim, Ahmad Sukari; Saad, Arman Zaharil Mat

    2016-01-01

    Biomaterials created 50 years ago are still receiving considerable attention for their potential to support development in the biomedical field. Diverse naturally obtained polysaccharides supply a broad range of resources applicable in the biomedical field. Lately, chitosan, a marine polysaccharide derived from chitins—which are extracted from the shells of arthropods such as crab, shrimp, and lobster—is becoming the most wanted biopolymer for use toward therapeutic interventions. This is a general short review of chitosan, highlighting the history, properties, chemical structure, processing method, and factors influencing the usage of chitosan derivatives in the biomedical field. PMID:27041872

  13. The framework of polysaccharide monooxygenase structure and chemistry.

    PubMed

    Span, Elise A; Marletta, Michael A

    2015-12-01

    Polysaccharide monooxygenases, or PMOs (also known as lytic PMOs or LPMOs), are a group of enzymes discovered in recent years to catalyze the oxidative degradation of carbohydrate polymers. The PMO catalytic domain has a β-sandwich fold that bears a strong resemblance to both immunoglobulin (Ig) and fibronectin type III (FnIII) domains. PMOs are secreted by fungi and bacteria, and there is recent evidence for their roles in pathogenesis, in addition to biomass processing. This review addresses the biological origins and functions of emerging PMO families, as well as describes the aspects of PMO structure that support the chemistry of copper-catalyzed, oxidative polysaccharide degradation.

  14. [Immunostimulating action of polysaccharides (heteroglycans) from higher plants].

    PubMed

    Wagner, H; Proksch, A; Riess-Maurer, I; Vollmar, A; Odenthal, S; Stuppner, H; Jurcic, K; Le Turdu, M; Fang, J N

    1985-01-01

    From the water or alcaline-water extracts of Echinacea purpurea (L.) Moench. and -angustifolia DC., Eupatorium cannabium L. and -perfoliatum L., Chamomilla recutita L. Rauscher, Calendula officinalis L., Baptisia tinctoria (L.) R. B., Achyrocline satureioides DC., Arnica montana L., Sabal serrulata Roem. et Schult., and Eleutherococcus (Acanthopanax) senticosus Maxim. polysaccharide fractions with molecular weights in the range of 25 000 to 500 000 and higher have been isolated, which, according to the granulocytes- and carbon clearance tests, showed significant immunostimulating activities. The isolated compounds belong to the group of watersoluble, acidic branched-chain heteroglycans. Their immunostimulating activity is compared and discussed with respect to other polysaccharides of biological activity.

  15. Impact of immobilized polysaccharide chiral stationary phases on enantiomeric separations.

    PubMed

    Ali, Imran; Aboul-Enein, Hassan Y

    2006-04-01

    Immobilized polysaccharide-based chiral stationary phases (CSPs) are gaining importance in the resolution of racemic compounds due to their stable nature on working with normal solvents and those prohibited for use with coated phases (tetrahydrofuran, chloroform, dichloromethane, acetone, 1,4-dioxane, ethyl acetate, and certain other ethers). This review discusses the use of immobilized polysaccharide CSPs in the chiral resolution of various racemates by liquid chromatography. The discussion includes immobilization methodologies, enantioselectivities, efficiencies, and a comparison of chiral recognition capabilities of coated vs. immobilized CSPs. Some applications of immobilized CSPs to the chiral resolution of racemic compounds are also presented. PMID:16830488

  16. Chitosan: A Promising Marine Polysaccharide for Biomedical Research.

    PubMed

    Periayah, Mercy Halleluyah; Halim, Ahmad Sukari; Saad, Arman Zaharil Mat

    2016-01-01

    Biomaterials created 50 years ago are still receiving considerable attention for their potential to support development in the biomedical field. Diverse naturally obtained polysaccharides supply a broad range of resources applicable in the biomedical field. Lately, chitosan, a marine polysaccharide derived from chitins-which are extracted from the shells of arthropods such as crab, shrimp, and lobster-is becoming the most wanted biopolymer for use toward therapeutic interventions. This is a general short review of chitosan, highlighting the history, properties, chemical structure, processing method, and factors influencing the usage of chitosan derivatives in the biomedical field.

  17. Evidence for microbial polysaccharide preparations containing polyester substituents.

    PubMed

    Giammatteo, P J; Stipanovic, A J; Nero, V P; Robison, P D

    1990-10-01

    CPMAS 13C-n.m.r. spectroscopy was employed to characterize the composition and solid phase morphology of gellan, welan, rhamsan and NW11. Spectra indicated that commercial preparations of these polysaccharides, which share a similar molecular backbone, contain a non-carbohydrate component exhibiting four inequivalent carbon atoms. Isolation of this component, followed by 13C-n.m.r. in CHCl3 and MS analysis, revealed its structure to be poly(beta-hydroxybutyrate). Evidence is presented which suggests that this polyester may be a covalent adduct to the above polysaccharides, although this cannot be unambiguously determined at this time. Further experimentation is in progress.

  18. Chitosan: A Promising Marine Polysaccharide for Biomedical Research.

    PubMed

    Periayah, Mercy Halleluyah; Halim, Ahmad Sukari; Saad, Arman Zaharil Mat

    2016-01-01

    Biomaterials created 50 years ago are still receiving considerable attention for their potential to support development in the biomedical field. Diverse naturally obtained polysaccharides supply a broad range of resources applicable in the biomedical field. Lately, chitosan, a marine polysaccharide derived from chitins-which are extracted from the shells of arthropods such as crab, shrimp, and lobster-is becoming the most wanted biopolymer for use toward therapeutic interventions. This is a general short review of chitosan, highlighting the history, properties, chemical structure, processing method, and factors influencing the usage of chitosan derivatives in the biomedical field. PMID:27041872

  19. Increasing stringiness of low-fat mozzarella string cheese using polysaccharides.

    PubMed

    Oberg, E N; Oberg, C J; Motawee, M M; Martini, S; McMahon, D J

    2015-07-01

    When fat content of pasta filata cheese is lowered, a loss of fibrous texture occurs and low-fat (LF) mozzarella cheese loses stringiness, making it unsuitable for the manufacture of string cheese. We investigated the use of various polysaccharides that could act as fat mimetics during the stretching and extruding process to aid in protein strand formation and increase stringiness. Low-fat mozzarella cheese curd was made, salted, and then 3.6-kg batches were heated in hot (80°) 5% brine, stretched, and formed into a homogeneous mass. Hot (80°C) slurries of various polysaccharides were then mixed with the hot cheese and formed into LF string cheese using a small piston-driven extruder. Polysaccharides used included waxy corn starch, waxy rice starch, instant tapioca starch, polydextrose, xanthan gum, and guar gum. Adding starch slurries increased cheese moisture content by up to 1.6% but was not effective at increasing stringiness. Xanthan gum functioned best as a fat mimetic and produced LF string cheese that most closely visually resembled commercial string cheese made using low-moisture part skim (LMPS) mozzarella cheese without any increase in moisture content. Extent of stringiness was determined by pulling apart the cheese longitudinally and observing size, length, and appearance of individual cheese strings. Hardness was determined using a modified Warner-Bratzler shear test. When LF string cheese was made using a 10% xanthan gum slurry added at ~1%, increased consumer flavor liking was observed, with scores after 2wk of storage of 6.44 and 6.24 compared with 5.89 for the LF control cheese; although this was lower than an LMPS string cheese that scored 7.27. The 2-wk-old LF string cheeses containing xanthan gum were considered still slightly too firm using a just-about-right (JAR) test, whereas the LMPS string cheese was considered as JAR for texture. With further storage up to 8wk, all of the LF string cheeses softened (JAR score was closer to 3

  20. Definition and characterization of enzymes for maximal biocatalytic solubilization of prebiotic polysaccharides from potato pulp.

    PubMed

    Thomassen, Lise V; Larsen, Dorte M; Mikkelsen, Jørn D; Meyer, Anne S

    2011-08-10

    Potato pulp is a high-volume co-processing product resulting from industrial potato starch manufacturing. Potato pulp is particularly rich in pectin, notably galactan branched rhamnogalacturonan I polysaccharides, which are highly bifidogenic when solubilized. The objective of the present study was to characterize and compare four homogalacturonan degrading enzymes capable of catalyzing the required solubilization of these pectinaceous polysaccharides from potato pulp in a 1 min reaction. An additional purpose was to assess the influence of the pH and the potential buffer chelating effects on the release of these polysaccharides from the potato pulp. The pH and temperature optima of two selected pectin lyases from Emericella nidulans (formerly known as Aspergillus nidulans) and Aspergillus niger were determined to 8.6 and 4.0, respectively, at ≥100 °C within 1 min of reaction. The optima for the two selected polygalacturonases from E. nidulans and Aspergillus aculeatus were determined to pH 4.4 and 46 °C, and pH 3.7 and ≥80 °C, respectively. The polygalacturonase from A. aculeatus was 4-42 times more heat-resistant at 50 °C than the other enzymes. The difference in pH optima of the pectin lyases and the exceptional thermal stabilities of some of the enzymes are proposed to be related to specific amino acid substitutions, stabilizing hydrogen bonding and structural traits of the enzymes. The K(M) and V(max) values ranged from 0.3-0.6g/L and 0.5-250.5 U/mg protein, respectively. Phosphate buffer induced release of a higher amount of dry matter than Tris-acetate buffer at pH 6, indicating a chelating effect of the phosphate. Moreover, the phosphate had a higher chelating effect at pH 6 than at pH 4. The optimal conditions for a high yield of polysaccharides from potato pulp were therefore: 1% (w/w) potato pulp treated with 1% (w/w) enzyme/substrate (E/S) pectin lyase from E. nidulans and 1% (w/w) E/S polygalacturonase from A. aculeatus at pH 6.0 and 60 °C for

  1. Increasing stringiness of low-fat mozzarella string cheese using polysaccharides.

    PubMed

    Oberg, E N; Oberg, C J; Motawee, M M; Martini, S; McMahon, D J

    2015-07-01

    When fat content of pasta filata cheese is lowered, a loss of fibrous texture occurs and low-fat (LF) mozzarella cheese loses stringiness, making it unsuitable for the manufacture of string cheese. We investigated the use of various polysaccharides that could act as fat mimetics during the stretching and extruding process to aid in protein strand formation and increase stringiness. Low-fat mozzarella cheese curd was made, salted, and then 3.6-kg batches were heated in hot (80°) 5% brine, stretched, and formed into a homogeneous mass. Hot (80°C) slurries of various polysaccharides were then mixed with the hot cheese and formed into LF string cheese using a small piston-driven extruder. Polysaccharides used included waxy corn starch, waxy rice starch, instant tapioca starch, polydextrose, xanthan gum, and guar gum. Adding starch slurries increased cheese moisture content by up to 1.6% but was not effective at increasing stringiness. Xanthan gum functioned best as a fat mimetic and produced LF string cheese that most closely visually resembled commercial string cheese made using low-moisture part skim (LMPS) mozzarella cheese without any increase in moisture content. Extent of stringiness was determined by pulling apart the cheese longitudinally and observing size, length, and appearance of individual cheese strings. Hardness was determined using a modified Warner-Bratzler shear test. When LF string cheese was made using a 10% xanthan gum slurry added at ~1%, increased consumer flavor liking was observed, with scores after 2wk of storage of 6.44 and 6.24 compared with 5.89 for the LF control cheese; although this was lower than an LMPS string cheese that scored 7.27. The 2-wk-old LF string cheeses containing xanthan gum were considered still slightly too firm using a just-about-right (JAR) test, whereas the LMPS string cheese was considered as JAR for texture. With further storage up to 8wk, all of the LF string cheeses softened (JAR score was closer to 3

  2. Extraction and purification of polysaccharides from pine medicinal mushroom, Tricholoma matsutake (higher Basidiomycetes) fruit bodies.

    PubMed

    You, Qinghong; Yin, Xiulian; Zhao, Yuping; Jiang, Zhonghai

    2014-01-01

    Ultrasound-assisted extraction, purification, and characterization of antioxidant polysaccharides from Tricholoma matsutake were carried out. On the basis of the results of a single-factor test, three independent and main variables, including extraction time (X1: 130-170 s), ultrasonic power (X2: 340-380 W), and ratio of water to raw material (X3: 45-55 mL/g) were studied by Box-Behnken design. The optimum extraction conditions are as follows: extraction time 160 s, ultrasonic power 365 W, and ratio of water to raw material 53.5 mL/g. Under the optimized conditions, the yield of T. matsutake polysaccharides (TMP) was 7.97±0.31%. The crude TMP was purified by DEAE-cellulose 52 chromatography and Sephadex G-100 chromatography to afford two fractions, TMP-1 and TMP-2. The crude TMP contained 85.76% carbohydrates, 3.57% proteins, and 0.13% uronic acids. The constituent monosaccharides were predominantly glucose, galactose, mannose, xylose, fucose, and glucuronic acid residues.

  3. Development of capsular polysaccharide-based glycoconjugates for immunization against melioidosis and glanders.

    PubMed

    Burtnick, Mary N; Heiss, Christian; Roberts, Rosemary A; Schweizer, Herbert P; Azadi, Parastoo; Brett, Paul J

    2012-01-01

    Burkholderia pseudomallei and Burkholderia mallei, the etiologic agents of melioidosis and glanders, respectively, cause severe disease in humans and animals and are considered potential agents of biological warfare and terrorism. Diagnosis and treatment of infections caused by these pathogens can be challenging and, in the absence of chemotherapeutic intervention, acute disease is frequently fatal. At present, there are no human or veterinary vaccines available for immunization against these emerging/re-emerging infectious diseases. One of the long term objectives of our research, therefore, is to identify and characterize protective antigens expressed by B. pseudomallei and B. mallei and use them to develop efficacious vaccine candidates. Previous studies have demonstrated that the 6-deoxy-heptan capsular polysaccharide (CPS) expressed by these bacterial pathogens is both a virulence determinant and a protective antigen. Consequently, this carbohydrate moiety has become an important component of the various subunit vaccines that we are currently developing in our laboratory. In the present study, we describe a reliable method for isolating CPS antigens from O-polysaccharide (OPS) deficient strains of B. pseudomallei; including a derivative of the select agent excluded strain Bp82. Utilizing these purified CPS samples, we also describe a simple procedure for covalently linking these T-cell independent antigens to carrier proteins. In addition, we demonstrate that high titer IgG responses can be raised against the CPS component of such constructs. Collectively, these approaches provide a tangible starting point for the development of novel CPS-based glycoconjugates for immunization against melioidosis and glanders.

  4. Strong cellulase inhibition by Mannan polysaccharides in cellulose conversion to sugars.

    PubMed

    Kumar, Rajeev; Wyman, Charles E

    2014-07-01

    Cellulase enzymes contribute a major fraction of the total cost for biological conversion of lignocellulosic biomass to fuels and chemicals. Although a several fold reduction in cellulase production costs and enhancement of cellulase activity and stability have been reported in recent years, sugar yields are still lower at low enzyme doses than desired commercially. We recently reported that hemicellulose xylan and its oligomers strongly inhibit cellulase and that supplementation of cellulase with xylanase and β-xylosidase would significantly reduce such inhibition. In this study, mannan polysaccharides and their enzymatically prepared hydrolyzates were discovered to be strongly inhibitory to fungal cellulase in cellulose conversion (>50% drop in % relative conversion), even at a small concentration of 0.1 g/L, and inhibition was much greater than experienced by other known inhibitors such as cellobiose, xylooligomers, and furfural. Furthermore, cellulase inhibition dramatically increased with heteromannan loading and mannan substitution with galactose side units. In general, enzymatically prepared hydrolyzates were less inhibitory than their respective mannan polysaccharides except highly substituted ones. Supplementation of cellulase with commercial accessory enzymes such as xylanase, pectinase, and β-glucosidase was effective in greatly relieving inhibition but only for less substituted heteromannans. However, cellulase supplementation with purified heteromannan specific enzymes relieved inhibition by these more substituted heteromannans as well, suggesting that commercial preparations need to have higher amounts of such activities to realize high sugar yields at the low enzyme protein loadings needed for low cost fuels production.

  5. pH-responsive self-assembly of polysaccharide through a rugged energy landscape

    PubMed Central

    Morrow, Brian H.; Payne, Gregory F.

    2015-01-01

    Self-assembling polysaccharides can form complex networks with structures and properties highly dependent on the sequence of triggering cues. Controlling the emergence of such networks provides an opportunity to create soft matter with unique features; however, it requires a detailed understanding of the subtle balance between the attractive and repulsive forces that drives the stimuli-induced self-assembly. Here we employ all-atom molecular dynamics simulations on the order of 100 ns to study the mechanisms of the pH-responsive gelation of the weakly basic aminopolysaccharide chitosan. We find that low pH induces a sharp transition from gel to soluble state, analogous to pH-dependent folding of proteins, while at neutral and high pH self-assembly occurs via a rugged energy landscape, reminiscent of RNA folding. A surprising role of salt is to lubricate conformational search for the thermodynamically stable states. Although our simulations represent the early events in the self-assembly process of chitosan, which may take seconds or minutes to complete, the atomically-detailed insights are consistent with recent experimental observations and provide a basis for understanding how environmental conditions modulate the structure and mechanical properties of the self-assembled polysaccharide systems. The ability to control structure and properties via modification of process conditions will aid in the technological efforts to create complex soft matter with applications ranging from bioelectronics to regenerative medicine. PMID:26383701

  6. Therapeutic efficacy of a polysaccharide isolated from Cordyceps sinensis on hypertensive rats.

    PubMed

    Xiang, Feixiang; Lin, Liming; Hu, Min; Qi, Xiangqian

    2016-01-01

    This study was aimed to investigate the antihypertensive effect of a polysaccharide fraction from Cordyceps sinensis on spontaneously hypertensive rats (SHR). The CSP1, one component of Cordyceps sinensis polysaccharides (CSP), was obtained after water extraction, deproteinization, de-colorization and purification with DEAE-cellulose 52. And a more homogeneous component CSP1-2 was obtained using Sepharose CL-6B chromatography. CSP1-2 mainly consisted of mannose, glucose and galactose in a molar ratio of about 2:2:1 and its average molecular weight was approximately 2.70×10(4)Da. Pharmacological tests showed that CSP1, in which the CSP1-2 was its main component, had antihypertensive effect by stimulating the secretion of vasodilator NO, decreasing the level of ET-1, epinephrine, noradrenaline and angiotensin II, inhibiting the increase of transforming growth factor β1 (TGF-β1) and lowering the level of inflammatory mediator of C-reactive protein (CRP). These results suggested that CSP1 may possess high potential in treating hypertension. PMID:26432374

  7. Purification and characterization of a novel polysaccharide-peptide complex from Clinacanthus nutans Lindau leaves.

    PubMed

    Huang, Danmin; Li, Yunhong; Cui, Fengjie; Chen, Jun; Sun, Jiamin

    2016-02-10

    A novel polysaccharide-peptide complex CNP-1-2 with molecular weight of 9.17 × 10(4) Da was obtained from Clinacanthus nutans Lindau leaves by hot water extraction, ethanol precipitation, and purification with Superdex 200 and DEAE-Sepharose Fast Flow column chromatography. CNP-1-2 exhibited the highest growth inhibitory effect on human gastric cancer cells SGC-7901 with inhibition ratio of 92.34% and stimulated activation of macrophages with NO secretion level of 47.53 μmol/L among the polysaccharide fractions. CNP-1-2 comprised approximately 87.25% carbohydrate and 9.37% protein. Monosaccharide analysis suggested that CNP-1-2 was composed of L-rhamnose, l-arabinose, D-mannose, D-glucose and D-galactose with a molar ratio of 1.30:1.00:2.56:4.95:5.09. Methylation analysis, FT-IR, and (1)H NMR spectroscopy analysis revealed that CNP-1-2 might have a backbone consisting of 1,4-linked Glcp, 1,3-linked Glcp, 1,3-linked Manp, 1,4-linked Galp, 1,2,6-linked Galp and 1,2,6-linked Galp. Its side chain might be composed of 1-linked Araf, 1,6-linked Galp and 1-linked Rhap residues. AFM (atomic force micrograph) analysis revealed that CNP-1-2 had the molecular aggregation along with branched and entangled structure.

  8. Radical scavenging and antioxidant effects of Matricaria chamomilla polyphenolic-polysaccharide conjugates.

    PubMed

    Kolodziejczyk-Czepas, Joanna; Bijak, Michal; Saluk, Joanna; Ponczek, Michal B; Zbikowska, Halina M; Nowak, Pawel; Tsirigotis-Maniecka, Marta; Pawlaczyk, Izabela

    2015-01-01

    Matricaria chamomilla L. (MC), a member of the Asteraceae family, is one of the oldest medicinal plants, widely used worldwide for a variety of healing applications. Its recommendations, derived from both traditional and modern medicine, include numerous disorders such as inflammation, ulcers, wounds, gastrointestinal disorders, stomach ache, pharyngitis, rheumatic pain, as well as the other ailments. This work is focused on another aspect of the biological activity of chamomile polyphenolic-polysaccharide conjugates--their antioxidant properties in the protection of blood plasma components against in vitro oxidative stress. Measurements of DPPH and ABTS radical scavenging indicated considerable anti-free radical action of MC. Pre-incubation of blood plasma with MC considerably diminished the extent of ONOO(-)-induced oxidative modifications such as protein carbonyl groups, SH groups, 3-nitrotyrosine, as well as the formation of lipid hydroperoxides. The analysis of the FRAP assay result shows a considerable increase of ferric reducing ability of blood plasma in the presence of MC. The results obtained in this study indicate that polyphenolic-polysaccharide conjugates isolated from M. chamomilla substances possess antioxidant properties. The M. chamomilla macromolecular glycoconjugates may be useful in the creation of new natural-based medications or dietary supplements, helpful in the prevention and treatment of oxidative stress-mediated disorders. PMID:25285848

  9. Separation, characterization and anticancer activities of a sulfated polysaccharide from Undaria pinnatifida.

    PubMed

    Han, Yun; Wu, Jun; Liu, Tingting; Hu, Youdong; Zheng, Qiusheng; Wang, Binsheng; Lin, Haiyan; Li, Xia

    2016-02-01

    The purpose of this paper was to investigate separation, characterization and anticancer activities of a sulfated polysaccharide (SPUP) from Undaria pinnatifida. Firstly, polysaccharide from U. pinnatifida was separated by DEAE-52 cellulose and Sephacryl S-400 column chromatography. As results, SPUP was obtained with the yield of 19.42%. Then, SPUP was characterized using chemical analysis, gas chromatography, size-exclusion HPLC chromatography, UV-vis spectra and FT-IR spectrum. The content of total sugar, uronic acid, protein and sulfate radical were 80.48%, 3.21%, 7.12% and 29.14%, respectively. SPUP was a heteropolysaccharide composed of fucose, glucose and galactose in a molar percentage of 27.15:19.34:53.51 with molecular weight of 97.9 kDa. Finally, the strongly against breast cancer activity of SPUP was confirmed by DMBA-induced breast cancer rats model. AS results, SPUP can significantly restrain breast abnormal enlargement, prolong tumor latency and reduced tumor incidence. Immunomodulatory activity and regulating abnormal sex hormones level might contribute to its anticancer activities. PMID:26616455

  10. The Dendrobium catenatum Lindl. genome sequence provides insights into polysaccharide synthase, floral development and adaptive evolution

    PubMed Central

    Zhang, Guo-Qiang; Xu, Qing; Bian, Chao; Tsai, Wen-Chieh; Yeh, Chuan-Ming; Liu, Ke-Wei; Yoshida, Kouki; Zhang, Liang-Sheng; Chang, Song-Bin; Chen, Fei; Shi, Yu; Su, Yong-Yu; Zhang, Yong-Qiang; Chen, Li-Jun; Yin, Yayi; Lin, Min; Huang, Huixia; Deng, Hua; Wang, Zhi-Wen; Zhu, Shi-Lin; Zhao, Xiang; Deng, Cao; Niu, Shan-Ce; Huang, Jie; Wang, Meina; Liu, Guo-Hui; Yang, Hai-Jun; Xiao, Xin-Ju; Hsiao, Yu-Yun; Wu, Wan-Lin; Chen, You-Yi; Mitsuda, Nobutaka; Ohme-Takagi, Masaru; Luo, Yi-Bo; Van de Peer, Yves; Liu, Zhong-Jian

    2016-01-01

    Orchids make up about 10% of all seed plant species, have great economical value, and are of specific scientific interest because of their renowned flowers and ecological adaptations. Here, we report the first draft genome sequence of a lithophytic orchid, Dendrobium catenatum. We predict 28,910 protein-coding genes, and find evidence of a whole genome duplication shared with Phalaenopsis. We observed the expansion of many resistance-related genes, suggesting a powerful immune system responsible for adaptation to a wide range of ecological niches. We also discovered extensive duplication of genes involved in glucomannan synthase activities, likely related to the synthesis of medicinal polysaccharides. Expansion of MADS-box gene clades ANR1, StMADS11, and MIKC*, involved in the regulation of development and growth, suggests that these expansions are associated with the astonishing diversity of plant architecture in the genus Dendrobium. On the contrary, members of the type I MADS box gene family are missing, which might explain the loss of the endospermous seed. The findings reported here will be important for future studies into polysaccharide synthesis, adaptations to diverse environments and flower architecture of Orchidaceae. PMID:26754549

  11. Data supporting the role of enzymes and polysaccharides during cassava postharvest physiological deterioration.

    PubMed

    Uarrota, Virgílio Gavicho; Moresco, Rodolfo; Schmidt, Eder Carlos; Bouzon, Zenilda Laurita; da Costa Nunes, Eduardo; de Oliveira Neubert, Enilto; Peruch, Luiz Augusto Martins; Rocha, Miguel; Maraschin, Marcelo

    2016-03-01

    This data article is referred to the research article entitled The role of ascorbate peroxidase, guaiacol peroxidase, and polysaccharides in cassava (Manihot esculenta Crantz) roots under postharvest physiological deterioration by Uarrota et al. (2015). Food Chemistry 197, Part A, 737-746. The stress duo to PPD of cassava roots leads to the formation of ROS which are extremely harmful and accelerates cassava spoiling. To prevent or alleviate injuries from ROS, plants have evolved antioxidant systems that include non-enzymatic and enzymatic defence systems such as ascorbate peroxidase, guaiacol peroxidase and polysaccharides. In this data article can be found a dataset called "newdata", in RData format, with 60 observations and 06 variables. The first 02 variables (Samples and Cultivars) and the last 04, spectrophotometric data of ascorbate peroxidase, guaiacol peroxidase, tocopherol, total proteins and arcsined data of cassava PPD scoring. For further interpretation and analysis in R software, a report is also provided. Means of all variables and standard deviations are also provided in the Supplementary tables ("data.long3.RData, data.long4.RData and meansEnzymes.RData"), raw data of PPD scoring without transformation (PPDmeans.RData) and days of storage (days.RData) are also provided for data analysis reproducibility in R software.

  12. Physico-chemical properties of Salmonella typhi Vi polysaccharide-diphtheria toxoid conjugate vaccines affect immunogenicity.

    PubMed

    An, So Jung; Yoon, Yeon Kyung; Kothari, Sudeep; Kothari, Neha; Kim, Jeong Ah; Lee, Eugene; Kim, Deok Ryun; Park, Tai Hyun; Smith, Greg W; Carbis, Rodney

    2011-10-13

    In this study it was demonstrated that the immunogenicity of Vi polysaccharide-diphtheria toxoid conjugates was related to the physical and chemical structure of the conjugate. Conjugates were prepared in two steps, firstly binding adipic acid dihydrazide (ADH) spacer molecules to diphtheria toxoid (DT) carrier protein then secondly binding varying amounts of this derivatized DT to a fixed amount of Vi capsular polysaccharide purified from Salmonella enterica Serovar Typhi. As the amount of DT bound to the Vi increased the size of the conjugate increased but also the degree of cross-linking increased. The immunogenicity of the conjugates was tested in mice and measured by ELISA for anti Vi and anti DT IgG responses, and the results revealed a trend that as the amount of DT bound to the Vi increased the anti Vi responses increased. This study establishes a correlation between physico-chemical characteristics of the conjugate and the magnitude of the anti Vi and anti DT responses. PMID:21843575

  13. Polysaccharide chemistry regulates kinetics of calcite nucleation through competition of interfacial energies

    PubMed Central

    Hamm, Laura M.; Han, Nizhou; De Yoreo, James J.; Dove, Patricia M.

    2013-01-01

    Calcified skeletons are produced within complex assemblages of proteins and polysaccharides whose roles in mineralization are not well understood. Here we quantify the kinetics of calcite nucleation onto a suite of high-purity polysaccharide (PS) substrates under controlled conditions. The energy barriers to nucleation are PS-specific by a systematic relationship to PS charge density and substrate structure that is rooted in minimization of the competing substrate–crystal and substrate–liquid interfacial energies. Chitosan presents a low-energy barrier to nucleation because its near-neutral charge favors formation of a substrate–crystal interface, thus reducing substrate interactions with water. Progressively higher barriers are measured for negatively charged alginates and heparin that favor contact with the solution over the formation of new substrate–crystal interfaces. The findings support a directing role for PS in biomineral formation and demonstrate that substrate–crystal interactions are one end-member in a larger continuum of competing forces that regulate heterogeneous crystal nucleation. PMID:23690577

  14. Structural characterization and emulsifying properties of polysaccharides of Acacia mearnsii de Wild gum.

    PubMed

    Grein, Aline; da Silva, Bruno C; Wendel, Cinthia F; Tischer, Cesar A; Sierakowski, Maria Rita; Moura, Angela B Dewes; Iacomini, Marcello; Gorin, Philip A J; Simas-Tosin, Fernanda F; Riegel-Vidotti, Izabel C

    2013-01-30

    Polysaccharides (GNF) from Acacia mearnsii de Wild gum exudates, collected from trees growing in the south of Brazil, were characterized ((13)C and HSQC NMR, GC-MS, colorimetric assays). A commercial gum arabic (GAC) was analyzed similarly and compared with GNF. There were differences, consistent with distinct behavior in tensiometry tests and as emulsion stabilizer. GNF had a higher protein content than GAC, with small differences in the monosaccharide composition, the greater one being the lower uronic acid content of GNF (4%), compared with GAC (17%). GNF had a much broader molecular mass distribution, M(w)/M(n), and a lower M(w). GNF was more efficient in lowering the surface tension of water and saline solutions and was more efficient in emulsifying castor oil droplets. Results were discussed taking into account structural and molecular differences between the studied gums. It was concluded that polysaccharides from A. mearnsii de Wild are candidates as substitutes of currently commercialized arabic gums (Acacia senegal and Acacia seyal) having, depending on their application, improved properties.

  15. Structure and bioactivity of a polysaccharide extracted from protocorm-like bodies of Dendrobium huoshanense.

    PubMed

    Li, Fan; Cui, Shao-Hua; Zha, Xue-Qiang; Bansal, Vibha; Jiang, Yu-Lan; Asghar, Muhammad-Naeem; Wang, Jun-Hui; Pan, Li-Hua; Xu, Bing-Fa; Luo, Jian-Ping

    2015-01-01

    The crude polysaccharides of Dendrobium huoshanense were fractionated by anion-exchange chromatography and gel permeation chromatography, giving one homogeneous fraction DHP-4A with molecular weight of 2.32 × 10(5)Da. UV spectrum indicated that there was no existence of proteins and nucleic acids in DHP-4A. Monosaccharide analysis revealed that DHP-4A was made up of glucose, arabinose, mannose and rhamnose with a molar ratio of 13.8:3.0:6.1:2.1. T