ABSTRACT
We introduced a strategy for preparing a carbohydrate microarray and demonstrated its utility for characterizing carbohydrate binding and activities. We isolated the lipopolysaccharide (LPS) components from different bacteria and explored the possibility of immobilizing these glycoconjugates on a high-binding polystyrene plate. Carbohydrate-specific combination was examined by observing the binding of the blood group B analogic LPS O-polysaccharide from Escherichia coli on the high-binding polystyrene plate and anti-B from a broad spectra antibody of human blood serum. Strong binding of antibodies was screened, as it was evident that relative response value is two times higher than control. The hybridization results indicated that this method is a reliable technique for the detection of human intestinal bacteria and is expected to be applied in diagnostics and seroepidemiology.
Subject(s)
Lipopolysaccharides , Serum , Humans , Lipopolysaccharides/chemistry , Polystyrenes , Seroepidemiologic Studies , Carbohydrates/chemistry , Escherichia coli , ImmunoglobulinsABSTRACT
The antioxidant activities of polysaccharide from balsam pear were studied. It was determined by gel permeation chromatography that the molecular weight distribution of purified polysaccharide was concentrated near 33582â¯Da. It indicated that the monosaccharide components were rhamnose, galacturonic acid, galactose, xylose and arabinose. Fourier transform infrared spectroscopy, nuclear magnetic resonance and Congo red experiments showed that there are C1, C2, C3, C5 links, and stable ß-triple helix conformation in aqueous solution. It was found that polysaccharide had good scavenging effect on free radicals. So, balsam pear polysaccharide should be a potential antioxidant.
Subject(s)
Antioxidants/pharmacology , Plant Extracts/pharmacology , Polysaccharides/pharmacology , Pyrus/chemistry , Antioxidants/chemistry , Antioxidants/isolation & purification , Chemical Fractionation , Free Radical Scavengers/chemistry , Free Radical Scavengers/pharmacology , Molecular Conformation , Molecular Weight , Monosaccharides/analysis , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Spectrum AnalysisABSTRACT
The aim of this study was to investigate the effect of phosphorylation on the antioxidant activity of (1â¯ââ¯3)-ß-d-glucan from yeast cell wall. Alkali-insoluble (1â¯ââ¯3)-ß-d-glucan was extracted from yeast cell wall by an acid-base method. It was found that the purity of the sample was greatly improved after the precipitation was treated with alkali at 90 â and then by acetic acid, which was about 96.5%. Phosphorylated (1â¯ââ¯3)-ß-d-glucan was prepared. Infrared (IR) spectra and nuclear magnetic resonance spectra (NMR) confirmed the successful introduction of phosphate into glucan. The substitution degree of phosphate was 0.18. The phosphorylated (1â¯ââ¯3)-ß-d-glucan could significantly increase SOD and CAT contents in serum, liver and brain of mice, and reduce MDA level in serum, liver and brain to a certain extent in vivo. This lays a solid foundation for the research and development of phosphorylated (1â¯ââ¯3)-ß-d-glucan antioxidant.
Subject(s)
Antioxidants/metabolism , beta-Glucans/chemistry , Animals , Antioxidants/chemistry , Catalase/blood , Catalase/metabolism , Cell Wall/chemistry , Cell Wall/metabolism , Magnetic Resonance Spectroscopy , Malondialdehyde/blood , Malondialdehyde/metabolism , Mice , Phosphorylation , Saccharomyces cerevisiae/metabolism , Spectrophotometry, Infrared , Superoxide Dismutase/blood , Superoxide Dismutase/metabolismABSTRACT
BACKGROUND: In recent years, the antioxidant effects of polysaccharides have become a hot spot in the field of polysaccharide research. METHOD: Herein, the action mechanisms of polysaccharide antioxidation and scavenging free radicals were analyzed. The research progresses on the preparation methods and antioxidant properties of polysaccharides and their derivatives were summarized. CONCLUSION: Investigating the antioxidant activities of polysaccharides and their derivatives can find useful polysaccharides and their derivatives, which have great potential as natural antioxidants used in functional foods or medicines.
Subject(s)
Antioxidants/chemistry , Polysaccharides/chemistry , Althaea/chemistry , Althaea/metabolism , Antioxidants/isolation & purification , Ficus/chemistry , Ficus/metabolism , Plant Extracts/chemistry , Polysaccharides/isolation & purification , Reactive Oxygen Species/metabolismABSTRACT
BACKGROUND: The natural polysaccharides contain plant polysaccharides, animal polysaccharides and microbial polysaccharides. They are a kind of biological macromolecules with immune regulation, anti-tumor, anti-radiation, anti-inflammation, anti-fatigue and anti-aging effects. OBJECTIVE: These effects are related to their antioxidant properties. RESULTS: The action mechanisms of antioxidation and scavenging free radicals for natural polysaccharides were reviewed. The recent research progresses and our work on antioxidant properties of polysaccharides and their derivatives were summarized. At last, the existing problems of antioxidant polysaccharides were analyzed, and the development prospects were also presented. CONCLUSION: It is important to study the antioxidant activities of polysaccharides and their derivatives for the development of natural antioxidants.
Subject(s)
Antioxidants/pharmacology , Biological Products/pharmacology , Polysaccharides/pharmacology , Animals , Bacteria/metabolism , Plants/metabolismABSTRACT
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.
Subject(s)
Antineoplastic Agents/administration & dosage , Chitosan/chemistry , Dextrans/chemistry , Drug Carriers/chemistry , Hyaluronic Acid/chemistry , Antineoplastic Agents/chemistry , Drug Delivery Systems , Drug Liberation , Hydrophobic and Hydrophilic Interactions , Molecular Structure , Nanoparticles/chemistryABSTRACT
Glycochips can be used to map the carbohydrate-protein interactions in high-throughput manner. Carbohydrates are immobilized on the support surface by covalent or noncovalent binding. The immobilization strategies for glycochips were summarized herein. In addition, some recently developed techniques for detection of carbohydrate-protein interactions were also involved.
Subject(s)
Carbohydrates/chemistry , Protein Array Analysis/methods , Humans , Microchemistry/methods , Protein Array Analysis/instrumentation , Proteins/chemistry , Proteins/metabolism , Surface PropertiesABSTRACT
The pseudo-trisaccharide allosamidin 1 is a potent inhibitor of all family-18 chitinases, and it is confirmed to have insecticidal and antifungal activities. But the synthesis of allosamidins is very difficult, and it is a challengeable subject. Allosamidins were synthesized in solid-liquid phase, total solid-phase and total liquid-phase, respectively. Solid-liquid phase method realizes the partial solid-phase synthesis of allosamidins. Total solid-phase method greatly simplifies the purification process. Total liquid-phase method shortens the synthetic steps of allosamidins. The insecticidal and antifungal activities of allosamidins were also reported herein.
Subject(s)
Acetylglucosamine/analogs & derivatives , Antifungal Agents/pharmacology , Enzyme Inhibitors/pharmacology , Insecticides/pharmacology , Trisaccharides/chemical synthesis , Trisaccharides/pharmacology , Acetylglucosamine/chemical synthesis , Acetylglucosamine/chemistry , Acetylglucosamine/pharmacology , Animals , Antifungal Agents/chemical synthesis , Antifungal Agents/chemistry , Bombyx/drug effects , Carbohydrate Conformation , Chitinases/antagonists & inhibitors , Chitinases/metabolism , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Fungi/drug effects , Insecticides/chemical synthesis , Insecticides/chemistry , Spodoptera/drug effects , Trisaccharides/chemistryABSTRACT
Many natural products contain sugar residues, which are essential components for great medicinal importance. The sugar moieties can improve water-solubility of natural products and decrease their toxicity. At the same time, the glycosidic residues are crucial for the activities of natural products. Much effort has been expended over the past decades in developing novel and efficient methodologies to synthesize the glycosylated natural products. This review highlights recent developments in the synthesis of glycosylated natural products. The structure-activity relationships of some of these glycosylated natural products, together with the structure characteristics of their interaction with the biological targets, are also involved.
Subject(s)
Biological Products/chemistry , Biological Products/pharmacology , Glycosylation , Structure-Activity RelationshipABSTRACT
The penta-N-acetyl-chitopentaose 2 has been prepared by using recombinant E. coli strains harboring the nodC gene (encoding chitooligosaccharide synthase) from Azorhizobium caulinodans. Then, the deacetylase NodB removed the N-acetyl moiety from the nonreducing terminus of 2 to give tetra-N-acetyl-chitopentaose 3. N-Acylation of 3 with stearyl chloride was performed in DMF containing water and provided the corresponding lipo-chitopentaose nodulation factor 4. A binding chitinase assay indicated that 4 was much more stable than 3.
Subject(s)
Chitinases/chemistry , Oligosaccharides/biosynthesis , Oligosaccharides/chemistry , Acylation , Amidohydrolases/chemistry , Azorhizobium caulinodans/enzymology , Azorhizobium caulinodans/genetics , Bacterial Proteins/chemistry , Escherichia coli/genetics , Genetic Engineering/methods , Glucosides/biosynthesis , N-Acetylglucosaminyltransferases/genetics , N-Acetylglucosaminyltransferases/metabolismABSTRACT
Fluorophore-assisted carbohydrate electrophoresis (FACE) is a straightforward, sensitive method for determining the presence and relative abundance of individual (oligo)saccharide in a(n) (oligo)saccharide mixture. The single terminal aldehydes of (oligo)saccharides were tagged with the charged fluorophore 8-aminonaphthalene-1,3,6-trisulfonate (ANTS), and separated with high resolution on the basis of size by polyacrylamide gel electrophoresis. ANTS fluorescence labeling is not biased by (oligo)saccharide length. Therefore, band fluorescence intensity is directly related to the relative abundance of individual (oligo)saccharide moieties in heterogeneous sample. In the same time, it also indicates that FACE can be used to investigate the interactions of carbohydrates and proteins.
Subject(s)
Electrophoresis, Polyacrylamide Gel/methods , Oligosaccharides/analysis , Oligosaccharides/chemistry , Proteins/chemistry , Concanavalin A/chemistry , Fluorescent Dyes , Hydrolysis , NaphthalenesABSTRACT
A new compound 2, possessing a tetra-N-acetyl-chitotetraosyl moiety as a constituent, was synthesized by bacterial fermentation, which used allosamizoline 1 as the initial acceptor. A 2-binding chitinase assay, indicated that the chitinase was inactivated by 2 with IC50 = 0.03 microg/mL.
Subject(s)
Fermentation , Glucosamine/analogs & derivatives , Carbohydrate Conformation , Chitinases/antagonists & inhibitors , Chitinases/metabolism , Enzyme Activation/drug effects , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/metabolism , Enzyme Inhibitors/pharmacology , Glucosamine/biosynthesis , Glucosamine/chemistry , Glucosamine/pharmacology , Protein BindingABSTRACT
A sensitive,specific, and rapid method for the detection of carbohydrate-protein interactions is demonstrated by fluorophore-assisted carbohydrate electrophoresis (FACE). The procedure is simple and the cost is low. The advantage of this method is that carbohydrate-protein interactions can be easily displayed by FACE, and the carbohydrates do not need to be purified.
Subject(s)
Carbohydrates , Electrophoresis/methods , Fluorescent Dyes/metabolism , Naphthalenes/metabolism , Proteins/metabolism , Electrophoresis/economics , Electrophoresis/instrumentation , Sensitivity and SpecificityABSTRACT
A new compound 7, possessing a tetra-N-acetyl-chitotetraosyl moiety as a constituent, was synthesized by bacterial fermentation which used allosamizoline 6 as the initial acceptor.
Subject(s)
Acetylcholinesterase/metabolism , Bacterial Proteins/metabolism , Glucosamine/analogs & derivatives , N-Acetylglucosaminyltransferases/metabolism , Animals , Electrophorus , Glucosamine/chemical synthesis , Glucosamine/chemistry , Glucosamine/metabolism , Molecular StructureABSTRACT
The epoxyalkyl (1-->3)-beta-D-pentaglucosides 2 and 3 were synthesized in order by acetylation, glycosidation, oxidation, and deacetylation of 1. The immunological activities (superoxide anion production activity, phagocytic activity, and lymphocyte proliferation) and scavenging ability toward superoxide anion of (1-->3)-beta-D-pentaglucoside (1) and its epoxyalkyl derivatives (2 and 3) were investigated. Superoxide anion released from human blood monocytes was measured by the reduction of ferricytochrome c. Phagocytosis by peritoneal macrophages was detected through a teal ingesting that measured the chicken red blood cells (CRBC). Lymphocyte proliferation was determined by the MTT method. The scavenging ability of 1, 2, and 3 toward superoxide anions was evaluated by means of chemiluminescence (CL). The results showed that 2 and 3 had a little higher immunological activity and scavenging ability toward superoxide anion than 1, which indicated that the reducing end of the oligoglucosides was quite important for maximum biological activity.
Subject(s)
Glucosides/chemical synthesis , Macrophages, Peritoneal/drug effects , Monocytes/drug effects , Oligosaccharides/chemical synthesis , Animals , Cell Proliferation/drug effects , Cells, Cultured , Chickens , Glucosides/immunology , Humans , Lymphocyte Activation/drug effects , Monocytes/immunology , Oligosaccharides/immunology , Oligosaccharides/pharmacology , Phagocytosis/drug effects , Structure-Activity Relationship , Superoxides/metabolismABSTRACT
A sensitive, specific, and rapid method for the detection of carbohydrate-protein interactions was demonstrated using quantum dots (QDs) as a fluorescence label coupled with protein. 1,3-Dipolar cycloaddition between azide and alkyne was exploited to attach alpha-d-glucopyranoside to a C(14) hydrocarbon chain that noncovalently binds to the microtiter well surface, and the product formation was detected by both electrospray ionization-mass spectrometry (ESI-MS) and QD- (or fluorescein isothiocyanate (FITC))-conjugated lectin binding. It indicated that the peak intensity of the fluorescence emission was proportional to the initial concanavalin A (Con A) concentration in the range of 2 x 10(-3) micromol/L to 2 x 10(-2)mmol/L with a detection limit at least 100 times lower than that of the FITC-based method.
Subject(s)
Carbohydrate Metabolism , Microarray Analysis/methods , Quantum Dots , Staining and Labeling/methods , Adsorption , Carbohydrates/chemistry , Concanavalin A/chemistry , Concanavalin A/metabolism , Fluorescein-5-isothiocyanate , Fluorescence , Microarray Analysis/instrumentation , Molecular Structure , Protein Binding , Spectrometry, Mass, Electrospray IonizationABSTRACT
The title pentasaccharide was synthesized via a 2+3 strategy. The disaccharide donor, 3-O-acetyl-2-O-benzoyl-4,6-O-benzylidene-beta-D-glucopyranosyl-(1-->3)-2-O-benzoyl-4,6-O-benzylidene-alpha-D-glucopyranosyl trichloroacetimidate (8), was obtained by selective coupling of allyl 2-O-benzoyl-4,6-O-benzylidene-alpha-D-glucopyranoside with 3-O-acetyl-2-O-benzoyl-4,6-O-benzylidene-alpha-D-glucopyranosyl trichloroacetimidate (4), followed by deallylation, and trichloroacetimidation. Meanwhile, the trisaccharide acceptor, allyl 2-O-benzoyl-4,6-O-benzylidene-beta-D-glucopyranosyl-(1-->3)-2-O-benzoyl-4,6-O-benzylidene-beta-D-glucopyranosyl-(1-->3)-2-O-benzoyl-4,6-O-benzylidene-beta-D-glucopyranoside (12), was prepared by coupling of allyl 2-O-benzoyl-4,6-O-benzylidene-beta-D-glucopyranosyl-(1-->3)-2-O-benzoyl-4,6-O-benzylidene-beta-D-glucopyranoside with 4, followed by deacetylation. Condensation of 8 with 12, followed by epoxidation, and deprotection, gave the target pentaoside.
Subject(s)
Glucosides/chemical synthesis , Oligosaccharides/chemical synthesis , Carbohydrate Conformation , Carbohydrate Sequence , Magnetic Resonance Spectroscopy , Molecular Conformation , Molecular Structure , Oligosaccharides/chemistry , Oxidation-ReductionABSTRACT
The (1-->3)-beta-D-pentaglucoside was synthesized as its (R)-2,3-epoxypropyl glycoside via 2+3 strategy. The disaccharide donor 8 was obtained by 3-selective coupling of 2 with 4, followed by deallylation, and trichloroacetimidation. Meanwhile, the trisaccharide acceptor 12 was prepared by coupling of 10 with 4, followed by deacetylation. Condensation of 8 with 12, followed by epoxidation, and deprotection, gave the target pentaoside. The results of these bioassays demonstrated that the (1-->3)-beta-D-glucanase was obviously inactivated by 15 with k(app)=3.79 x 10(-4) min(-1). At the same time, we found that the 15 was more active as compared to the laminaripentaose in eliciting phytoalexin accumulation in tobacco cotyledon tissue, and it could be kept longer time than laminaripentaose, which indicated it is much more stable than laminaripentaose.
Subject(s)
Epoxy Compounds/chemical synthesis , Epoxy Compounds/pharmacology , Glucan 1,3-beta-Glucosidase/drug effects , Glucosides/chemical synthesis , Glucosides/pharmacology , Oligosaccharides/chemical synthesis , Oligosaccharides/pharmacology , Plant Extracts/metabolism , Biological Assay , Carbohydrate Sequence , Cotyledon/drug effects , Cotyledon/metabolism , Epoxy Compounds/chemistry , Epoxy Compounds/metabolism , Glucan 1,3-beta-Glucosidase/metabolism , Glucans , Glucosides/chemistry , Molecular Sequence Data , Molecular Structure , Oligosaccharides/metabolism , Plant Extracts/chemistry , Polysaccharides/pharmacology , Protein Binding , Sesquiterpenes , Terpenes , Nicotiana/drug effects , Nicotiana/metabolism , PhytoalexinsABSTRACT
We describe a approach for the synthesis of (1-->3)-beta-D-oligosaccharide derivatives 10-18. 1-9 were synthesized by treating peracetylated (1-->3)-beta-D-oligosaccharides with the corresponding alkenyl alcohols and Lewis acid (SnCl(4)) catalyst. Epoxidation of the corresponding alkenyl oligoglucosides took place by m-CPBA. NaOMe in dry methanol was used for the deacetylation of the blocked derivatives, to give 10-18 in an overall yields of 25-32%. In subsequent glucan-binding protein of soybean assays, we found that 16 was most active, with an IC(50) value of 9 mM. However, the activities of 17, 18, 13, 14, 15, 10, 11, and 12 were gradually decreased. At the same time, we found 16 was most active as compared to the other (1-->3)-beta-D- oligoglucoside derivatives in eliciting phytoalexin accumulation in soybean cotyledon tissue, and 16 was kept longer time than (1-->3)-beta-D-glucohexaose, which indicated 16 is much more stable than (1-->3)-beta-D-glucohexaose.
Subject(s)
Epoxy Compounds/chemical synthesis , Glucosides/chemical synthesis , Oligosaccharides/chemical synthesis , Plant Extracts/biosynthesis , Plant Proteins/metabolism , Epoxy Compounds/metabolism , Epoxy Compounds/pharmacology , Glucosides/metabolism , Glucosides/pharmacology , Oligosaccharides/metabolism , Oligosaccharides/pharmacology , Sesquiterpenes , Terpenes , PhytoalexinsABSTRACT
We describe a approach for the synthesis of a mixture of 3,4-epoxybutyl (1-->3)-beta-D-oligoglucosides. The particular (1-->3)-beta-D-glucan isolated from the cell walls of Saccharomyces cerevisiae was recovered from the aqueous medium as water-insoluble particles by the spray drying (GS) method, and it was characterized by FTIR spectroscopy. The acid-solubilized (1-->3)-beta-D-oligoglucosides were prepared by partial acid hydrolysis of glucan particles, which were qualitatively analyzed by fluorophore-assisted carbohydrate electrophoresis (FACE). The peracetylated 3-butenyl (1-->3)-beta-D-oligoglucosides were synthesized by treating peracetylated (1-->3)-beta-D-oligoglucosides with the 3-butenyl alcohols and a Lewis acid (SnCl4) catalyst. Epoxidation of the peracetylated 3-butenyl oligoglucosides took place with m-chloroperoxybenzoic acid (m-CPBA). NaOMe in dry methanol was used for the deacetylation of the blocked derivatives, to give the 3,4-epoxybutyl (1-->3)-beta-D-oligoglucoside mixture in an overall yield of 21%. The sample was analyzed by positive-ion electrospray ionization mass spectrometry (ESIMS). In a 3,4-epoxybutyl (1-->3)-beta-D-oligoglucoside-binding (1-->3)-beta-D-glucanase assay, we found that the (1-->3)-beta-D-glucanase was obviously inactivated by the 3,4-epoxybutyl (1-->3)-beta-D-oligoglucosides. At the same time, we found the 3,4-epoxybutyl (1-->3)-beta-D-oligoglucoside mixture was more active as compared to the underivatized oligoglucoside mixture in eliciting phytoalexin accumulation in tobacco cotyledon tissue. Furthermore, it could be kept for a longer time than a (1-->3)-beta-D-oligoglucoside mixture, which indicated it is much more stable than (1-->3)-beta-D-oligoglucosides.