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Synthetic, multivalent, carbohydrato assemblies are important tools in studying the avidity of many naturally occuring lectins for their ligands. This report details a simple, high-yielding three-step procedure to convert unprotected carbohydrates into N-allylglycosides. This method compliments the reductive amination procedure but allows the reducing-end pyranose ring to remain intact. No carbohydrate protecting groups are needed, and the resulting N-allylglycosylamide can be easily linked to other molecules. Two examples of analogs of silyl Lewis(x) and sulfo Lewis(X) have been derivatized by this process.

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We studied the stereoselective synthesis of several beta-N-aryl-glycosides by glycosylation of aromatic primary amines using unprotected carbohydrates in aqueous solution. This was the first report showing an efficient method for the synthesis with one step of beta-N-glycosyl-para-amino-phenyl alanine building blocks for the tumor-associated carbohydrate antigen (TACA) glycopeptides synthesis. Analysis of products by 1H and 13C NMR indicated that the Amadori rearrangement had not occurred after formation of the stereoselective beta-N-glycoside bond (natural N-glycoprotein linkage). The study of the chemical and enzymatic stability in aqueous media of beta-N-aryl-glycosides synthesized was also investigated. For the first time we have shown that the N-glycosidic bond was relatively stable at pH near to 7 and more stable than the O-glycosidic bond to enzymatic hydrolysis. This higher enzymatic and chemical stability of the N-glycosidic bond is essential to envisage further development of stable TACA building blocks.

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Moringa oleifera seed cake, which is the coproduct of oil extraction, may be a source of lectin. This study reports the purification of a lectin from M. oleifera cake (WSMoLC) and the determination of its effect on larvae, eggs and oviposition of Aedes aegypti. WSMoLC was isolated (purification fold: 3573) by treatment of water extract with ammonium sulphate (60%) and chromatography of lectin-rich fraction on chitin column. In gel trypsin, digestion followed by mass spectrometry revealed the similarity of WSMoLC peptides with other M. oleifera seed proteins. WSMoLC exerted deleterious effects on larvae (LC50: 0.89 mg/mL) and eggs (EC50: 0.14 mg/mL) and served as an oviposition-stimulant at the concentration of 0.1 mg/mL. Larvicidal activity may be linked to lectin resistance to digestion and stimulatory effect on the activities of protease and alpha-amylase at larvae gut. Furthermore, the larvae did not start to express different enzymes in response to lectin effect. This study adds value to M. oleifera seed cake as a source of insecticidal lectin with high potential to be used in control of A. aegypti because, at the same time that it stimulates the oviposition by females, it could kill the eggs or larvae that may arise from them.

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Isolation and structure elucidation of a new moenomycin antibiotic (C1, 1e) that lacks the branching methyl group in the 4-position of unit F are reported.The smallest antibiotically active degradation product of 1e is the trisaccharide derivative 3.This observation is in contrast to structure activity relations in the moenomycin A series where it was found that disaccharide 4a is fully active.

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Using a crude beta-N-acetylhexosaminidase from Aspergillus oryzae both tri-N-acetylchitotriose (GlcNAc)3 (1,n=1) and tetra-N-acetylchitotetraose (GlcNAc4) (1,n=2) act respectively as both glycosyl donor and glycosyl acceptor to give product mixtures containing significant quantities of the corresponding penta- and hexasaccharides <1 (n=3) and 1 (n=4), respectively> which are readily isolated and purified by charcoal-Celite chromatography.

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N(OMe)-linked disaccharide analogues, isosteric to the corresponding natural disaccharides, have been synthesized by chemoselective assembly of unprotected natural monosaccharides with methyl 6-deoxy-6-methoxyamino-alpha -D-glucopyranoside in an aqueous environment. The coupling reactions were found to be chemo- and stereoselective affording beta-(1?6) disaccharide mimics when using Glc and GlcNAc; in the case of Gal, the beta-anomer was prevalent (beta:alpha = 7:1). An iterative method for the synthesis of linear N(OMe) oligosaccharide analogues was demonstrated, based on the use of an unprotected monosaccharide building block in which an oxime functionality at C-6 is converted during the synthesis into the corresponding methoxyamino group. The conformational analysis of these compounds was carried out by using NMR spectroscopy, ab initio, molecular mechanics, and molecular dynamics methods. Optimized geometries and energies of fourteen conformers for each compound have been calculated at the B3LYP/631G* level. Predicted conformational equilibria were compared with the results based on NMR experiments and good agreement was found. It appears that N(OMe)-linked disaccharide analogues exhibit a slightly different conformational behavior to their parent natural disaccharides.

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Glycosyl dithiocarbamates (GDTCs) have been synthesized directly from the corresponding unprotected sugars and dithiocarbamate salts in good yields by using 2-chloro-1,3-dimethylimidazolinium chloride (DMC) as condensing agent in aqueous media. The three-component one-pot synthesis of GDTCs starting from unprotected sugars, carbon disulfide, and secondary amines has also been successfully demonstrated. This is the first report on the direct synthesis of GDTCs from unprotected sugars without using any protecting groups.

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A lectin was purified from the cotyledons of Luetzelburgia auriculata (Fr. All) Ducke by affinity chromatography on agarose-N-acetyl-D-galactosamine. The lectin is a potent agglutinin for rabbit erythrocytes, reacts with human red cells, but is inactive against cow, sheep, and goat erythrocytes. Hemagglutination of rabbit erythrocytes was inhibited by either 0.39 mM N-acetyl-neuraminic acid or N-acetyl-D-galactosamin, 12.5 mM D-lactose or D-melibiose, 50 mM D-galactose or raffinose. Its hemagglutinating activity was lost at 80C, 5 min, and the activation energy required for denaturation was 104,75 kJ mol-1. Chromatography on Sephadex G-100, at pH 7.6, showed that at this hydrogenic ionic concentration the native lectin was a homotetramer (123.5 kDa). By denaturing SDS-PAGE, LAA seemed to be composed of a mixture of 29 and 15 kDa polypeptide subunits. At acidic and basic pHs it assumed different conformations, as demonstrated by exclusion chromatography on Superdex 200 HR 10/30. The N-terminal sequence of the 29 kDa band was SEVVSFSFTKFNPNQKDII and the 15 kDa band contained a mixture of SEVVSFSFTKFNPNQKDII and KFNQIVAVEEDTDXESQPQ sequences, indicating that these bands may represent full-length and its endogenous fragments, respectively. The lectin is a glycoprotein having 3.2% neutral carbohydrate, with a pI of 5.8, containing high levels of Asp + Asn and Glu + Gln and hydroxy amino acids, and low amount or absence of sulfur amino acids, Its absorption spectrum showed a maximum at 280 nm and a epsilon1cm1% of 5.2. Its CD spectrum was characterized by minima near 228 nm, maxima near 196 nm and a negative to positive crossover at 210 nm. The secondary structure content was 6% alpha-helix, 8% parallel beta-sheet, 38% antiparallel beta-sheet, 17% beta-turn, 31% unordered and others contribution, and 1% RMS (root mean square). In the fluorescence spectroscopy, excitation of the lectin solution at 280 nm gave an emission spectrum in the 285-445 nm range. The wavelength maximum emission was in 334.5 nm, typical for tryptophan residues buried inside the protein.

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Novel heterobranched cyclodextrins (CDs), N-acetylglucosaminyl-cyclodextrins (GlcNAc-CD), were synthesized from a mixture of GlcNAc and alpha, beta, or gamma CD by the reverse reaction of N-acetylhexosaminidase from jack bean. Optimum pH and temperature for the production of GlcNAc-alphaCD by N-acetylhexosaminidase were pH 4.9 and 50-70C, respectively. The maximum yield of GlcNAc-alphaCD was 17.5% (mol/mol) at the concentration of 1 M GlcNAc and 0.25 M alphaCD. The reverse reaction product, GlcNAc-alphaCD, was separated into two peaks by HPLC analysis on the ODS column. Their structures were identified as 6-O-beta-D-N-acetylglucosaminyl-alphaCD and 2-O-beta-D-N-acetylglucosaminyl-alphaCD by FAB-MS and NMR spectroscopies. N-Acetylhexosaminidase from jack bean also synthesized N-acetylgalactosaminyl-alphaCD from N-acetylgalactosamine and alphaCD.

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Galactokinases (GalKs) have attracted significant research attention for their potential applications in the enzymatic synthesis of unique sugar phosphates. The galactokinase (GalKSpe4) cloned from Streptococcus pneumoniae TIGR4 presents a remarkably broad substrate range including 14 diverse natural and unnatural sugars. TLC and MS studies revealed that GalKSpe4 had relaxed activity towards galactose derivatives with modifications on the C-6, 4- or 2-positions. Additionally, GalKSpe4 can also tolerate glucose while glucose derivatives with modifications on the C-6, 4- or 2-positions were unacceptable. More interestingly, GalKSpe4 can phosphorylate l-mannose in moderate yield (43%), while other l-sugars such as l-Gal cannot be recognized by this enzyme. These results are very significant because there is rarely enzyme reported that can phosphorylate such uncommon substrates as l-mannose.

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