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A new multi-enzyme system for a one-pot synthesis of sialyl oligosaccharides: Combined use of beta-galactosidase and alpha(26)-sialyltransferase coupled with regeneration in situ of CMP-sialic acid

An irreversible one-pot enzymatic synthesis of sialyl oligosaccharides has been achieved with a beta-galactosidase-catalyzed galactosylation of an acceptor followed by a sialyltransferase-catalyzed sialylation with regeneration in situ of CMP-sialic acid.

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Lectin-like protein fractions in lactic acid bacteria isolated from chickens

Agglutination of fifty bacterial strains isolated from gastric- intestinal tract of chickens with yeasts treated with glutaraldehyde and glycine was assayed to study possible adhesion mechanisms to epithelia. It was found that Lactobacillus, fermentum, Lactobacillus fermentum subsp. cellobiosus and Lactobacillus animalis glutinated yeasts massively and this agglutination was inhibited in each strain by different representative sugars: agglutination was inhibited in three selected strains with 0.2 M mannose. The three strains agglutinated chicken red blood cells, but did not agglutinate human red blood cells, whereas the agglutinating capacity for other animal erythrocytes was variable. The lectins were partially purified on a Sephadex G-75 column and affinity chromatography. The molecular weight of each of the protein subunits was estimated as about 50-67 kDa using electrophoretic techniques. After treatment with lipase and metaperiodate modifications bacterial superficies and/or liberation of some structures produced an unspecific interaction with increase in the yeast agglutination and hemagglutination. The optimal conditions of work were: pH 6-7, Ca2+ 1 mM, Mg2+ 2 mM and 37-42 C.

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Metal chelation by the common 2-amino-2-deoxy-, 2-N-acetylamino-2-deoxy-, and 2-deoxy-hexoses

The chelating properties of the common aldohexoses d-glucose, d-mannose, and d-galactose are characteristically modified in 2-substituted derivatives. The 2-amino-2-deoxy-aldohexoses provide mono- and bis-metallisable anionic ligands after their reaction with metal probes of the PdIIN 2 type (N2 = bidentate nitrogen ligand). The 2-amino function reliably participates in metal binding of the, mostly pyranoidic, carbohydrate chelators. Acetylation of the amino function yields the biologically important 2-N-acetylamino-2-deoxy-hexoses (GlcNAc, ManNAc, and GalNAc). On reaction with the palladium probe, the metal-binding properties of the deprotonated acetylamino function depends on the steric requirements introduced by the acetyl residue which is forced into a coplanar arrangement with the chelate ring. In the two 2-deoxy-aldohexoses, 2-deoxy-arabino-d-hexose (the 2-deoxy derivative of both d-glucose and d-mannose, ‘2-deoxy-glucose’) and 2-deoxy-lyxo-d-hexose (‘2-deoxy galactose’), the 2-position cannot contribute to metal binding. As a result, furanose-1,3 chelation becomes an important metal-binding mode. Due to the decreased acidity of the 2-deoxy-glycose’s 1-hydroxy function, monometallation also takes place at the pyranose’s 3,4-site.

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Efficient preparation of three building blocks for the synthesis of heparan sulfate fragments: Towards the combinatorial synthesis of oligosaccharides from hypervariable regions

New, multigram routes to suitably protected L-iduronyl monosaccharide donors 4 and 5 and 2-azidoglucose acceptors 6 and 7 are described. The L-iduronyl and D-glucuronyl disaccharides 1-3 were then prepared from these compounds, by means of efficient and regioselective protective group manipulations. These disaccharides form the basis of a combinatorial approach toward the synthesis of heparan sulfate fragments representative of the heterogeneous regions of this polysaccharide. Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)Introduction.

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Temperature dependencies of amide1H- and15N-chemical shifts in hyaluronan oligosaccharides

Temperature coefficients (Deltadelta/DeltaT) of amide chemical shifts of N-acetylglucosamine residues have been measured in a range of oligosaccharides of the important vertebrate polysaccharide hyaluronan. Odd- and even-numbered oligosaccharides with glucuronic acid, A-4,5-unsaturated glucuronic acid and N-acetylglucosamine at the termini were investigated. All amide proton temperature coefficients were only slightly less negative (-6.9 to -9.1 ppb/C) than those of amide protons in free exchange with water (?-11 ppb/C), indicating an absence of persistent intramolecular hydrogen bonds. With the exception of amide groups in reducing-terminal N-acetylglucosamine rings, all amide proton environments have the same temperature coefficient (-6.9 ppb/C), irrespective of differences in amide group chemical shifts and 3JHH coupling constants, i.e. they do not sense subtle differences in orientation of the amide group. Amide nitrogen temperature coefficients report the same phenomena but with greater sensitivity. These data provide a set of reference values for temperature coefficients measured in other carbohydrates with acetamido sugars. Copyright

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High-performance liquid chromatographic determination of chitin in the snow crab, Chionoecetes opilio

A HPLC method was developed for the determination of glucosamine in crab shells. The amount of glucosamine residue can be used to estimate the amount of chitin in analyzed material. Crab samples were hydrolysed with 6 M HCl at 100 C for 13 h and the released glucosamine was determined by HPLC using refractive index (RI) detection. Separations were performed using an ODS-2 column with an isocratic mobile phase consisting of 10% methanol, 90% hexanesulfonate, pH 2.1. Glucosamine identification was carried out not only on basis of retention time, but also with respect to mass spectra, which were acquired by HPLC-MS.

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ARYNIC CONDENSATION OF NITRILE ANIONS IN THE PRESENCE OF THE COMPLEX BASE NaNH2-CH3CH2(OCH2CH2)2ONa

Nitrile anions are easily condensed with aryl halides in the presence of Complex Base NaNH2-CH3CH2(OCH2CH2)2ONa via the intermediate arynes.

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Palladium-catalyzed homocoupling of aryl halides in the presence of fluoride

This report describes the fluoride-mediated homocoupling of aryl iodides and bromides catalyzed by palladium(0). This coupling protocol is tolerant of electron-donating and electron-withdrawing substitutents on the aryl halide, as well as ortho substitution. Optimum reaction conditions entail 10 mol% Pd(dba)2, 3 equiv of tetrabutyl ammonium fluoride (TBAF) in DMF at 90C.

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Open versus Closed Polyaromatic Nanocavity: Enhanced Host Abilities toward Large Dyes and Pigments

Host functions of polyaromatic nanocavities were revealed by using an M2L4 molecular cage and capsule. On the basis of the previously reported M2L4 capsule with a closed polyaromatic cavity, a new M2L4 cage (as a mixture of the isomers) was prepared by the quantitative assembly of two metal ions and four desymmetrized bispyridine ligands with a single polyaromatic panel. The obtained, open nanocavity of the cage exhibited enhanced binding abilities toward large dyes and pigments in water. For example, two molecules of coumarin dyes were bound in the nanocavity and showed strong whitish emission (up to PhiF=34 %). Furthermore, metallopigments, the sizes of which are larger than the inner cavities of the cage and capsule, were bound only in the open polyaromatic nanocavity of the cage to give water-soluble 1:1 host?guest complexes.

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Chemo-enzymatic synthesis of 1,4-oxazepanyl sugar as potent inhibitor of chitinase

N-Acetyl glucosamine 1 is selectively converted into 2 without protection of the other hydroxyl groups by allylation of the anomeric alkoxide in N,N-dimethylformamide containing lithium bromide. We use cell density cultures to produce the allylated derivative of penta-N-acetyl-chitopentaose by using 2 as the initial acceptor for the synthesis of 3 in vivo. Upon periodate oxidation, 3 is transferred to 4. Compound 4 is quickly subjected to sodium borohydride reduction and NH3 amination, which afforded the target compound 5. In 5-binding chitinase assay, it indicates that the chitinase is obviously inactivated by 5 with IC50 = 4.7 mumol/L.

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