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The oligosaccharide products from the successive reactions were analyzed by normal-phase high performance liquid chromatography, mass spectrometry and nuclear magnetic resonance. The effect of difucosylation on sensitivity to glycosidase digestion was also investigated.RESULTS Both FUT8 and NodZ are able to further fucosylate the monofucosylated chitooligosaccharides. Structural analyses of the resulting oligosaccharides showed that the reducing terminal GlcNAc residue and the third GlcNAc residue from the non-reducing end are fucosylated via α1,6-linkages. The difucosylation protected the oligosaccharides from extensive degradation to GlcNAc by hexosamidase and lysozyme, and also even from defucosylation by fucosidase.CONCLUSIONS The sequential actions of FUT8 and NodZ on common substrates effectively produce site-specific-difucosylated chitooligosaccharides.<br /><br /> This modification confers protection to the oligosaccharides against various GENERAL SIGNIFICANCE The action of a combination of eukaryotic and bacterial α1,6-fucosyltransferases on chitooligosaccharides results in the formation of difucosylated products, which serves to stabilize chitooligosaccharides against Studies on the effect of inflammation on the synthesis of glucosylated intermediates of glycoprotein biosynthesis in rat liver microsomes.Glycoprotein biosynthesis is substantially increased in liver during experimental inflammation. This study describes the effect of inflammation on the incorporation of labelled Glc from UDP-Glc into glucosylated lipid-linked intermediates of glycoprotein biosynthesis in liver microsomes. Maximum incorporation of Glc into lipid sugar and lipid oligosaccharide fractions occurred using microsome fractions from rats suffering from inflammation for 48-72 hr. Incorporation of Glc into lipid-sugar fractions was increased about three-fold over controls and incorporation into lipid-oligosaccharide fractions was increased about four-fold compared to controls. Maximum incorporation of Glc into lipid-sugar and lipid-oligosaccharide occurred at pH 6 and pH 5, 5627. Preparation of oligosaccharides from human milk.<br /><br />In Nishihara S, Angata K, Aoki-Kinoshita KF, Hirabayashi J, editors. Glycoscience Protocols (GlycoPODv2) [Internet]. [http://allinno.com/product/healthcare/671.html 2'-fucosyllactose] (JP) Japan Consortium for Glycobiology and Glycotechnology; 21–.Labeling of oligosaccharides for quantitative mass spectrometry.Quantitative analysis of oligosaccharide mixtures and their derivatives by electrospray ionization mass spectrometry (ESI MS) is challenging, for example, due to different affinities of the analytes to alkali ions. To overcome this source of discrimination and to enhance signal intensity, labeling studies with cellobiose as model compound were performed with the goal to develop a rapid, easy, and robust method. Hydrazone formation with the permanently charged Girard's T reagent as well as reductive amination with five different charge providing amines were studied under various conditions.<br /><br /> In both reaction types, the removal of water turned out to be the critical step because only under these conditions are the reactions pushed to completion. By working with only a slight excess of reagents, no purification is necessary to achieve excellent signalnoise ratios, avoiding further sources of discrimination. Comparing various reducing agents with respect to their selectivity and stability in the acidic reaction medium, 2-picoline borane turned out to be superior to the commonly used sodium cyanoborohydride. Thus, by replacement of the toxic NaCNBH(3) by the more selective and stable, non-toxic 2-picoline borane, complete reductive amination with low amounts of reagents and without unlabeled alditol formation was achieved with o-aminobenzoic acid, a useful reagent for ESI MS in negative mode. For MS in positive mode Girard's T derivatization was Elsevier Ltd. [https://en.wikipedia.org/wiki/2%27-Fucosyllactose 2'-fucosyllactose] reserved.Structural characterization of carbohydrate sequence, linkage, and branching in a quadrupole Ion trap mass spectrometer neutral oligosaccharides and N-linked Several oligosaccharide and N-linked glycan samples have been utilized to evaluate structural detail obtained with an ion trap mass spectrometer (ITMS).<br /><br /> Using multistage MSMS (MSn) in a commercial instrument, linear sequence, linkage, and branching have been studied, as well as the positional isomers within branched glycans.