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Preparation, conformation, and mild hydrolysis of 1-glycosyl-2- acetylhydrazines of the hexoses, pentoses, 2-acetamido-2-deoxyhexoses, and fucose

The title compounds were prepared and their conformations studied by 1H-NMR. Their acid hydrolysis under mild conditions was monitored by 1H- NMR.

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Selective pivaloylation of 2-acetamido-2-deoxy sugars.

Selective pivaloylation of 2-acetamido-2-deoxy-D-glucose, its methyl alpha- and beta-glycosides, and the methyl alpha-glycoside of N-acetyl-D-muramic acid under various conditions has been studied. The structures of the products were established by 1H-n.m.r. spectroscopy and acetylation. The orders of acylation, HO-6 greater than HO-3 greater than HO-1 greater than HO-4 for 2-acetamido-2-deoxy-D-glucose and HO-6 greater than HO-3 greater than HO-4 for its methyl glycosides, were established. Methyl 2-acetamido-2-deoxy-3,6-di-O-pivaloyl-alpha- and -beta-D-glucopyranosides and 2-acetamido-2-deoxy-1,3,4,6-tetra-O-pivaloyl-D-glucopyranose were hydrolysed by rabbit serum esterases.

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Formation of LacNAc mimetics employing novel donor substrates for enzymatic beta1?4 galactosylation

The formation of LacNAc mimetics employing novel donor substrates for enzymatic beta1?4 galactosylation is discussed. In examining C-6 modified 4-nitrophenyl beta-D-galacto-pyranosides as donor structures the beta-galactosidase revealed a broad substrate specificity which allowed synthesis of various disaccharide components. The important role of carbohydrates in vital biological recognition processes has increasingly stimulated efforts in glycoconjugate research. Based on the findings, the primary alcohol function in galactosides does not seem to be crucial for recognition.

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A novel rhamnose-binding lectin family from eggs of steelhead trout (Oncorhynchus mykiss) with different structures and tissue distribution.

An L-rhamnose-binding isolectin named STL3 (subunit Mr, 21.5 k) was isolated from eggs of the steelhead trout (Oncorhynchus mykiss) in addition to STL1 (subunit Mr, 31.4 k) and STL2 (subunit Mr, 21.3 k) that had been already isolated. STLs were composed of noncovalently linked subunits. The primary structures of STL1 and STL3 were analyzed by the combined use of protein sequencing and cDNA sequencing. A cDNA encoding STL2, of which the protein sequence had been previously studied, was also analyzed. The STL1 subunit (289 amino acid residues) had different structural properties compared to those of the STL2 subunit (195 amino acid residues) and the STL3 subunit (195 amino acid residues); e.g., the number of repeated domain (three for STL1, and two for STL2 and STL3), although all of them were composed of tandemly repeated homologous domains (40 to 53% identities). The lectin levels in various tissues and during the embryonic development showed that STL1 had different distribution and expression profiles from those of STL2 and STL3. Although STL1 could be detected in several tissues and serum of both male and female steelhead trout, STL2 and STL3 were only abundant in the ovary. STL2 and STL3 levels dramatically decreased just after hatching, however, the STL1 level increased temporarily. These results indicate that the multiple lectins from eggs of the steelhead trout form a novel rhamnose-binding lectin family with different structures and tissue distribution to share distinct functions in eggs.

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Design, synthesis and biological evaluation of carbohydrate-functionalized cyclodextrins and liposomes for hepatocyte-specific targeting

Targeting glycan-binding receptors is an attractive strategy for cell-specific drug and gene delivery. The C-type lectin asialoglycoprotein receptor (ASGPR) is particularly suitable for liver-specific delivery due to its exclusive expression by parenchymal hepatocytes. In this study, we designed and developed an efficient synthesis of carbohydrate-functionalized beta-cyclodextrins (betaCDs) and liposomes for hepatocyte-specific delivery. For targeting of ASGPR, rhodamine B-loaded betaCDs were functionalized with glycodendrimers. Liposomes were equipped with synthetic glycolipids containing a terminal d-GalNAc residue to mediate binding to ASGPR. Uptake studies in the human hepatocellular carcinoma cell line HepG2 demonstrated that betaCDs and liposomes displaying terminal d-Gal/d-GalNAc residues were preferentially endocytosed. In contrast, uptake of betaCDs and liposomes with terminal d-Man or D-GlcNAc residues was markedly reduced. The d-Gal/d-GalNAc-functionalized betaCDs and liposomes presented here enable hepatocyte-specific targeting. Gal-functionalized betaCDs are efficient molecular carriers to deliver doxorubicin in vitro into hepatocytes and induce apoptosis.

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Kinetics of acid hydrolysis of acetylglucosamine

The kinetics of acid hydrolysis of N-acetylglucosamine at different temperatures and reagent concentrations was studied. A mathematical model of the hydrolysis was proposed. The rate constant and activation energy of deacetylation were calculated.

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Evaluation of Natural and Synthetic Phosphate Donors for the Improved Enzymatic Synthesis of Phosphate Monoesters

Undesired product hydrolysis along with large amounts of waste in form of inorganic monophosphate by-product are the main obstacles associated with the use of pyrophosphate in the phosphatase-catalyzed synthesis of phosphate monoesters on large scale. In order to overcome both limitations, we screened a broad range of natural and synthetic organic phosphate donors with several enzymes on a broad variety of hydroxyl-compounds. Among them, acetyl phosphate delivered stable product levels and high phospho-transfer efficiency at the lower functional pH-limit, which translated into excellent productivity. The protocol is generally applicable to acid phosphatases and compatible with a range of diverse substrates. Preparative-scale transformations using acetyl phosphate synthesized from cheap starting materials yielded multiple grams of various sugar phosphates with up to 433 g L?1 h?1 space-time yield and 75% reduction of barium phosphate waste. (Figure presented.).

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Efficient synthesis of globoside and isogloboside tetrasaccharides by using beta(1?3) N-acetylgalactosaminyltransferase/UDP-N-acetylglucosamine C4 epimerase fusion protein

The beta(1?3) N-acetylgalactosaminyltransferase/UDP-N-acetylglucosamine C4 epimerase fusion protein was constructed and used in coupled enzymatic reactions to synthesize a variety of globotetraose and isoglobotetraose derivatives from the corresponding lactoside acceptors.

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A SHORT BIOMIMETIC SYNTHESIS OF LEPTOSPHAERIN

(+)-Leptosphaerin, a metabolite of the marine Ascomycete Leptosphaeria oraemaris Linder, was synthesized in a standard six-step procedure from N-acetyl-D-glucosamine.

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Enzymic transfer of 6-modified D-galactosyl residues: Synthesis of biantennary penta- and hepta-saccharides having two 6-deoxy-D-galactose residues at the nonreducing end and evaluation of 6-deoxy-D-galactosyl transfer to glycoprotein using bovine beta-(1 4)-galactosyltransferase and UDP-6-deoxy-D-galactose

UDP-6-Deoxy-D-galactose and UDP-6-deoxy-6-fluoro-D-galactose were synthesized and their transfer to 2-acetamido-2-deoxy-D-glucose (N-acetyl-D-glucosamine) by beta-(1 ? 4)-galactosyltransferase was examined. The transfer rates of 6-deoxy-D-galactose and 6-deoxy-6-fluoro-D-galactose were 1.3 and 0.2% of that of D-galactosyl transfer, respectively. The 2-acetamido-4-O-(6-deoxy-beta-D-galactopyranosyl)-2-deoxy-D-glucopyranos e (6′-deoxy-N-acetyllactosamine) and methyl 2-acetamido-4-O-(6-deoxy-6-fluoro-beta-D-galactopyranosyl)-2-deoxy-D-glu copyranoside (6′-deoxy-6′-fluoro-N-acetyllactosamine) were synthesized enzymatically in 30 and 59% yields, respectively. Further, 6-deoxy-D-galactose could be completely transferred to N-linked type biantennary oligosaccharides having two N-acetyl-D-glucosaminyl residues at the nonreducing end to give the corresponding penta- and hepta-saccharides in 55 and 57% yields, respectively. An assay of 6-deoxy-D-galactosyl transfer using asialo agalacto alpha1-acid glycoprotein as an acceptor suggested that 6-deoxy-D-galactose was transferred to about 30% of the N-acetyl-D-glucosaminyl residues in the N-linked oligosaccharides of the glycoprotein.

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