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Purification and Characterization of &beta-D-Galactosidase and &alpha-D-Mannosidase from Papaya (Carica papaya) Seeds

beta-D-Galactosidase was purified 115-fold from a saline extract of papaya seeds by fractionation with ammonium sulfate, DEAE-Sephadex chromatography and gel-filtration on Sephadex G-75, G-150, and G-100.The purified beta-D-galactosidase (MW, 56,000 daltons) had an isoelectric point (pI) at pH 8,4 and the optimal pH for its activity was 3.5 to 4.5.The enzyme activity was inhibited by Cu(2+), Ag(1+), Hg(2+), Pb(2+), NaAsO2 and p-chloromercuribenzoate at concentrations of 1E-3 M.Among the various mono- and oligosaccharides tested, D-galactose, D-galacturonic acid, D-galactono-gamma-lactone and melibiose significantly inhibited the enzyme activities at concentrations of 2E-3 to 1E-2 M.The purified enzyme hydrolyzed p-nitrophenyl beta-D-galactoside (Km = 1.0E-3 M), methyl beta-D-galactoside (Km = 1.6E-2 M), aminoethyl beta-D-galactoside (Km = 3.3E-2 M) and lactose (Km = 9.1E-2 M). beta-(1–>3)-Linked galactotetraosyl-erythritol and asialo-glycopeptide isolated from fetuin were also hydrolyzed to the extent of 78 and 75percent, respectively, on the basis of their galactose contents. alpha-D-Mannosidase from papaya seeds was also purified 130-fold by ammonium sulfate fractionation, DEAE-Sephdex chromatography, gel-filtration on Sephadex G-150 and hydroxylapatite chromatography.The purified enzyme (MW, 156,000 daltons), consisting of two subunits (78,000 x 2), was inhibited by Hg(2+), Ag(1+), Cu(2+), p-chloromercuribenzoate, D-glucose, D-glucosamine and D-mannose at concentration of 1E-3 to 1E-2 M.The alpha-D-mannosidase hydrolyzed p-nitrophenyl alpha-D-mannoside (Km = 5.6E-3 M), methyl alpha-D-mannoside (Km = 2.8E-2 M), alpha-D-mannosyl-D-mannitol (Km = 2.2E-2 M), alpha (1–>2)-linked D-mannobiosyl-D-mannitol (Km = 6.3E-3 M) and D-mannotriosyl-D-mannitol (Km = 5.3E-3 M).

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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One-step synthesis of beta-C-glycosidic ketones in aqueous media: The case of 2-acetamido sugars

The one step synthesis of beta-D-C-glycosidic ketones by condensation of pentane-2,4-dione with unprotected N-acetyl-D-gluco-, manno-, and galactosamine in alkaline aqueous media has been explored. N-Acetyl-D-gluco- and mannosamine gave, in good yield, a mixture of the two gluco and manno C-glycosidic ketones, which were separated by crystallization after acetylation, whereas N-acetyl-D-galactosamine afforded the galacto C-glycosidic ketone which was isolated as its acetylated derivative in 50% yield.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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The isoforms of rat natural killer cell receptor NKR-P1 display a distinct binding of complex saccharide ligands

Lectin-like receptors of natural killer cells are critical for the regulation of their effector function. When these receptors are crosslinked with antibodies or multivalent ligands, they transmit either activating or inhibitory signals. Here we describe binding and inhibition experiments with recombinant extracellular ligand-binding domains of the rat NKR-P1A (activating) and NKR-P1B (inhibitory) receptors. We did not observe any difference in the binding of monosaccharide ligands by these receptors, but revealed dramatic differences in the binding of oligosaccharides and glycodendrimers. Our results explain the immunostimulatory effects of the glycodendrimers, which bind exclusively to the activating NKR-P1A isoform.

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Reference£º
Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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A highly selective route to beta-C-glycosides via nonselective samarium iodide induced coupling reactions.

Stereoselective preparation of beta-C-glycosides has been developed from acetylated glycopyranosyl 2-pyridyl sulfones, involving a samarium-Barbier coupling procedureoxidation-isomerization sequence.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Core-sheath nanostructured chitosan-based nonwovens as a potential drug delivery system for periodontitis treatment

Core-sheath nanofibers were successfully prepared via coaxial electrospinning by using chitosan with well-defined structural characteristics as the shell layer and poly (vinyl alcohol) (PVA) containing tetracycline hydrochloride (TH) as the core layer. The effects of the average degree of deacetylation (DD?) of chitosan and the post-electrospinning genipin crosslinking on physicochemical and biological properties of resulting nonwovens were evaluated. Defect-free and geometrically uniform nanofibers with diameters predominantly in the range of 100?300 nm were prepared, and transmission electron microscopy (TEM) revealed the core-sheath structures and its preservation after crosslinking. The mechanical properties, as well as the stability of nonwovens in aqueous medium, were greatly improved by genipin-crosslinking, which enabled a sustained release of TH over 14 days. Results also revealed that the release profile of TH in the presence of lysozyme was affected by the composition of the shell layer, as the TH release rate increases with decreasing of DD?. Further in vitro antimicrobial activity demonstrated that the cross-linked nonwovens containing TH showed strong activity against bacterial strains associated with periodontal disease. Additionally, the nonwovens did not demonstrate cytotoxic toward fibroblast (HDFn) cells, hence showing their potential for applications as a novel drug delivery platform for periodontitis treatment.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Combining Click Reactions for the One-Pot Synthesis of Modular Biomolecule Mimetics

Here, we report on the first combined one-pot use of the two so-called “click reactions”: The thiol-ene coupling and the copper-catalyzed alkyne-azide cycloaddition. These reactions were employed in an alternating and one-pot fashion to combine appropriately functionalized monomeric carbohydrate building blocks to create mimics of trisaccharides and tetrasaccharides as single anomers, with only minimal purification necessary. The deprotected oligosaccharide mimics were found to bind both plant lectins and human galectin-3.

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Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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Polarized light-stimulated enzymatic hydrolysis of chitin and chitosan

Illumination with white linearly polarized light (WLPL) stimulated chitinase and chitosanase in their degradation of chitin and chitosan, respectively. Enzymes were illuminated at room temperature in separate vessels, then admixed in reactors containing polysaccharides. Hydrolysis of chitosan to glucosamine followed first order kinetics whereas hydrolysis of chitin to N-acetylglucosamine deviated from the first order kinetics. In both cases, an increase in the rate of hydrolysis depended on the illumination time. Efficient degradation required up to 60 min exposure of the enzyme to WLPL.

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Reference£º
Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics

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A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline

Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. To construct homogeneous glycoconjugates from natural resource of homogeneous glycans is therefore a practical approach to solve this problem. We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates.

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Tetrahydropyran – Wikipedia,
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Datura stramonium agglutinin: Cloning, molecular characterization and recombinant production in Arabidopsis thaliana

Datura stramonium seeds contain at least three chitin-binding isolectins [termed Datura stramonium agglutinin (DSA)] as homo- or heterodimers of A and B subunits. We isolated a cDNA encoding isolectin B (DSA-B) from an immature fruit cDNA library; this contained an open reading frame encoding 279 deduced amino acids, which was confirmed by partial sequencing of the native DSA-B peptide. The sequence consisted of: (i) a cysteine (Cys)-rich carbohydrate-binding domain composed of four conserved chitin-binding domains and (ii) an extensin-like domain of 37 residues containing four SerPro4-6 motifs that was inserted between the second and third chitin-binding domains (CBDs). Although each chitin-binding domain contained eight conserved Cys residues, only the second chitin-binding domain contained an extra Cys residue, which may participate in dimerization through inter-disulfide bridge formation. Using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry, the molecular mass of homodimeric lectin composed of two B-subunits was determined as 68,821 Da. The molecular mass of the S-pyridilethylated B-subunit were found to be 37,748 Da and that of the de-glycosylated form was 26,491 Da, which correlated with the molecular weight estimated from the deduced sequence. Transgenic Arabidopsis plants overexpressing the dsa-b demonstrated hemagglutinating activity. Recombinant DSA-B was produced as a homodimeric glycoprotein with a similar molecular mass to that of the native form. Moreover, the N-terminus of the purified recombinant DSA-B protein was identical to that of the native DSA-B, confirming that the cloned cDNA encoded DSA-B.

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Tetrahydropyran – Wikipedia,
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Amphiceramide A and B, novel glycosphingolipids from the marine sponge Amphimedon compressed

Glycolipid analysis of the Caribbean sponge Amphimedon compressa has shown it to contain two novel glycosphingoli- pids, amphiceramide A (1a) andB(2a), which possess an unusual A6-phytosphingosine. The saccharide chain of amphiceramide A is composed of a B-glucose residue glycosylated at the 6-position by an N-acetyl-B-glucosamine and has never been found before in a natural product. The saccharide chain of amphiceramide B consists of an allolactose [Gal(1p 6)Glc] residue p-linked to the ceramide and is found here for the first time in a natural glycosphingolipid. In addition, the sponge contains a new molecular species, acetamidoglucosyl ceramide (3a), and the known glucosyl ceramide 4a (halicerebroside A) and melibiosyl ceramide 5a (amphimelibioside C).

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Reference£º
Tetrahydropyran – Wikipedia,
Tetrahydropyran – an overview | ScienceDirect Topics