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Biotechnology Journal Dec 2010Disaccharide phosphorylases are glycosyltransferases (EC 2.4.1.α) of specialized carbohydrate metabolism in microorganisms. They catalyze glycosyl transfer to phosphate... (Review)
Review
Disaccharide phosphorylases are glycosyltransferases (EC 2.4.1.α) of specialized carbohydrate metabolism in microorganisms. They catalyze glycosyl transfer to phosphate using a disaccharide as donor substrate. Phosphorylases for the conversion of naturally abundant disaccharides including sucrose, maltose, α,α-trehalose, cellobiose, chitobiose, and laminaribiose have been described. Structurally, these disaccharide phosphorylases are often closely related to glycoside hydrolases and transglycosidases. Mechanistically, they are categorized according the stereochemical course of the reaction catalyzed, whereby the anomeric configuration of the disaccharide donor substrate may be retained or inverted in the sugar 1-phosphate product. Glycosyl transfer with inversion is thought to occur through a single displacement-like catalytic mechanism, exemplified by the reaction coordinate of cellobiose/chitobiose phosphorylase. Reaction via configurational retention takes place through the double displacement-like mechanism employed by sucrose phosphorylase. Retaining α,α-trehalose phosphorylase (from fungi) utilizes a different catalytic strategy, perhaps best described by a direct displacement mechanism, to achieve stereochemical control in an overall retentive transformation. Disaccharide phosphorylases have recently attracted renewed interest as catalysts for synthesis of glycosides to be applied as food additives and cosmetic ingredients. Relevant examples are lacto-N-biose and glucosylglycerol whose enzymatic production was achieved on multikilogram scale. Protein engineering of phosphorylases is currently pursued in different laboratories with the aim of broadening the donor and acceptor substrate specificities of naturally existing enzyme forms, to eventually generate a toolbox of new catalysts for glycoside synthesis.
Topics: Biotechnology; Disaccharides; Glycosyltransferases; Models, Molecular
PubMed: 21154671
DOI: 10.1002/biot.201000217 -
Food Chemistry Apr 2021Disaccharides are sugars composed of two monosaccharides joined by a glycosidic linkage. The specific properties of a disaccharide depend on the type of the glycosidic...
Disaccharides are sugars composed of two monosaccharides joined by a glycosidic linkage. The specific properties of a disaccharide depend on the type of the glycosidic linkage and the identity of the two component monosaccharides. In this work, seven disaccharide isomers (gentiobiose, isomaltose, melibiose, lactose, maltose, cellobiose, and sucrose) were analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) using a graphene oxide matrix. Each disaccharide was identified by its unique cleavage pattern. To determine the feasibility of quantitative analyses based on specific fragment patterns, mixtures of sucrose with cellobiose or maltose were prepared at different ratios and analyzed by MALDI-MS, where a strong linear correlation was observed between the relative peak intensity of the sucrose fragment peak at m/z 185 and the amount of sucrose in the mixture. The calibration curve was successfully applied to obtain the relative amount of maltose and sucrose in four different honey samples.
Topics: Disaccharides; Graphite; Honey; Isomerism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 33071193
DOI: 10.1016/j.foodchem.2020.128356 -
Analytica Chimica Acta May 2022Glycans are ubiquitous, structurally diverse molecules that have specific and general roles involving metabolism, structure, and cell-to-cell signaling. Functional...
Glycans are ubiquitous, structurally diverse molecules that have specific and general roles involving metabolism, structure, and cell-to-cell signaling. Functional specificity depends strongly on the complexity of structures that polysaccharides can adopt based on their subunit composition, length, extent of branching, glycosidic bond connectivity and anomeric configuration. However, a rapid and comprehensive characterization of glycan isomers can be challenging owing to limitations associated with their separation. Here, ten composition, anomeric and connectivity disaccharide isomers were separated and detected using high-resolution differential ion mobility-mass spectrometry (DMS-MS, also known as FAIMS). Focus was primarily directed to compositional isomers corresponding to epimers that differ by the axial or equatorial position of a single hydroxyl group. DMS resolving power was enhanced 14-fold primarily by increasing the fraction of helium in the ion carrier gas and lowering the flow rate. At relatively high disaccharide concentrations, DMS-MS of each disaccharide resulted in complex and unique multi-peak spectra with up to ten fully and partially resolved peaks for β-1,4-mannobiose (Man-1,4β-Man), which can be attributed to the DMS separation and subsequent dissociation of ionic non-covalently bound oligomers into monomer ions. Each DMS spectrum has at least one differentiating peak that is not in the other spectra, indicating that DMS can be used to fully or partially resolve composition, configuration and connectivity isomers. At relatively low disaccharide concentrations, mixtures of disaccharide epimers can also be readily separated by DMS. The integration of high-resolution, ambient pressure DMS with complementary reduced-pressure ion mobility and MS-based glycomics and glycoproteomics workflows may be useful for improving the characterization of glycans and glycosylated biomolecules.
Topics: Disaccharides; Humans; Ions; Isomerism; Mass Spectrometry
PubMed: 35473855
DOI: 10.1016/j.aca.2022.339783 -
Organic & Biomolecular Chemistry May 2020The uptake and metabolism of the disaccharide trehalose by Mycobacterium tuberculosis is essential for the virulence of this pathogen. Here we describe the...
The uptake and metabolism of the disaccharide trehalose by Mycobacterium tuberculosis is essential for the virulence of this pathogen. Here we describe the chemoenzymatic synthesis of new azido-functionalised asymmetric trehalose probes that resist degradation by mycobacterial enzymes and are used to probe trehalose processing pathways in mycobacteria.
Topics: Carbohydrate Conformation; Disaccharides; Microscopy, Fluorescence; Mycobacterium tuberculosis; Trehalose
PubMed: 32350493
DOI: 10.1039/d0ob00253d -
Glycobiology May 2022Bifidobacterium pseudocatenulatum grows well in the early stages of cultivation in medium containing sucrose (Suc), whereas its growth in medium containing the analogue...
Growth of Bifidobacterium pseudocatenulatum in medium containing N-acetylsucrosamine: enzyme that induces the growth of this bacterium via degradation of this disaccharide.
Bifidobacterium pseudocatenulatum grows well in the early stages of cultivation in medium containing sucrose (Suc), whereas its growth in medium containing the analogue disaccharide N-acetylsucrosamine (SucNAc) tends to exhibit a considerable delay. To elucidate the cause of this phenomenon, we investigated the proliferation pattern of B. pseudocatenulatum in medium containing D-glucose (Glc) and SucNAc and identified the enzyme that degrades this disaccharide. We found that B. pseudocatenulatum initially proliferates by assimilating Glc, with subsequent growth based on SucNAc assimilation depending on production of the β-fructofuranosidase, which can hydrolyze SucNAc, after Glc is completely consumed. Thus, B. pseudocatenulatum exhibited a diauxic growth pattern in medium containing Glc and SucNAc. In contrast, when cultured in medium containing Glc and Suc, B. pseudocatenulatum initially grew by degrading Suc via the phosphorolysis activity of Suc phosphorylase, which did not react to SucNAc. These observations indicate that B. pseudocatenulatum proliferates by assimilating Suc and SucNAc via different pathways. The β-fructofuranosidase of B. pseudocatenulatum exhibited higher hydrolytic activity against several naturally occurring Suc-based tri- or tetrasaccharides than against Suc, suggesting that this enzyme actively catabolizes oligosaccharides other than Suc.
Topics: Bifidobacterium pseudocatenulatum; Disaccharides; Oligosaccharides; Sucrose; beta-Fructofuranosidase
PubMed: 35138388
DOI: 10.1093/glycob/cwac001 -
Organic & Biomolecular Chemistry May 2022Heparan sulfate (HS), a glycosaminoglycan related to heparin, is a linear polysaccharide, consisting of repeating disaccharide units. This compound is involved in...
Heparan sulfate (HS), a glycosaminoglycan related to heparin, is a linear polysaccharide, consisting of repeating disaccharide units. This compound is involved in multiple biological processes such as inflammation, coagulation, angiogenesis and viral infections. Our work focuses on the synthesis of simple HS analogs for the study of structure-activity relationships, with the aim of modulating these biological activities. Thioglycoside analogs, in which the interglycosidic oxygen is replaced by a sulfur atom, are very interesting compounds in terms of therapeutic applications. Indeed, the thioglycosidic bond leads to an improvement of their stability and can allow the inhibition of enzymes involved in physiological and pathological processes. In our previous work, we developed a synthetic sequence which led to a non-sulfated thiodisaccharide analog of HS. In this paper, we report our results of the development of a new synthetic method allowing access to the novel sulfated -disaccharide, as well as to their oxygenated analogues (-disaccharide and sulfated -disaccharide). These 4 compounds were also tested for the inhibition of heparanase, an enzyme involved in biological processes like tumor growth and inflammation. The obtained IC values in the micromolar range showed the impact of the interglycosidic sulfur atom and the 6-sulfate group.
Topics: Disaccharides; Glucuronidase; Heparitin Sulfate; Humans; Inflammation; Sulfur
PubMed: 35388870
DOI: 10.1039/d2ob00250g -
The Journal of the Arkansas Medical... Jan 1966
Topics: Carbohydrate Metabolism, Inborn Errors; Disaccharides; Humans; Infant; Infant Nutritional Physiological Phenomena; Infant, Newborn
PubMed: 4221401
DOI: No ID Found -
Analytical Chemistry Jan 2023Carbohydrates play critically important roles in energy supply and biological functions in living systems. However, it has been a great challenge to identify saccharides...
Carbohydrates play critically important roles in energy supply and biological functions in living systems. However, it has been a great challenge to identify saccharides and distinguish their isomers because they have highly similar structures and many possible positions for glycosidic linkages. In this work, an ambient ionization tandem mass spectrometry method was developed to characterize disaccharide structural isomers with in situ methylation. The direct analysis in real time ion source can be used to facilitate the methylation reaction of disaccharides with tetramethylammonium hydroxide. The hydroxyl groups of disaccharides can be methylated instantaneously, and the products can be ionized at the same time. The methylated product ions from full scan mass spectrometry (MS) and tandem MS can be used to distinguish a variety of disaccharide structural isomers with different glycosidic linkages, compositions, and configurations. Characteristic marker ions were discovered, and they can be used for the assignment of linkage type and identification of specific isomeric forms. The method was used for the direct identification of disaccharide isomers from real commercial products such as honey, wine, and milk without complex sample pretreatment or chromatographic separation.
Topics: Disaccharides; Tandem Mass Spectrometry; Methylation; Carbohydrates; Ions; Isomerism; Glycosides; Spectrometry, Mass, Electrospray Ionization
PubMed: 36635092
DOI: 10.1021/acs.analchem.2c03485 -
Molecules (Basel, Switzerland) Apr 2024Rare sugars are known for their ability to suppress postprandial blood glucose levels. Therefore, oligosaccharides and disaccharides derived from rare sugars could...
Rare sugars are known for their ability to suppress postprandial blood glucose levels. Therefore, oligosaccharides and disaccharides derived from rare sugars could potentially serve as functional sweeteners. A disaccharide [α-d-allopyranosyl-(1→2)-β-d-psicofuranoside] mimicking sucrose was synthesized from rare monosaccharides D-allose and D-psicose. Glycosylation using the intermolecular aglycon delivery (IAD) method was employed to selectively form 1,2- α-glycosidic linkages of the allopyranose residues. Moreover, β-selective psicofuranosylation was performed using a psicofuranosyl acceptor with 1,3,4,6-tetra--benzoyl groups. This is the first report on the synthesis of non-reducing disaccharides comprising only rare d-sugars by IAD using protected ketose as a unique acceptor; additionally, this approach is expected to be applicable to the synthesis of functional sweeteners.
Topics: Disaccharides; Sucrose; Glycosylation; Sweetening Agents; Fructose; Glucose
PubMed: 38675593
DOI: 10.3390/molecules29081771 -
Carbohydrate Research Jun 2020Bacterial glycosyltransferases are potential targets for the development of novel antibiotics and anti-virulence agents. We report a novel inhibitor design for the...
Bacterial glycosyltransferases are potential targets for the development of novel antibiotics and anti-virulence agents. We report a novel inhibitor design for the retaining α-1,4-galactosyltransferase LgtC from Neisseria meningitidis. Our design is based on the installation of an electrophilic warhead on the LgtC acceptor substrate and targeted at a non-catalytic cysteine residue in the LgtC active site. We have successfully synthesised two prototype inhibitors in four steps from lactulose. The key step in our synthesis is a Heyns rearrangement, during which we observed the formation of a hitherto unknown side product. While both lactosamine derivatives behaved as moderate inhibitors of LgtC, they also retained residual substrate activity. These results suggest that in contrast to our original design, these inhibitors do not act via a covalent mode of action, but are most likely non-covalent inhibitors.
Topics: Bacteria; Bacterial Proteins; Carbohydrate Conformation; Disaccharides; Drug Design; Galactosyltransferases; Molecular Docking Simulation
PubMed: 32402851
DOI: 10.1016/j.carres.2020.108017