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D-Allose and its derivatives play important roles in the field of health care and food nutrition. Pure and well-defined D-allose derivatives can facilitate the elucidation of their structure-activity relationship as an essential step for drug design. The Lattrell-Dax epimerization, refers to the triflate inversion using nitrite reagent, is known as valuable method for the synthesis of rare D-allose derivatives. Here, the influence of protecting group patterns on the transformation efficiency of D-glucose derivatives into synthetically useful D-alloses and D-allosamines via the Lattrell-Dax epimerization was studied. For C3 epimerization of D-glucose derivatives bearing O2-acyl group, an anomeric configuration-dependent acyl migration from O2 to O3 was found. In addition, a neighbouring group participation effect-mediated SN1 nucleophilic substitution of the D-glucosamine bearing C2 trichloroacetamido (TCA) group in the Lattrell-Dax epimerization was dependent upon anomeric configuration. Thus, the effect of anomeric configuration on the Lattrell-Dax epimerization of D-glucose suggests that β-D-glucosides with low steric hindrance at C2 should be better substrates for the synthesis of D-allose derivatives. Significantly, the efficient synthesis of the orthogonally protected D-allose 13 and D-allosamine 18 will serve well for further assembly of complex glycans.

The galectins are a family of β-galactoside-specific animal lectins, and have attracted much attention as novel regulators of the immune system. Galectin-10 is well-expressed in eosinophils, and spontaneously forms Charcot-Leyden crystals (CLCs), during prolonged eosinophilic inflammatory reactions, which are frequently observed in eosinophilic diseases. Although biochemical and structural characterizations of galectin-10 have been done, its biological role and molecular mechanism are still unclear, and few X-ray structures of galectin-10 in complex with monosaccharides/oligosaccharides have been reported. Here, X-ray structures of galectin-10 in complexes with seven monosaccharides are presented with biochemical analyses to detect interactions of galectin-10 with monosaccharides/oligosaccharides. Galectin-10 forms a homo-dimer in the face-to-face orientation, and the monosaccharides bind to the carbohydrate recognition site composed of amino acid residues from two galectin-10 molecules of dimers, suggesting that galectin-10 dimer likely captures the monosaccharides in solution and in vivo. d -Glucose, d -allose, d -arabinose, and D-N-acetylgalactosamine bind to the interfaces between galectin-10 dimers in crystals, and they affect the stability of molecular packing in crystals, leading to easy-dissolving of CLCs, and/or inhibiting the formation of CLCs. These monosaccharides may serve as effectors of G10 to form CLCs in vivo.

Ribose-5-phosphate isomerase B is of great importance for biocatalysis and biosynthesis, but the multifunctional residues in active sites hinder the research efforts. This study employed rational design strategies to locate the key residues of RpiB from Ochrobactrum sp. CSL1 (OsRpiB). A single-mutant S9T of a noncontact residue showed 80% activity improvement toward d-allose. A double-mutant S98H/S134H further increased the activity to 3.6-fold. The mutations were analyzed by kinetics and molecular dynamics analyses, indicating that S9T might enhance the substrate binding and catalysis by inducing a steric effect, and S98H/S134H could strengthen both ring opening and binding of d-allose. Though S98H/S134H showed low temperature stability, its potential was explored by isomerizing d-allose to d-psicose with higher conversion and in less reaction time. The findings of this study were beneficial for illustrating the complex functions of key residues in RpiBs and applying OsRpiB in preparing rare sugars.

The nematocidal activities of the fatty acid esters of d-allose were examined using the larvae of C. elegans. Among the fatty acid esters, 6-O-octanoyl-d-allose (3) showed significant activity. 6-O-octanoyl-d-glucose (5) showed no activity, indicating that the D-allose moiety is essential for the nematocidal activity of 3. A nonhydrolyzable alkoxy analog 6-O-octyl-d-allose (6) also showed activity equivalent to that of 3.

D-Allose (D-All), C-3 epimer of D-glucose, is a rare sugar known to suppress reactive oxygen species generation and prevent hypertension. We previously reported that D-allulose, a structural isomer of D-All, prolongs the lifespan of the nematode Caenorhabditis elegans. Thus, D-All was predicted to affect longevity. In this study, we provide the first empirical evidence that D-All extends the lifespan of C. elegans. Lifespan assays revealed that a lifespan extension was induced by 28 mM D-All. In particular, a lifespan extension of 23.8 % was achieved (p < 0.0001). We further revealed that the effects of D-All on lifespan were dependent on the insulin gene daf-16 and the longevity gene sir-2.1, indicating a distinct mechanism from those of other hexoses, such as D-allulose, with previously reported antiaging effects.

In this study, gum of Araucaria heterophylla was collected. The collected gum was subjected for extraction of polysaccharide using solvent extraction system. Thus, extracted polysaccharide was further purified using solvent method and was characterized using UV-Vis spectroscopy, Phenol sulfuric acid assay, FTIR, TGA, TLC and GC-MS. The gum derived polysaccharide was found to have the following sugars Rhamnose, Allose, Glucosinolate, Threose, Idosan, Galactose and Arabinose. The extracted polysaccharide was tested for various in-vitro bioactive studies such as antibacterial activity, antioxidant activity and anticancer activity. The polysaccharide was found to have antioxidant and anticancer activity. Further, the polysaccharide was subjected for carboxymethylation to favor the nanocarrier synthesis, where it was chelated using Sodium Tri Meta Phosphate (STMP) to form nanocarriers. The nanocarriers so formed were loaded with curcumin and were characterized using FTIR, SEM, EDX and AFM. Both the loaded and unloaded nanocarriers were studied for its in-vitro cytotoxic effect against the MCF7 human breast cancer cell lines. The nanocarriers were found to deliver the drug efficiently against the cancer cell line used in this study.

Oxazolidinones are an important class of heterocyclic compounds that are used as chiral auxiliaries in asymmetric synthesis and as biologically active pharmaceutical agents. Moreover, carbohydrates are ideal scaffolds to generate libraries of bioactive compounds due the presence of defined configuration. We report here asymmetric alkylation studies on N-derivatized glucose-and allose-based spirooxazolidinones which do act as chiral auxiliaries.

Rare sugars are regarded as functional biological materials due to their potential applications as low-calorie sweeteners, antioxidants, nucleoside analogs, and immunosuppressants. D-Allose is a rare sugar that has attracted substantial attention in recent years, owing to its pharmaceutical activities, but it is still not widely available. To address this limitation, we continuously produced D-allose from D-allulose using a packed bed reactor with commercial glucose isomerase (Sweetzyme IT). The optimal conditions for D-allose production were determined to be pH 8.0 and 60°C, with 500 g/L D-allulose as a substrate at a dilution rate of 0.24/h. Using these optimum conditions, the commercial glucose isomerase produced an average of 150 g/L D-allose over 20 days, with a productivity of 36 g/L/h and a conversion yield of 30%. This is the first report of the successful continuous production of D-allose from D-allulose by commercial glucose isomerase using a packed bed reactor, which can potentially provide a continuous production system for industrial applications of D-allose.

Rare sugars are monosaccharides with tremendous potential for applications in pharmaceutical, cosmetics, nutraceutical, and flavors industries. The four rare sugars, including; gulose, allose, altrose and talose are stereoisomers that are different in the hydroxyl group orientation (axial or equatorial) on the C2-4 atoms. The DFT, AIM, and, NBO calculations were used to probe the probability of formation of internal H-bonds in four rare sugars. The AIM analysis identified that altrose and talose can form three predominantly intramolecular H-bonds, whereas gulose and allose revealed one and two H-bonds, respectively and these normal intramolecular H-bonds are mostly closed-shell interactions. The theoretical calculated O-H stretching FT-IR vibrational frequencies confirmed that the intramolecular H-bonds shifted toward low frequencies in comparison to the free hydroxyl group, which caused the red-shift. Also, the lowest IR frequency in each sugar was related to the structure with the highest stabilization energy and the most strongest intramolecular H-bonds.

Methylotrophs utilizes cheap, abundant one-carbon compounds, offering a promising green, sustainable and economical alternative to current sugar-based biomanufacturing. However, natural one-carbon assimilation pathways come with many disadvantages, such as complicated reaction steps, the need for additional energy and/or reducing power, or loss of CO2, resulting in unsatisfactory biomanufacturing performance. Here, we predicted eight simple, novel and carbon-conserving formaldehyde (FALD) assimilation pathways based on the extended metabolic network with non-natural aldol reactions using the comb-flux balance analysis (FBA) algorithm. Three of these pathways were found to be independent of energy/reducing equivalents, and thus chosen for further experimental verification. Then, two novel aldol reactions, condensing D-erythrose 4-phosphate and glycolaldehyde (GALD) into 2R,3R-stereo allose 6-phosphate by DeoC or 2S,3R-stereo altrose 6-phosphate by TalBF178Y/Fsa, were identified for the first time. Finally, a novel FALD assimilation pathway proceeding via allose 6-phosphate, named as the glycolaldehyde-allose 6-phosphate assimilation (GAPA) pathway, was constructed in vitro with a high carbon yield of 94%. This work provides an elegant paradigm for systematic design of one-carbon assimilation pathways based on artificial aldolase (ALS) reactions, which could also be feasibly adapted for the mining of other metabolic pathways.

The concentrations effects of sugars (trehalose and allose) and oils (canola and coconut oil) on the characteristics of rice starch suspension and mechanical properties of rice starch film were studied. The samples were prepared using 3% (w/w) rice starch, with 10% or 30% (w/w) sugar (trehalose or allose) added and 10% or 30% (w/w) oil (canola or coconut). The droplet size of the film suspension increased with increasing oil concentration both in trehalose and allose, which blended with oil. The flow behavior of the film suspensions showed shear-thinning behavior as calculated by the Power Law model. The apparent viscosity tended to increase with the addition of sugar and oil. The breaking stress of the films blended with sugar and oil was less than that of control. On preparation day and after 7 days' storage, the breaking strain tended to increase more with the addition of coconut oil than with that of canola oil. However, breaking stress and breaking strain decreased after 28 days' storage. Adding sugar had correlation with mechanical properties whereas adding oil had correlation with film suspension characteristics, allowed the sugar and oil to interact and inhibited starch chain mobility due to concentration, sugar type, and oil type. PRACTICAL APPLICATION: Trehalose, allose, canola oil, and coconut oil could be used as a plasticizer in a starch edible/biodegradable film system. The preparation process of filmogenic solution was depended on the combination of sugar and oil that could change the flow behavior and affected the mechanical properties of the edible film. The sugar and oil might improve the mechanical properties of the film by a hydroxyl group of sugar and lubricating properties of the oil.

Two rarely phenolic acid-substituted alloses (1, 2) and one new glucoside (3), as well as nine known compounds (4-12) were isolated from rhizomes of Cibotium barometz (L.) J. Sm. Structures of 1-3 were established by extensively spectroscopic analyses (NMR, MS, etc.) and acid hydrolysis. All compounds were evaluated for the hepatoprotective activities against APAP-induced HepG2 cell damage. Compounds 1, 4-7, 10 exhibited significant hepatoprotective activities, even more strongly than positive control, bicycol. In addition, compounds 1 and 9 could reduce PC12 cell death induced by serum deprivation.

The mechanism of d-allose in reducing the allergenicity and digestibility of ultrasound-pretreated α-lactalbumin (α-LA) was studied. The intensity reduction and peak red shift occurred in fluorescence spectra of glycated samples. Enzyme-linked immunosorbent assay and basophil degranulation analysis showed that d-allose significantly reduced the allergenicity of α-LA, and ultrasound-pretreated α-LA showed the lowest allergenicity after glycation. Compared with α-LA, the degree of hydrolysis decreased in glycated samples. Size-exclusion high-performance liquid chromatography showed that the glycated α-LA was resistant to digestive enzymes. The glycated sites and average degree of substitution per peptide molecule were determined using LC Orbitrap MS/MS. These results suggested that the masking of linear allergenic epitopes by glycation could reduce the allergenicity. Therefore, the combination of ultrasound pretreatment and glycation is a potential method to reduce protein allergenicity in food processing and provides a useful approach for the application of rare sugars in food processing.

The replacement of hydroxyl groups by fluorine atoms on hexopyranose scaffolds may allow access to the discovery of new chemical entities possessing unique physical, chemical and ultimately even biological properties. The prospect of significant effects generated by such multiple and controlled substitutions encouraged us to develop diverse synthetic routes towards the stereoselective synthesis of polyfluorinated hexopyranoses, six of which are unprecedented. Hence, we report the synthesis of heavily fluorinated galactose, glucose, mannose, talose, allose, fucose, and galacturonic acid methyl ester using a Chiron approach from inexpensive levoglucosan. Structural analysis of single-crystal X-ray diffractions and NMR studies confirm the conservation of favored 4C1 conformation for fluorinated carbohydrate analogs, while a slightly distorted conformation due to repulsive 1,3-diaxial F···F interaction is observed for the trifluorinated talose derivative. Finally, the relative stereochemistry of multi-vicinal fluorine atoms has a strong effect on the lipophilicities (logP)., Polyfluorinated hexopyranoses display unique physical, chemical and biological properties, however their stereoselective synthesis is highly challenging. Here, the authors report a synthetic approach based on the chemical manipulation of inexpensive levoglucosan to obtain heavily fluorinated monosaccharides stereoselectively.

The mechanism of the thermal degradation and the toxicity of the thermal degradation products of agar were studied using TG/DTA, Fourier-transform infrared spectroscopy and pyrolysis gas chromatography/mass spectrometry. It was found that the thermal degradation of agar is a single-step reaction, the thermal degradation temperature (T0, Tp, Tf) increases with increasing gel strength (P) and the influence of P on the thermal degradation rate is modest. The thermal degradation of agar is an exothermic reaction, and the activation energy of the reaction increases with increasing P. In the thermal degradation, agar is first decomposed into 3,6-anhydropyran galactopyranose and galactopyranose, then 3,6-anhydropyran galactopyranose, and finally furyl hydroxymethyl ketone, through loop opening, dehydration and hydrogen transfer. Galactopyranose follows three degradation pathways, and its final degradation products are 3,4-atrosan, d -allose, furfural and 5-(hydroxymethyl)-2-furancarboxaldehyde. Of the degradation products, furyl hydroxymethyl ketone, furfural, and 5-(hydroxymethyl)-2-furancarboxaldehyde show some toxicity to humans.

1,2,6-Trideoxy-6-amido-d-allose derivative was synthesized and found to exhibit higher growth-inhibitory activity against plants than the corresponding deoxy-d-allose ester, which indicates that an amide group at C-6 of the deoxy-d-allose amide enhances inhibitory activity. In addition, the mode of action of the deoxy-d-allose amide was significantly different from that of d-allose which inhibits gibberellin signaling. Co-addition of gibberellin GA3 restored the growth of rice seedlings inhibited by the deoxy-d-allose amide, suggesting that it might inhibit biosynthesis of gibberellins in plants to induce growth inhibition.

D-Allose is a potential medical sugar because it has anticancer, antihypertensive, anti-inflammatory, antioxidative, and immunosuppressant activities. Allose production from fructose as a cheap substrate was performed by a one-pot reaction using Flavonifractor plautiiD-allulose 3-epimerase (FP-DAE) and Clostridium thermocellum ribose 5-phosphate isomerase (CT-RPI). The optimal reaction conditions for allose production were pH 7.5, 60°C, 0.1 g/l FP-DAE, 12 g/l CT-RPI, and 600 g/l fructose in the presence of 1 mM Co2+. Under these optimized conditions, FP-DAE and CT-RPI produced 79 g/l allose for 2 h, with a conversion yield of 13%. This is the first biotransformation of fructose to allose by a two-enzyme system. The production of allose by a one-pot reaction using FP-DAE and CT-RPI was 1.3-fold higher than that by a two-step reaction using the two enzymes.

A linear tetramer of β-(1 → 6)-linked 3-azido-3-deoxy- d -allose containing glycosyl donor and glycosyl acceptor functions in the terminal monosaccharide units was prepared starting from 3-azido-3-deoxy-1,2:5,6-di-O-isopropylidene-α- d -allofuranose. Cyclization of the linear tetramer under glycosylation conditions afforded the corresponding cyclic tetrasaccharide in 77% yield; its deprotection and reduction of the azido groups resulted in the formation of the cyclic tetramer of 3-amino-3-deoxy- d -allose with axial amino groups, a potential scaffold for the synthesis of tetravalent functional clusters.

The pear quantity is not consistent with the quality of pear paste products due to different proportions of edible cane sugar (sucrose) added during food processing. To determine sucrose addition rate in pear paste, a mathematical model was established to quantify soluble solid concentration, sucrose-adding ratio and d -allose concentration. Gas chromatography-mass spectrometer (GC-MS) results showed that 18 compounds were detected in pear paste. When the total soluble solids (TSS) concentration of pear paste was constant, there was a negative linear relationship between d -allose content and the Ratio of Sucrose Added (RSA) (0.97 ≤R2 ≤ 0.99)., Accordingly, the mathematical model of TSS-RSA-Allose concentration was fitted. It was found that the theoretical values of the pear paste at varied RSAs were comparable with the actual values under different conditions: 85% of the verified values differed by ≤ 10%. This study provides some reference for product testing departments to analyze products and establish the evaluation standard of sucrose ratioin pear paste.

A rapid method for the quantitation of sixteen neutral and acidic monosaccharides, from both animal and plant sources was developed using ultra-high performance liquid chromatography triple quadrupole mass spectrometry (UHPLC/QqQ-MS) in dynamic multiple reaction monitoring (dMRM) mode. Monosaccharides including three pentoses (ribose, xylose, arabinose), two deoxyhexoses (rhamnose, fucose), five hexoses (fructose, mannose, allose, glucose, galactose), two hexuronic acids (glucuronic acid, galacturonic acid), and two N-acetyl-hexosamines (GlcNAc, GalNAc), were derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP), while underivatized sialic acids, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), were simultaneously analyzed with a 10-minute run. With the optimized UHPLC conditions, baseline separations of the isomers were achieved. The sensitivity and calibration ranges of this method were determined. The limits of detection were between femtomoles and attomoles with linear ranges spanning four to six orders of magnitude and CVs ≤ 7.2%. Spiking experiments performed on a pooled fecal sample demonstrated the high accuracy of this method even when applied to samples with complicated matrices. The validated method was applied to fecal samples from an infant transitioning from breast milk to weaning foods. Major milk monosaccharides including galactose, fucose, glucose, GlcNAc, and Neu5Ac were found to be the most abundant components in the feces of milk-fed infants. PMP-derivatives of other monosaccharides including apiose, lyxose, altrose, talose, gulose, glucosamine, galactosamine, mannosamine, and N-acetylmannosamine (ManNAc) were also tested and could be added to the quantitation method depending on the need. The speed and sensitivity of the method makes it readily adaptable to rapid throughput analysis of monosaccharides in biological samples.

d-Allose is the C3 epimer of d-glucose and has been reported to have beneficial health effects. The transporter mediating intestinal transport of d-allose is unknown. We examined whether d-allose is absorbed via sodium-dependent glucose cotransporter 1 (SGLT1) as well as via glucose transporter type 5 (GLUT5) using rats. For examination of absorption via SGLT1, KGA-2727, an SGLT1-specific inhibitor, and d-allose were orally administered. KGA-2727 blocked the increase of plasma d-allose levels and suppressed them throughout the experiment (0–180 min), whereas without KGA-2727, the plasma d-allose levels peaked at around 60–90 min. For examination of absorption via GLUT5, rats were fed a high-fructose diet for 3weeks to increase the abundance and activity of GLUT5 in the small intestine. High-fructose diet-fed rats did not exhibit significant changes in the plasma d-allose levels compared to control rats fed a high-glucose diet. These results indicate that SGLT1 but not GLUT5 mediates the intestinal absorption of d-allose., Highlights ● Plasma d-allose levels were blocked by KGA-2727, an SGLT1-specific inhibitor. ● Fructose-fed rats which highly express intestinal GLUT5 did not exhibit significant changes in the plasma d-allose levels. ● SGLT1 clearly mediates intestinal d-allose transport.

Diacetone- d -allose is an important building block for the synthesis of pharmaceutical ingredients. The current batch production process involves a chemical oxidation and a reduction step that include hazardous reagents, produce a large amount of waste and require an extensive downstream processing. Current investigations aim to develop a catalytic reduction process of the intermediate diacetone- d -ketoglucose (DAKG) with molecular hydrogen, a green solvent, and a heterogeneous metal catalyst. Improvement of productivity and simultaneous reduction of the environmental impact shall be achieved by transforming the conventional batch procedure into a continuous process with a stationary catalyst packing. The screening studies presented in this work revealed that high reaction rates and a high selectivity (>98%) can be achieved with a commercial platinum catalyst under mild conditions in an aqueous buffer solution. Transition from batch into a continuous process was realized in several mini-scale fixed bed reactors to provide a fast screening and high mass transfer rates. A high selectivity (99%) and a conversion of 30% was achieved. The results indicate that the reactor performance was influenced by internal diffusion limitations and also by external mass transfer. To attain full conversion of DAKG, the reaction was performed in a random packed bed reactor and a selectivity of >96% was achieved.

d-Allose, a C-3 epimer of d-glucose, is a naturally occurring rare monosaccharide that shows anti-proliferative activity against several human cancer cell lines. However, d-allose requires a relatively high concentration for the activity to be observed. Thus, developing more potent derivatives is needed for application. In cells, d-allose is converted to d-allose-6-phosphate (A6P), which is responsible for the anti-proliferative activity of d-allose. In this study, we synthesized A6P derivative 1 with biodegradable protecting groups, which showed higher anti-proliferative activity than A6P against a MOLT-4F human leukemia cell line. Similarly protected derivative of d-glucose-6-phosphate (G6P) (2) and tetraacetyl-A6P (3) showed weaker and less activity compared with 1, respectively, suggesting that both A6P moiety and the protecting group on the phosphate group are responsible for the activity. In addition, significantly weaker induction of thioredoxin-interacting protein (TXNIP) expression by 1 compared with d-allose suggests that 1 exhibited cytotoxicity through the synergetic effect of inducing TXNIP expression and other mechanisms.

d -Allose was considered as a kind of rare sugars with testified potential medicinal and agricultural benefits. l -Rhamnose isomerase (L-RI, EC 5.3.1.14), an aldose-ketose isomerase, played a significant part in producing rare sugar. In this article, a thermostable d -allose-producing L-RI was characterized from a thermotolerant bacterium, Thermobacillus composti KWC4. The recombinant L-RI was activated obviously in the presence of Mn2+ with an optimal pH 7.5 and temperature 65 °C. The Michaelis-Menten constant (Km), turnover number (kcat) and catalytic efficiency (kcat/Km) for l -rhamnose were 33.8 mM, 1189.8 min−1 and 35.2 min−1 mM−1, respectively. At a higher temperature, Mn2+ played a pivotal role in strengthening the thermostability of T. composti L-RI. The differential scanning calorimetry (DSC) results showed the denaturing temperature (Tm) of T. composti L-RI was increased by 3 °C in presence of Mn2+. Although the T. composti L-RI displayed the optimum substrate as l -rhamnose, it could also effectively catalyze the isomerization between d -allulose and d -allose. When the reaction reached equilibrium, the sole product d -allose was produced from D-alluose by T. composti L-RI.

Palladium-catalyzed oxidation can single out the secondary hydroxyl group at C3 in glucose, circumventing the more readily accessible hydroxyl at C6 and the more reactive anomeric hydroxyl. Oxidation followed by reduction results in either allose or allitol, each a rare sugar that is important in biotechnology. Also, N-acetylglucosamine is selectively oxidized at C3. These results demonstrate that glucose and N-acetylglucosamine, the most readily available chiral building blocks, can be versatile substrates in homogeneous catalysis.

Interactions between proteins and carbohydrates are ubiquitous in biology. Therefore, understanding the factors that determine their affinity and selectivity are correspondingly important. Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide–aromatic interactions. A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate–aromatic interaction. Our data demonstrate that the carbohydrate–aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. Galactose seems to make the weakest and allose the strongest sugar–aromatic interactions, with glucose, N-acetylglucosamine (GlcNAc) and mannose in between. The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide–aromatic interaction energies. Peracetylation of the monosaccharides substantially increases the strength of the sugar–aromatic interaction in the context of our N-glycoprotein host. Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH–π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes.

We compared the growth inhibitory effects of all aldohexose stereoisomers against the model animal Caenorhabditis elegans. Among the tested compounds, the rare sugars d-allose (d-All), d-talose (d-Tal), and l-idose (l-Ido) showed considerable growth inhibition under both monoxenic and axenic culture conditions. 6-Deoxy-d-All had no effect on growth, which suggests that C6-phosphorylation by hexokinase is essential for inhibition by d-All.

Abnormal sialylation of proteins is highly associated with many major diseases, such as cancers and neurodegenerative diseases. However, this study is challenging owing to the difficulty in enriching trace sialylated glycopeptides (SGs) from highly complex biosamples. The key to solving this problem relies strongly on the design of novel SG receptors to capture the sialic acid (SA) moieties in a specific and tunable manner. Inspired by the saccharide-saccharide interactions in life systems, here we introduce saccharide-based SG receptors into this study. Allose (a monosaccharide) displays specific and pH-sensitive binding toward SAs. Integrating allose units into a polyacrylamide chain generates a saccharide-responsive smart copolymer (SRSC). Such design significantly improves the selectivity of SA binding; meanwhile, this binding can be intelligently triggered in a large extent by solution polarity and pH. As a result, SRSC exhibits high-performance enrichment capacity toward SGs, even under 500-fold interference of bovine serum albumins digests, which is notably higher than conventional materials. In real biosamples of HeLa cell lysates, 180 sialylated glycosylation sites (SGSs) have been identified using SRSC. This is apparently superior to those obtained by SA-binding lectins including WGA (18 SGSs) and SNA (22 SGSs). Furthermore, lactose displays good chemoselectivity toward diverse disaccharides, which indicated the good potential of lactose-based material in glycan discrimination. Subsequently, the lactose-based SRSC facilitates the stepwise isolation of O-linked or N-linked SGs with the same peptide sequence but varied glycans by CH3CN/H2O gradients. This study opens a new avenue for next generation of glycopeptide enrichment materials.

We describe the syntheses of three different deoxy-D-allose analogs [2-deoxy-D-allose (2-DOAll), 1,2-dideoxy-D-allose (1,2-DOAll), and 1,2-didehydro-1,2-dideoxy-D-allose (1,2-DHAll)] and their fatty acid esters via regioselective lipase-catalyzed transesterification. Among them, 2-DOAll and its decanoate (2-DOAll-C10) showed higher inhibitory activity on plant growth, which is similar to D-allose (All) and its decanoate (All-C10). Bioassay results of deoxy-All-C10 on four plant species suggest that the hydroxy group at the C-1 position might be important showing growth inhibitory activity. In addition, co-addition of gibberellin (GA3) with 1,2-DHAll-C10 and 2-DOAll-C10 recovered plant growth, suggesting that they might mainly inhibit biosynthesis of gibberellin.

d -allose is one kind of rare sugar and its biological functions and metabolic pathways in several organisms have been reported, but its function in plants’ resistance has not been well understood. In this research, we analyzed the function of d -allose in plant immunity in tomato. The plants which were pretreated with 5 mM d -allose increased the resistance to Botrytis cinerea and Pseudomonas syringae pv. tomato (Pst) DC3000 compared with the plants treated with water, with lighter symptom. The d -allose treatment induced the resistance to B. cinerea and Pst DC3000 maybe through regulating reactive oxygen species (ROS) accumulation and the expression levels of defense-related genes.

Fluorescence resonance energy transfer (FRET)-based sensors transduce ligand recognition into a change in the fluorophore spectrum, as ligand binding alters the distance between and orientation of two fluorescent proteins. Here, we report a dramatic increase in the signal intensity of FRET-based sugar sensors with bacterial periplasmic binding proteins (PBPs) in the binding moiety, by increasing the analysis temperature, usually higher than 50 °C. The increased signal intensity results from a sudden decrease in background signal at critical temperatures, while recovering the maximum FRET ratios in the presence of ligands. When tested with a maltose sensor using a maltose-binding protein as the binding moiety, the FRET ratio at the critical temperature, 55 °C, was 17-fold higher than at ambient temperatures. Similar effects were observed using analogous sensors for allose, arabinose, and glucose, providing highly dynamic and quantitative ratio changes at the critical temperatures. The proposed mechanism underlying the signal improvement is thermal relaxation of the binding proteins at the critical temperature; this hypothesis was supported by the results of intrinsic tryptophan fluorescence and circular dichroism experiments. In summary, this study shows that the conformational relaxation of proteins under specific conditions can be leveraged for highly sensitive and rapid measurements of ligands using FRET-based sensors.

Ketone derived from diacetone-α-D-glucose is a suitable starting material for the synthesis of 3-C-linked glycohybrids containing 1,2,3-triazole moiety as an intersugar linkage. The pyranose tautomers of 3-deoxy-3-(1,2,3-1H-triazol-1-yl)glucose, 3-C-[(1,2,3-1Htriazol-1-yl)methyl]allose and 3-C-[(1,2,3-1H-triazol-1-yl)methyl]gulose moieties are released upon the acidic hydrolysis of the corresponding O-isopropylidene-protected furanosyl-type synthetic intermediates. Some of the title compounds show a rare property of activating β-galactosidase from Escherichia coli.

The solubility and ice melting point of a binary system containing water and D-allose (D-All), the C3 epimer of D-glucose, were investigated and, for the first time, a binary phase diagram was cons...

To characterize l-rhamnose isomerase (l-RI) from the thermophilic bacterium Clostridium stercorarium and apply it to produce d-allose from d-allulose. A recombinant l-RI from C. stercorarium exhibited the highest specific activity and catalytic efficiency (k cat/K m) for l-rhamnose among the reported l-RIs. The l-RI was applied to the high-level production of d-allose from d-allulose. The isomerization activity for d-allulose was maximal at pH 7, 75 °C, and 1 mM Mn2+ over 10 min reaction time. The half-lives of the l-RI at 65, 70, 75, and 80 °C were 22.8, 9.5, 1.9, and 0.2 h, respectively. To ensure full stability during 2.5 h incubation, the optimal temperature was set at 70 °C. Under the optimized conditions of pH 7, 70 °C, 1 mM Mn2+, 27 U l-RI l−1, and 600 g d-allulose l−1, l-RI from C. stercorarium produced 199 g d-allose l−1 without by-products over 2.5 h, with a conversion yield of 33% and a productivity of 79.6 g l−1 h−1. To the best of our knowledge, this is the highest concentration and productivity of d-allose reported thus far.

Rare sugars are defined as monosaccharides that exist in nature but are only present in limited quantities. d-Allose is a rare sugar that has been reported to have some unique physiological effects. The present study describes suitable synthetic procedures for novel rare sugars of d-allose that are 18F-labeled at the C-3 and C-6 positions and the preparation of the appropriate labeling precursors. The goal is to facilitate in vivo, noninvasive positron emission tomography (PET) investigation of the behavior of rare sugar analogs of d-allose in organs. We found five precursors that were practical for labeling, three for 3-deoxy-3-[18F]fluoro-d-allose ([18F]3FDA) and two for 6-deoxy-6-[18F]fluoro-d-allose ([18F]6FDA). With manual operation synthesis, the highest radiochemical conversion rates were 75% for [18F]3FDA with a precursor of 1,2,4,6-tetra-O-acetyl-3-O-trifluoromethanesulfonyl-β-d-glucopyranose and 69% for [18F]6FDA with a precursor of 1,2,3,4-tetra-O-acetyl-6-O-trifluoromethanesulfonyl-β-d-allopyranose. Furthermore, the practical yields of [18F]3FDA and [18F]6FDA using an automated synthesizer were also investigated. Radiochemical yields of 67% and 49% were obtained for [18F]3FDA and [18F]6FDA, respectively, in an automated synthesizer. As basic assessment of stability for use in PET scanning, high performance liquid chromatography analysis showed no decomposition of [18F]3FDA and [18F]6FDA after up to 6 h in rabbit blood plasma.

The anti-proliferative activities of the 6-O-acyl derivatives of d-allose against the human leukemia MOLT-4F cell line were examined. The activity of the 6-O-dodecanoyl derivative (3) was approximately 30 times stronger than that of d-allose. An evaluation of the derivatives of 3 that occurred in a furanose form revealed the pyranose forms of 3 to be important for the anti-proliferative activity.

Various glycosyl hydrolase enzymes of the clan GH-H (http:// www.cazy.org/Glycoside-Hydrolases.html) catalyse transglycosylation reactions, mostly by using starch (e.g. , CGTase, family GH13) or sucrose (e.g. , glucansucrase, family GH70) as the donor substrate, thus transferring glucose. Some alternative biocatalysts for glycosyl transfer reactions have also been discovered; these include glycosynthases and glycosyltransferases. In recent years we have introduced the concept of sucrose analogues. Sucrose analogues such as a-d-glycopyranosyl bd-fructofuranosides have been reported as promising tools for the synthesis of functional fructo-oligosaccharides. In recent review articles we have suggested that these analogues can be used as glycopyranosyl donors for artificial transglycosylation reactions, which are not performed in nature. 9] Sucrose analogues are readily available from the cheap and abundant starting material sucrose. They are obtained through a transfructosylation reaction catalysed by levansucrase from Bacillus subtilis or Bacillus megaterium, by transferring fructose from sucrose to mannose, galactose, xylose or fucose. a-d-Allopyranosyl b-d-fructofuranoside (1, All-Fru, Scheme 1) is an allose that contains a sucrose analogue derived from sucrose by dehydrogenisation to 3-ketosucrose and subsequent hydrogenation to All-Fru. This synthesis has been established at an industrial scale. Allose is a rare sugar in nature; only a few plant species, for example, Passiflora edulis, Sideritis grandiflora and Protea rubropilosa, are known to contain glycoconjugates with blinked allose residues. Allose and allosylated glycoconjugates show promising properties for pharmaceutical use in cancer therapy, in immune suppression, for inhibition of neutrophil production and as antioxidants. To the best of our knowledge no a-d-allosyltransferase is known, however, research by Thorson and co-workers indicated a possible b-allosyl transfer performed by the glycosyltransferase OleD enzyme (family GT1). In recent work, we sought to identify enzymes able to transfer d-allose from the sucrose analogue All-Fru as the donor substrate. For this, All-Fru was incubated with various enzymes from the glycosyl hydrolase clan GH-H. As this sucrose analogue is a-linked and enzymes of the GH-H clan generally retain the configuration at the anomeric centre, the formed products should exhibit a-linkage. The sucrose isomerase (SI, GH13) from Protaminobacter rubrum showed no activity towards All-Fru. In contrast, incubation of All-Fru (292 mm) with amylosucrase (NpAs, GH13) from Neisseria polysaccharaea at pH 6.6 and 30 8C showed slow hydrolysis : even after 80 h, consumption of All-Fru was incomplete. Glucansucrases from Lactobacillus reuteri strains 121 (GTFA) and 180 (GTF180), and GTFR from Streptococcus oralis (all GH70) were able to hydrolyse 292 mm All-Fru. As hydrolysis activity on All-Fru was highest for GTFA (complete hydrolysis after 80 h), this enzyme was used in further studies. The glucansucrase GTFA from L. reuteri 121 is an extracellular enzyme that produces the polymer reuteran (mainly a(1!4)-linked glucan, with minor a(1!6) and branches) from sucrose. In contrast, with sucrose as donor, no formation of allose oligoor polysaccharides was observed by TLC. The transferase activity of GTFA-DN-CHis (a N-truncated version was used in all experiments) with All-Fru was observed with several acceptor substrates (note: concentration depended on solubility in the solvent system), thus obtaining a variety of functionalities (Table 1) such as sugars (methyl a-d-glucopyranoside (a-d-Me-Glcp, 2) and methyl 6-O-p-toluenesulfonyl-ad-glucopyranoside (a-d-Me-6-Ts-Glcp, 4)), amino acids (e.g. , N-(tert-butoxycarbonyl)-d-serine methyl ester (N-Boc-d-serineOMe, 6)) and ( )-epicatechin (8). Incubating GTFA with All-Fru (292 mm) and a-d-Me-Glcp (772 mm, 2) as the acceptor resulted in methyl a-d-allopyranosyl-(1!4)-a-d-glucopyranoside (3) as the main product (50% yield). The a-configuration of the glycosidic bond was determined by H NMR (H-1’: d= Scheme 1. The sucrose analogue All-Fru.

Ultrasound-assisted extraction was employed to extract polysaccharide from Virginia (flue-cured) tobacco flowers. The orthogonal matrix method (L9(3)(4)) was used to determine the optimal extraction conditions as to ultrasound power, extraction time, ratio of solvent to solid, and extraction temperature at 300 W, 4 min, 35 (mL/g), and 70 °C respectively. The crude extract was successively purified by chromatography, yielding two major polysaccharide fractions, termed Fr-I and Fr-II. Both fractions are heteropolysaccharides, mainly containing glucose, mannose, and allose with an a-configuration. Thermo gravimetric analysis (TGA) indicated that the degradation temperatures (Td) of Fr-I and Fr-II were 185 °C and 190 °C respectively. The preliminary antioxidant activity test in vitro showed both fractions could potentialize the scavenging effect on hydroxyl and DPPH radicals in a dose-dependent manner. In conclusion, the two polysaccharides may be useful as naturally potential antioxidant agents for application in food and medicinal fields.

1,6-Anhydrohexopyranose, which is a major pyrolysis product of cellulose and other carbohydrates, has a unique 1C4 chair conformer. Thus, during pyrolysis, the molecule must undergo a conformational transformation from the 4C1 to the 1C4 geometry, and this interconversion is expected to be influenced by the sugar's hydroxyl group configuration. In this article, the influences of the C2-, C3-, and C4-OH configuration on the 1,6-anhydrohexopyranose yield were evaluated under vacuum pyrolysis conditions for five hexoses (glucose, mannose, allose, galactose, and gulose). Furthermore, the results are discussed within the context of theoretical results that indicate the relative stability of two types of the conformers of glycopyranosyl oxocarbenium ions with C6-pseudo-equatorial and C6-pseudo-axial orientations, which were calculated with density functional theory (DFT). The latter conformation would lead to the formation of the 1,6-anhydrohexopyranose with a 1C4 conformation. The theoretical results were in good agreement with the experimental yields; C2-OH (both axial and equatorial) and C3-OH (equatorial) enhance 1,6-anhydrohexopyranose formation, while C4-OH (axial) inhibits the formation (axial/equatorial: in 4C1 form). These results also support the heterolysis mechanism for this transformation, and such conformational change was suggested to be more effective in the evaporation process from liquid to gaseous substance.

Stable amides and oligopeptides derived from methyl trans,trans -3-azido-4-hydroxymethyl- l -azetidine carboxylate, prepared in 19% yield from diacetone allose, is the first example of a β-amino-azetidine carboxylic acid incorporated into peptidomimetics and provides a scaffold for investigating secondary structure induced by a novel β-amino acid. A number of azetidine iminosugars containing a NHAc substituent were prepared but none of them were β- N -acetylhexosaminidase inhibitors.

The preparation of the δ-heptonolactones, 4- O -benzyl- D - glycero - D - D - gulo -heptono-1,5-lactoneand 4- O -benzyl- D - glycero - D - allo -heptono-1,5-lactone, by Kiliani reactions on 3- O -benzylglucose and 3- O -benzylallose are reported; acetonides of the two lactones are described.

The Kiliani synthesis on the 4 readily accessible ketohexoses ( d -fructose, d -tagatose, l -sorbose, d -psicose) allows access to 4 diastereomeric 2-C-methyl-aldohexoses (2-C-methyl- d -mannose, 2-C-methyl- d -talose, 2-C-methyl- l -gulose, 2-C-methyl- d -allose) and 4 diastereomeric 2-C-methyl-alditols (2-C-methyl- d -mannitol, 2-C-methyl- d -talitol, 2-C-methyl- l -gulitol, 2-C-methyl- d -allitol). Microbial oxidation of 2-C-methyl- d -mannitol and 2-C-methyl- l -gulitol gave 5-C-methyl- d -fructose; microbial oxidation of 2-C-methyl- d -talitol afforded 5-C-methyl- d -psicose, whereas 2-C-methyl- d -allitol formed 5-C-methyl- l -psicose. Both enantiomers of 5-C-methyl-fructose were equilibrated by d -tagatose-3-epimerase (DTE) with both enantiomers of 5-C-methyl-psicose. These transformations demonstrate that polyol dehydrogenases and DTE act on branched synthetic sugars. Full NMR analyses show that 5-C-methyl- d -fructose is present as the β-pyranose and β-furanose forms in a ratio of 90:10; all pyranose and furanose forms of 5-C-methyl- d -psicose are present in solution. The combination of chemical and biological procedures allows the environmentally friendly generation of a new family of branched monosaccharides.

Recently, monosaccharide-triazole conjugates have proved to possess a large variety of useful biological activities. This paper describes synthesis of a new series of 3-deoxy-3-C-triazolylmethyl-allose derivatives. These new compounds are obtained from acetonide-protected 3-deoxy-3-azidomethyl allose and commercial alkynes via Cu(I) catalyzed 1,3-dipolar cycloaddition. The obtained molecular scaffolds differ from those described earlier by the presence of a methylene linker (-CH2-) between the C(3) of allose and the triazole moiety. It was demonstrated that acetonide-protected monosaccharide, 3-deoxy-3-C-(4-phenyl-1H-1,2,3-triazol-1-yl)methyl-1,2:5,6-di-O-isopropylidene-α-d-allofuranose, inhibited α-L-fucosidase for 26% at 0.1 mM concentration, but a deprotected analog, 3-deoxy-3-C-(4-(4-tert-butylphenyl)-1H-1,2,3-triazol-1-yl)methyl-β-d-allofuranose, showed 15% inhibition of β-glucosidase at 1 mM concentration.

Ruthenium-catalysed epimerisation of secondary carbohydrate alcohols was used to determine the thermodynamic equilibrium between non-anomeric epimers of partially protected glucose, mannose and allose derivatives. A cyclopentadienylruthenium catalyst was used to epimerise each of two pure epimeric alcohols in two separate experiments. The epimerisation reactions were run until the same ratio of epimers was obtained from the two experiments.