Synthesis Of Oligosaccharides Research Articles

Harnessing Free Sulfate Groups in Glycosylation Reactions.

General strategies for synthesizing sulfated oligosaccharides employ the protection of sulfate groups for glycosylation or post-sulfation after the synthesis of oligosaccharides. However, the chemical behavior of free sulfate groups in glycosylation reactions has not been thoroughly studied. We examined several glycosyl donors with free sulfate groups, but neither glycosyl imidates nor thioglycosides achieved products. Conversely, activating 6-sulfated GalNAc donors with either a 2-NTroc group or 2-O-acetyl group using an ortho-hexynylbenzoate group at the anomeric position yielded β-glycosides in good yield. These results indicate that glycosylations with a free sulfate group can be performed and that the neighboring group participation of 2-NHTroc and 2-O-acetyl groups works even with an unprotected sulfate group at 6-position. Ab initio calculations supported the formation of acyloxionium-cation via 2-O-acyl participation. Additionally, glycosylation reactions under various counter cations of the sulfate group, such as Na+, Li+, K+, Ba2+ and pyridinium were examined. Results showed that sodium and lithium salt donors yielded the products in good yield. These results were also supported by ab initio calculations. Practical glycosylation reactions between disaccharide donor-acceptor pairs with free sulfate groups successfully yielded the sulfated tetrasaccharides. This study also discusses how a sodium salt acts as a protecting group during glycosylation.

Catalyst-free and wavelength-tuned glycosylation based on excited-state intramolecular proton transfer

The chemoselectivity of organic reactions is a fundamental topic in organic chemistry. In the long history of chemical synthesis, achieving chemoselectivity is mainly limited to thermodynamic conditions by an exogenous activation strategy. Here, we design an endogenous activation method, which can be used to control the chemoselectivity of phenol and naphthol through the photo-induced excited-state intramolecular proton transfer (ESIPT). A wavelength-tuned glycosylation is developed to showcase the penitential of this new strategy. Traditionally, an exogenous activator (electrophilic promoters) is essential to induce the cleave of a polar single bond, and this strategy has been extensively studied and used in the glycosylation chemistry, for the formation of oxocarbenium cation intermediate. In our systems, the oxocarbenium cation intermediates can be selectively formed from glycosyl donors bearing tunable chromophoric groups under mild conditions of acid-base free and redox neutrality, which enables continuous synthesis of oligosaccharides.

Photocatalyst-free light-promoted carbohydrate synthesis and modification

Photoredox catalysis has recently emerged as a powerful approach for preparing oligosaccharides because it uses mild conditions, is compatible with partially or completely unprotected carbohydrate substrates, and exhibits impressive regio‐ and stereo‐selectivity and high functional group tolerance. However, most catalytic photoredox reactions require an external photocatalyst (organic dye or expensive transition-metal complex) to deliver key glycosyl radicals. Several photocatalyst-free photocatalytic reactions that avoid the use of expensive metal salts or organic-dye additives have received significant attention. In this review, we highlight the most recent developments in photocatalyst-free light-promoted carbohydrate synthesis and modification, which is expected to inspire broad interest in further innovations in the green synthesis of saccharides.

Controllable Enzymatic Synthesis of Natural Asymmetric Human Milk Oligosaccharides.

Among human milk oligosaccharides (HMOs), linear HMOs are synthesized through mature but varied routes. Although branched HMOs can be synthesized by chemical, enzymatic, or chemoenzymatic methods, these methods cannot be easily applied to the synthesis of asymmetric multiantennary oligosaccharides. Herein, we developed a controllable method to synthesize asymmetric biantennary HMOs. In our synthetic route, GlcNAcβ1,3(GlcN3β1,6)Glaβ1,4Glc was first chemically synthesized as the core tetrasaccharide, which contains β1,6GlcN3 as the "stop" sugar in transferase-catalyzed glycosylation. The desired sugars at the GlcNAcβ1-3Gal arm can be assembled using galactosyltransferase, N-acetylglucosaminyltransferase, and fucosyltransferase. Then, the Staudinger reduction and acetylation were used to transform GlcN3 to GlcNAc and assemble sugars by initiating the "go" process. By manipulating transferase-catalyzed glycosylations, 22 natural asymmetric biantennary oligosaccharides were synthesized.

Mechanistic impact of Gracilaria bailinae extracts on photosynthesis and metabolism in Phaeodactylum tricornutum

In the intricate realm of aquatic ecosystems, biotic interactions play pivotal roles in shaping the physiological responses and survival strategies of microorganisms. This study investigates the effects of Gracilaria bailinae on photosynthesis and metabolism on the diatom Phaeodactylum tricornutum and the ecological significance. Our results reveal considerable suppression by G. bailinae on both its light-dependent and light-independent reactions of photosynthesis in P. tricornutum. A pronounced decline in carbon fixation was observed causing. the diatom to prioritize its carbon flux towards carbohydrate synthesis for its cellular energy needs. At high G. bailinae concentrations a marked reduction in lipid content indicated their importance as emergency energy sources. This response in lipid mobilization under photosynthetic stress is an evolutionary strategy for environmental adaptation. In addition, G. bailinae-induced stress amplified lysosomal activity in the diatom. Such an upsurge in oxidative stress appears to fast-track cellular death. We conclude that the ROS production, induced by G. bailinae, acts as a linchpin in mediating stress responses, thereby significantly reconfiguring the metabolism in the diatom. This study not only elucidates the physiological countermeasures of microalgae against biotic stressors but it also underscores the complex interactions between aquatic microorganisms.

Distinct exposure impact of non-degradable and biodegradable microplastics on freshwater microalgae (Chlorella pyrenoidosa): Implications for polylactic acid as a sustainable plastic alternative

Microplastics (MPs) are increasingly recognized as significant sources of harm to biota in various environments. However, the detrimental impacts of aged MPs with different structures and degradability remain poorly understood. In this study, aged MPs from polylactic acid (PLA), polyethylene (PE), and polystyrene (PS), representing biodegradable, aliphatic, and aromatic plastics, respectively, were prepared to examine their effects on microalgae (Chlorella pyrenoidosa). Structural and property analyses indicated the presence of aging and oxygen-containing functional groups on the surfaces of the MPs, which correlated with an increase in negative electrical charge (i.e., aged PLA > aged PE ≈ aged PS). Aged PLA MPs affected microalgae biomass, promoted protein synthesis, and elevated mild oxidative stress. In contrast, aged PE and PS MPs not only affected biomass, protein, and carbohydrate synthesis but also inhibited photosynthetic pigment production and activity, resulting in intracellular oxidative stress. Excitation-emission-matrix spectra analysis showed that PLA induced microalgae to secrete large amounts of humic acid-like extracellular polymers, whereas aged PE and aged PS groups contained only small amounts of them and proteins. This study addresses critical knowledge gaps in the toxicology of various aged MPs on microalgae and provides valuable insights into the potential of PLA as a sustainable alternative to conventional plastics in microalgae culture industry.

Investigation of Neighboring Group Participation in 3,4-Diacetylated Glycosyl Donors in the Gas Phase.

A key challenge in oligosaccharide synthesis is the stereoselective installation of glycosidic bonds. Each glycosidic linkage has one of two possible stereo-chemical geometries, α/β or 1,2-cis/trans. An established approach to install 1,2-trans glycosidic bonds is neighboring group participation (NGP), mediated by a 2-O-acyl group. Extension of this intramolecular stabilization to nucleophilic groups located at more remote positions has also been suggested, but remains poorly understood. Previously, we employed infrared ion spectroscopy to characterize the molecular ions of monoacetylated sugar donors and showed how the strength of the stabilizing effect depends on the position of the participating ester group on the glycosyl donor ring as well as on its relative stereochemistry. In this work, we investigated glycosyl donors carrying two acyl groups. Using isotope labelling and isomer population analysis we were able to resolving spectra of isomeric mixtures and establish the relative contribution of individual species. We conclude that 3,4-diacetyl mannosyl donors exclusively form a dioxanium ion as a result of C-3 acyl stabilization. In contrast, the glucosyl and galactosyl cations form mixtures of C-3 and C-4 acyl participation products. Hence, the combination of isotope labeling and population analysis allows for the study of increasingly complex glycosyl cations.

Catalytic Activation of Thioglycosides with Copper-Carbenes for Stereoselective 1,2-Cis-Furanosylations.

Thioglycoside activation, crucial for oligosaccharide synthesis, faces challenges with the need for stoichiometric promoters, additives, and cryogenic conditions, particularly in stereoselective 1,2-cis-linkage formation. This study introduces a carbene-based catalytic method using Cu(OTf)2 for thioglycoside activation, enabling efficient 1,2-cis-furanosylation in ribose and arabinose. The method is orthogonal to conventional thioglycoside and alkyne donors, accommodates sterically demanding acceptors, and achieves stereoselectivity independent of the donor anomeric configuration and C2-protecting groups, with copper chelation playing a key role.

Zinc deficiency in ruminants and its management: A brief review

Minerals are organic substances found in nature and living things. In ruminants, minerals are one of the crucial components of the body. One of the mineral elements that is very important for the body is Zinc (Zn). The prominent role of Zn in ruminants is the synthesis and degradation of carbohydrates, fats, proteins, and nucleic acids. Ruminants can experience Zn deficiency by showing clinical symptoms such as dermatitis, anorexia and parakeratosis, stunted growth, and nail damage. In contrast, ruminants can also experience Zn poisoning with various clinical symptoms such as general weakness, diarrhea, hemolytic anemia, and decreased body weight. Current Zn deficiency can be overcome by adding commercial supplement products containing Zn. This paper aims to add brief insights into the benefits and treatment of Zn deficiency in ruminants.

Microfluidics-Based Ionic Catch and Release Oligosaccharide Synthesis (ICROS-Microflow) to Expedite Glycosylation Chemistry.

A continuous microfluidic glycosylation strategy that requires no chromatography between steps and significantly expedites glycosylation chemistry is described. This practical approach incorporates the advantages of imidazolium-based chromatography-free purification and in situ mass spectrometry reaction monitoring to achieve fast reaction optimization and shorter reaction times. We demonstrate the utility of this strategy in the synthesis of a series of glycoside targets, including an α-(1,6)-trimannoside and a branched Man5 oligomannoside, within 1 day.

Regulatory role of microcystin in the response of microcystin-producing cyanobacteria to elevated CO2: Insights from metabolic profiling

The regulatory role of microcystin in the response of microcystin-producing cyanobacteria to elevated CO2 remains poorly understood. To address this gap, this study compared the responses of wild-type toxic Microcystis PCC 7806 and its mcyB-knockout mutant to elevated CO2 using metabolomic profiling under nitrogen (N)-rich and N-poor conditions. Under N-poor conditions, elevated CO2 promoted carbohydrate synthesis and tricarboxylic acid cycle in both strains, without affecting their growth. Under N-rich conditions, both strains exhibited increased biomass with rising CO2 levels, attributed to enhanced carbohydrate synthesis, tricarboxylic acid cycle, glutamate-glutamine cycle, purine synthesis, and arginine synthesis. However, compared to the mutant, the proliferation of wild-type toxic Microcystis was less stimulated by elevated CO2. The difference was associated with its reduced activity in the pentose phosphate pathway, likely linked to microcystin synthesis. Besides, the difference was related to higher utilization of glutamate, arginine, and aspartate due to increased microcystin production, indicating the regulatory role of microcystin in the response of microcystin-producing cyanobacteria to elevated CO2. Importantly, elevated CO2 could enhance microcystin synthesis by promoting the production of carbon backbones (malonyl CoA), amino acids (including arginine, glutamate and aspartate) and methyl donors (S-adenosylmethionine) of the wild-type toxic Microcystis PCC 7806. Notably, sufficient nitrogen sources were required for increased amino acid and methyl donors synthesis at high CO2 concentration. The discovery revealed underlying mechanisms behind the potential for elevated CO2 levels to increase toxicity risk associated with Microcystis blooms.

The response mechanism of alkalophilic Nitzschia sp. NW129 to low alkalinity-A study combining physiological and transcriptional analysis

Global aquatic acidification significantly threatens alkaline lake ecosystems. The mechanisms by which alkaliphilic microalgae, the key producers in these ecosystems, respond to reduced environmental alkalinity remain poorly understood. Here, we investigated the responses of alkalophilic Nitzschia sp. NW129 to low alkalinity (pH 9.2) through integrated physiological-biochemical and transcriptomic analyses. Relative to the control (pH 11.5), we observed a 60.1 % decrease in polysaccharide content, while total lipids and proteins increased by 1.74-fold and 2-fold, respectively. Transcriptome analysis revealed up-regulation of genes encoding carbonic anhydrase (CA) and malic enzyme (ME), along with those involved in glycolysis and fatty acid (FA) synthesis, compensating for carbon supply and shifting carbon flux from carbohydrate synthesis to lipid accumulation. Enhanced expression of TCA cycle genes and those encoding F-ATP synthase and inorganic pyrophosphatase (PPase) provided sufficient energy for cellular homeostasis, further facilitated by the up-regulated expression of ATP-dependent V-ATPase and ABC transporter genes. Temporal analysis revealed that the expression of genes involved in protein synthesis pathways was up-regulated on days 1 and 4 but notably down-regulated on day 2, suggesting protein degradation at this time to balance energy supply for adaptation. Despite these coping shifts, impairments in photosynthetic energy dissipation and electron transport, along with transcriptional changes including down-regulating cell cycle and inducing apoptotic pathways, ultimately caused a substantial reduction in biomass. These findings provide a basic understanding of the response mechanisms of alkalophilic microalgae to low alkalinity stress, which should aid to develop strategies to improve microalgal tolerance against acidification.

Immobilization and Kinetic Properties of ß-N-Acetylhexosaminidase from Penicillium oxalicum

The application of immobilized enzymes often plays a key role in successfully implementing an economically feasible biocatalytic process at an industrial scale. Designing an immobilized biocatalyst involves solving several tasks, from the selection of the carrier and immobilization method to the characterization of the kinetic properties of the immobilized enzyme. In this study, we focused on the kinetic properties of free and immobilized ß-N-acetylhexosaminidase (Hex), a promising enzyme for application in the field of biotechnology, especially for the synthesis of bioactive carbohydrates. Hex was immobilized via covalent binding in methacrylate particles. The effect of immobilizing Hex from Penicillium oxalicum into porous particles on kinetic properties was investigated, and mathematical and experimental modeling showed that the kinetic behavior of the enzyme was significantly influenced by diffusion in the particles. Along with the study on kinetics, a simple method was developed to investigate the reversible inhibition of the immobilized enzyme in a continuous-flow system. The method is suitable for application in cases where a chromogenic substrate is used, and here it was applied to demonstrate the inhibitory effects of N-acetyl-glucosaminyl thiazoline (NAG-thiazoline) and O-(2-Acetamido-2-deoxy-D-glucopyranosylidene)amino N-phenyl carbamate ((Z)-PugNAc) on Hex.

Photolabile ortho‐Nitro‐Benzyl Carbonate as a Permanent Hydroxyl Protecting Group for the Synthesis of Digalactosyl Diacylglycerol

Comprehensive SummaryTraditional protecting groups are often removed under harsh conditions with potentially hazardous reagents, thereby impeding the convenient synthesis of oligosaccharides and glycosides. Herein, we present to utilize the photolabile ortho‐nitro‐benzyl carbonate (oNBC) as a permanent hydroxyl protecting group for stereocontrolled synthesis of glycosides. The Ph3PO‐modulated glycosylation with strongly disarmed per‐O‐oNBC‐protected glycosyl ynenoates preferred to afford glycosides with excellent α‐selectivities via the β‐phosphonium transition state. Based on the oNBC‐mediated galactosylation, synthesis of the glycolipid digalactosyl diacylglycerol (DGDG) containing six double bonds and two esters was achieved in a straightforward manner.

The Role of PLATZ6 in Raffinose Family Oligosaccharides Loading of Leaves via PLATZ Family Characterization in Cucumber

The plant AT protein and zinc-binding protein (PLATZ) genes, a novel cluster of plant-specific zinc-finger-dependent DNA-binding proteins, play a crucial role in regulating stress response and plant development. However, there has been little study focus on the role of the cucumber PLATZ family in assimilating loading in leaves. (1) In this study, a total of 12 PLATZ genes were identified from the cucumber genome. The cucumber PLATZ genes were clustered into five groups, and unevenly distributed on five chromosomes. A single pair of cucumber PLATZ genes underwent segmental duplication. (2) The results of genome-wide expression analysis suggested that the cucumber PLATZ genes were widely expressed in a wide range of cucumber tissues, with three PLATZ (PLATZ2, PLATZ6, and PLATZ12) genes exhibiting high expression in the vascular tissues of cucumber leaves. PLATZ2, PLATZ6, and PLATZ12 proteins were primarily located in cytomembrane and nucleus. (3) In VIGS-PLATZ6 plants, the expression of Galactinol synthase 1 (GolS1) and STACHYOSE SYNTHASE (STS), two genes involved in the synthesis of raffinose family oligosaccharides (RFOs) were observed to be decreased in cucumber leaves. In conclusion, the comprehensive analysis of the cucumber PLATZ family and the preliminary functional verification of PLATZ6 lay the foundation for the molecular and physiological functions of cucumber PLATZ genes.

Recent Progress in the Synthesis and Glycosylation of Rare Sugars

AbstractOut of 42 naturally occurring monosaccharides, only seven are abundant in Nature (glucose, galactose, mannose, fructose, xylose, ribose, and l-arabinose), while the others have been classified as ‘rare sugars’. Fungi and bacteria use a wide range of monosaccharides, in contrast to mammals, reflected in their glycosylated metabolites, as well as the cellular machineries that are involved in their sugar metabolism. Recognition of the microbiome’s impact on human health has led to increased interest in microbial glycans, as they often mediate interaction between host and microbes. Efficient access to rare sugars and oligosaccharides is necessary to study their roles in Nature, which can provide new pharmacological leads. Furthermore, it enables the synthesis of bioactive glycosylated natural products and congeners. This short review highlights recent progress in the synthesis and the efficient, site- and stereoselective glycosylation of rare sugars. Finally, it provides a recent example where synthetic access to rare sugars has enabled biochemical studies to better understand and interfere with processes in Nature.1 Introduction2 Synthesis of Rare Sugars2.1 Syntheses from Renewable Feedstock2.2 De Novo Syntheses3 Glycosylation3.1 Catalyst-Controlled Glycosylation3.2 One-Pot Iterative Oligosaccharide Synthesis in Solution4 Application in Biochemical Research5 Conclusion

ZnI2-Mediated cis-Glycosylations of Various Constrained Glycosyl Donors: Recent Advances in cis-Selective Glycosylations.

An efficient and versatile glycosylation methodology is crucial for the systematic synthesis of oligosaccharides and glycoconjugates. A direct intermolecular and an indirect intramolecular methodology have been developed, and the former can be applied to the synthesis of medium-to-long-chain glycans like that of nucleotides and peptides. The development of a generally applicable approach for the stereoselective construction of glycosidic bonds remains a major challenge, especially for the synthesis of 1,2-cis glycosides such as β-mannosides, β-L-rhamnosides, and β-D-arabinofuranosides with equatorial glycosidic bonds as well as α-D-glucosides with axial ones. This review introduces the direct formation of cis-glycosides using ZnI2-mediated cis-glycosylations of various constrained glycosyl donors, as well as the recent advances in the development of stereoselective cis-glycosylations.

Local adaptation of Dromiciops marsupials (Microbiotheriidae) from southern South America: Implications for species management facing climate change.

The two species of the microbiotheriid marsupial genus Dromiciops (Dromiciops bozinovici: "Panchos's monito del monte" and Dromiciops gliroides: "monito del monte") exhibit a marked latitudinal genetic differentiation. Nevertheless, it is unclear whether this differentiation results from neutral processes or can be explained, to some extent, by local adaptation to different environmental conditions. Here, we used an SNP panel gathered by Rad-seq and searched for footprints of local adaptation (putative loci under selection) by exploring genetic associations with environmental variables in the two species of Dromiciops in Chilean and Argentinean populations. We applied three methods for detecting outlier SNPs and two genotype-environment associations approaches to quantify associations between allelic frequencies and environmental variables. Both species display strong genetic structure. D. bozinovici exhibited three distinct genetic groups, marking the first report of such structuring in this species using SNPs. In contrast, D. gliroides displayed four genetic clusters, consistent with previous studies. Both species exhibited an association of their genetic structure with environmental variables. D. bozinovici exhibited significant associations of allelic frequencies with elevation, precipitation during the warmest periods, and seasonality in the thermal regime. For D. gliroides, genetic variation appeared to be associated with more variables than D. bozinovici, including precipitation and temperature-related variables, isothermality, and elevation. All the outlier SNPs were mapped to the D. gliroides reference genome to explore if they fell within functionally known genes. These results represent a necessary first step toward identifying the genome regions that harbor genes associated with climate adaptations in Dromiciops. Notably, we identified genes involved in various functions, including carbohydrate synthesis (ALG8), muscle and neuronal regulation (MEF2D), and stress responses (PTGES3). Ultimately, this study contributes valuable insights that can inform targeted conservation strategies aimed at preserving the genetic diversity of Dromiciops in the face of environmental challenges.

Acid-Catalyzed Transformation of Pyranosides into Furanosides as a Tool for Preparation of Furanoside Synthetic Blocks.

The importance of natural glycoconjugates containing furanoside residues causes a continued demand for the development of efficient methods for the synthesis of corresponding oligosaccharide derivatives to be used as molecular probes in glycobiological studies. Currently, the chemical synthesis of furanose-containing oligosaccharides often represents a significant challenge because of the lack of short, efficient, and reliable methods for the preparation of selectively substituted furanoside blocks. Herein, we report an easy protocol toward galactofuranose-containing molecules based on the unusual equilibrium between pyranoside and furanoside forms observed for a series of substituted galactosides. The method's utility is illustrated by the syntheses of furanoside-containing oligosaccharides related to the antigenic polysaccharides of Aspergillus fumigatus and Klebsiella pneumoniae O2ac.

Divergent Synthesis and Antigenicity Evaluation of Core Oligosaccharides of the Lipopolysaccharides from Acinetobacter baumannii SMAL and ATCC 19606.

Acinetobacter baumannii poses a serious threat to human health. Pathogenic bacterial lipopolysaccharides (LPSs) are potent immunogens for the development of antibacterial vaccines. To investigate the antigenic properties of A. baumannii LPS, five well-defined core oligosaccharide fragments from the LPS of A. baumannii SMAL and ATCC 19606 were synthesized. A divergent synthesis strategy based on orthogonally protected α-(2 → 5)-linked Kdo dimer 6 was developed. Selective exposure of different positions in this key precursor and then elongation of sugar chains via stereocontrolled formation of both 1,2-trans and 1,2-cis-2-aminoglycosidic linkages permitted the efficient synthesis of the targets. The synthetic route also highlights a 4-O and then 7-O glycosylation sequence for assembly of the novel 4,7-branched Kdo framework. Antigenicity assay using the glycan microarray technique disclosed that tetrasaccharide 3 featuring both 4,7-branch and α-(2 → 5)-Kdo-Kdo structural elements was a potential antigenic determinant.