Number Of Sugar Chains Research Articles

Multidirectional pharmacological effects of saponins in light of clinical trials: a systematic review of literature

Introduction: Saponins are secondary metabolites widely distributed in higher plants with multidirectional pharmacological effects. We distinguish between steroidal and triterpenoidal saponins, which are composed of glycone and aglycone. The varied medicinal properties of saponins are influenced by the presence of different substituents in the polycyclic aglycone, e.g. hydroxyls, hydroxymethyls, carboxyls and acyl groups, as well as the difference in bonding, composition and number of sugar chains. It is noteworthy that foods rich in these compounds can reduce the risk of developing various diseases, and the isolation of these compounds from the plant matrix provides new opportunities to create potential and effective drugs.
 Purpose of the work: Conduct a literature review on the basis of available clinical studies on the pharmacological effects of saponins extracted from medicinal plants and their potential application in medicine.
 Materials and methods: The work was based on a literature review of available medical publications from clinical trials conducted for the multidirectional therapeutic effects of saponins. Reports available in publications and scientific studies posted in medical information databases such as Pubmed and Google Scholar were summarised.
 Results: Clinical studies confirm the pharmacological effect of saponins in the treatment of sports injuries and trauma, type II diabetes, abnormal glycemic state fasting glucose, ischemic heart disease, pancytopenia and aplastic anemia.
 Conclusions: With the advancement of scientific research on saponins, it is possible to use them to produce potential drugs and dietary supplements with pharmacological effects. However, before saponins can be used for medical treatment, further research on their efficacy and safety is needed.

Various Multicharged Anions of Ginsenosides in Negative Electrospray Ionization with QTOF High-Resolution Mass Spectrometry.

When characterizing components from ginseng, we found a vast number of multicharged anions presented in the liquid chromatography-mass spectrometry (LC-MS) chromatograms. The source of these anions is unclear yet, while ginsenosides, the major components of ginseng, are the main suspected type of molecules because of their sugar moiety. Our investigation using 14 pure ginsenosides affirmed that the multicharged anions were formed by ginsenosides rather than other types of ingredients in ginseng. Various anions could be observed for each ginsenoside. These anions contain ions ([M-2H]2-, [M+Adduct]2-), as well as those formed by polymerization of at least two ginsenosides, such as [nM-2H]2-, [nM-H+Adduct]2-, and [nM-3H]3-. The presence of so different types of ions from a ginsenoside explains the reason for the large number of anions in the LC-MS analysis of ginseng. We further found that formation of [nM-2H]2- ions was influenced by the number of sugar chains: ginsenosides containing two sugar chains produced all [nM-2H]2- ion types, whereas ginsenosides containing one sugar chain did not produce [2M-2H]2-. Thus, [2M-2H]2- and [3M-2H]2- can be utilized to rapidly identify monodesmosidic and/or bidesmosidic ginsenosides as joint diagnostic anions. The position of the glycosyl radical might be the key factor affecting the formation of multicharged multimer ions from monodesmosidic ginsenosides. Consequently, three groups of ginsenoside isomers were differentiated by characteristic [nM-2H]2- anions. Using concentration-dependent characteristics and collision-induced dissociation (CID), we confirmed that [nM-2H]2- ions are non-covalently bound multimers whose aggregation has marked distinction between monodesmosidic and bidesmosidic ginsenosides, accounting for the differentiated formation of [nM-2H]2- between them. Graphical Abstract.

Surface properties of adsorption layers formed from triterpenoid and steroid saponins

Abstract Saponins are natural surfactants with non-trivial surface and aggregation properties which find numerous important applications in several areas (food, pharma, cosmetic and others). In the current paper we study the surface properties of ten saponin extracts, having different molecular structure with respect to the type of their hydrophobic fragment (triterpenoid or steroid aglycone) and the number of sugar chains (1 to 3). We found that the triterpenoid saponins Escin, Tea Saponin and Ginsenosides have area per molecule in the range between 0.5 and 0.7 nm2, and the adsorbed molecules are orientated perpendicularly to the interface. The comparison of the experimentally measured surface elasticities with theoretically estimated ones shows that the saponins with very high dilatational and shear elasticities (up to 2000 mN/m) have molecular interaction parameter in the adsorption layers which is above the threshold value for two-dimensional phase transition. In other words, the highly elastic layers are in surface condensed state, due to strong attraction between the adsorbed molecules. Furthermore, these adsorption layers have non-linear rheological response upon expansion and contraction, even at relatively small deformation. Layers from the other studied saponins (steroids and crude mixtures of triterpenoid saponins), which are unable to form strong intermolecular bonds within the adsorption layer, have zero shear elasticity and viscosity and low dilatational elasticity and viscosity, comparable in magnitude to those reported in literature for protein adsorption layers. The comparison of the results, obtained by several independent experimental methods, allowed us to formulate the conditions under which the results from different interfacial rheology tests could be compared, despite the complex non-linear response of the saponin adsorption layers.

Identification of ginsenoside markers from dry purified extract of Panax ginseng by a dereplication approach and UPLC–QTOF/MS analysis

A dry purified extract of Panax ginseng (PEG) was prepared using a manufacturing process that includes column chromatography, acid hydrolysis, and an enzyme reaction. During the manufacturing process, the more polar ginsenosides were altered into less polar forms via cleavage of their sugar chains and structural modifications of the aglycones, such as hydroxylation and dehydroxylation. The structural changes of ginsenosides during the intermediate steps from dried ginseng extract (DGE) to PEG were monitored by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectroscopy (UPLC–QTOF/MS). 22 ginsenosides isolated from PEG were used as the reference standards for determining of unknown ginsenosides and further suggesting of the metabolic markers. The elution order of 22 ginsenosides based on the type of aglycones, and the location and number of sugar chains can be used for the structural elucidation of unknown ginsenosides. This information could be used in a dereplication process for quick and efficient identification of ginsenoside derivatives in ginseng preparations. A dereplication approach helped the identification of the metabolic markers in the UPLC–QTOF/MS chromatograms during the conversion process with multivariate analyses, including principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) plots. These metabolic markers were identified by comparing with the dereplication information of the reference standards of 22 ginsenosides, or they were assigned using the pattern of the MS/MS fragmented ions. Consequently, the developed metabolic profiling approach using UPLC–QTOF/MS and multivariate analysis represents a new method for providing quality control as well as useful criteria for a similarity evaluation of the manufacturing process of ginseng preparations.

Solution Properties of Amphiphilic Telomers with Multiple Sugar Chains and Terminal Alkyl Chain

Amphiphilic telomers with multiple sugar chains and a terminal undecyl or heptadecyl chain (i.e., C(n)Am-mGEMA, where n and m represent alkyl chain lengths of 11 or 17 with a degree of polymerization of 2.0 or 3.0 for glucosyloxyethyl methacrylate (GEMA) units, respectively) were synthesized via monomeric radical telomerization in the presence of 2-aminoethanethiol hydrochloride. Surface tension, pyrene fluorescence, and dynamic light scattering were measured to characterize the solution properties of the synthesized telomers. In addition, the effects of alkyl chain length and degree of polymerization of hydrophilic GEMA units on the measured properties were evaluated by comparison with those of conventional polyoxyethylene dodecyl ether nonionic surfactants. C(n)Am-mGEMA telomers exhibited higher critical micelle concentration (CMC) values than polyoxyethylene dodecyl ether surfactants with similar number of hydrophilic groups did. The synthesized telomers are highly efficient in reducing the surface tension of water, despite the relatively large hydrophilic structures within the sugar units (GEMA). A unique behavior was observed in that adsorption at the air-water interface and solution aggregation occurred simultaneously at a concentration below CMC (as determined by the surface tension method). This suggests that aggregate formation occurs readily in solution along with the adsorption at the interface because of strong attractive interactions between multiple sugar GEMA chains. Further, aggregates formed by C(n)Am-mGEMA telomers differ depending on the number of sugar chains, i.e., an increase in the degree of polymerization of the telomers increases the size of the aggregates. This indicates that it is easier for telomers with more sugar GEMA chains to form large aggregates due to the interactions between their hydroxyl groups.

Exfoliation of Helicobacter pylori from Gastric Mucin by Glycopolypeptides from Buttermilk

In the human stomach, Helicobacter pylori, an ulcer pathogenic bacterium, colonizes the gastric mucosal layer primarily. The ability of glycopolypeptides (GPP) prepared from buttermilk to exfoliate H. pylori bound to gastric mucin was investigated. The GPP were prepared from buttermilk by digestion with trypsin, papain, pancreatin, bromelain, or pepsin. Helicobacter pylori ATCC 43504T and 43579 adhered more strongly to all of the GPP tested than to whole buttermilk, the soluble fraction of buttermilk, gastric mucin prepared from mouse stomach, or commercial pig gastric mucin. The GPP digested with trypsin, papain, or pancreatin were significantly more adherent. When the GPP concentration was 10mg/mL, bound H. pylori ATCC 43504T, 43579, and 5 clinical isolates were exfoliated markedly from immobilized porcine gastric mucin following treatment with GPP digested with trypsin or pancreatin. This ability of GPP did not correlate with sialic acid content, indicating that sialic acid content is not important in the exfoliation of this microorganism. Such an ability may depend on the structure or number of sugar chains, or the position of sialic acid. We conclude that GPP promote the exfoliation of H. pylori bound to gastric mucin and prevent the de novo adherence of this microorganism. As such, GPP are a promising food material for preventing H. pylori infection.

Expression of Pichia anomala INV1 gene in Saccharomyces cerevisiae results in two different active forms of hypoglycosylated invertase.

The Pichia anomala invertase gene (INV1) was introduced at different copy numbers into a sucrose-nonfermenting mutant of Saccharomyces cerevisiae and expressed from its own promoter sequences. The level reached in the production of invertase by the transformants (up to 540 units/10(10) cells) was in agreement with the INV1 gene dosage. Two forms of multimeric active and glycosylated invertase displaying different subcellular locations and molecular masses could be detected in the transformants. One was found to be present in the culture medium and in the periplasm, and the other could only be detected inside the cell. Each of the two heterologous forms of invertase was shown to be an oligomer composed of identical subunits. The difference found in the apparent molecular masses of their monomers (81.5 and 78.3 kDa, respectively) seems to be due to the size of their N-linked oligosaccharide chains (on average 2.4 and 1.9 kDa, respectively), since the number of sugar chains (9) and the molecular mass of the protein moiety (60.5 kDa) are identical in both forms. The shorter size of their oligosaccharides must also be the reason for the lower apparent molecular masses of the heterologous invertases when compared with the enzyme purified from P. anomala. The hypoglycosylated invertase accumulated within the cells of the transformants to an unusual level (up to 130 units/10(10) cells). Such accumulation of active enzyme inside the cells, as well as its underglycosylation, could be due to intrinsic properties of the P. anomala invertase that are determined by the particular primary structure of its protein moiety.

COUNTERCURRENT CHROMATOGRAPHIC SEPARATION OF SUGARS AND THEIR p-NITROPHENYL DERIVATIVES BY CROSS-AXIS COIL PLANET CENTRIFUGE

Sugars and their p-nitrophenyl (PNP) derivatives were separated by high-speed countercurrent chromatography using a cross-axis coil planet centrifuge (cross-axis CPC) equipped with a pair of eccentric coil assemblies. A polar two-phase solvent system composed of 1-butanol/acetic acid/water (4:1:5) was used for the separation of sucrose and fucose, and glucuronic acid (lactoid form) and galacturonic acid, while 1-butanol/ethanol/water (4:1:4) was used for the separation of free and lactoid forms of glucuronic acid. PNP-sugar derivatives such as neutral sugars, uronic acids, and amino sugars were separated with a less polar solvent system composed of n-hexane/ethyl acetate/1-butanol/methanol/water at various volume ratios. PNP-glucose derivatives were further separated according to the number of sugar chains, and five PNP-neutral sugars were resolved by adding 0.1 M sodium tetraborate in the two-phase solvent system. Overall results of experiments revealed that the cross-axis CPC is useful for the separation of polar compounds such as sugars.

Estimation of the number of O-linked oligosaccharides per heavy chain of human serum IgA1 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS) analysis of the hinge glycopeptide.

In our previous study, gas-phase hydrazinolysis was used to analyze the glycoform of the O-linked oligosaccharide of human serum IgA1. All O-linked oligosaccharide chains are known to be present in the hinge portion. However, the number of O-linked oligosaccharide chains on IgA1 remained unclear. In order to determine the number of linked sugar chains, we applied matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS) to the hinge glycopeptide prepared from human serum IgA1. MALDI-TOFMS did not show clear peaks, probably due to the microheterogeneity of the structure of each sugar chain. However, elimination of peripheral sialic acid and galactose residues by sequential treatment with neuraminidase and beta-galactosidase gave clear mass spectra with several sharp peaks. On the basis of these spectra, we conclude that IgA1 prepared from normal human serum carries different numbers of sugar chains. There are two major populations, one contains five GalNAc residues and the other four GalNAc residues. On the other hand, the hinge glycopeptide prepared from myeloma IgA1 was composed mainly of one population containing four GalNAc residues. Earlier, we reported incomplete glycosylation of IgA1 isolated from the serum of an IgA1 myeloma patient. In this experiment, the presence of four O-linked oligosaccharides per heavy chain of IgA1 from a myeloma patient was found. The reason why only four out of five sites on the hinge glycopeptide were fully glycosylated in the IgA1 from the IgA1 myeloma patient is not clear.

The glycoprotein allergen Ag-54 (Cla h II) from Cladosporium herbarum. Structural studies of the carbohydrate moiety.

The carbohydrate moiety of an important allergen, Ag-54 in Cladosporium herbarum was studied by alkaline-borohydride treatment, gel filtration, high-performance liquid chromatography, methylation analysis, gas liquid chromatography and mass spectometry. The Ag-54 protein core possessed a very limited number of sugar chains. The carbohydrate moiety consisted mainly of one large highly branched polysaccharide chain which accounted for nearly 75% of the total molecular weight of the glycoprotein. The carbohydrate moiety is made up of D-mannose and D-glucose units in pyranose form having D-galactofuranose side chains attached. Mannose is both 1,2- and 1,6-linked, while glucose is 1,4- and 1,6-linked. Some of the 1,6-linked galactofuranose side chains are bound through C-2 of the 1,6-linked mannose units, and the rest to C-3 of 1,6-linked mannose and 1,2-linked mannose units. A few oligoglucosidic chains of approximately 4 glucose units are also attached to the protein.

γ‐グルタミルトランスペプチダーゼの糖鎖構造 そのがん性変化

The structures of sugar chains of γ-glutamyl transpeptidases (γ-GTP) purified from rat liver and rat ascites AH-66 hepatoma were comparatively studied. The number of sugar chains in one molecule of hepatoma γ-GTP was approximately four times higher than that of liver γ-GTP. Only acidic sugar chains were found in liver γ-GTP, while hepatoma enzyme contained approximately 30% of neutral sugar chains which were high mannose type and complex type. The acidic sugar chains of liver γ-GTP were sialylated bi-, tri-, tetraantennary sugar chains and tetraantennary sugar chains with Galβ1→GlcNAc repeating outer chains. In contrast, no Galβ1→GlcNAc repeating outer chain was found in the acidic sugar chains of hepatoma enzyme. No bisect N-acetylglucosamine residue was found in the sugar chains of liver enzyme like liver enzymes of other animals, while some of hepatoma enzyme did contain this residue. Furthermore, some of the outer chain moieties of the complex type sugar chains of hepatoma γ-GTP did not contain galactose residue indicating that the deletion phenomena as found in the glycolipids of cancerous tissues were included in the biosynthesis of these sugar chains. These results indicate that the glycosylation of hepatoma γ-GTP is not strictly controlled as in the case of liver γ-GTP.