High-performance Gel Permeation Chromatography Research Articles

Systematic Chemical Analysis ofCrude Glycan Isolates from the Seven‑herb Decoction Quanzhenyiqitang with Anti-COPD Activity.

The classical Chinese Medicine prescription, Quanzhenyiqitang (QZYQT), containing seven tonic herbs (Shudi, Dangshen, Maidong, Baizhu, Niuxi, Fuzi, and Wuweizi) is clinically used to treat chronic obstructive pulmonary disease (COPD). Although there are studies on the pharmacological effects of QZYQT, little attention has been paid to its active carbohydrate ingredients. We performed a systematic chemical analysis of the crude glycan isolates from the seven-herb decoction (GI-QZYQT) after confirming its anti-COPD activity. GI-QZYQT could enhance lung function, reduce lung damage, and alleviate inflammatory response in mice with COPD. Moreover, two monosaccharides (fructose and glucose) and six oligosaccharides (sucrose, melibiose, 1-kestose, raffinose, mannotriose, and stachyose), accounting for 40.23% of GI-QZYQT, were discovered using hydrophilic interaction liquid chromatography-evaporative light-scattering detection. Inulin-type fructan with an average molecular weight of 2112 Da was identified using high-performance gel-permeation chromatography in combination with monosaccharide mapping analysis, accounting for 20.10% of GI-QZYQT in mass. The comparison study showed that the identified monosaccharides, oligosaccharides, and the inulin-type fructan of GI-QZYQT were mainly derived from herbs of Shudi, Dangshen, Maidong, Baizhu, and Niuxi. These findings provide crucial information on the chemical composition of GI-QZYQT, which is vital for the in-depth understanding of its bioactivity, mechanism, and product development.

Ultrasound assisted hot water extraction of polysaccharides from Taraxacum mongolicum: Optimization, purification, structure characterization, and antioxidant activity.

Ultrasound assisted hot water extraction (UAHWE) was applied to extraction of polysaccharides from Taraxacum mongolicum with hot water as extract solvent. Experimental factors in UAHWE process were optimized by response surface methodology. The optimal extraction parameters to achieve the highest Taraxacum mongolicum polysaccharides (TMPs) yield (12.08±0.14)% by UAHWE were obtained under the ultrasound power of 200W, extraction temperature of 62°C, solid-to-liquid ratio of 1:20g/mL, and extraction time of 40min, and then the crude TMPs were further purified by DEAE-52 and Sephadex G-100 chromatography to obtain a homogenous polysaccharide fraction (TMPs-1-SG). Subsequently, the structure of TMPs-1-SG was characterized by UV-vis, Fourier transform infrared spectroscopy (FT-IR), high performance gel permeation chromatography (HPGPC), high performance liquid chromatography (HPLC), scanning electron microscope (SEM), transmission electron microscopy (TEM), and Congo red test. The results display that TMPs-1-SG with an average molecular weight of 5.49 × 104Da was comprised of mannose (Man), galactose (Gal), xylose (Xyl), and arabinose (Ara) with a molar ratio of 39.85:52.61:27.14:6.30. Moreover, TMPs-1-SG did not contain a triple helix structure. Furthermore, TMPs-1-SG and TEM presented a sheet-like, rod-shaped, and irregular structure. Finally, the antioxidant activity of TMPs-1-SG was evaluated by in vitro experiment. The IC50 values of scavenging DPPH and OH radicals for TMPs-1-SG achieved 0.71mg/mL and 0.75mg/mL, respectively. The findings can provide an effective method for extracting polysaccharides from natural resources.

Characterization and anti-aging activities of polysaccharide from Rana dybowskii Guenther.

Introduction: Rana dybowskii Guenther (RDG), as a traditional Chinese medicine, has been shown to have antioxidant effects. However, studies on the anti-aging effect of RDG are still limited. Methods: In this study, we prepared polysaccharides from the skin of RDG (RDGP) by hot water extraction, alcohol precipitation, ion-exchange chromatography and gel chromatography. The proteins were removed using the Sevage method in combination with an enzymatic method. The structural features were analyzed using high-performance gel permeation chromatography, β-elimination reaction and Fourier transform infrared spectra. The anti-aging effect of RDGP was investigated by using D-Gal to establish an aging model in mice, and pathological changes in the hippocampus were observed under a microscope. Results: We obtained the crude polysaccharide DGP from the skin of RDG, with a yield of 61.8%. The free protein was then removed by the Sevage method to obtain DGPI and deproteinated by enzymatic hydrolysis combined with the Sevage method to further remove the bound protein to obtain the high-purity polysaccharide DGPII. Then, DGPIa (1.03 × 105Da) and DGPIIa (8.42 × 104Da) were obtained by gel chromatography, monosaccharide composition analysis showed that they were composed of Man, GlcA, GalNAc, Glc, Gal, Fuc with molar ratios of 1: 4.22 : 1.55: 0.18 : 8.05: 0.83 and 0.74 : 1.78: 1: 0.28: 5.37 : 0.36, respectively. The results of the β-elimination reaction indicated the presence of O-glycopeptide bonds in DGPIa. The Morris water maze test indicated that mice treated with DGPIIa exhibited a significantly shorter escape latency and increased time spent in the target quadrant as well as an increase in the number of times they traversed the platform. Pathologic damage to the hippocampus was alleviated in brain tissue stained with hematoxylin-eosin. In addition, DGPIIa enhanced the activities of SOD, CAT, and GSH-Px and inhibited the level of MDA in the serum and brain tissues of aging mice. Discussion: These results suggest that RDGP has potential as a natural antioxidant and provide useful scientific information for anti-aging research.

The Physicochemical Properties and Antioxidant Activities of the Hawthorn Pectin Extracted Using Ultra-High Pressure Assisted Acid Extraction (UPAAE).

This study aims to investigate the positive effects of ultra-high pressure assisted acid extraction (UPAAE) on both physicochemical properties and antioxidant activities of hawthorn pectin. The basic indicators, structure characterization, and antioxidant activities were measured, which could indicate the disadvantages and advantages among traditional water extraction (WE), acid extraction (AE), and UPAAE. The results show that the hawthorn pectin of UPAAE has a decrease in esterification degree, protein content, and total polyphenols, but has an increase in total galacturonic acid aldehyde compared to the hawthorn pectin of AE. In the Fourier Transform Infrared Spectroscopy (FT-IR) and Scanning Electron Microscopy (SEM) analyses, the hawthorn of UPAAE has typical pectin absorption peaks in the FT-IR spectrum and a distinct layered structure in the SEM surface image. The ion chromatography profiles show that the molar ratio of galacturonic acid to arabinose in the hawthorn pectin of UPAAE increases and 5.50 μg/mg ribose appears compared to the pectin of AE and WE. The high performance gel permeation chromatography (HPGPC) profile indicates that the molecular weight distribution of hawthorn pectin of UPAAE is more concentrated and has the highest molecular weight compared to the pectin of the other two extraction methods. In the vitro antioxidant activity analysis, the pectin of UPAAE exhibits the highest scavenging rate against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals (93.70%), which is close to the scavenging rate of vitamin C (96.30%). These findings demonstrated that UPAAE is a more efficient and environmentally friendly method for pectin extraction from hawthorn. It is also an effective way to enhance its antioxidant activity, which has great application prospects in the food industries.

Effects of Auricularia auricula-judae (Bull.) Quél. polysaccharide acid hydrolysate on glucose metabolism in diabetic mice under oxidative stress

BackgroundOxidative stress can lead to uncontrolled glucose metabolism and, thus, diabetes. Auricularia auricula-judae (Bull.) Quél. polysaccharides possess biological activities, such as antioxidant and hypoglycemic effects, but their mechanism of their acid hydrolysates on oxidative stress-injured glucose metabolism disorders is unclear. PurposeUsing diabetic mice, we investigated the effect of the acid hydrolysate of polysaccharides from Auricularia auricula-judae (Bull.) Quél. on improving diabetes. Study design and MethodsThe structural information of sample polysaccharides was measured by high performance gel permeation chromatography, nuclear magnetic resolution, and high performance liquid chromatography. The diabetic model was established by intraperitoneal injection of streptozotocin. For eight consecutive weeks, the mice were orally administered sample polysaccharides (100, 200, and 300 mg/kg b.w. per day) for intervention. The improvement effect of the samples on diabetes was explored by detecting the changes in biochemical indicators in mice, and the underlying mechanism was studied by transcriptomic and metabolomic analysis. ResultsThe results showed that acid hydrolysate of Auricularia auricula-judae (Bull.) Quél. polysaccharides consisted mainly of mannose, xylose, glucuronic acid, and glucose; its weight-averaged molecular weight was 6.3842 × 104 Dalton, its number average molecular weight was 2.9594 × 104 Dalton; and the molecule contained α-Glc(1→4)-, β-Glc(1→3)-, and β-Man(1→4)-linked glycosidic bonds. A total of 100 mg/kg b.w. per day sample was the best intervention concentration. After eight weeks of intervention, the sample polysaccharides significantly reduced dynamic blood glucose and serum lipids, enhanced antioxidant enzyme activities, promoted glucagon like peptide-1 and insulin secretion, improved insulin sensitivity and alleviated insulin resistance in diabetic mice. Transcriptomic and metabolomic analyses showed that sample polysaccharides was able to ameliorate disorders of glucose metabolism by modulating gene expression such as glucokinase; and modulate the state of oxidative stress in mice in vivo by regulating the glutathione metabolism pathway. ConclusionAcid hydrolysate of Auricularia auricula-judae (Bull.) Quél. polysaccharides improved glucose metabolism disorders by slowing down the oxidative stress injury in mice, thereby alleviating diabetes. This study provided a basis for determining the underlying mechanism of the antidiabetic effect of Auricularia auricula-judae (Bull.) Quél. polysaccharides, which would significantly improve the deep development and application of these materials in diabetes control.

A chondroitin sulfate purified from shark cartilage and bovine serum albumin interaction activity

Chondroitin sulfate (CS) was extracted and purified from shark cartilage, and its interaction with bovine serum albumin (BSA) were studied. The content of chondroitin sulfate in shark cartilage was 29.97 % using the 1,9-dimethyl-methylene blue method. The molecular weight of CS was determined to be 62.464 kDa by high-performance gel permeation chromatography. UV and FT-IR spectroscopy identified the characteristics of CS and its functional group information. NMR spectroscopy and disaccharide derivatization revealed that CS was predominantly composed of disulfated disaccharides, specifically ΔDi4,6S. Fluorescence quenching experiments indicated that the interaction between CS and BSA exhibited static quenching, with a binding site number of 1. The binding process was primarily mediated by van der Waals forces and hydrogen bonds. Furthermore, synchronous and 3D fluorescence spectroscopy demonstrated that CS had minimal impact on the polarity and hydrophobicity of the microenvironment surrounding Tyr and Trp residues. UV–vis absorption and circular dichroism (CD) spectroscopy demonstrated the altered structure of BSA. The molecular docking analysis revealed that CS formed hydrogen bonds and salt bridges with BSA, predominantly binding to the IIA substructure domain of BSA. Investigating the interaction between CS and BSA holds the potential for enhancing its applications in drug delivery and tissue engineering endeavors.

Structure characterization and cryoprotective activity of an exopolysaccharide from Pseudoalteromonas sp. LP6-12-2

Non-toxic natural antifreeze agents are urgently needed in many fields. Exopolysaccharides produced by psychrophilic bacteria from harsh habitats, such as Antarctica, to battle cold environments are a significant source of biological antifreeze agents. In this study, a homogeneous exopolysaccharide (EPS0-1) was extracted from Antarctica Pseudoalteromonas sp. LP6-12-2. Using High-performance gel permeation chromatography and High-efficiency anion chromatogram, respectively, the homogeneity, molecular weight, and monosaccharide composition of EPS0-1 were examined. EPS0-1 with a molecular weight (Mw) of 4351.31 kDa was an α-configuration polysaccharide containing uronic acid, amino and sulfuric acid groups, and composed of galactosamine hydrochloride, rhamnose, glucosamine hydrochloride, galactose, glucose, mannose, galacturonic acid, and glucuronic acid with rates of 1.40:9.40:4.90:32.0:37.2:6.50:4.10:4.40, respectively. Congo red test and I2-KI reaction found that the triple helix structure of EPS0-1 polysaccharide chain was weakly bound internally, with a longer side chain and/or more branches. The smooth and dense surface of EPS0-1 under Atomic force microscopy and Scanning electron microscopy revealed that it was large polysaccharide particles. Freeze-thaw cycle and freezing point determination experiment showed that EPS0-1 may significantly increase E. Coli's freezing resistance and decrease the solution's freezing point, respectively. EPS0-1 was capable of increasing the viscosity of the solution and displaying a high degree of thermal stability, which was proved by the rheological properties. And it was not hazardous to human normal liver cell line (LO2) in a specific concentration range. Therefore, the findings of this investigation established the basis for EPS0-1 as a potential antifreeze and food functional component.

Polysaccharide extraction from Myxopyrum smilacifolium trunk and its antioxidant capacity

As a traditional medicinal plant in Vietnam, Myxopyrum smilacifolium has been used for a long history to treat cough, nervous disorders, numbness, rheumatism, and cephalalgia. Nevertheless, reports on the antioxidant activity and extraction of M. smilacifolium polysaccharides are still very rare. This study was designed to extract a high yield of polysaccharides from the M. smilacifolium trunk and characterize it. As a result, the maximum yield of the polysaccharides of 5.13 ± 0.05% was obtained with an extraction time of three hours, extraction temperature of 100 oC, the ratio of water to sample of 1:50 as extraction solution, extraction number of times 3, and the ratio of ethanol to extract volume 5:1 (v/v). Polysaccharides was characterized by Fourier transform infrared spectroscopy (FT-IR) and high-performance gel-permeation chromatography (HPGPC). The average molecular weight of extracted polysaccharides was around 3.78 × 105 Da. In vitro assays dan impressive antioxidant activities of the extracted polysaccharides, containing 0.2423 ± 0.0028 mg GA/g or 0.2142 ± 0.0007 μmol AS/g. The IC50 values of polysaccharides in the DPPH and ABTS methods were 0.89 mg/mL and 3.85 mg/mL, respectively. These findings exhibited the potential for application or further research and development of polysaccharides from Myxopyrum smilacifolium.

Optimized Extraction and Fermentation of Antioxidant Polysaccharides from Polygonatum odoratum

Purpose: We wished to determine the optimal method for the extraction of polysaccharides with antioxidant activities from <i>Polygonatum odoratum</i>.Methods: Single-factor and orthogonal tests were applied to identify the best parameters for water extraction and fermentation. The solid-liquid ratio, temperature, and duration of water extraction were determined according to the total polysaccharide content. The fermentation temperature, agitation speed, inoculation amount, and fermentation time were investigated according to the optical density of Saccharomyces cerevisiae. The molecular weight distributions of water- and fermentation-extracted polysaccharides were determined by high performance gel permeation chromatography. The antioxidant activities of the polysaccharides were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) ABTS radical scavenging capacities, and ferric reducing antioxidant power (FRAP).Results: The optimum water extraction parameters were 80°C for 2.5 h with a 1:60 solid-liquid ratio. For fermentation, an 8% extraction volume was inoculated with <i>S. cerevisiae</i>. Optimal fermentation occurred at 28°C for 28 h, with an agitation speed of 180 r/min. The molecular weights of water- and fermentation-extracted polysaccharides were 2.067×10<sup>4</sup> and 9.475×10<sup>3</sup> Da, respectively. The DPPH, ABTS, and FRAP results demonstrated an obvious increase in antioxidant activity after fermentation.Conclusion: The fermentation extraction of <i>P. odoratum</i> may provide valuable materials for future cosmetic use.

Integrating multi-index determination coupled with hierarchical cluster analysis to evaluate the quality consistency of PVE30, an anti-HSV "glycoprotein" macromolecule of Prunellae Spica.

Prunellae Spica (PS), derived from the dried fruit spikes of Prunella vulgaris L., is a traditional Chinese medicinal herb. Our previous studies found that PVE30, a water-extracting ethanol-precipitating "glycoprotein" macromolecule of PS, was a potential anti-herpes simplex virus (HSV) candidate. However, due to the complex structure and diverse bioactivity of the "glycoprotein", ensuring its quality consistency across different batches of PVE30 becomes particularly challenging. This poses a significant hurdle for new drug development based on PVE30. Our study aimed to integrate multi-index determination coupled with hierarchical cluster analysis (HCA) to holistically profile the quality consistency of "glycoprotein" in PVE30. High-performance gel permeation chromatography with refractive index detector (HPGPC-RID) was used to characterise the molecular weight (Mw) distribution, HPLC-PDA was used to quantitatively analyse the composed monosaccharides and amino acids, and UV-VIS was used to quantify the contents of polysaccharides and proteins. Qualitative and quantitative consistency was analysed for each single index in 16 batches of PVE30, and a 16 × 38 data matrix, coupled with HCA, was used to evaluate the holistic quality consistency of PVE30. The newly developed and validated methods were exclusive, linear, precise, accurate, and stable enough to quantify multi-indexes in PVE30. Single-index analysis revealed that 16 batches of PVE30 were qualitatively consistent in Mw distribution, polysaccharides and proteins, and the composition of composed monosaccharides and amino acids but quantitatively inconsistent in the relative contents of some "glycoprotein" macromolecules, as well as the composed monosaccharides/amino acids. HCA showed that the holistic quality of PVE30 was inconsistent, the inconsistency was uncorrelated with the regions where PS was commercially collected, and the contents of 17 amino acids and 2 monosaccharides contributed most to the holistic quality inconsistency. Multi-index determination coupled with HCA was successful in evaluating the quality consistency of PVE30, and the significant difference in quantitative indices was not caused by the origin of PS. The cultivating basis should be confirmed for PVE30-based new drug development.

Preparation, structural characterization and in vitro activity of ginger polysaccharide

AbstractA polysaccharide was extracted from the fresh ginger by water extraction and alcohol precipitation. A homogeneous polysaccharide was obtained by Sevage deproteinization, dialysis, DEAE-52 ion exchange chromatography and Sephadex G-100 gel column chromatography. The molecular weight of the ginger polysaccharide was determined by high performance gel permeation chromatography. The monosaccharide composition, scanning electron microscope and other physicochemical parameters of the ginger polysaccharide were further determined. The spectral properties of the ginger polysaccharide were investigated by ultraviolet and infrared spectroscopy. On this basis, methylation analysis, 1D and 2D NMR analysis were used to investigate its possible chemical structure. The results showed that the ginger polysaccharide group was divided into neutral polysaccharide components with high homogeneity, Mw = 35.52 kDa, Mn = 216.56 kDa and the sugar content was 93.5%. The ginger polysaccharide is mainly composed of Fuc, Ara, Rha, Gal, Glc, Xyl, Man, Gal-UA and Glc-UA, the amount of substance ratio was 1.01:1.96:9.54:14.82:64.20:3.84:2.07:1.41:1.16. To study the structure–activity relationship of ginger polysaccharide, acetylated ginger polysaccharide and phosphate ginger polysaccharide were prepared by chemical methods in this study. The results of antioxidant activity in vitro showed that the ginger polysaccharide and its derivatives had good biological activity in freeing ABTS + radical scavenging, freeing hydroxyl radical scavenging, freeing DPPH radical scavenging and reducing ability. Interestingly, the results showed that the ginger polysaccharide had a certain inhibitory effect on the growth of E. coli in vitro. In general, this study provides a certain experimental basis and reference for the further research on the activity of ginger polysaccharide and its derivatives. Graphical Abstract

Degradation Product of Sea Cucumber Polysaccharide by Dielectric Barrier Discharge Enhanced the Migration of Macrophage In Vitro.

This study investigated the effect of dielectric barrier discharge (DBD) on sea cucumber polysaccharide (SP-2) and evaluated its anti-inflammatory properties. The SP-2 was depolymerized by applying an input voltage of 60~90 V for 3~9 min. The features of the products were examined using high-performance gel permeation chromatography, HPLC-PAD-MS, and the Fourier transform infrared (FTIR) spectrum. The anti-inflammatory properties of the product were investigated by measuring nitric oxide (NO) release, ROS accumulation, and cell migration using RAW264.7 cells (LPS-induced or not-induced). The results showed SP-2 depolymerized into homogeneous and controllable-size oligosaccharide products. The depolymerized ratio can reach 80%. The results of the measurement of reducing sugars indicate that SP-2 was cleaved from within the sugar chain. The SP-2 was deduced to have a monosaccharide sequence of GlcN-Man-Man-Man-Man-Man based on the digested fragment information. The depolymerization product restrained the release of NO and the accumulation of ROS. By testing the RAW264.7 cell scratch assay, it was found that it enhances the migration of immune cells. DBD degradation of SP-2 leads to homogeneous and controllable-size oligosaccharide products, and this technique can be used for polysaccharide structure analysis. The depolymerized product of SP-2 has an anti-inflammatory capability in vitro.

Structural characterization of a low-molecular-weight polysaccharide from Plumula Nelumbinis and evaluation of its antioxidant and anti-inflammatory activity

Plumula Nelumbinis polysaccharides (PNP) exhibit remarkable bioactivities, however their structure-activity relationship is still not clear. Herein, a novel low-molecular-weight polysaccharide (LPNP) was isolated and refined from Plumula Nelumbinis. The structure of LPNP was characterized using High-performance gel permeation chromatography (HPGPC), monosaccharide composition analysis, NMR spectroscopy, and methylation analysis. The results revealed that LPNP had a molecular weight of 11.6 kDa, consisting six monosaccharides. Glycosidic linkages →3)-β-L-Araf-(1→, →5)-β-L-Araf-(1→, T-β-L-Araf-(1→, →3)-β-D-Galp-(1→, →4)-α-D-Glcp-(1→, →4)-α-D-Xylf-(1→, T-α-D-Xylp-(1→, and T-α-D-Glcp-(1→ were the main types of linkages in LPNP. LPNP could strongly protect RAW264.7 cells from H2O2-induced damage by preserving stable cell viability, reducing reactive oxygen species (ROS) and malondialdehyde (MDA) production and lactate dehydrogenase (LDH) secretion, enhancing catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activity, and increasing glutathione (GSH) content. LPNP inhibited the activation of the MAPK/NF-κB cell pathway, thereby decreasing the expression of NO, INF-γ, TNF-α, IL-6, and IL-1β. LPNP could be exploited as a prospective source of natural substances in functional foods or pharmaceuticals.

Structure characterization of novel heteropolysaccharides from Pteridium revolutum with antioxidant and antiglycated activities.

This study aims to analysis the structures of polysaccharides isolated from Pteridium revolutum and their antioxidant and antiglycated activities. Three novel water-soluble heteropolysaccharides, named PRP0, PRP1, and PRP2, were isolated from P. revolutum. The average molecular weight was determined by high performance gel permeation chromatography analysis as 1.04×106, 8.39×105, and 7.37×105 Da, respectively. Their structures were characterized using physicochemical and spectroscopic methods. The antioxidant and antiglycated activities were assayed in vitro. PRP0, PRP1, and PRP2 consist of l-Ara, l-Rha, d-Man, d-Xyl, d-Fuc, d-Gal, and d-Glc in different proportions. PRP1 mainly has a backbone of (1→3,6)-linked d-Man and (1→3)-linked d-Gal on main chain. PRP2 is mainly composed of (1→2,4)-linked d-Man and (1→3)-linked d-Gal on main chain. All polysaccharides have strong scavenging power on 2,2-difenil-1-picril-hidrazil and hydroxyl radicals and significantly antiglycated activity in Bovine serum albumin-Glucose model, which showing that the polysaccharides have potential application value on the functional food.

Structural characterization, antioxidant and antimicrobial activities of polysaccharide from Akebia trifoliata (Thunb.) Koidz stem

Polysaccharides have a variety of beneficial pharmacological impact on human health. Akebia trifoliata (Thunb.) Koidz. has promising development prospects as a food resource with medicinal value. The aim of this study was to investigate the structural characterization, antioxidant, and antibacterial properties of A. trifoliata (Thunb.) Koidz polysaccharides (ATKPs). ATKP-II was purified from ATKP by DEAE-cellulose column with NaCl solution as eluent. ATKP and ATKP-II structures were characterized by high performance gel permeation chromatography, gas chromatography, ultraviolet-visible, Fourier transform infrared spectroscopy, thermogravimetry analysis and scanning electron microscopy. ATKP and ATKP-II were primarily composed of rhamnose, arabinose, xylose, mannose, glucose, and galactose in a molar percent of 1.6: 22.1: 3.6: 6.3: 55.7: 10.7, and 0.5: 22.1: 3.7: 10.2: 42.1: 21.4, respectively. Their structure may contain β-D-glucopyranose. The thermogravimetry analysis showed that ATKP and ATKP-II have good thermal stability at 230 °C and 200 °C, respectively. ATKP had the best antioxidant activities for 2, 2-diphenyl-1-picrylhydrazyl, hydroxyl, and superoxide free radical scavenging activities in vitro, and reducing ability than that of the purified polysaccharides. Moreover, ATKP was demonstrated an appreciable in vitro antibacterial activity, against Staphylococcus aureus, Bacillus subtilis, Salmonella, Penicillium italicum, Rhizopus and Aspergillus niger, but showed no activity against Escherichia coli and Saccharomycetes. These results demonstrated that ATKP displayed excellent antioxidant and antibacterial activities. This study provides a basis for the development and utilization in ATKP.

Inonotus obliquus polysaccharide are linear molecules that alter the abundance and composition of intestinal microbiota in Sprague Dawley rats.

The macromolecular polysaccharide Inonotus obliquus polysaccharide (IOP) is composed of various monosaccharides, and it could modulate the composition and diversity of intestinal flora. However, its impact on the intestinal flora in rats of different genders remains unclear. Therefore, this study aims to investigate the structural changes of IOP and its effects on the intestinal flora after administration in male and female rats. In this study, the molecular weight and purity of IOP were analyzed by high-performance gel permeation chromatography (HPGPC) and phenol sulfuric acid method, and NMR was used to confirm the chemical structure of IOP. Sex hormone [testosterone (T) and estradiol (E2)] levels and intestinal microbial changes were detected by enzyme-linked immunosorbent assay (ELISA) and 16S rRNA, respectively, after gavage of IOP (100 mg/kg) in male and female Sprague Dawley (SD) rats. HPGPC analysis showed that the average molecular weight (Mw) of IOP was 4,828 Da, and the total sugar content of the purified IOP was 96.2%, indicating that the polysaccharide is of high purity. NMR revealed that IOP is a linear macromolecule with an α-D-type glucose backbone. The results of ELISA and 16S rRNA showed that the IOP increased the abundance of beneficial bacteria, such as Clostridia_UCG-014 and Prevotellaceae_NK3B31, and reduced that of harmful bacteria, such as Colidextribacter and Desulfobacterota in the intestine of both male and female rats, and IOP changed the levels of sex hormones in male and female rats. Further analyses revealed that the increase in alpha diversity was higher in male than female rats. α diversity and β diversity revealed a significant difference in the composition of cecal microbiota between male and female rats in the control group, but IOP intake reduced this difference. Meanwhile, α analysis revealed a change in the composition of bacterial flora was more stable in male than female rats. This study enhances our comprehension of the IOP structure and elucidates the alterations in intestinal flora following IOP administration in rats of varying genders. Nonetheless, further investigation is warranted to explore the specific underlying reasons for these discrepancies.

The Separation, Purification, Structure Identification, and Antioxidant Activity of Elaeagnus umbellata Polysaccharides.

In order to investigate the antioxidant activity of Elaeagnus umbellata polysaccharides, the physicochemical characteristics of purified Elaeagnus umbellata polysaccharides (EUP, consisting of two fractions, EUP1 and EUP2) were investigated using UV spectrophotometry, high-performance liquid chromatography (HPLC), high-performance gel permeation chromatography (HPGPC), and Fourier transform infrared spectroscopy (FT-IR). This revealed that EUP1 and EUP2 were acidic polysaccharides with an average molecular weight (MW) of 63 and 38 kDa, respectively. EUP1 mainly consisted of L-rhamnose and D-galactose in a molar ratio of 2.05:1, and EUP2 consisted of D-mannose, L-rhamnose, D-galactose, and D-arabinose in a molar ratio of 2.06:1:2.78:1. Furthermore, EUP exhibited considerable antioxidant potential for scavenging hydroxyl, superoxide anion, DPPH, and ABTS radicals. Therefore, EUP can be developed as a potential antioxidant for the functional food or pharmaceutical field.

Characterization of structure and antioxidant activity of exopolysaccharides from endophytic Lysinibacillus sphaericus Ya6 under acid-base stress

In this study, the structure and antioxidant activity of exopolysaccharides (EPS) produced by Lysinibacillus sphaericus Ya6 under different acid-base stresses were investigated. The structure of EPS was analyzed by UV-visible and mid-IR, and the monosaccharide composition and the molecular weight were determined by High-performance liquid chromatography (HPLC) and high-performance gel permeation chromatography respectively. The findings indicated that EPS produced by Lysinibacillus sphaericus Ya6 was a homogenous polysaccharide with a molecular weight of 79.25 kDa that was composed of mannose and glucose. The molecular weight and monosaccharide composition of polysaccharides changed under acid-base stress conditions. With the addition of the acid-base stress level, the logarithmic growth period of Lysinibacillus sphaericus Ya6 was prolonged for a short while, but later growth rates and EPS production under stress conditions were all higher than under the control. In vitro determinations found that acid-base stress boosted the antioxidant activity of EPS. Particularly under conditions of alkaline stress, DPPH radical scavenging capacity and superoxide anion scavenging of EPS were higher than those of the control, and their capacities to scavenge DPPH was increased from 39.30% (the control) to 95.65% at pH 10.0, which was close to ascorbic acid's (95.77%) as a positive control.

Characterization of biologically active exopolysaccharide produced by Streptomyces sp. NRCG4 and its anti-Alzheimer efficacy: in-vitro targets

BackgroundExopolysaccharides are extremely powerful molecules with a wide range of uses in pharmaceuticals due to their structural and compositional complexity. Marine microorganisms often produce bioactive substances with novel functions and structures because of their special living conditions. Polysaccharides from marine microorganisms are of interest to new drug discovery. ResultsThe current research focused on the isolation of bacteria from Red Sea, Egypt, that have the ability to produce a new natural exopolysaccharide in order to be examined in treating Alzheimer’s illness to obviate side effects of synthetic drugs. Properties of exopolysaccharide (EPS) produced by an isolated Streptomyces strain were investigated for its capability to play as anti-Alzheimer. This strain was identified morphologically, physiologically, and biochemically and actually was confirmed by molecularly 16S rRNA analysis as Streptomyces sp. NRCG4 with accession number MK850242. The produced EPS was fractionated by precipitation 1:4 volumes of chilled ethanol and the third major fraction (1:3) listed as NRCG4, and then the functional groups, MW, and chemical evaluation have been detected via Fourier-transform infrared (FTIR), high-performance gel permeation chromatography (HPGPC), and high-performance liquid chromatography (HPLC). The findings showed that NRCG4 was an acidic EPS composed of mannuronic acid, glucose, mannose, and rhamnose in a molar ratio of 1.2:1.5:2.8:1.0, respectively. NRCG4 Mw was determined to be 4.25 × 105 gmol−1 and the Mn to be 1.97 × 105 gmol−1. Also, the NRCG4 included uronic acid (16.0%) and sulfate (0.0%), but no protein was found. In addition, antioxidant and anti-inflammation activity was measured through various methods. This study confirmed that NRCG4 exopolysaccharide exerted anti-Alzheimer’s characters via inhibition of cholinesterase and tyrosinase as well as anti-inflammatory and antioxidant abilities. Additionally, it occurred a potential role in the suppression of Alzheimer’s disease risk factors through its antioxidant (metal chelation, radical scavenging capability), anti-tyrosinase and anti-inflammatory characteristics. The anti-Alzheimer’s disease efficacy of NRCG4 exopolysaccharide may be assigned to its unique determined chemical composition. ConclusionsThe present study highlighted those exopolysaccharides could be harnessed to improve pharmaceutical industry (anti-Alzheimer, anti-tyrosinase, anti-inflammatory, and antioxidant agents).

Polysaccharide isolated from wax apple suppresses ethyl carbamate-induced oxidative damage in human hepatocytes.

Wax apple (Syzygium samarangense) has received growing research interest for its high nutritional and medicinal value due to its constituents such as polysaccharide, organic acids, flavonoids, minerals, and other substances. In this study, wax apple polysaccharide (WAP) was isolated from this plant and its protective effect against ethyl carbamate (EC)‍-induced oxidative damage was evaluated in human hepatocytes (L02 cells). Firstly, a series of analyses such as high-performance liquid chromatography (HPLC), high-performance gel permeation chromatography (HPGPC), Fourier transform infrared spectroscopy (FT-IR), gas chromatography/mass spectrometry (GC/MS), and 1H and 13C nuclear magnetic resonance (NMR) were conducted to identify the structure of WAP. Thereafter, in vitro cell experiments were performed to verify the protective effects of WAP against EC-induced cytotoxicity, genotoxicity, and oxidative damage in L02 cells. Our results revealed that WAP is composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, arabinose, and fucose in a molar ratio of 2.20:‍3.94:‍4.45:‍8.56:‍8.86:‍30.82:‍39.78:‍1.48. Using a combination of methylation and NMR spectroscopic analysis, the primary structure of WAP was identified as Araf-(1→, Glcp-(1→, →2)‍-Araf-(1→, →3)‍-Galp-(1→, →3)‍-Araf-‍(1→, and →6)‍-Galp-‍(1→. Cell experiments indicated that WAP exhibited significant protective effects on EC-treated L02 cells via suppressing cytotoxicity and genotoxicity, reducing reactive oxygen species (ROS) and O2•- formation, as well as improving mitochondrial membrane potential (MMP) and glutathione (GSH). In a nutshell, WAP has the potential as an important therapeutic agent or supplement for hepatic oxidative damage. Meanwhile, further studies are needed to prove the above effects in vivo at the biological and clinical levels.