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Human gut Bacteroides and Parabacteroides species play crucial roles in human health and are known for their capacity to utilize diverse polysaccharides. Understanding how these bacteria utilize medicinal polysaccharides is foundational for developing polysaccharides-based prebiotics and drugs. Here, we systematically mapped the utilization profiles of 20 different medicinal polysaccharides by 28 human gut Bacteroides and Parabacteroides species. The growth profiles exhibited substantial variation across different bacterial species and medicinal polysaccharides. Ginseng polysaccharides promoted the growth of multiple Bacteroides and Parabacteroides species; in contrast, Dendrobium polysaccharides selectively promoted the growth of Bacteroides uniformis. This distinct utilization profile was associated with genomic variation in carbohydrate-active enzymes, rather than monosaccharides composition variation among medicinal polysaccharides. Through comparative transcriptomics and genetical manipulation, we validated that the polysaccharide utilization locus PUL34_Bu enabled Bacteroides uniformis to utilize Dendrobium polysaccharides (i.e. glucomannan). In addition, we found that the GH26 enzyme in PUL34_Bu allowed Bacteroides uniformis to utilize multiple plant-derived mannan. Overall, our results revealed the selective utilization of medicinal polysaccharide by Bacteroides and Parabacteroides species and provided insights into the use of polysaccharides in engineering the human gut microbiome., Competing Interests: Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Natural polysaccharides attract scientists and industries' interest with diverse applications in biomaterials, immune regulation, gut microbiota regulation, food additives, and more. Nevertheless, the absence of standardized purity criteria created significant challenges in characterizing polysaccharides, leading to varied and complicated structures, and impeding progress in research and development. Consequently, it is essential to obtain samples that adhere to high-level and unified purity standards for effective polysaccharide research. Herein we review current methods for assessing polysaccharide purity, emphasizing a commonly neglected impurity: mixtures of polysaccharides with similar molecular sizes. Such a mixture, previously regarded as a single sample, can present a much more complex and ambiguous chemical structure. To address this challenge, we propose enhancing the purity criteria to identify these mixtures. Briefly, the molecular size changes are monitored following hydrolysis with diagnostic enzymes which are identified through methylation and monosaccharide analysis. A pure polymer will exhibit a complete shift in its HPGPC peak, while a polymer mixture will display residual peaks. Establishing standardized criteria for assessing polysaccharide quality represents a significant milestone in the industrialization of these important biomolecules. By enabling accurate, consistent, reliable characterization, this advancement lays the groundwork for broader polysaccharide applications in pharmaceuticals, food, materials, and beyond., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

This study investigated the structural composition and antioxidant properties of pectic polysaccharides extracted from Rohdea japonica (Thunb.) Roth. Pectins, which belong to a complex category of acidic polysaccharides, possess a wide range of biological effects stemming from their distinctive structural domains. The polysaccharides were extracted using water, and were subsequently purified through ion exchange and gel permeation chromatography. In order to elucidate their structural features, Fourier Transform Infrared Spectroscopy and Nuclear Magnetic Resonance techniques were applied. Two specific polysaccharides, WRJP-A2a and WRJP-A3b, with molecular weights of 42.7 kDa and 64.1 kDa, respectively, were identified to contain varying proportions of homogalacturonan, rhamnogalacturonan I, and rhamnogalacturonan II domains. Regarding antioxidant capacity, WRJP-A3b exhibited superior scavenging capabilities against DPPH, ABTS, and hydroxyl radicals, potentially attributed to its higher galacturonic acid content and abundance of homogalacturonan domains. These results enhance our comprehension of the structure-activity interplay of pectic polysaccharides sourced from Rohdea japonica (Thunb.) Roth and their potential utility in the healthcare and functional food sectors.

Autophagy is a vital physiological process that maintains intracellular homeostasis by removing damaged organelles and senescent or misfolded molecules. However, excessive autophagy results in cell death and apoptosis, which will lead to a variety of diseases. Galectins are a type of animal lectin that binds to β-galactosides and can bind to the cell surface or extracellular matrix glycans, affecting a variety of immune processes in vivo and being linked to the development of many diseases. In many cases, galectins and autophagy both play important regulatory roles in the cellular life course, yet our understanding of the relationship between them is still incomplete. Galectins and autophagy may share common etiological cofactors for some diseases. Hence, we summarize the relationship between galectins and autophagy, aiming to draw attention to the existence of multiple associations between galectins and autophagy in a variety of physiological and pathological processes, which provide new ideas for etiological diagnosis, drug development, and therapeutic targets for related diseases., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Two pectic polysaccharides (WRSP-A2b and WRSP-A3a) have been obtained from Radix Sophorae Tonkinensis and comparatively investigated in terms of their physical properties and antioxidant activities. Monosaccharide composition, FT-IR, NMR and enzymatic analyses indicate that both WRSP-A2b (13.6 kDa) and WRSP-A3a (44.6 kDa) consist of homogalacturonan (HG), rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II) domains, with mass ratios of 0.9:1.8:1 and 2.3:2.9:1, respectively. The RG-I domains were further purified and characterized. Results show that WRSP-A2b contains a highly branched RG-I domain, primarily substituted with α-(1→5)-linked arabinans, whereas WRSP-A3a contains a small branched RG-I domain mainly composed of β-(1→4)-linked galactan side chains. WRSP-A3a exhibits stronger antioxidant activity in scavenging different radicals than WRSP-A2b, a finding that may be due to its higher content of GalA residues and HG domains. Our results provide useful information for screening natural polysaccharide-based antioxidants from Radix Sophorae Tonkinensis., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Galectin-3 (Gal-3) has a long, aperiodic, and dynamic proline-rich N-terminal tail (NT). The functional role of the NT with its numerous prolines has remained enigmatic since its discovery. To provide some resolution to this puzzle, we individually mutated all 14 NT prolines over the first 68 residues and assessed their effects on various Gal-3-mediated functions. Our findings show that mutation of any single proline (especially P37A, P55A, P60A, P64A/H, and P67A) dramatically and differentially inhibits Gal-3-mediated cellular activities (i.e., cell migration, activation, endocytosis, and hemagglutination). For mechanistic insight, we investigated the role of prolines in mediating Gal-3 oligomerization, a fundamental process required for these cell activities. We showed that Gal-3 oligomerization triggered by binding to glycoproteins is a dynamic process analogous to liquid-liquid phase separation (LLPS). The composition of these heterooligomers is dependent on the concentration of Gal-3 as well as on the concentration and type of glycoprotein. LLPS-like Gal-3 oligomerization/condensation was also observed on the plasma membrane and disrupted endomembranes. Molecular- and cell-based assays indicate that glycan binding-triggered Gal-3 LLPS (or LLPS-like) is driven mainly by dynamic intermolecular interactions between the Gal-3 NT and the carbohydrate recognition domain (CRD) F-face, although NT-NT interactions appear to contribute to a lesser extent. Mutation of each proline within the NT differentially controls NT-CRD interactions, consequently affecting glycan binding, LLPS, and cellular activities. Our results unveil the role of proline polymorphisms (e.g., at P64) associated with many diseases and suggest that the function of glycosylated cell surface receptors is dynamically regulated by Gal-3., Competing Interests: The authors declare no competing interest.

Panax ginseng is a traditional medicine used in China to treat many diseases. Polysaccharides are primary active components and have many pharmacological effects. Gastric ulcer is a serious gastrointestinal disease. However, whether polysaccharides influence gastric ulcers is unclear. In this study, the effective gastroprotective impacts and potential mechanisms of Panax ginseng polysaccharides (GPS) on gastric damage induced by ethanol in rats were investigated by macroscopically evaluating gastric mucosal injuries (improved ulcer index (UI)), histopathological staining (H&E and PAS), increased NO and PGE2 levels, and suppression of oxidative stress (increased superoxide dismutase (SOD) and catalase (CAT) and decreased malondialdehyde (MDA)) and inflammation (reduced tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and myeloperoxidase (MPO)). Pretreatment with GPS ameliorated the expression of I-κB/NF-κB and JAK/STAT proteins in the rat stomach exposed to ethanol. The results indicated that GPS prevent ethanol-induced gastric injuries in rats by predominantly suppressing gastric inflammation and oxidative stress through NF-κB and STAT inhibition.

A neutral polysaccharide fraction (WGFPN) was isolated from Panax ginseng flowers. Monosaccharide composition and HPSEC-MALLS-RI (high-performance size exclusion chromatography coupled with multi-angle laser light scattering detector and refractive index detector) analyses showed WGFPN was a heterogalactan with a molecular weight of 11.0 kDa. Methylation, 1D/2D NMR (nuclear magnetic resonance) spectra and enzymatic hydrolysis methods were used to characterize the structure of WGFPN. It possessed a less branched (1 → 4)-β-D-galactan and a significantly branched (1 → 6)-β-D-galactan. The side chains of (1 → 6)-β-D-galactan were branched with α-L-1,5-Araf and t-α-L-Araf residues at O-3. Trace amount of 1,4-linked Glcp, terminal Galp, terminal Glcp and terminal Manp residues might attached to the 1,6-linked galactan through O-3 or 1,4-linked galactan through O-6 as side chains. WGFPN could activate RAW264.7 macrophages through increasing macrophage phagocytosis, releasing NO and secreting TNF-α, IL-6, IFN-γ and IL-1β in vitro. Moreover, WGFPN could enhance the immunity of cyclophosphamide (CTX)-induced immunosuppressed mice in vivo. Hence, WGFPN might be a potential natural immunomodulatory agent., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Polysaccharides are essential macromolecules that are present in all living organisms. They have a range of biological activities, such as antiviral, antioxidant, immunity-enhancing, and anticancer activities. In this study, a polysaccharide (PCPS) was separated and extracted from dry mycelium of Penicillium chrysogenum by a boiling water step and gel-filtration chromatography. Its structure was characterized by high performance gel-permeation chromatography, chemical derivative, and nuclear magnetic resonance analyses. The results showed that PCPS is a neutral galactomannan with an apparent molecular weight of 19.5 kDa. We evaluated the antiviral activity of PCPS. In half-leaf assays of tobacco plants, the protective effect of PCPS against Tobacco mosaic virus (TMV) was stronger than the protective effects of ningnanmycin and oligosaccharins. Electron microscopy analyses showed that PCPS can directly inactivate viral particles. The mechanism of the antiviral activity of PCPS was explored in a preliminary study. PCPS induced the production of NO and H 2 O 2 to initiate an early defense response. Treatment with PCPS resulted in increased transcript levels of the genes PAL, 4CL, LPO, and increased activities of phenylalanine lyase and peroxidase, which improved the TMV resistance of Nicotiana glutinosa. Expression of the PR-1b gene was also activated during the defense response., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Panax notoginseng is a widely used traditional Chinese medicine and has extensive pharmacological effects. In this work, water-soluble polysaccharides from Panax notoginseng were isolated and fractionated. One starch-like polysaccharide (PNPN) and six pectin fractions (PNPA-1A, PNPA-1B, PNPA-2A, PNPA-2B, PNPA-3A and PNPA-3B) were obtained. Monosaccharide composition, enzymatic hydrolysis, nuclear magnetic resonance and methylation analysis were combined to characterize their structures. PNPA-1A and PNPA-2A mainly contained 1,4-β-D-galactans, 1,5-α-L-arabinan and arabinogalactan II (AG-II). PNPA-3A was a typical rhamnogalacturonan I (RG-I) type pectin with 1,4-β-D-galactan and 1,5/1,3,5-α-L-arabinan side chains. PNPA-1B, PNPA-2B and PNPA-3B consisted of homogalacturonan (HG) as major domains, together with different ratios of RG-I and rhamnogalacturonan II (RG-II) domains. These results will provide basis for further investigation of structure-activity relationships of Panax notoginseng polysaccharides and be useful for the application of Panax notoginseng., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Insulin resistance is the main cause of type 2 diabetes. Polysaccharide is one of the main active components in mushrooms. Some mushroom polysaccharides have been reported to have beneficial effects against type 2 diabetes. However, the structural features and mechanisms of polysaccharides with hypoglycemic activity have not been elucidated. In this paper, six alkali-soluble total polysaccharides, six neutral polysaccharides and six acidic polysaccharides were extracted and purified from mushrooms fruiting bodies of different species. The effects of these polysaccharides on improving insulin resistance were compared using high fatty acids and glucose-induced hepatocytes. Among them, the neutral polysaccharide AAMP-N of Armillaria mellea fruiting body showed the most significant activity, indicating that mannogalactoglucan is the main active domain. AAMP-N enhanced insulin sensitivity in vitro. Also AAMP-N lowered blood glucose, and modulated lipid metabolism in db/db mice. In addition, AAMP-N protected damaged pancreatic islets in mice. Our results demonstrated the role of natural polysaccharides from mushrooms in the improvement of insulin sensitivity and lipid metabolism, and provided basis for the development of mushroom polysaccharides as hypoglycemic healthy food., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Four water-soluble polysaccharides were extracted from Pleurotus eryngii, Flammulina velutipes, Pleurotus ostreatus and white Hypsizygus marmoreus. Using anion exchange and gel permeation chromatography, a neutral and an acidic fraction were purified from each water-soluble polysaccharide. Their molecular weights were all around 20 kDa except that the acidic polysaccharide from Pleurotus ostreatus (named WPOPA) had a lower molecular weight of 5 kDa. Four neutral polysaccharides were mainly composed of galactose (42.7%-69.1%), followed by Man (19.4%-39.3%) and Glc (1.1%-15.9%). Four acidic polysaccharides contained glucose (59.0%-81.8%) as major sugar and minor glucuronic acid (4.5%-9.5%). Acidic polysaccharides exhibited stronger antioxidant activities than neutral fractions, and WPOPA showed the best antioxidant effects. Structural analysis indicated WPOPA had β-(1 → 6)-glucan backbone branched at O-3 by β-1,3-d-Glcp, t-β-d-Glcp and t-β-d-GlcpA. This investigation would be useful for screening natural antioxidants and significant in developing mushroom polysaccharides as functional foods., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Response surface methodology (RSM) using a Box-Behnken design (BBD) was applied to optimize the extraction of Paxillus involutus polysaccharides. The optimum conditions included an extraction time of 3h, extraction temperature of 79°C and a ratio of liquid to raw material of 43.1mL/g. Under the optimized conditions, the polysaccharides yield was 12.25%. Then, the polysaccharides were purified with DEAE-Cellulose 52 and Sephadex G-100 gel columns, and the fraction denoted as PIP2-1 with a molecular weight of 32kDa was obtained. PIP2-1 was composed of mannose, glucose, galactose, fucose with the mole percentages of 2.8%, 62.2%, 25.4% and 9.6%. The PIP2-1 possessed typical Fourier Transform infrared spectroscopy (FTIR) characterization of polysaccharides. The methylation analysis showed that the PIP2-1 mainly included 1-linked-fuc, 1,3-linked-man, 1-linked-glc, 1,4-linked-glc, 1,6-linked-glc, 1-linked-gal, 1,6-linked-gal, 1,4,6-linked-gal and 1,2,6-gal glycosidic bonds. Furthermore, PIP2-1 showed significant antioxidant activity against hydroxyl radicals (OH), 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and superoxide radicals. Finally, PIP2-1 significantly enhanced the release of TNF-α and IL-6 in RAW264.7 cells. The results indicated that PIP2-1 could be exploited as a natural antioxidant and immunomodulator for functional food and medical applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)

From the bee pollen of Nelumbo nucifera, we used hot water extraction and chromatographic fractionation on DEAE-cellulose and Sepharose CL-6B to purify polysaccharides (WNPP) to homogeneity. WNPP-2-RG (3.8 × 10 5  Da) consisted primarily of Rha (11.5%), GalA (12.0%), Gal (41.2%) and Ara (29.7%). Structure analysis showed that this polysaccharide is a RG-I type pectin containing AG-I and AG-II side chains comprised of 1,5-L-Ara (25.6%), t-D-Gal (12.0%), and 1,6-D-Gal (18.3%) as the primary linkage types. Immunological activity assays demonstrated that WNPP-2-RG increased lymphocyte proliferation and macrophage phagocytosis, and induced production of NO. Compared to WNPP-2-RG, its hydrolysates (RG-8H-P) had no effect on lymphocyte proliferation, although they did promote macrophage phagocytosis and decreased NO production. Our results suggest that Gal side chain residues may play a crucial role in macrophage phagocytosis, whereas Ara is unimportant for NO production and neither Gal nor Ara impact lymphocyte proliferation., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Despite it is used as a nutraceutical against diabetes and obesity, the mechanism of action of bee pollen is still unclear. Pectic bee pollen polysaccharide (RBPP-P) was isolated from Rosa rugosa , and its structure was characterized by 13 C-NMR and Fourier transform-infrared spectroscopy (FT-IR). Using high glucose and fatty acids-treated HepG2 cells and high fat diet (HFD)-induced obesity mice, we detected its effect on insulin function and lipid metabolism based on autophagy. RBPP-P contained arabinogalactan, rhamnogalacturonan I, and homogalacturonan domains. In vivo studies demonstrated that RBPP-P markedly ameliorated insulin resistance, glucose intolerance, and liver steatosis in obese mice. The suppressive effects of RBPP-P on liver steatosis and triglyceride content were mediated by increased autophagy and lipase expression in liver. In AMPK knockdown cells ( prkaa 1/2 -/- MEF) and HFD-fed mice tissues (liver, gonadal white adipose, and inguinal white adipose), RBPP-P enhanced autophagy in AMPK/mTOR-dependent way in liver, but not in adipose tissue. These findings demonstrated that bee pollen polysaccharide alleviated liver steatosis and insulin resistance by promoting autophagy via an AMPK/mTOR-mediated signaling pathway, suggesting that RBPP-P could be a novel therapeutic agent used for the treatment of obesity and diabetes.

This study was aimed at investigating the immunomodulating activity of Hericium erinaceus polysaccharide (HEP) in mice, by assessing splenic lymphocyte proliferation (cell-mediated immunity), serum hemolysin levels (humoral immunity), phagocytic capacity of peritoneal cavity phagocytes (macrophage phagocytosis), and NK cell activity. ELISA of immunoglobulin A (SIgA) in the lamina propria, and western blotting of small intestinal proteins were also performed to gain insight into the mechanism by which HEP affects the intestinal immune system. Here, we report that HEP improves immune function by functionally enhancing cell-mediated and humoral immunity, macrophage phagocytosis, and NK cell activity. In addition, HEP was found to upregulate the secretion of SIgA and activate the MAPK and AKT cellular signaling pathways in the intestine. In conclusion, all these results allow us to postulate that the immunomodulatory effects of HEP are most likely attributed to the effective regulation of intestinal mucosal immune activity.

In this paper, α-amylase-assisted extraction was used to isolate the polysaccharide that remained in hot water-extracted ginseng. The yield of the polysaccharide was 9.0%, almost equal to that of the hot water-extracted polysaccharide. Using anion exchange and gel permeation chromatography, the polysaccharide was fractionated into a neutral polysaccharide fraction and six pectic fractions. The neutral fraction accounted for 76% of the polysaccharide and contained both amylopectin and amylose. The pectic polysaccharide fractions were identified to be arabinogalactan, type-I rhamnogalacturonan and homogalacturonan-type pectin by high-performance liquid chromatography, Fourier transform-infrared and nuclear magnetic resonance analysis. Structural and lymphocyte proliferation activity results showed that these polysaccharides were different from those extracted by hot water, indicating that ginseng contains complex polysaccharides with diverse structures, which results in its diverse pharmacological activities. The α-amylase-assisted extraction is a novel method for preparing ginseng polysaccharides and could be applied toward the further study and exploration of ginseng. These findings provide technical and theoretical support for ginseng pharmacology., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Neutral (WGPN) and acidic (WGPA) polysaccharides were fractionated from ginseng polysaccharide. WGPN and WGPA decreased fasting blood glucose by different manners of administration. Intra-gastric administration of WGPA showed a marked hypoglycemic effect, which may be related to its anti-oxidative activity. The results indicated that WGPA may have anti-diabetic potential.

Ginseng acidic polysaccharide WGPA isolated from the root of Panax ginseng C. A. Meyer was fractionated into WGPA-A and WGPA-N by anion-exchange chromatography. The antifatigue activity of ginseng acidic polysaccharide WGPA has been reported in our previous research. This present study was designed to identify its active component and elucidate the mechanism for preventing chronic fatigue syndrome (CFS). WGPA, WGPA-A and WGPA-N were orally administered to mice once daily for 15 days. The effects of these compounds on physiological biomarkers of oxidative stress and on the morphology of the mitochondria in striated skeletal muscle were assessed. The results of forced swimming test-induced indicated that WGPA and WGPA-A could lengthen the swimming time, while WGPA-N could not. In addition, malondialdehyde and lactate dehydrogenase levels in serum were enhanced; while those of superoxide dismutase and glutathione peroxidase were lowered. Interestingly, the structural degeneration of mitochondria were all ameliorated. These findings suggested that WGPA-A is the active component of WGPA, it might have potential therapeutic effects for CFS and the oxidative stress might be involved in the pathogenesis. Our results also provided essential data for a better understanding of the antifatigue effects of P. ginseng extracts.

The non-lichenized ascomycete Bulgaria inquinans (Fries), growing in the Changbai Mountain of China, has been used as medicinal diet for many years. In a previous study, we have reported that a heteropolysaccharide BIWS-4b from the fruit bodies of B. inquinans (Fries) exhibited markedly antimalarial and immunostimulating activities. In this paper, the structural features and antioxidant activity of BIWS-4b were investigated. The results showed that BIWS-4b contains an α-(1→2), (1→6)-mannan core to which the glucogalactan chains are attached. The glucogalactan chains were composed of (1→6)-, (1→5)- and (1→5,6)-linked β-Galf, (1→4)-linked and non-reducing terminal β-Glcp units, and might be attached to the mannan core at the O-2 positions of α-Manp units. The antioxidant assays showed that BIWS-4b exhibited good activities, including free radicals scavenging effects, ferrous ion-chelating ability and reducing power. Thus, BIWS-4b could be used as a natural antioxidant agent for food and pharmaceutical industries., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

In the present study, bee pollen polysaccharides from Rosa rugosa (WRPP) were extracted and fractionated. WRPP were purified to neutral (WRPP-N) and acidic polysaccharides (WRPP-1, WRPP-2) with DEAE-Cellulose. WRPP-N were mainly composed of glucose, mannose, arabinose and galactose, indicating the existence of glucan, arabinogalactan (AG) and mannoglucan. WRPP-1 mainly consisted of rhamnose (3.0%), galacturonic acid (12.4%), galactose (24.7%) and arabinose (53.9%), and contained a large proportion of AGs. WRPP-2 consisted of rhamnose (7.8%), galacturonic acid (23.0%), galactose (15%) and arabinose (48.7%), while WRPP-2 contained more galacturonic acid compared to WRPP-1. WRPP-1 and WRPP-2 were composed by type I rhamnogalacturonan (RG-I), homogalacturonan (HG) and AG fragments, while WRPP-2 contained more HG and RG-I. All the fractions had significant anti-proliferative activity in HT-29 and HCT116 cells; the neutral and acidic fractions were shown to have significant synergistic effects which accounted for the antitumor activity of bee pollen polysaccharides from Rosa rugosa in vitro.

A mannogalactoglucan, named LE-MGG, was isolated from the basidiocarps of Lentinus edodes by hot water-extraction, ethanol precipitation anion exchange chromatography, and further purified by gel-permeation chromatography (GPC). Its structural features were investigated by high performance liquid chromatography (HPLC), high performance gel-permeation chromatography (HPGPC), methylation analysis, periodate oxidation-Smith degradation, and by IR and NMR spectroscopy, including two-dimensional (2D) NMR. HPLC analysis revealed that LE-MGG contained mannose-galactose-glucose in the molar ratio of 10:18:72. GPC and HPGPC showed that LE-MGG was a homogeneous fraction (d=1.34) and its molecular weight was estimated to be 18kDa. Chemical and spectroscopic studies indicated that LE-MGG consists of (1→6)-, (1→4)- and (1→3)-linked β-d-glucopyranosyl residues, (1→6)-linked α-d-galactopyranosyl residues, (1→3,6)- and (1→2,4)-linked α-d-mannopyranosyl residues and terminal residues of β-d-glucopyranosyl. Cytotoxicity assay showed that LE-MGG presented higher antitumor activities against S-180 cell with a dose-dependent manner, and exhibited lower cytotoxicity to carcinoma HCT-116 and HT-29 cells. Our studies showed also that LE-MGG presented antitumor bioactivities on Sarcoma 180 solid tumor cell implanted in Kunming mice. This finding suggests that mannogalactoglucan should be explored as potential antitumor agents and could be potentially applied as a natural antitumor drug., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The pH-modified citrus pectin (MCP) has been demonstrated to inhibit galectin-3 in cancer progression. The components and structures of MCP related to this inhibition remained unknown. In this paper, we fractionated MCP on DEAE-cellulose column into a homogenous neutral fraction MCP-N (about 20 kDa) and a pectin mixture fraction MCP-A (wide molecular distribution on Sepharose CL-6B chromatography). Both MCP-N and MCP-A inhibited hemagglutination mediated by galectin-3 with minimum inhibition concentration (MIC) 625 and 0.5 μg/ml, respectively. MCP-N was identified to be a type I arabinogalactan (AG-I) with a main chain of β-1→4-galactan. MCP-N was digested by α-L-arabinofuranosidase to give its main chain structure fraction (M-galactan, around 18 kDa), which was more active than the original molecule, MIC 50 μg/ml. The acidic degradation of M-galactan increased the inhibitory activity, MIC about 5 times lower than M-galactan. These results above showed that the functional motif of the β-1→4-galactan fragment might lie in the terminal residues rather than in the internal region of the chain. Therefore, MCP-N and its degraded products might be developed to new potential galectin-3 inhibitors. This is the first report concerning the fractionation of MCP and its components on galectin-3 inhibition. The information provided in this paper is valuable for screening more active galectin-3 inhibitors from natural polysaccharides.

A range of molecular probes for cell wall polysaccharides has been used to explore the structure and location of water-extracted pectic polysaccharides occurring in fractions isolated from ginseng roots. The LM19 homogalacturonan (HG) epitope was abundant in an HG fraction and analysis of LM19 binding to a rhamnogalacturonan-I (RG-I) rich-fraction indicated that the LM19 epitope is sensitive to acetylation. A specific RG-I epitope (LM16), four arabinogalactan-protein (AGP) epitopes (LM2, LM14, JIM16, MAC207) and an extensin epitope (JIM20) were found to be abundant and co-located in several isolated polysaccharide fractions including an arabinogalactan fraction and two RG-I fractions. Detection of the RG-I, AGP and extensin epitopes identified in isolated polysaccharide fractions in sections of ginseng roots indicated that they were most abundant in secretory cavities found in the cortical regions of ginseng roots. In addition, the immunocytochemical study indicated that polysaccharide epitope masking is a widespread phenomenon in the primary cell walls of ginseng roots.

Ginseng polysaccharide has anticancer activity. However, the structure-activity relationship and the activity mechanism are still unclear. Therefore, it is necessary to study the anticancer activity of structurally different ginseng polysaccharide fractions and their potential mechanisms. Ginseng polysaccharide fractions and their temperature-modified products were assayed for their effects on HT-29 cell proliferation by MTT assay, on cell cycle progression by flow cytometry, and on caspase-3 activation by western blot analysis. The HG-rich ginseng pectin inhibited cell proliferation and induced cell cycle arrest in the G2/M phase. The temperature-modified HG-rich pectin had dramatically increased antiproliferative effect and induced apoptosis accompanied by the activation of caspase-3. Starch-like glucan and arabinogalactan of ginseng exhibited no antiproliferative effects. Even after temperature modification, their inhibitory effects either remained unchanged or increased slightly. The HG-rich pectin exerts its antiproliferative effect via cell cycle arrest and the temperature modification markedly increased the antiproliferative effect.

Aim of the Study: Panax ginseng C. A. Meyer (ginseng) is a well-known Chinese herb often used in Asian countries for physical strength development. Ginseng polysaccharides are its active component and have a lot of pharmaceutical activities. However, anti-fatigue activity of ginseng polysaccharides has not yet been tested. The current study was designed to evaluate the anti-fatigue activity of ginseng polysaccharides (WGP) in an animal test for fatigue and compare the activities between the neutral (WGPN) and acidic (WGPA) portion in an attempt to determine whether the medicinal uses are supported by pharmacological effects., Materials and Methods: WGP, WGPN and WGPA were orally administrated to mice once daily for 15 days. Anti-fatigue activity was assessed using the forced swim test (FST) and serum biochemical parameters were determined by autoanalyzer and commercially available kits., Results: While all compounds were found to reduce immobility in the FST, the effect of WGPA was demonstrated in lower doses compared with WGP and WGPN. Moreover, the FST-induced reduction in glucose (GLU) and glutathione peroxidase (GPx) and increase in creatine phosphokinase (CK), lactic dehydrogenase (LDH) and malondialdehyde (MDA) levels, all indicators of fatigue, were inhibited by the corresponding doses of WGP, WGPN and WGPA., Conclusions: Ginseng polysaccharides have anti-fatigue activity, also reflected in the effects on the physiological markers for fatigue. The acidic polysaccharide is more potent than the neutral polysaccharide., (Copyright 2010 Elsevier Ireland Ltd. All rights reserved.)

A neutral polysaccharide fraction (WGPN) prepared from Panax ginseng C.A. Meyer by hot water extraction and DEAE-cellulose chromatography was tested for its anticancer activity alone and in combination with 5-fluorouracil (5-FU) in Sarcoma-180 (S180) tumor-bearing mice by intragastric administration. WGPN alone inhibited S180 tumor growth in a bell-shaped dose-response curve, and the combination with 5-FU showed a synergistic effect. Studies of various immunological activities in S180-bearing mice revealed that WGPN stimulated the proliferation of lymphocytes, increased natural killer cell cytotoxicity, enhanced the phagocytosis and nitric oxide production by macrophages, and increased the level of tumor necrosis factor-alpha in serum. In combination with 5-FU, WGPN mitigated damage to the immune system caused by 5-FU in S180-bearing mice. These results suggest that WGPN might be a potential adjuvant for chemotherapeutic drugs.

An alkali-soluble β-glucan (AHEP-A-b, 20 kDa) purified from Hericium erinaceus fruiting bodies, was structurally characterized and examined for antioxidant activity. Methylation analysis and NMR spectroscopy show that the backbone of AHEP-A-b is composed of (1→6)-linked-D-β-glucopyran residues, branched at O-3 of glucopyranose (Glcp) residues with [→3)-β-D-Glcp-(1→] oligosaccharides or single unit of β-Glcp. Periodate oxidation analysis and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) indicate that the degree of polymerization (DP) of [→3)-β-D-Glcp-(1→] side chains is 2 to 8. Functionally, AHEP-A-b is a relatively strong antioxidant as demonstrated by using 2, 2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radical (ABTS·+), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, and hydroxyl radicals scavenging assays. The present study lays the foundation for further studies into structure-activity relationships of polysaccharides from H. erinaceus. [ABSTRACT FROM AUTHOR]

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Influenza virus is prone to seasonal spread and widespread outbreaks, which pose important challenges to public health security. Therefore, it is important to effectively prevent and treat influenza virus infection. Schisandra polysaccharide (SPJ) is a polysaccharide derived from the fruit of Schisandra chinensis (Turcz.) Baill. In this study, we evaluated the antiviral activity of SPJ in vitro and in vivo, especially against influenza A virus (IAV) infection. By analyzing SPJ structure and monosaccharide composition, the molecular weight of SPJ was determined to be 115.5 KD, and it is composed of galacturonic acid (89.4%), rhamnose (0.8%), galactose (4.4%), arabinose (3.8%), and glucose (1.7%). Immunofluorescence analysis showed that SPJ treatment reduced the positive rate of viral nucleoproteins in cells, indicating that the compound had an inhibitory effect on influenza virus replication. Furthermore, SPJ therapy improved the survival of infected mice. Lung virus titer assays indicated that SPJ treatment significantly reduced viral loading in the lung tissue of infected mice and alleviated the pathological damage caused by influenza virus infection. Moreover, SPJ reduced cytokine expression during influenza virus challenge. In conclusion, SPJ has anti‐influenza virus effects and may have potential as an anti‐influenza drug candidate in further clinical studies. [ABSTRACT FROM AUTHOR]

Galectin-3 is crucial to many physiological and pathological processes. The generally accepted dogma is that galectins function extracellularly by binding specifically to β(1→4)-galactoside epitopes on cell surface glycoconjugates. Here, we used crystallography and NMR spectroscopy to demonstrate that negatively charged homogalacturonans (HG, linear polysaccharides of α(1→4)-linked- D -galacturonate (GalA)) bind to the galectin-3 carbohydrate recognition domain. The HG carboxylates at the C6 positions in GalA rings mandate that this saccharide bind galectin-3 in an unconventional, "topsy-turvy" orientation that is flipped by about 180o relative to that of the canonical β-galactoside lactose. In this binding mode, the reducing end GalA β-anomer of HGs takes the position of the nonreducing end galactose residue in lactose. This novel orientation maintains interactions with the conserved tryptophan and seven of the most crucial lactose-binding residues, albeit with different H-bonding interactions. Nevertheless, the HG molecular orientation and new interactions have essentially the same thermodynamic binding parameters as lactose. Overall, our study provides structural details for a new type of galectin–sugar interaction that broadens glycospace for ligand binding to Gal-3 and suggests how the lectin may recognize other negatively charged polysaccharides like glycoaminoglycans (e.g. heparan sulfate) on the cell surface. This discovery impacts on our understanding of galectin-mediated biological function. [ABSTRACT FROM AUTHOR]

This study reports on in vivo immunomodulatory activities mediated by WPEP-N-b, a heterogalactan from Pleurotus eryngii. Using cyclophosphamide (CTX)–induced immunosuppressed mice, we demonstrate here that WPEP-N-b enhances immunity as determined by the immune organ index, peripheral blood immune cell content, splenocyte proliferation, NK cell activity and T lymphocyte subpopulations. WPEP-N-b prevented apoptosis of bone marrow cells induced by CTX. The level of cytokines (i.e. TNF-α, IL-6 and IL-1β) and macrophage activity in these immunocompromised mice were restored upon treated with WPEP-N-b. Mechanistically, it appears that WPEP-N-b enhances splenocyte proliferation and NK cell activity might through the Toll-like receptor 4 (TLR4)-PKC signaling axis, and increases macrophage activity by activating JNK, p38 and NF-κB signaling pathways and Toll-like receptor 2 (TLR2) is the possible receptor of WPEP-N-b in macrophages. Our findings indicate that WPEP-N-b may function as a natural immune stimulant. • WPEP-N-b enhances the overall health of immunodeficient mice. • WPEP-N-b restores immune abnormalities induced by cyclophosphamide. • WPEP-N-b increases splenocyte proliferation and NK cell activity through the PKC signaling pathway. • WPEP-N-b promotes macrophage activity via JNK, p38 and NF-κB signaling pathways. [ABSTRACT FROM AUTHOR]

Water-soluble polysaccharides were isolated from the leaves and roots of Isatis indigotica Fort., and their structural features were studied and compared. One neutral polysaccharide fraction (WFIP-N) and three pectin fractions (WFIP-A-A, WFIP-A-B and WFIP-A-C) were obtained from the leaves, and one neutral polysaccharide fraction (WRIP-N) and two pectin fractions (WRIP-A-A and WRIP-A-B) were obtained from the roots. WFIP-A-B (Mw = 34.6 kDa) and WRIP-A-B (Mw = 29.9 kDa) were the major pectic polysaccharides. Monosaccharide composition, FT-IR, enzymatic hydrolysis, NMR and methylation analysis indicated that both WFIP-A-B and WRIP-A-B are composed of rhamnogalacturonan I (RG-I), rhamnogalacturonan II (RG-II) and homogalacturonan (HG) domains with mass ratios of 1.5:1.0:0.4 and 0.3:1.0:1.7, respectively. WFIP-A-B and WRIP-A-B were found to be rich in RG-I and HG domains, respectively, and mainly contained type II arabinogalactan (AG-II) and α-L-1,5-arabinan side chains, but those in WRIP-A-B were more numerous and longer. Our results provide structural features and differences between these polysaccharides which will help to elucidate their functional differences. [ABSTRACT FROM AUTHOR]

Here, we compare the content and composition of polysaccharides derived from the mycelium (40.4 kDa intracellular polysaccharide, IPS) and culture (27.2 kDa extracellular polysaccharide, EPS) of Penicillium oxalicum. Their chemical structures investigated by IR, NMR, enzymolysis and methylation analysis indicate that both IPS and EPS are galactomannans composed of α-1,2- mannopyranose (Man p) and α-1,6-Man p in a backbone ratio of ~3:1, respectively, both decorated with β-l,5-galactofuranose (Gal f) side chains. A few β-l,6-Gal f residues were also detected in the IPS fraction. EPS and IPS have different molecular weights (Mw) and degrees of branching. IPS obtained by alkaline extraction of P. oxalicum have been reported to be galactofuranans, a composition different from our IPS. Up to now, there have been no reports on the fine structure of EPS. Our results of galectin-mediated hemagglutination demonstrate that IPS exhibits greater inhibitory effects on five galectins compared with EPS. In addition, we find that Gal f , a five-membered ring form of galactose, can also inhibit galectins. IPS may provide a new source of galectin inhibitors. These results increase our understanding of structure-activity relationships of polysaccharides as galectin inhibitors. • The structures of IPS and EPS were comparatively investigated. • IPS and EPS contain mannan composed of α-1,2-Man p and α-1,6-Man p as backbone. • IPS and EPS contain different galactofuranan as side chains. • IPS had stronger inhibitory effects on five galectins compared with EPS. [ABSTRACT FROM AUTHOR]

In the manuscript, water-soluble polysaccharides WPNI was extracted from notopterygium incisum roots and separated into two homogeneous fractions WPNI-A-a and WPNI-A-b. WPNI-A-a was an arabinogalactan (AG). WPNI-A-b belonged to HG type pectin. The structure of WPNI-A-b was analyzed by FT-IR, NMR, enzymatic hydrolysis (Endo-PG) and UPLC-FLD-MSn. WPNI-A-b was dominated by HG domain, covalently linked with AG and RG-II domains. Oligogalacturonides produced by Endo-PG from HG domain were non-, mono-, di- or tri-methyl esterified with degree of polymerization (DP) from 1 to 6. The distribution of methyl-ester groups was in a block-wise manner. The interaction of WPNI-A-b and its enzymatic hydrolysis products with galectin-1, galectin-3, galectin-7 and galectin-8 showed that AG domain exhibited stronger binding avidity to galectins than RG-II and HG domain, while oligogalacturonides showed no binding activities to galectins. The results would be useful for the application of the pectin from notopterygium incisum. [ABSTRACT FROM AUTHOR]

• Crosslinked nanofibers of neutral polysaccharides were prepared. • The nanofibers show excellent water resistance and biocompatibility. • The nanofibers are expected to seek extensive biomedical applications. Neutral polysaccharides such as konjac glucomannan, starch and pullulan are abundant in nature and have unique property. Their nanofibers hold great potential for biomedicine, which however, are seldom applied in the field due to the lack of crosslinking method. In this work, we report a periodate oxidation - adipic acid dihydrazide (ADH) crosslinking strategy to prepare robust and biocompatible neutral polysaccharide nanofibers. Neutral polysaccharides with adjacent dihydroxyl groups are firstly partially oxidized with periodate to give dialdehyde polysaccharides, and their electrospun nanofibers are then crosslinked with ADH to form dihydrazone crosslinkers. The resulting crosslinked neutral polysaccharide nanofibers exhibit high water resistance and excellent mechanical properties because of the high reactivity of Schiff base crosslinking reaction. Moreover, the crosslinked neutral polysaccharide nanofibers show good biocompatibility due to the low toxicity of ADH. These robust and biocompatible neutral polysaccharide nanofibers are expected to seek extensive applications in a variety of biomedical fields. [ABSTRACT FROM AUTHOR]

Highlights • A novel linear 3- O -methylated galactan (WCCP-N-b) from C. cibarius was identified. • WCCP-N-b increased phagocytosis and secretion of NO, TNF-α, IL-6 and IL-1β. • WCCP-N-b activated macrophages through Akt/NF-κB, MAPKs via TLR2 receptor. • The molecular weight of WCCP-N-b impacted the activation. Abstract A novel polysaccharide (WCCP-N-b) with a molecular weight of 18 kDa was isolated and purified from the fruiting bodies of Cantharellus cibarius. Monosaccharide composition, methylation analysis and NMR spectra indicated that WCCP-N-b was a linear α-1,6-galactan, partially methylated at O-3 of galactose. The molar ratio of Gal, 3-methylated-Gal, Glc and Man was 14.4:4.6:1.0:1.2. WCCP-N-b could significantly increase macrophage phagocytosis, release of NO and secretion of TNF-α, IL-6 and IL-1β. On a cellular mechanistic level, WCCP-N-b activated MAPKs and NF-κB signaling pathway via Toll-like receptor 2 (TLR2). To further elucidate the structure-function relationship, WCCP-N-b was hydrolyzed by acid. Four degraded fragments were obtained, with molecular weights of 16.1 kDa, 11.2 kDa, 5 kDa and 3.5 kDa, respectively. Their macrophage activation effects were significantly decreased along with the molecular weight decrease. Collectively, WCCP-N-b could activate RAW264.7 cells, and the activation effect was related to its molecular weight. [ABSTRACT FROM AUTHOR]

Abstract Water-soluble pectic polysaccharides isolated from Panax ginseng flower buds (WGFPA) were completely fractionated into six homogeneous fractions (WGFPA-1a, WGFPA-2a, WGFPA-3a, WGFPA-1b, WGFPA-2b and WGFPA-3b) by a combination of ion-exchange and size exclusion chromatographies. Monosaccharide composition, enzymatic hydrolysis and 13C nuclear magnetic resonance (NMR) spectra analysis were combined to characterize their structural features. Furthermore, the interactions between these polysaccharides and galectin-3 were evaluated by biolayer interferometry assay. The results showed that WGFPA-1a, WGFPA-2a and WGFPA-3a were rhamnogalacturonan I (RG-I) type pectin with abundant side chains, including α-L-1,5-arabinan, β-D-1,4-galactan, arabinogalactan I (AG-I) and arabinogalactan II (AG-II), exhibiting strong binding activities to galectin-3 with apparent KD values 4.9 μM, 0.71 μM and 0.24 μM, respectively. WGFPA-1b, WGFPA-2b and WGFPA-3b were homogalacturonan (HG) type pectin covalently linked with different ratios of rhamnogalacturonan II (RG-II) domains, showing weaker or no interactions with galectin-3. This study provides useful structural information for further investigation on the structure-activity relationship of ginseng flower buds pectin. [ABSTRACT FROM AUTHOR]

Galactans exhibit many biological activities, including modulation of human gut microbiota. By optimizing hydrolysis conditions and fractionation protocols, we purified a series of galacto-oligosaccharides from potato galactan, with degrees of polymerization (DP) up to 15. These oligosaccharides were characterized by using HPAEC, MALDI-TOF, 1 H 13 C-NMR HSQC and HMBC experiments. Analysis of NMR data allowed chemical shift assignments to be made for oligos up to DP15. We used these NMR data to calculate intensity ratios of resonances from non-terminal to terminal residues, and correlated them with galactan chain length. Using this approach, we were able to estimate the length of galactan side chains in complex rhamnogalacturonan-I polysaccharides. Furthermore, we demonstrate here that prebiotic effects on Lactobacillus bacteria are related to galactan chain length, suggesting a potential application in the study of the human gut microbiome. [ABSTRACT FROM AUTHOR]

Dextran is a biocompatible glucose-based polymer that is widely used clinically as a plasma volume expander, anti-thrombotic agent, macromolecular carrier, peripheral blood flow enhancer, and artificial tears promoter. Because dextran has polydisperse molecular weights and tends to produce innumerable multi-charged precursor ions in the ion source, it is difficult to analysis this polymer using conventional liquid chromatography mass spectrometry. In this assay, all ion fragmentation strategy is used to solve this problem by allowing all dextran precursor ions generated in the ion source to enter the collision cell. Then serial dextran-specific fragments can be effectively generated from all precursor ions by higher energy collision dissociation and scanned by Orbitrap detector. These high resolution fragment ions provide high specificity and sensitivity for the quantitation of dextran in rat plasma. Here, we report a new quantitative method using size exclusion chromatography combined with Q Exactive mass spectrometry based on an all ion fragmentation strategy. Assay validation showed that this method is linear over the concentration range from 3 to 300 μg/mL. Our approach has been successfully applied to a pharmacokinetic study of dextran in rat and will promote the development of studies with other polysaccharides. [ABSTRACT FROM AUTHOR]

Our previous paper reported the structure of ginseng pectic polysaccharides related to the cell migration inhibitory effects, but the underlying mechanisms are poorly understood. In this manuscript, rhamnogalacturonan I (RGI)-rich pectins prepared from ginseng pectin were investigated for their effect on cell migration. The results indicated that the combination of homogalacturonan (HG) and RGI-rich pectins exerted stronger effects than either HG- or RGI-rich pectin alone. Further studies revealed that the effects of HG- and RGI-rich pectins were dependent on pretreatment, which caused alterations in cell morphologies such as cell size and shape, focal adhesion, and the organization of actin filaments, suggesting that HG and RGI pectins exert synergistic effects on cell migration, likely through different ways. Morphological data and quantitative cell adhesion and spreading assays showed that HG- and RGI-rich pectin treatment decreased cell adhesion and cell spreading on the substratum, suggesting that HG- and RGI-rich pectins may exert their effects on cell migration via decreasing cell adhesion and cell spreading. Additionally, we showed that L-929 cells expressed little galectin-3 (Gal-3) and that lactose, an inhibitor of Gal-3 did not block the activities of HG- and RGI-rich pectins, implicating that cell migration inhibited by pectin did not correlate to Gal-3. [ABSTRACT FROM AUTHOR]

Polysaccharides including pectin, alginate and chitosan are fabricated into dressings of micrometer-scaled architecture (micro- fiber or particle) and widely applied in wound treatments clinically. This work characterized and compared the properties of electrospun nanofibrous dressings of these polysaccharides. We found that although the three polysaccharide nanofiber mats had comparable mechanical strength and vapour permeability, the pectinate nanofiber mat could absorb 1.2 times and 3.6 times more exudates than the alginate and chitosan nanofiber mats, respectively, within less time. Moreover, the pectinate nanofiber mat showed much higher antibacterial activity (73.1%) than the alginate and chitosan nanofiber mats (11.8% and 17.1%, respectively). Further examinations demonstrated that the superior absorbency and antibacterial activity of the pectinate nanofiber mat were associated with the moderate extent of swelling of pectinate nanofibers under hydrated conditions. All these results suggest that the pectinate nanofiber mat might be a superior wound dressing to the alginate and chitosan nanofiber mats. [ABSTRACT FROM AUTHOR]

Commercially-supplied potato galactan (PG) is widely used as a model polysaccharide in various bioactivity studies. However, results using this galactan are not always consistent with the stated composition. Here, we assessed its composition by fractionating this commercial PG and purified its primary components: PG-A, PG-B and PG-Cp with weight-averaged molecular weights of 430, 93, and 11.3 kDa, respectively. PG-Cp consists of free β-1,4-galactan chains, whereas PG-A and PG-B are type I rhamnogalacturonans with long β-1,4-galactan side chains of up to 80 Gal residues and short β-1,4-galactan side chains of 0 to 3 Gal residues that display a "trees in lawn" pattern. Structures of these polysaccharides correlate well with their activities in terms of galectin-3 binding and gut bacterial growth assays. Our study clarifies the confusion related to commercial PG, with purified fractions serving as better model polysaccharides in bioactivity investigations. [Display omitted] • Commercial potato galactan was separated into sub-fractions PG-A, PG-B and PG-Cp. • PG-Cp consists of free β-1,4-galactan chains. • PG-A and PG-B are RG-I type polysaccharides with β-1,4-galactan side chains. • Their structures were characterized and a "trees in lawn" model is proposed. • Purified PG fractions are better model substances for structure-activity studies. [ABSTRACT FROM AUTHOR]

Although several approaches have been used to evaluate binding of carbohydrates to lectins, results are not always comparable, especially with larger polysaccharides. Here, we quantitatively assessed and compared binding of pectin-derived polysaccharides to galectin-3 (Gal-3) using five methods: surface plasmon resonance (SPR), bio-layer interferometry (BLI), fluorescence polarization (FP), competitive fluorescence-linked immunosorbance (cFLISA), and the well-known cell-based hemagglutination assay (G3H). Our studies revealed that whereas Gal-3-pectin binding parameters determined by SPR and BLI were comparable and correlated with inhibitory potencies from the G3H assay, results using FP and cFLISA assays were highly variable and depended greatly on the probe and mass of the polysaccharide. In the cFLISA assay, for example, pectins showed no inhibition when using the DTAF-labeled asialofetuin probe, but did when using a DTAF-labeled pectin probe. And the FP approach with the DTAF-lactose probe did not work on polysaccharides and large galactan chains, although it did work well with smaller galactans. Nevertheless, even though results derived from all of these methods are in general agreement, derived K D , IC 50 , and MIC values do differ. Our results reflect the variability using various techniques and therefore will be useful to investigators who are developing pectin-derived Gal-3 antagonists as anti-cancer agents. [ABSTRACT FROM AUTHOR]

Nanofibers of natural polymers represent an essential class of materials in biomedicine. Pectin is a plant-sourced anionic polysaccharide widely used in food products and biomedicine owning to its abundance, biocompatibility and inherent bioactivity. However, current electrospun pectin nanofibers are suffered from high content of carrier polymer, which may lead to low integrity and mechanical strength as well as in vivo toxicity. We report here a strategy to reduce the content of carrier polymer, polyethylene oxide (PEO) in our study, in pectin nanofibers, via electrospinning at low loading followed with selective washing. With improved electrospinning condition, we first enabled electrospinning of pectin nanofibers at low PEO loading. Then the PEO was removed by washing with a selective solvent to give pectin nanofibers containing only 1.5% PEO. The strategy was versatile to pectins from various sources and of various degree of esterification. The pectin nanofibers exhibited Young's modulus as high as 358.5 MPa. In view of their rich bioactivity, the pectin nanofibers of low content of carrier polymer are promising materials for a wide range of biomedical applications. [ABSTRACT FROM AUTHOR]

Polysaccharides from fungi have many bioactivities. Previous studies showed that galactomannans from Penicillium oxalicum antagonize galectin-8-mediated activity. Here, two intracellular and two extracellular galactomannans were purified and their structures were comparatively characterized by NMR, partial acid hydrolysis and methylation. All four of them were identified to be galactomannans with similar mannan backbones having 1,2-/1,6-linkages (~3:1) and various amounts of galactofuranan side chains. The interaction of those polysaccharides with galectin-8 was assessed by hemagglutination and biolayer interferometry. These results show that side chains are important for the interaction, and the more the side chains, the stronger the interaction. But the side chains alone did not show act on galectin-8, which indicated that the cooperation between backbone and side chains is another necessary factor for this interaction. Our findings provide important information about structure-activity relationships and the galactofuranose-containing galactomannans might be as potential therapeutic of galectin-8 related diseases. [Display omitted] [ABSTRACT FROM AUTHOR]

Abstract: A (1→6)-β-d-glucan (WPLE-N-1) and two mannogalactoglucans (WPLE-N-2 and WPLE-N-3) were isolated from the basidiocarps of Lentinus edodes by hot water extraction, ethanol precipitation, anion exchange chromatography, and further purified by gel-permeation chromatography (GPC). Their structural features were investigated by high performance liquid chromatography (HPLC), high performance gel-permeation chromatography (HPGPC), methylation analysis, periodate oxidation–Smith degradation, Fourier transform-infrared (FT-IR) and NMR spectroscopy, including two-dimensional (2D) NMR. HPLC analysis revealed that WPLE-N-1 was mainly composed of glucose (92%) with small amount of galactose (3.9%) and mannose (4.1%), WPLE-N-2 and WPLE-N-3 contained mannose–galactose–glucose in the molar ratio of 10:27:63 and 5:12:83, respectively. GPC and HPGPC showed that WPLE-N-1, WPLE-N-2 and WPLE-N-3 are homogeneous fractions and their molecular weights were estimated to be 757.5kDa, 20.9kDa and 4.7kDa, respectively. Chemical and spectroscopic studies indicated that WPLE-N-1 consisted of (1→6)-β-d-glucopyranosyl residues; while WPLE-N-2 and WPLE-N-3 were found to contain (1→6)-, (1→4)- and (1→3)-α-d-glucopyranosyl residues, (1→6)-α-d-galactopyranosyl residues, (1→3,6)- and (1→2,4)-α-d-mannopyranosyl residues and terminal residues of d-glucopyranosyl residues. On a preliminary bioactivity test, these three polysaccharides exhibited antitumor activity against Sarcoma S-180, Carcinoma HCT-116 and HT-29 in vitro. This finding suggests that mannogalactoglucan should be explored as potential antitumor agents and utilized as tumor cell growth inhibitors for food and pharmaceutical industries. [Copyright &y& Elsevier]

Abstract: In this paper, we further analysed the structure of a type I rhamnogalacturonan (RG-I) pectin (WGPA-2-RG) fractionated from ginseng polysaccharides. Methylation and periodate oxidation analyses showed that WGPA-2-RG has a backbone consisting of alternating rhamnose (Rha) and galacturonic acid (GalA) residues and side chains consisting of type II arabinogalactan (AG-II). Partial acidic hydrolysis for 6h completely removed arabinose (Ara), partial galactose (Gal), but little GalA and Rha. During partial hydrolysis, the molecular weight of WGPA-2-RG decreased smoothly, suggesting that the Ara and cleavable Gal residues exist on the surface of the molecule, while GalA and Rha residues exist in the core of the molecule. The bioactivity assay showed that the arabinogalactan side chains of WGPA-2-RG are essential structures for stimulating NO secretion and lymphocyte proliferation. However, removal of the Ara and Gal residues through hydrolysis did not appreciably affect the ability of WGPA-2-RG to enhance macrophage phagocytosis. [Copyright &y& Elsevier]

Abstract: A water-soluble polysaccharide (HA) was extracted from the Herba Asari root. HA was separated into a starch-like glucan fraction (HA1) and a pectin fraction (HA2) using DEAE-cellulose. HA2 was further fractionated into three pectic polysaccharides, HA2-a, HA2-b and HA2-c, using ion-exchange chromatography. NMR and sugar composition analyses demonstrated that HA2-a is an arabinogalactan (AG) and HA2-b and HA2-c are xylogalacturonans (XGA) with AG domains. Lymphocyte proliferation assays showed that both the neutral polysaccharide and acidic polysaccharide were potent B and T cell stimulators that may have two different modes of action. [Copyright &y& Elsevier]