Glucan polysaccharides isolated from Lactarius hatsudake Tanaka mushroom: Structural characterization and in vitro bioactivities.

Commercially available mushroom polysaccharides have found widespread use as adjuvant tumor treatments. However, the bioactivity of polysaccharides in Lactarius hatsudake Tanaka (L. hatsudake), a mushroom with both edible and medicinal uses, remains relatively unexplored. To address this gap, five L. hatsudake polysaccharides with varying molecular weights were isolated, named LHP-1 (898 kDa), LHP-2 (677 kDa), LHP-3 (385 kDa), LHP-4 (20 kDa), and LHP-5 (4.9 kDa). Gas chromatography-mass spectrometry, nuclear magnetic resonance, and atomic force microscopy, etc., were employed to determine their structural characteristics. The results confirmed that spherical aggregates with amorphous flexible fiber chains dominated the conformation of the LHP. LHP-1 and LHP-2 were identified as glucans with α-(1,4)-Glcp as the main chain; LHP-3 and LHP-4 were classified as galactans with varying molecular weights but with α-(1,6)-Galp as the main chain; LHP-5 was a glucan with ß-(1,3)-Glcp as the main chain and ß-(1,6)-Glcp connecting to the side chains. Significant differences were observed in inhibiting tumor cell cytotoxicity and the antioxidant activity of the LHPs, with LHP-5 and LHP-4 identified as the principal bioactive components. These findings provide a theoretical foundation for the valuable use of L. hatsudake and emphasize the potential application of LHPs in therapeutic tumor treatments.

Heteropolysaccharides from S. cerevisiae show anti-adhesive properties against E. coli associated with Crohn's disease.

The Saccharomyces cerevisiae CNCM I-3856 was previously reported to strongly inhibit adherent-invasive Escherichia coli (AIEC) adhesion to intestinal epithelial cells in vitro and to favor AIEC elimination from the gut in a murine model of Crohn's disease in vivo. In order to identify which cell wall components of yeast are responsible for AIEC elimination, constituent polysaccharides of yeast were isolated and their anti-adhesive ability against AIEC adhesion in vitro was screened. A fraction containing mannan, ß-glucan and α-glucan extracted from yeast cell-walls was shown to inhibit 95% of AIEC adhesion in vitro and was thus identified as the strongest anti-adhesive yeast cell wall component. Furthermore, this mannan-glucan-containing fraction was shown to accelerate AIEC decolonization from gut in vivo. This fraction could be proposed as a treatment to eliminate AIEC bacteria in patients with Crohn's disease, a microbial trigger of intestinal inflammation.

Structure and immunomodulatory activity of a water-soluble α-glucan from Hirsutella sinensis mycelia.

Hirsutella sinensis, the anamorph of Ophiocordyceps sinensis (syn. Cordyceps sinensis), is a great substitute for precious and rare wild Cordyceps sinensis to effectively treat a variety of lung and kidney diseases. In this study, an α-glucan (named as HSWP-2a) was obtained by hot water extraction, DEAE-cellulose separation, and Sepharose CL-6B purification from H. sinensis mycelia. Different from known α-glucans, HSWP-2a is an α-(1 â†’ 4)-D-glucan that branched at O-6, O-3, or O-2 with a terminal 1-linked α-D-Glcp as side chain, with an average molecular weight of 870.70 kDa. Immunological tests showed that HSWP-2a could remarkably enhance the phagocytosis of macrophages and increase the production of NO, IL-1ß, IL-6, and TNF-α, via activating the p38, JNK, and NF-κB signaling pathways. Moreover, HSWP-2a could significantly promote splenic lymphocyte proliferation. Taken together, HSWP-2a may be potentially utilized as a natural immunomodulatory agent.

Structure, size and aggregated morphology of a ß-D-glucan from Lignosus rhinocerotis as affected by ultrasound.

The effect of ultrasonic treatment on the structure, size and aggregated morphology of Lignosus rhinocerotis polysaccharide (LRP) was investigated. Ultrasonic treatment for 10 min has demonstrated to improve the aqueous solubility of LRP, leading to a uniform and narrow LRP particle size distribution. Meanwhile, short-time ultrasound was found to obviously decrease the molecular size parameters (Mw, Mn, z1/2, [η] and Rh) of LRP, and transform the hyperbranched LRP molecules into flexible and extended chains, which would reaggregate to form spherical aggregates under long-time ultrasonication. Additionally, Congo red experiment combined with CD analysis indicated the existence of triple helix structure in LRP, which was still retained after ultrasonic treatment. Furthermore, under short-time ultrasonication, the spherical aggregates with some branched chains in the native LRP solution could disaggregate and form triple helixes that could be further arranged to a dense network structure, but the untangled LRP chains would reaggregate after long-time ultrasonication. CHEMICAL COMPOUNDS STUDIED IN THIS ARTICLE: Congo red (PubChem CID: 11313); Sodium hydroxide (PubChem CID: 14798); Potassium bromide (PubChem CID: 253877).

[Fungal Beta-glucans: Structure and Effect on Host Immune Responses].

Fungi are eukaryotic microorganisms that show complex life cycles, including both anamorph and teleomorph stages. Beta-1,3-1,6-glucans (BGs) are major cell wall components in fungi. BGs are also found in a soluble form and are secreted by fungal cells. Studies of fungal BGs extensively expanded from 1960 to 1990 due to their applications in cancer immunotherapy. However, progress in this field slowed down due to the low efficacy of such therapies. In the early 21st century, the discovery of C-type lectin receptors significantly enhanced the molecular understanding of innate immunity. Moreover, pathogen-associated molecular patterns (PAMPs) and pattern recognition receptors (PRRs) were also discovered. Soon, dectin-1 was identified as the PRR of BGs, whereas BGs were established as PAMPs. Then, studies on fungal BGs focused on their participation in the development of deep-seated mycoses and on their role as a source of functional foods. Fungal BGs may have numerous and complex linkages, making it difficult to systematize them even at the primary structure level. Moreover, elucidating the structure of BGs is largely hindered by the multiplicity of genes involved in cell wall biosynthesis, including those for BGs, and by fungal diversity. The present review mainly focused on the characteristics of fungal BGs from the viewpoint of structure and immunological activities.

In vitro immunomodulatory activity of water-soluble glucans from fresh and dried Longan (Dimocarpus longan Lour.).

Longan (Dimocarpus longan Lour.) is a seasonal tropical fruit used by Chinese medicine in both fresh and dried pulp forms. Their polysaccharides have been reported to have biological activity. However, their composition and immune activity have not yet been disclosed. To fulfil this aim, hot water-soluble polysaccharides of fresh and dried longan pulp were fractionated according to their molecular weight by ultrafiltration (10, 50, 100 kDa cut off). The main polysaccharides recovered were 1,6-linked glucans branched at O-3 (4-8%), O-2 (1%), O-2,4 (0.1%), and O-3,4 (0.1%). The drying process promotes the solubility of the polysaccharides. These glucans from fresh and dried longan pulp have immunomodulatory activity, shown by in vitro phagocytosis, NO, TNF-α, and IL-6 macrophages production assays. They showed also to inhibit the inflammatory response induced by LPS. The immunological activity of these glucans seems to have different responses dependent on their molecular weight, related to the immune regulatory pathways.

Structural characterization and immunomodulatory mechanisms of two novel glucans from Morchella importuna fruiting bodies.

Two novel glucans named MIPB50-W and MIPB50-S-1 were obtained from edible Morchella importuna with molecular weights (Mw) of 939.2 kDa and 444.5 kDa, respectively. MIPB50-W has a backbone of α-(1 → 4)-d-glucan, which was substituted at O-6 position by α-d-Glcp-(1→. Moreover, MIPB50-S-1 has a backbone of α-(1 → 4)-d-glucan, which was substituted at O-6 position by α-d-Glcp-(1 → 6)-α-d-Glcp-(1→. This is the first report about glucan found in Morchella mushrooms. Furthermore, MIPB50-W and MIPB50-S-1 strengthened the phagocytosis function and the promoted secretion of interleukins (IL)-6/tumor necrosis factor-alpha (TNF-α) and nitric oxide (NO), which induced the activation of Toll-like receptor 2 (TLR2), TLR4 as well as mitogen activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways. Interestingly, MIPB50-S-1 performed the better immunomodulatory activity than that of MIPB50-W in almost all tests. Therefore, MIPB50-W and MIPB50-S-1 are potential immune-enhancing components of functional foods.

The antitumor role of a newly discovered α-d-glucan from Holotrichia diomphalia Bates as a selective blocker of aldolase A.

Aldolase A (ALDOA) facilitated aerobic glycolysis in cancer cells is a potential target in the treatment of hepatocellular carcinoma (HCC). However, only few effective inhibitors of ALDOA have been reported until now. In this research, we found a polysaccharide called HDPS-4II from Holotrichia diomphalia Bates, which can specifically bind to ALDOA with a dissociation constant of 2.86 µM. HDPS-4II with a molecular weight of 19 kDa was a linear triple-helix glucan composed of ɑ-d-1,4-Glcp and ɑ-d-1,6-Glcp in a ratio of 1.0:10.0. HDPS-4II significantly inhibited aldolase enzyme activity, glycolysis, and further inhibited the expression of phosphorylated AMPKα in HCC cells. Through analyzing ALDOA-overexpressing and -knockdown cells, it was confirmed that ALDOA mediated the viability and glycolysis inhibition of HDPS-4II. Moreover, HDPS-4II administration markedly inhibited tumor growth in mice xenografted with HCCs. These findings suggest that HDPS-4II, as an ALDOA antagonist, is a promising remedy in the treatment and prevention of HCC.

Biological Activity of High-Purity ß-1,3-1,6-Glucan Derived from the Black Yeast Aureobasidium pullulans: A Literature Review.

The black yeast Aureobasidium pullulans produces abundant soluble ß-1,3-1,6-glucan-a functional food ingredient with known health benefits. For use as a food material, soluble ß-1,3-1,6-glucan is produced via fermentation using sucrose as the carbon source. Various functionalities of ß-1,3-1,6-glucan have been reported, including its immunomodulatory effect, particularly in the intestine. It also exhibits antitumor and antimetastatic effects, alleviates influenza and food allergies, and relieves stress. Moreover, it reduces the risk of lifestyle-related diseases by protecting the intestinal mucosa, reducing fat, lowering postprandial blood glucose, promoting bone health, and healing gastric ulcers. Furthermore, it induces heat shock protein 70. Clinical studies have reported the antiallergic and triglyceride-reducing effects of ß-1,3-1,6-glucan, which are indicators of improvement in lifestyle-related diseases. The primary and higher-order structures of ß-1,3-1,6-glucan have been elucidated. Specifically, it comprises a single highly-branched glucose residue with the ß-1,6 bond (70% or more) on a backbone of glucose with 1,3-ß bonds. ß-Glucan shows a triple helical structure, and studies on its use as a drug delivery system have been actively conducted. ß-Glucan in combination with anti-inflammatory substances or fullerenes can be used to target macrophages. Based on its health functionality, ß-1,3-1,6-glucan is an interesting material as both food and medicine.

Structural studies of immunomodulatory (1 → 3)-, (1 → 4)-α glucan from an edible mushroom Polyporus grammocephalus.

A water soluble polysaccharide (PGPS) with molecular weight ~ 1.4 × 105 Da was isolated by alkali treatment from an edible mushroom Polyporus grammocephalus and purified by gel chromatography using sepharose-6B column. Monosaccharide analysis revealed that PGPS was made up of glucose only. PGPS contained (1 â†’ 3)-α-D-Glcp and (1 â†’ 4)-α-D-Glcp moieties in a molar ratio of nearly 1:2. Through a series of chemical and spectroscopic (1D/2D NMR) investigations, the repeating unit of the glucan was established as: →3)-α-D-Glcp(1 â†’ [4)-α-D-Glcp(1]2→ This α-glucan was observed to stimulate some prime components of immune system, namely, macrophages, splenocytes, and thymocytes.

Using Flammulina velutipes derived chitin-glucan nanofibrils to stabilize palm oil emulsion:A novel food grade Pickering emulsifier.

We herein report chitin-glucan nanofibrils from edible mushroom Flammulina velutipes (CGNFs) as a novel stabilizer for palm oil Pickering emulsion (o/w, 30:70, v:v). Generally, these CGNFs being composed of glucose and glucosamine, are threadlike with 4.9 ± 1.2 nm wide and 222.6 ± 91.9 nm long. They were easily absorbed on the oil-water interface to form a compact layer around the oil droplets referring to Pickering emulsion. This emulsion presented shear-thinning and gel-like behaviors, wherein CGNFs concentration had a profound influence on the emulsion volume, droplet size, and stabilization index. Moreover, CGNFs showed an ability to stabilize the emulsion with a minimum of surface coverage approximately 30%. It indicated that moderate concentration of NaCl improved the emulsification effect, and the emulsion were stable in a large range of pH. These CGNFs are easy to prepare, eco-friendly and sustainable, which provides a potential for large-scale application of Pickering emulsion in food and nutraceuticals fields.

Structural analysis of a glucoglucuronan derived from laminarin and the mechanisms of its anti-lung cancer activity.

Laminarin (LA), a storage glucan, was purified from the brown alga Sargassum thunbergii. After specific oxidation using the stable nitroxyl radical, 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), together with NaBr and NaClO, glucoglucuronan (LAO) was obtained. Compositional analysis of LAO showed a molar ratio of glucuronic acid (GlcA) to glucose (Glc) of 12.7: 1. Nuclear magnetic resonance (NMR) and mass spectroscopy (MS) showed LAO to have a backbone of (1 â†’ 3)-linked ß-D-GlcpA interspersed with (1 â†’ 3, 1 â†’ 6)-linked ß-D-Glcp, that was terminated with ß-D-GlcpA. LAO inhibited human lung cancer A549 cell proliferation in vitro. IC50 values at 12 h and 24 h were 2.70 mg/mL and 2.85 mg/mL, respectively. Western blotting showed that TSC2 was up-regulated at an LAO concentration of 3.80 mg/mL. FAK, PI3K, P-AKT and mTOR were down-regulated, indicating LAO inhibited cancer cell proliferation through the FAK/PI3K/AKT/mTOR pathway. Surface plasmon resonance (SPR) studies revealed that LAO showed an IC50 of 0.07 mg/mL inhibiting the binding of heparin to fibroblast growth factor 1 (FGF1) LAO inhibition of heparin binding to FGF2 fluctuated between 15% and 28%, suggesting that LAO inhibits A549 cell proliferation by selectively interacting with FGF1.

Synthesis of bioactive silver nanoparticles using alginate, fucoidan and laminaran from brown algae as a reducing and stabilizing agent.

In this report, polysaccharides - alginate, fucoidan, laminaran - were isolated from marine algae Saccharina cichorioides and Fucus evanescens and their activity as a reducing and stabilizing agents in the biogenic synthesis of silver nanoparticles was evaluated. The cytotoxic and antibacterial properties of obtained nanoparticles were also assessed. It was found that all tested polysaccharides could be used as a reducing agent; however, their catalytic activities varied significantly in the following range alginate < fucoidan < laminaran. Nanoparticles demonstrated cytotoxicity against rat C6 glioma cells. It was considerably higher for alginate- and laminaran-obtained nanosilver samples compared to fucoidan. Additionally, silver nanoparticles possessed considerable antibacterial properties more pronounced in fucoidan-obtained samples. Our data demonstrate that different algal polysaccharides can be used for the synthesis of silver nanoparticles with varying bioactivities.

Structural characterization of a safflower polysaccharide and its promotion effect on steroid-induced osteonecrosis in vivo.

A water-soluble polysaccharide (SPAW) was purified from Safflower and it was identified to be (1→3)-linked ß-d-Glucan. The therapeutic effect and underlying mechanism of SPAW on steroid-induced avascular necrosis of the femoral head (SANFH) in a rabbit model was performed here. The abnormal histopathologic changes and apoptosis of femoral head in model group were significantly reverted after SPAW (25, 100 and 200 mg/kg) administration for 60 days, as evidenced by the a decline of empty lacunae rate, the average bone marrow fat cell size and the proportion of apoptotic cells. Furthermore, administration of SPAW significantly decreased the Bax and caspase-3 protein expression, but increased the protein expression of Bcl-2 when compared these in model rabbits. Meanwhile, increased hydroxyproline (HOP) and decreased serum hexosamine (HOM) concentration in rabbit serum were turned to the opposite way. The present study suggested that SPAW may provide an alternative treatment for the treatment of SANFH.

Isolation and comparison of α- and ß-D-glucans from shiitake mushrooms (Lentinula edodes) with different biological activities.

A polysaccharide-enriched extract obtained from Lentinula edodes was submitted to several purification steps to separate three different D-glucans with ß-(1→6), ß-(1→3),(1→6) and α-(1→3) linkages, being characterized through GC-MS, FT-IR, NMR, SEC and colorimetric/fluorimetric determinations. Moreover, in vitro hypocholesterolemic, antitumoral, anti-inflammatory and antioxidant activities were also tested. Isolated glucans exerted HMGCR inhibitory activity, but only ß-(1→6) and ß-(1→3),(1→6) fractions showed DPPH scavenging capacity. Glucans were also able to lower IL-1ß and IL-6 secretion by LPS-activated THP-1/M cells and showed cytotoxic effect on a breast cancer cell line that was not observed on normal breast cells. These in vitro results pointed important directions for further in vivo studies, showing different effects of each chemical structure of the isolated glucans from shiitake mushrooms.

Composition, isolation, purification and biological activities of Sargassum fusiforme polysaccharides: A review.

Sargassum fusiforme polysaccharides, acidic water-soluble polysaccharides extract from Sargassum fusiforme, are mainly composed of alginic acid, fucoidan and laminaran. Alginic acid is carboxyl-containing polysaccharide formed by joining ß-D-mannuronic acid and α-L-guluronic acid through ß-(1→4)/α-(1→4) glycosidic bond. Fucoidan, a natural water-soluble sulfated heteropolysaccharide with fucose and sulfuric acid groups as the core structure, is mainly linked by L-fucose through α-(1→3) glycosidic bond and has the strongest biological activity. Laminaran is mainly composed of ß-D-glucose through ß-(1→3) glycosidic bond linkage. Sargassum fusiforme polysaccharides have a variety of pharmacological activities, including antioxidant, anti-tumor, promoting immunity, anti-aging, prompting bone growth, lowering blood glucose, anti-coagulation, anti-virus, anti-bacteria, anti-fatigue, promoting growth and development, and skin protection. These activities are closely related to the functions of fucoidan in Sargassum fusiforme polysaccharides, which fucoidan is able to strengthen immune system and antioxidation in human body. In this review, the composition, the isolation and purification, and the biological activities of Sargassum fusiforme polysaccharides are discussed and can bereference for further study.

Immunomodulating Effects Exerted by Glucans Extracted from the King Oyster Culinary-Medicinal Mushroom Pleurotus eryngii (Agaricomycetes) Grown in Substrates Containing Various Concentrations of Olive Mill Waste.

We have recently demonstrated that we could enhance glucan content in Pleurotus eryngii following cultivation of the mushrooms on a substrate containing different concentrations of olive mill solid waste (OMSW). These changes are directly related to the content of OMSW in the growing substrate. Using dextran sulfate sodium (DSS)-in-flammatory bowel disease (IBD) mice model, we measured the colonic inflammatory response to the different glucan preparations. We found that the histology damaging score (HDS) resulting from DSS treatment reach a value of 11.8 ± 2.3 were efficiently downregulated by treatment with the fungal extracted glucans. Glucans extracted from stalks cultivated at 20% OMSW downregulated to a HDS value of 6.4 ± 0.5 whereas those cultivated at 80% OMSW showed the strongest effects (5.5 ± 0.6). Similar downregulatory effects were obtained for expression of various intestinal cytokines. All tested glucans were equally effective in regulating the number of CD14/CD16 monocytes from 18.2 ± 2.7% for DSS to 6.4 ± 2.0 for DSS + glucans extracted from stalks cultivated at 50% OMSW. We tested the effect of glucans on lipopolysaccharide-induced production of TNF-α, which demonstrated that stalk-derived glucans were more effective than caps-derived glucans. Isolated glucans competed with anti-Dectin-1 and anti-CR3 antibodies, indicating that they contain ß-glucans recognized by these receptors. In conclusion, the most effective glucans in ameliorating IBD-associated symptoms induced by DSS treatment in mice were glucan extracts prepared from the stalk of P. eryngii grown at higher concentrations of OMSW. We conclude that these stress-induced growing conditions may be helpful in selecting more effective glucans derived from edible mushrooms.

Structural characterization of novel comb-like branched α-d-glucan from Arca inflata and its immunoregulatory activities in vitro and in vivo.

In the current study, we identified and characterized a novel water-soluble polysaccharide (JNY2PW) with significant immunoregulatory effects and no apparent overall toxicity. JNY2PW, which was isolated from Arca inflata, belongs to a novel class of α-glucans with a molecular weight of 5.25 × 107 Da. Its backbone is composed of (1 → 4)-linked α-d-glucopyranosyl residues and a single (1 → 6)-α-d-glucopyranosyl branched unit for every five α-d-glucopyranosyl residues, showing a comb-like α-d-glucan with intensive short branches. Using in vitro models, we demonstrated that JNY2PW exerts significant immunoregulatory effects by promoting the production of nitric oxide, interleukin-6, and tumor necrosis factor α. The pathway involves the activation of the TLR4-MAPK/NF-κB signaling cassette in murine RAW264.7 macrophages. In an in vivo immunosuppressive mice model induced by cyclophosphamide treatment, we found that the JNY2PW treatment produced good antitumor activity, comparable to that of chemotherapy by doxycycline in murine breast carcinoma 4T1-bearing mice, but devoid of any observable side effects (e.g. weight loss) related with doxycycline treatment. The anti-tumor mechanism of JNY2PW may involve an overall enhancement in the immune responses of the mice to tumors. These results indicate that JNY2PW possesses potential as an adjuvant to existing chemotherapy and current immune-oncology treatment.

Peyer's patch-immunomodulating glucans from sugar cane enhance protective immunity through stimulation of the hemopoietic system.

The aim of the present study was chemical clarification of in vitro Peyer's patch-immunomodulating polysaccharides in sugar cane molasses, and evaluation of in vivo modulating activity on immune function of T lymphocytes in Peyer's patches and on microenvironment of hemopoietic system. Five kinds of glucans, comprising of dextranase-sensitive and activity-related d-glucosyl moieties, were purified as in vitro Peyer's patch-immunomodulating polysaccharides from the molasses. Oral administration of a glucan-enriched subfraction induced IL-2 and GM-CSF-producing T lymphocytes in Peyer's patches, resulting in enhancement of IL-6 production in a hemopoietic microenvironment to boost neutrophil numbers in the peripheral blood stream. Oral administration of purified glucan or glucan-enrich sub-fraction of sugar cane reduced the number of Plasmodium berghei- or P. yoelii-infected erythrocytes in a murine infection model, using polysaccharide alone or via co-administration with the antimalarial drug, artesunate. These results suggested that Peyer's patch-immunomodulating glucans enhanced protective immunity through axis of Peyer's patches-hemopoietic system.

The edible mushroom Albatrellus ovinus contains a α-l-fuco-α-d-galactan, α-d-glucan, a branched (1 → 6)-ß-d-glucan and a branched (1 → 3)-ß-d-glucan.

Albatrellus ovinus, the sheep polypore, is a large, dense mushroom being rich in cell wall material. Polysaccharides were isolated by sequential extraction, enzymatic treatment and analyzed with respect to monosaccharide composition, glycosidic linkages by methylation and GC-MS as well as NMR spectroscopy. A fucogalactan composed of an (1 → 6)-α-d-galactan backbone with single α-l-Fucp residues attached at O-2 was identified in the hot water extract obtained after treatment with a protease and size exclusion chromatography. Both the hot water extract and the hot alkali extract contained an (1 → 4)-α-d-glucan whereas ß-d-glucans were mainly present in the latter. Structural analysis suggested the presence of two different ß-d-glucan backbone structures; a (1 → 6)-linked ß-d-glucan with single ß-d-Glcp residues at O-3 and also a (1 → 3)-linked ß-d-glucan with branches in O-6. In addition there were identified short (1 → 2)-linked ß-d-xylan and (1 → 3)-α-d-mannan chains.