ABSTRACT
We developed an efficient mixed-strain co-fermentation method to increase the yield of quinoa ß-glucan (Q+). Using a 1:1 mass ratio of highly active dry yeast and Streptococcus thermophilus, solid-to-liquid ratio of 1:12 (g/mL), inoculum size of 3.8 % (mass fraction), fermentation at 32 °C for 27 h, we achieved the highest ß-glucan yield of (11.13 ± 0.80)%, representing remarkable 100.18 % increase in yield compared to quinoa ß-glucan(Q-) extracted using hot water. The structure of Q+ and Q- were confirmed through Fourier Transform Infrared (FTIR) and Nuclear Magnetic Resonance (NMR) spectroscopies. Q+ contained 41.66 % ß-glucan, 3.93 % protein, 2.12 % uronic acid; Q- contained 37.21 % ß-glucan, 11.49 % protein, and 1.73 % uronic acid. The average molecular weight of Q+(75.37 kDa) was lower than that of Q- (94.47 kDa). Both Q+ and Q- promote RAW264.7 cell proliferation without displaying toxicity. They stimulate RAW264.7 cells through the NF-κB and MAPK signaling pathways, primarily inducing NO and pro-inflammatory cytokines by upregulating CD40 expression. Notably, Q+ exhibited stronger immunostimulatory activity compared to Q-. In summary, the fermentation enrichment method yields higher content of quinoa ß-glucan with increased purity and stronger immunostimulatory properties. Further study of its bioimmunological activity and structure-activity relationship may contribute to the development of new immunostimulants.
Subject(s)
Chenopodium quinoa , Fermentation , beta-Glucans , Chenopodium quinoa/chemistry , Mice , beta-Glucans/chemistry , beta-Glucans/pharmacology , beta-Glucans/isolation & purification , Animals , RAW 264.7 Cells , Adjuvants, Immunologic/pharmacology , Adjuvants, Immunologic/chemistry , Cell Proliferation/drug effects , Molecular Weight , Streptococcus thermophilus/chemistryABSTRACT
Emerging evidence links dietary fiber with altered gut microbiota composition and bile acid signaling in maintaining metabolic health. Yeast ß-glucan (Y-BG) is a dietary supplement known for its immunomodulatory effect, yet its impact on the gut microbiota and bile acid composition remains unclear. This study investigated whether dietary forms of Y-BG modulate these gut-derived signals. We performed 4-wk dietary supplementation in healthy mice to evaluate the effects of different fiber composition (soluble vs. particulate Y-BG) and dose (0.1% vs. 2%). We found that 2% particulate Y-BG induced robust gut microbiota community shifts with elevated liver Cyp7a1 mRNA abundance and bile acid synthesis. These diet-induced responses were notably different when compared with the prebiotic inulin, and included a marked reduction in fecal Bilophila abundance which we demonstrated as translatable to obesity in population-scale American Gut and TwinsUK clinical cohorts. This prompted us to test whether 2% Y-BG maintained metabolic health in mice fed 60% HFD over 13 wk. Y-BG consistently altered the gut microbiota composition and reduced Bilophila abundance, with trends observed in improvement of metabolic phenotype. Notably, Y-BG improved insulin sensitization and this was associated with enhanced ileal Glpr1r mRNA accumulation and reduced Bilophila abundance. Collectively, our results demonstrate that Y-BG modulates gut microbiota community composition and bile acid signaling, but the dietary regime needs to be optimized to facilitate clinical improvement in metabolic phenotype in an aggressive high-fat diet animal model.NEW & NOTEWORTHY The study shows that dietary Y-BG supplementation modulated gut microbiota, bile acid metabolism and associated signaling pathways. Y-BG significantly reduced Bilophila abundance which is associated with obesity in human cohorts. Correlation analysis confirmed functional interactions between bile acid composition, gut microbiota, and metabolic phenotype, although clinical benefit did not reach significance in an aggressive obesity model. Gut microbiota and bile acids correlated with metabolic parameters, indicating future potential of dietary Y-BG modulation of metabolic pathways.
Subject(s)
Bile Acids and Salts/metabolism , Bilophila/growth & development , Dietary Fiber/administration & dosage , Gastrointestinal Microbiome , Intestine, Small/microbiology , Liver/metabolism , Obesity/diet therapy , Yeasts/metabolism , beta-Glucans/administration & dosage , Animals , Bilophila/genetics , Cholesterol 7-alpha-Hydroxylase/genetics , Cholesterol 7-alpha-Hydroxylase/metabolism , Diet, High-Fat , Disease Models, Animal , Glucagon-Like Peptide-1 Receptor/genetics , Glucagon-Like Peptide-1 Receptor/metabolism , Insulin Resistance , Intestine, Small/metabolism , Inulin/administration & dosage , Male , Mice, Inbred C57BL , Obesity/metabolism , Obesity/microbiology , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Cytoplasmic and Nuclear/metabolism , Signal Transduction , beta-Glucans/isolation & purificationABSTRACT
Dendritic cells (DCs) are recognized as the most potent antigen-presenting cells, capable of priming both naïve and memory T cells. Thus, tumor-resident DCs (tumor-associated DCs: TADCs) play a crucial role in the immune response against tumors. However, TADCs are also well known as a "double-edged sword" because an immunosuppressive environment, such as a tumor microenvironment, maintains the immature and tolerogenic properties of TADCs, resulting in the deterioration of the tumor. Therefore, it is essential to maintain and enhance the anti-tumoral activity of TADCs to aid tumor elimination. This study demonstrated the potential for tumor growth inhibition of Aureobasidium pullulan-derived ß-glucan (AP-BG). Administration of AP-BG dramatically limited the development of different types of tumor cell lines transplanted into mice. Examination of the tumor-infiltrating leukocytes revealed that AP-BG caused high expression of co-stimulatory molecules on TADCs and enhanced the production of cytolytic granules as well as pro-inflammatory cytokines by the tumor-resident T cells. Furthermore, the syngeneic mixed lymphoid reaction assay and popliteal lymph node assay showed the significant ability of AP-BG to improve DCs' antigen-specific priming of T cells in vitro and in vivo. Taken together, ß-glucan might be an immune-potentiating adjuvant for cancer treatment. This highly widely-used reagent will initiate a new way to activate DC-targeted cancer immune therapy.
Subject(s)
Adjuvants, Immunologic/pharmacology , Aureobasidium/chemistry , Dendritic Cells/drug effects , Neoplasms/drug therapy , beta-Glucans/pharmacology , Adjuvants, Immunologic/isolation & purification , Adjuvants, Immunologic/therapeutic use , Animals , Cell Line, Tumor/transplantation , Dendritic Cells/immunology , Disease Models, Animal , Drug Screening Assays, Antitumor , Humans , Lymphocyte Activation/drug effects , Lymphocytes, Tumor-Infiltrating/drug effects , Lymphocytes, Tumor-Infiltrating/immunology , Mice , Mice, Transgenic , Neoplasms/immunology , Neoplasms/pathology , T-Lymphocytes/drug effects , T-Lymphocytes/immunology , Tumor Microenvironment/drug effects , Tumor Microenvironment/immunology , beta-Glucans/isolation & purification , beta-Glucans/therapeutic useABSTRACT
Inflammatory bowel disease (IBD), which significantly affects human health, has two primary presentations: Crohn's disease and ulcerative colitis (UC). Highland barley is the most common food crop for Tibetans and contains much more ß-glucan than any other crop. Highland barley ß-glucan (HBBG) can relieve the gastrointestinal dysfunction and promote intestines health. This study aimed to evaluate whether HBBG can relieve UC in mice. A mouse model of UC was established by adding 2% dextran sulfate sodium (DSS) to drinking water for 1 week. UC was alleviated after the introduction of the HBBG diet, as indicated by reductions in the disease activity index (DAI) score, histopathological damage, and the concentration of colonic myeloperoxidase (MPO), along with an improvement in colonic atrophy. Furthermore, we found that HBBG can increase the relative transcriptional levels of genes encoding ZO-1, claudin-1, occludin, and mucin2 (MUC2), thereby reducing intestinal permeability. Additionally, HBBG maintained the balance of proinflammatory and anti-inflammatory cytokines and modulated the structure of the intestinal flora.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Colitis, Ulcerative/drug therapy , Dextran Sulfate/toxicity , Hordeum/chemistry , Plant Extracts/pharmacology , beta-Glucans/pharmacology , Animals , Anti-Inflammatory Agents/isolation & purification , Colitis, Ulcerative/chemically induced , Colitis, Ulcerative/pathology , Cytokines/metabolism , Male , Mice , Mice, Inbred C57BL , beta-Glucans/isolation & purificationABSTRACT
Highland barley grains were roller-milled to produce five different fractions (B-1, B-2, B-3, B-4, and B-5). The distribution and physicochemical properties of ß-glucans from five roller-milled fractions were investigated. The B-4 fraction contained the highest concentration of ß-glucan (4.40%), and the outermost bran (B-1) had the lowest ß-glucan content (1.01%). Besides, ß-glucans from inner core B-5 (BG-5) had higher Mw (6.482 × 105 g/mol), whereas ß-glucans from outer bran B-1 (BG-1) showed lower Mw (5.859 × 104 g/mol) than those from other fractions. Accordingly, the viscosity of BG-5 was highest (0.038-0.365 Pa·s), and the water solubility index of BG-1 was highest (50.43-90.71%). BG-5 showed stronger foam stability and emulsifying properties but weaker foaming capability, while BG-1 exhibited stronger foaming capability. The foaming capability and emulsifying properties of ß-glucan samples were better under the neutral condition (pH = 7). The foam capabilities of all ß-glucan samples displayed higher values at 65 °C, and emulsifying properties exhibited higher values at 45 °C. This study is expected to promote the application of highland barley ß-glucans in food industry.
Subject(s)
Chemical Phenomena , Hordeum/chemistry , Seeds/chemistry , beta-Glucans/chemistry , beta-Glucans/isolation & purification , Emulsions/chemistry , Hydrogen-Ion Concentration , Molecular Weight , Rheology , Solubility , Spectroscopy, Fourier Transform Infrared , Temperature , Water/chemistryABSTRACT
This present work evaluated the effect of heat fluidization, microwave roasting and baking treatment of highland barley (HB) on the molecular, structural, thermal and antioxidant characteristics of ß-glucan. Fluorescence microscopy results showed that heat fluidization exhibited the greatest disruption effect on endosperm cell walls, resulting in the highest extractability (3.35 ± 0.06 g/100 g flour) and purity (92.67 ± 0.73%) of ß-glucan. After HB thermal processing, the molecular weight and polydispersity index of ß-glucan were respectively reduced by 3.68%-90.35% and 26.45%-39.83%, and its microscopic molecular morphology transformed from large sphere aggregate to alveolate gel network structure. Meanwhile, the structural elucidation by X-ray diffraction and infrared spectroscopy revealed that thermal processing induced the scission of polymeric chain and formation of lattice-type microgels without changing the primary functional groups of ß-glucan. Furthermore, thermogravimetry and antioxidant results indicated the thermal stability and antioxidant activity of ß-glucan were enhanced by thermal processing.
Subject(s)
Free Radical Scavengers/chemistry , Hordeum/chemistry , beta-Glucans/chemistry , Cell Wall/chemistry , Endosperm/chemistry , Flour , Free Radical Scavengers/analysis , Free Radical Scavengers/isolation & purification , Heating , Molecular Weight , Solid Phase Extraction/methods , beta-Glucans/analysis , beta-Glucans/isolation & purificationABSTRACT
To overcome the limitations of the Limulus amebocyte lysate (LAL) assay method for the diagnosis of invasive fungal infection, we applied a reaction system combining recombinant ß-glucan binding proteins and a scanning single-molecule counting (SSMC) method. A novel (1â3)-ß-D-glucan recognition protein (S-BGRP) and a (1â6)-ß-glucanase mutant protein were prepared and tested for the binding of (1â6)-branched (1â3)-ß-D-glucan from fungi. S-BGRP and (1â6)-ß-glucanase mutant proteins reacted with ß-glucan from Candida and Aspergillus spp. Although LAL cross-reacted with plant-derived ß-glucans, the new detection system using the SSMC method showed low sensitivity to plant (1â3)-ß-D-glucan, which significantly improved the appearance of false positives, a recognized problem with the LAL method. Measurement of ß-glucan levels by the SSMC method using recombinant ß-glucan-binding proteins may be useful for the diagnosis of fungal infections. This study shows that this detection system could be a new alternative diagnostic method to the LAL method.
Subject(s)
Biosensing Techniques , Endotoxins/isolation & purification , Mycoses/diagnosis , beta-Glucans/isolation & purification , Aspergillus/chemistry , Aspergillus/isolation & purification , Aspergillus/pathogenicity , Candida/chemistry , Candida/isolation & purification , Candida/pathogenicity , Endotoxins/chemistry , Humans , Mycoses/microbiology , Single Molecule Imaging , beta-Glucans/chemistryABSTRACT
Ganoderma lucidum (G. lucidum) polysaccharides and triterpenoids are the major bioactive compounds and have been used as traditional medicine for ancient times. Massive demands of G. lucidum have fascinated the researchers towards its application as functional food, nutraceutical and modern medicine owing to wide range of application in various diseases include immunomodulators, anticancer, antiviral, antioxidant, cardioprotective, hepatoprotective. G. lucidum polysaccharides exhibit immunomodulatory properties through boosting the action of antigen-presenting cells, mononuclear phagocyte system, along with humoral and cellular immunity. ß-Glucans isolated from G. lucidum are anticipated to produce an immune response through pathogen associated molecular patterns (PAMPs). ß-Glucans after binding with dectin-1 receptor present on different cells include macrophages, monocytes, dendritic cells and neutrophils produce signal transduction that lead to trigger the mitogen-activated protein kinases (MAPKs), T cells and Nuclear factor-κB (NF-κB) that refer to cytokines production and contributing to immune response. While triterpenoids produce antiviral effects through inhibiting various enzymes like neuraminidase, HIV-protease, DENV2 NS2B-NS3 protease and HSV multiplication. Polysaccharides and triterpenoids adjunct to other drugs exhibit potential action in prevention and treatment of various diseases. Immunomodulators and antiviral properties of this mushroom could be a potential source to overcome this current pandemic outbreak.
Subject(s)
Antiviral Agents/pharmacology , Immune System/drug effects , Immunomodulating Agents/pharmacology , Reishi , Triterpenes/pharmacology , Virus Diseases/drug therapy , beta-Glucans/pharmacology , Animals , Antiviral Agents/isolation & purification , Host-Pathogen Interactions , Humans , Immune System/immunology , Immune System/metabolism , Immunomodulating Agents/isolation & purification , Molecular Structure , Reishi/chemistry , Signal Transduction , Structure-Activity Relationship , Triterpenes/isolation & purification , Virus Diseases/immunology , Virus Diseases/metabolism , Virus Diseases/virology , beta-Glucans/isolation & purificationABSTRACT
An endo-ß-1,3(4)-glucanase AnENG16A from Aspergillus nidulans shows distinctive catalytic features for hydrolysis of ß-glucans. AnENG16A hydrolyzed Eisenia bicyclis laminarin to mainly generate 3-O-ß-gentiobiosyl-d-glucose and hydrolyzed barley ß-glucan to mainly produce 3-O-ß-cellobiosyl-d-glucose. Using molecular exclusion chromatography, we isolated and purified 3-O-ß-cellobiosyl-d-glucose and 3-O-ß-gentiobiosyl-d-glucose, respectively, from AnENG16A-hydrolysate of barley ß-glucan and E. bicyclis laminarin. Further study reveals that 3-O-ß-cellobiosyl-d-glucose had 8.99-fold higher antioxidant activity than barley ß-glucan and 3-O-ß-gentiobiosyl-d-glucose exhibited 43.0% higher antioxidant activity than E. bicyclis laminarin. Notably, 3-O-ß-cellobiosyl-d-glucose and 3-O-ß-gentiobiosyl-d-glucose exhibited 148.9% and 116.0% higher antioxidant activity than laminaritriose, respectively, indicating that ß-1,4-linkage or -1,6-linkage at non-reducing end of ß-glucotrioses had enhancing effect on antioxidant activity compared to ß-1,3-linkage. Furthermore, 3-O-ß-cellobiosyl-d-glucose showed 237.9% higher antioxidant activity than cellotriose, and laminarin showed 5.06-fold higher antioxidant activity than barley ß-glucan, indicating that ß-1,4-linkage at reducing end of ß-glucans or oligosaccharides resulted in decrease of antioxidant activity compared to ß-1,3-linkage.
Subject(s)
Antioxidants/pharmacology , Aspergillus nidulans/enzymology , Cellobiose/analogs & derivatives , Glucan Endo-1,3-beta-D-Glucosidase/metabolism , Glucans/metabolism , Glucose/pharmacology , Hordeum , beta-Glucans/metabolism , Antioxidants/metabolism , Biphenyl Compounds/chemistry , Catalysis , Cellobiose/biosynthesis , Glucose/analogs & derivatives , Glucose/metabolism , Hordeum/chemistry , Hydrolysis , Molecular Structure , Picrates/chemistry , Structure-Activity Relationship , Substrate Specificity , beta-Glucans/isolation & purificationABSTRACT
ß-D-glucans are proposed to have many health benefits. It is therefore important to have methods which can distinguish these from other carbohydrates present in natural products, as well as giving glucan content and structural information. Correlations between features in the CP/MAS spectra of ß-D-glucans and enzyme assay determined ß-D-glucan content were generally found to be poor. The ß-D-glucan in dry and hydrated forms of the mushroom Ganoderma lucidum was investigated in detail by spectral peak fitting to the anomeric carbon C1 region in CP/MAS NMR spectra. Hydrated samples gave spectra with enhanced resolution and suggested that a clear distinction between ß-D-glucans and other carbohydrates could be possible in the anomeric carbon C1 region. Chemical shift values for a range of carbohydrate polymers, which can be found alongside ß-D-glucans, as well as the values for various linkages are given. Contamination by other carbohydrates and buffer salts is discussed.
Subject(s)
Agaricales/chemistry , Edible Grain/chemistry , beta-Glucans/analysis , beta-Glucans/chemistry , Avena/chemistry , Carbohydrate Sequence , Hordeum/chemistry , Magnetic Resonance Spectroscopy , Triticum/chemistry , beta-Glucans/isolation & purificationABSTRACT
ß-1,3-D-glucan (BG) activates innate immunity and enhances immune responses. Fungi, such as mushrooms, produce a relatively large amount of BG, the structure and molecular weight of which varies depending on the species of fungi. This study was conducted to develop a detection probe for quantifying or detecting BG from fungi using BG-binding proteins. The binding properties of a new ß-glucan recognition protein (BGRP) against various BGs were compared. With reference to the amino acid sequences of BGRP in insects, an artificial BGRP (supBGRP) was designed with higher production efficiency using gene recombination technology. SupBGRP was produced in Escherichia coli with high efficiency, and its reactivity with BG from fungi was the highest among the BG-binding proteins examined. SupBGRP exhibited high reactivity with 1,6-branched BG and will be useful for the quantification and detection of fungal BG.
Subject(s)
Agaricales/chemistry , beta-Glucans/isolation & purification , beta-Glucans/metabolism , Amino Acid Sequence , Base Sequence , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , beta-Glucans/chemistryABSTRACT
Beta glucan (ß-glucan) has promising bioactive properties. Consequently, the use of ß-glucan as a food additive is favored with the dual-purpose potential of increasing the fiber content of food products and enhancing their health properties. Our aim was to evaluate the biological activity of ß-glucan (antimicrobial, antitoxic, immunostimulatory, and anticancer) extracted from Saccharomyces cerevisiae using a modified acid-base extraction method. The results demonstrated that a modified acid-base extraction method gives a higher biological efficacy of ß-glucan than in the water extraction method. Using 0.5 mg dry weight of acid-base extracted ß-glucan (AB extracted) not only succeeded in removing 100% of aflatoxins, but also had a promising antimicrobial activity against multidrug-resistant bacteria, fungi, and yeast, with minimum inhibitory concentrations (MIC) of 0.39 and 0.19 mg/mL in the case of resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa, respectively. In addition, AB extract exhibited a positive immunomodulatory effect, mediated through the high induction of TNFα, IL-6, IFN-γ, and IL-2. Moreover, AB extract showed a greater anticancer effect against A549, MDA-MB-232, and HepG-2 cells compared to WI-38 cells, at high concentrations. By studying the cell death mechanism using flow-cytometry, AB extract was shown to induce apoptotic cell death at higher concentrations, as in the case of MDA-MB-231 and HePG-2 cells. In conclusion, the use of a modified AB for ß-glucan from Saccharomyces cerevisiae exerted a promising antimicrobial, immunomodulatory efficacy, and anti-cancer potential. Future research should focus on evaluating ß-glucan in various biological systems and elucidating the underlying mechanism of action.
Subject(s)
Liquid-Liquid Extraction , Saccharomyces cerevisiae/chemistry , beta-Glucans/isolation & purification , beta-Glucans/pharmacology , Acids , Anti-Infective Agents/chemistry , Anti-Infective Agents/isolation & purification , Anti-Infective Agents/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/isolation & purification , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Chromatography, High Pressure Liquid , Dose-Response Relationship, Drug , Humans , Immunologic Factors/chemistry , Immunologic Factors/isolation & purification , Immunologic Factors/pharmacology , Immunomodulation/drug effects , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Liquid-Liquid Extraction/methods , Microbial Sensitivity Tests , Spectroscopy, Fourier Transform Infrared , beta-Glucans/chemistryABSTRACT
Elucidating the structure-activity relationship of curdlan is hampered by a lack of characterization with unique specific conformations (i.e., single- or triple-helix). In this study, single-helical curdlan is generated in dilute NaOH solutions at 35-50 °C, and characterized with NMR, SAXS, and GPC. The conformational transition from coil to single-helix and the intramolecular hydrogen bond interaction are explored using NMR. It is found that the two aforementioned types of curdlan interact with Congo Red in very different ways. Single-helical curdlan can encapsulate Congo Red to form a stable, supramolecular dye assembly, which is demonstrated by the shortest distance between the H3 of curdlan and the phenyl groups of Congo Red, and also the same self-diffusion coefficients of Congo Red and curdlan. In contrast, random-coil curdlan interacts weakly with Congo Red and cannot enwrap it. This study offers insight into the specific structure-activity relationship of beta-(1,3)-glucans.
Subject(s)
Congo Red/chemistry , beta-Glucans/chemistry , beta-Glucans/isolation & purification , Chromatography, Gel , Hydrogen Bonding , Magnetic Resonance Spectroscopy , Molecular Conformation , Scattering, Small Angle , Sodium Compounds , Spectrophotometry, Ultraviolet , Structure-Activity Relationship , TemperatureABSTRACT
Two polysaccharide fractions (GLSB50 and GLSB70) with total sugar content of 82.07 wt% and 53.79 wt%, respectively, were obtained from the water extracts of unbroken Ganoderma lucidum spores by sequential ethanol precipitation treatment. Compared with GLSB70, GLSB50 exhibited better activity on stimulation of humoral immune responses in immunosuppressed mice. A novel ß-D-glucan (GLSB50A-III-1) with weight average molecular weight (Mw) of 1.93 × 105 g/mol was purified from GLSB50 through chromatography separation. The exponent α value of Mark-Houwink-Sakurada equation was calculated to be 0.13, indicating that GLSB50A-III-1 presented globular spheres conformation in aqueous solution. Structural analysis showed that GLSB50A-III-1 mainly consisted of (1 â 3), (1 â 4), (1 â 6)-linked ß-d-glucose residues in the backbone, with two single ß-D-Glcp attached at O-6 of ß-(1 â 3) and ß-(1 â 4)-linked residues separately as side chains. The repeat unit of GLSB50A-III-1 was deduced as follows.
Subject(s)
Polysaccharides/chemistry , Reishi/chemistry , Spores, Fungal/chemistry , beta-Glucans/chemistry , Animals , Carbohydrate Sequence , Gas Chromatography-Mass Spectrometry , Magnetic Resonance Spectroscopy , Methylation , Mice, Inbred ICR , Molecular Conformation , Molecular Sequence Data , Molecular Structure , Molecular Weight , Spectroscopy, Fourier Transform Infrared , Water/chemistry , beta-Glucans/isolation & purificationABSTRACT
Agrocybe cylindracea is a common source of active polysaccharides, but their fine structures are not clearly elucidated. In the present study, four fractions were purified from the alkaline extract of A. cylindracea (JACP), and their chemical components and structures were compared by HPAEC-PAD, methylation combined with GC-MS, and 1D/2D NMR analysis. Results showed the purified fractions' physicochemical properties, including monosaccharide compositions, molecular weights, viscosities and surface morphology considerably varied. JACP-30 was identified as a fucoglucogalactan with a α-(1 â 6)-galactopyranosyl as main chain. JACP-50p and JACP-80r were characterized as ß-(1 â 6)-glucans with side chains composed of terminal and 3-substituted ß-glucopyranosyl residues attached at O-3 for every three residues. Similarly, the backbone of JACP-80 was ß-(1 â 6)-linked glucopyranosyl and ß-(1 â 3,6)-linked glucopyranosyl residues at a ratio of 4:1. This work provides more information to the understanding of polysaccharides from A. cylindracea, further guiding its biological researches and developing the application in food and biomedicine industries.
Subject(s)
Agrocybe/chemistry , Fungal Polysaccharides/chemistry , Liquid-Liquid Extraction/methods , Monosaccharides/chemistry , beta-Glucans/chemistry , Carbohydrate Sequence , Fungal Polysaccharides/classification , Fungal Polysaccharides/isolation & purification , Humans , Methylation , Molecular Weight , Monosaccharides/classification , Monosaccharides/isolation & purification , Sodium Hydroxide/chemistry , beta-Glucans/classification , beta-Glucans/isolation & purificationABSTRACT
Four types of mycelial extracts were derived from the airlift liquid fermentation (ALF) of Pleurotus flabellatus, namely exopolysaccharide (EX), endopolysaccharide (EN), hot water (WE), and hot alkali (AE) extracts. Such extracts were screened for their active components and biological potential. EN proved to be most effective in inhibition of lipid peroxidation (EC50 = 1.71 ± 0.02 mg/mL) and in Cupric ion reducing antioxidant capacity (CUPRAC) assay (EC50 = 2.91 ± 0.01 mg TE/g). AE exhibited most pronounced ability to chelate ferrous ions (EC50 = 4.96 ± 0.08 mg/mL) and to scavenge ABTS radicals (EC50 = 3.36 ± 0.03 mg TE/g). ß-glucans and total phenols contributed most to the chelating ability and quenching of ABTS radicals. Inhibition of lipid peroxidation correlated best with total glucans, total proteins, and ß-glucans. Total proteins contributed most to CUPRAC antioxidant capacity. Antifungal effect was determined against Candida albicans ATCC 10231 (MIC: 0.019-0.625 mg/mL; MFC: 0.039-2.5 mg/mL), and towards C. albicans clinical isolate (MIC and MFC: 10.0-20.0 mg/mL). Comparison of cytotoxicity against colorectal carcinoma HCT 116 cells (IC50: 1.8 ± 0.3-24.6 ± 4.2 mg/mL) and normal lung MRC-5 fibroblasts (IC50: 17.0 ± 4.2-42.1 ± 6.1 mg/mL) showed that EN, and especially AE possess selective anticancer activity (SI values 3.41 and 9.44, respectively). Slight genotoxicity was observed only for AE and EX, indicating the low risk concerning this feature. Notable antioxidative and anticandidal activities, selective cytotoxicity against colorectal carcinoma cells, and absence/low genotoxicity pointed out that ALF-cultivated P. flabellatus mycelium could be considered as a valuable source of bioactive substances.
Subject(s)
Antifungal Agents/isolation & purification , Antineoplastic Agents/isolation & purification , Biological Factors/isolation & purification , Bioreactors/microbiology , Pleurotus/growth & development , Antifungal Agents/chemistry , Antifungal Agents/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Biological Factors/chemistry , Biological Factors/pharmacology , Candida albicans/drug effects , Candida albicans/growth & development , Cell Proliferation/drug effects , Cell Survival/drug effects , Fermentation , Fungal Polysaccharides/isolation & purification , Fungal Polysaccharides/pharmacology , HCT116 Cells , Humans , Lipid Peroxidation/drug effects , Microbial Sensitivity Tests , Pleurotus/chemistry , beta-Glucans/isolation & purification , beta-Glucans/pharmacologyABSTRACT
Beta-glucans are polysaccharides of D-glucose monomers linked by (1-3) beta-glycosidic bonds, are found to have a potential immunogenicity risk in biotherapeutic products, and are labeled as process contaminants. A common source of beta-glucans is from the cellulose found in traditional depth filter media. Typically, beta-glucan impurities that leach into the product from the primary clarification depth filters can be removed by the subsequent bind-and-elute affinity chromatography capture step. Beta-glucans can also be removed by a bind-and-elute cation exchange chromatography step, which is useful for removing beta-glucans introduced by a post-Protein A depth filtration step. However, the increasing prevalence of flowthrough polishing chromatography poses a challenge for beta-glucan removal due to the lack of any bind-and-elute chromatography steps after the post-Protein A depth filter. In this work, a depth filter flush strategy was developed to control beta-glucan leaching into the product pool. Different loading conditions for the depth filtration and subsequent chromatography steps were evaluated to determine the robustness of the optimized flush strategy. Carry through runs demonstrated greater than two-fold reduction in beta-glucan levels using the optimized wash as compared to standard filter flush conditions.
Subject(s)
Antibodies, Monoclonal/chemistry , Chromatography, Affinity/methods , Filtration/methods , Immunoglobulin G/immunology , Membranes, Artificial , beta-Glucans/isolation & purification , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/metabolism , HumansABSTRACT
Polysaccharides from P. eryngii mushroom were selectively extracted using low-cost technologies (water at different conditions of temperature and pressure). Mannogalactan was the main polysaccharide in cold-water extracted fraction (CWEF), while a linear (1â6)-ß-d-glucan was the main polymer in hot-water extracted fraction (HWEF). Autoclave-extracted fraction (AEF) contained a mixture of at least four different α- and ß-glucans. The report of linear (1â6)-ß-glucan and linear (1â3)-ß-glucan is a new finding for P. eryngii fruiting bodies. The immunostimulatory properties of the fractions on THP-1 macrophages were studied. All fractions at 50, 250 and 500 µg/mL were not cytotoxic and produced different stimulus on NO, IL-1ß and IL-10 secretion by the cells. Thus, our results showed that it is possible to concentrate different P. eryngii polysaccharides in selected fractions using a simple and low-cost procedure. Since biological effects depends on the polysaccharide structure, this technique allows the obtainment of fractions with distinct immunomodulatory activities.
Subject(s)
Fruiting Bodies, Fungal/chemistry , Fungal Polysaccharides/pharmacology , Immunologic Factors , Macrophages/drug effects , Pleurotus/chemistry , Polysaccharides , beta-Glucans , Complex Mixtures/chemistry , Complex Mixtures/isolation & purification , Complex Mixtures/pharmacology , Fungal Polysaccharides/isolation & purification , Humans , Immunologic Factors/isolation & purification , Immunologic Factors/pharmacology , Immunomodulation , Molecular Structure , Polysaccharides/isolation & purification , Polysaccharides/pharmacology , Structure-Activity Relationship , THP-1 Cells , beta-Glucans/isolation & purification , beta-Glucans/pharmacologyABSTRACT
BACKGROUND: ß-glucan from Lentinus edodes (LNT) is a plant-derived medicinal fungus possessing significant bioactivities on anti-tumor. Both hypoxia-induced factor-1α (HIF)-1α and Nur77 have been shown to be involved in the development of breast cancer. However, there is yet no proof of Nur77/HIF-1α involvement in the process of LNT-mediated tumor-inhibition effect. METHODS: Immunohistochemistry, immunofluorescence and Hematoxylin-Eosin staining were used to investigate tumor growth and metastasis in MMTV-PyMT transgenic mice. Proliferation and metastasis-associated molecules were determined by Western blotting and reverse transcription-quantitative PCR. Hypoxic cellular model was established under the exposure of CoCl2. Small interference RNA was transfected using Lipofectamine reagent. The ubiquitin proteasome pathway was blunted by adding the proteasome inhibitor MG132. RESULTS: LNT inhibited the growth of breast tumors and the development of lung metastases from breast cancer, accompanied by a decreased expression of HIF-1α in the tumor tissues. In in vitro experiments, hypoxia induced the expression of HIF-1α and Nur77 in breast cancer cells, while LNT addition down-regulated HIF-1α expression in an oxygen-free environment, and this process was in a manner of Nur77 dependent. Mechanistically, LNT evoked the down-regulation of HIF-1α involved the Nur77-mediated ubiquitin proteasome pathway. A strong positive correlation between Nur77 and HIF-1α expression in human breast cancer specimens was also confirmed. CONCLUSION: Therefore, LNT appears to inhibit the progression of breast cancer partly through the Nur77/HIF-1α signaling axis. The findings of the present study may provide a theoretical basis for targeting HIFs in the treatment of breast cancer.
Subject(s)
Antineoplastic Agents/pharmacology , Breast Neoplasms/drug therapy , Fungal Polysaccharides/pharmacology , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Nuclear Receptor Subfamily 4, Group A, Member 1/metabolism , Shiitake Mushrooms , beta-Glucans/pharmacology , Adult , Aged , Animals , Antineoplastic Agents/isolation & purification , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Disease Progression , Female , Fungal Polysaccharides/isolation & purification , Gene Expression Regulation, Neoplastic , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Lung Neoplasms/genetics , Lung Neoplasms/metabolism , Lung Neoplasms/prevention & control , Lung Neoplasms/secondary , MCF-7 Cells , Mice, Transgenic , Middle Aged , Nuclear Receptor Subfamily 4, Group A, Member 1/genetics , Proteasome Endopeptidase Complex/metabolism , Shiitake Mushrooms/chemistry , Signal Transduction , Tumor Burden/drug effects , Ubiquitination , beta-Glucans/isolation & purificationABSTRACT
INTRODUCTION: acrylamide is formed in food through Maillard's reaction during thermal processing, and has been shown to be neurotoxic in humans, and a possible carcinogen. Studies have shown that β-glucans from Pleurotus ostreatus have diverse biological properties such as antioxidant and anticancer activities. OBJECTIVE: the aim of this work was to evaluate the protective effect of β-glucans from Pleurotus ostreatus against the harmful effects of acrylamide consumption in mice. METHODS: β-glucans were obtained by alkaline-acid hydrolysis of Pleurotus ostreatus, and the content was characterized by liquid chromatography. To evaluate the effect of β-glucans on the expression of glutathione, Balb/c mice were used, and 4 test groups were established. All groups were fed normally, and the groups treated with acrylamide were administered the compound intragastrically at a concentration of 50 μg/mL; β-glucans were administered at a concentration of 50 μg/mL. RESULTS: mice exposed to acrylamide showed a marked variation in the activity of glutathione enzymes in the liver. Significant differences (p < 0.05) were only found in the expression of glutathione transferase, which was increased almost 3 times in the group treated with β-glucans as compared with the control group, and 1.5 times as compared with the group treated with acrylamide. CONCLUSIONS: the results show that β-glucans could act by increasing the activity of enzymes involved in xenobiotic detoxification, thus protecting the biological system against the harmful effects caused by acrylamide intake
INTRODUCCIÓN: la acrilamida se forma en los alimentos a través de la reacción de Maillard durante el proceso térmico, y ha demostrado ser neurotóxica en humanos y un posible carcinógeno. Algunos estudios han demostrado que los β-glucanos de Pleurotus ostreatus tienen diversas propiedades biológicas, como actividades antioxidantes y anticancerígenas. OBJETIVO: el objetivo de este trabajo fue evaluar el efecto protector de los β-glucanos de Pleurotus ostreatus contra los efectos nocivos por consumo de acrilamida en ratones (prueba in vivo). MÉTODOS: los β-glucanos se obtuvieron por hidrólisis ácido-alcalina de Pleurotus ostreatus y su contenido se caracterizó por cromatografía líquida. La oxidación de los lípidos se evaluó mediante el método de TBARS, y para evaluar el efecto de los β-glucanos en la expresión de glutatión se usaron ratones Balb/c, y se establecieron 4 grupos de prueba. Todos los grupos fueron alimentados normalmente; a lo grupos tratados con acrilamida, esta se les administró intragástricamente en una concentración de 50 μg/ml, y los β-glucanos en una concentración de 50 μg/ml. RESULTADOS: en el presente trabajo, los ratones expuestos a acrilamida mostraron una marcada variación en la actividad de las enzimas de glutatión determinadas en el hígado. Solo se encontraron diferencias significativas (p < 0,05) en la expresión de glutatión-transferasa, que aumentó casi 3 veces en el grupo tratado con β-glucano en comparación con el grupo de control, y 1,5 veces con respecto al grupo tratado con acrilamida. CONCLUSIONES: los resultados muestran que los β-glucanos podrían actuar como agentes antioxidantes que protegen el hígado contra el estrés oxidativo causado por la ingesta de acrilamida