Tea made from the leaves of Eryngium foetidum is traditionally consumed by the Amazonian population for treatment of pain and inflammation. This study was carried out with a polysaccharide fraction (EFS100R) obtained from the hot aqueous extract of E. foetidum leaves. Structural analysis revealed that the polysaccharides of EFS100R are predominantly constituted by galactose, galacturonic acid and arabinose monosaccharide units. Methylation and NMR spectroscopy further identified the fraction as primarily composed of type II arabinogalactan, with smaller proportions of homogalacturonan and type I rhamnogalacturonan. To evaluate the potential antinociceptive and anti-inflammatory properties of EFS100R, the abdominal writhing model induced by acetic acid in mice was used. Results demonstrated that the E. foetidum polysaccharide fraction significantly reduced the number of writhing episodes, leukocyte migration and plasma extravasation when previously administrated to the acetic acid intraperitoneally, indicating that EFS100R has therapeutic potential as an anti-inflammatory and analgesic agent. Thus, this study shows that water-soluble polysaccharides contribute to the effects related to the E. foetidum tea.

Mushroom-derived β-D-glucans are structurally diverse bioactive polysaccharides with promising applications in functional foods, nutraceuticals, and health-related products. Despite extensive research, establishing clear structure–function relationships remains a major challenge due to the intrinsic complexity of these polymers and the lack of standardized characterization approaches. Therefore, this review provides a critical overview of methodologies used for the characterization of mushroom β-D-glucans, with particular emphasis on advanced analytical strategies for purification assessment and structural elucidation, including nuclear magnetic resonance (NMR), methylation analysis, and complementary physicochemical methods. We discuss the limitations of commonly used approaches, particularly the risks associated with relying on isolated or limited analytical methodologies, and propose an integrated workflow for the structural characterization of mushroom-derived β-D-glucans. Furthermore, we systematically correlate key structural features, such as molecular weight, branching degree, and solubility, with reported biological effects, providing a conceptual framework to guide future investigations. By integrating chemical, physicochemical, and biological perspectives, this review aims to improve reproducibility, minimize misinterpretation, and support the rational development of β-D-glucan-based applications. Overall, this work provides critical insights and practical recommendations to advance the understanding and translational potential of mushroom-derived β-D-glucans.

Background: Ganoderma lucidum is a medicinal mushroom widely recognized for its high content of bioactive polysaccharides, particularly β-glucans with immunomodulatory properties. This study aimed to optimize polysaccharide extraction conditions to maximize yield and glucan content, and to evaluate the biological activity of the obtained fractions in an experimental model of intestinal mucositis. Methods: Polysaccharides were extracted using a combination of hot-water extraction and ethanol precipitation, optimized by response surface methodology. Optimal conditions (121 °C for 120 min followed by 90% ethanol precipitation) yielded a crude polysaccharide fraction (Poli-GL). A subsequent freeze–thaw process generated a soluble fraction (S-Poli-GL). Structural and compositional characterization was performed using enzymatic assays, monosaccharide profiling, and NMR spectroscopy. The biological effects of Poli-GL and S-Poli-GL were evaluated in a 5-fluorouracil-induced intestinal mucositis model following oral administration at doses of 30, 100, and 300 mg/kg. Results: The optimized extraction protocol enabled efficient recovery of polysaccharides enriched in glucans. S-Poli-GL exhibited a high total glucan content, including 43.3% β-glucans and 3.45% α-glucans, along with minor amounts of galactose and mannose. Structural analysis confirmed the predominance of branched β-(1→3),(1→6)-D-glucans. While Poli-GL did not prevent mucositis development, S-Poli-GL significantly reduced the disease activity index and attenuated intestinal inflammation, indicating enhanced biological activity associated with the soluble glucan-rich fraction. Conclusions: Optimization of extraction and fractionation improves the functional properties of G. lucidum polysaccharides. The soluble glucan-enriched fraction (S-Poli-GL) demonstrated significant protective effects in intestinal mucositis, supporting its potential as a therapeutic candidate and warranting further investigation for clinical application.

Mushroom-derived (1 → 3),(1 → 6)-β-D-glucans exhibit diverse biological activities, including effects relevant to cancer research, which may be modulated through chemical modification. In the present study, sequential hydroxypropylations were performed on a highly purified (1 → 3),(1 → 6)-β-D-glucan (NM-Glc) from the mushroom Lactarius quieticolor to generate derivatives with distinct structural characteristics (HP1 and HP2). The derivatives were characterized by NMR, GC–MS, and SEC-MALS-RI, which suggested preferential hydroxypropyl substitution at the O-6 and O-4 positions. Each sample (NM-Glc (unmodified glucan; Mw 1.26 × 105 g.mol−1), HP1 (Mw 4.02 × 104 g.mol−1, MS: 0.70, DSFrac: 0.38, DSSub: 0.57), and HP2 (Mw 5.66 × 104 g.mol−1, MS: 0.94, DSFrac: 0.47, DSSub: 0.83) was evaluated for its biological effects on B16-F10 murine melanoma cells at 10, 100, and 1000 μg.mL−1. NM-Glc did not exhibit significant cytotoxicity toward tumor cells under the evaluated conditions, whereas the hydroxypropyl-modified glucans exhibited cytotoxicity at 100 and 1000 μg.mL−1 and reduced cell adhesion to the substrate (observed only for HP2 at 1000 μg.mL−1). Acridine orange/ethidium bromide (AO/EB) staining revealed an increased proportion of apoptotic-like cells following treatment with HP1 and HP2, with HP2 producing the most pronounced effects. Therefore, hydroxypropylation is a promising strategy to generate (1 → 3),(1 → 6)-β-D-glucans capable of reducing melanoma cell viability in vitro.

Tropical fruits are an important and yet underexplored source of dietary fibers, and biribá (Annona mucosa), a fruit native to the Amazon region, remains poorly characterized in this regard. In this study, soluble (SDF) and insoluble dietary fiber (IDF) fractions from biribá fruit pulp were extracted, fractionated, and structurally characterized using a combination of enzymatic–gravimetric methods, chromatographic techniques, monosaccharide composition, linkage analysis, and NMR spectroscopy. Six distinct polysaccharides were identified. The SDF fraction yielded a highly methyl-esterified (DE = 68.67 ± 0.43%) homogalacturonan, a type II arabinogalactan (Mw of 62 kDa), and a highly branched 4-O-methyl-glucuronoxylan. The SDF fraction was evaluated as a macrophage modulator in vitro, and was non-cytotoxic at concentrations up to 1000 μg/mL, while stimulating macrophages to secrete nitric oxide, a classical marker of the M1 phenotype. The IDF fraction contained a low-substituted acidic xylan, a xyloglucan/mannan complex (Mw of 36 kDa) and a cellulosic residue. Overall, this study provides the first detailed characterization of biribá pulp polysaccharides, revealing marked structural diversity across dietary fiber fractions. The observed increase in nitric oxide production by macrophages provides initial evidence of the biological activity of the soluble dietary fiber fraction and supports further investigation of the relationship between its structural features and biological properties.

Natural molecules with biological properties are widely investigated to better understand their broad range of functions. Among these, (1→3),(1→6)-β-D-glucans are extensively studied due to their remarkable biological activities, particularly their immunomodulatory effects. In this study, we extracted and purified a (1→3),(1→6)-β-D-glucan (Mw 6.02 × 105 g.mol-1) from the mushroom Amanita semigemmata. The polysaccharide (AS-P) was chemically characterized by 1H, 13C and HSQC-NMR, methylation, monosaccharide composition, homogeneity and molecular weight analysis. Additionally, sugar and protein content were quantified. In vitro experiments demonstrated that AS-P is non-cytotoxic at concentrations up to 500 μg.mL-1 and modulates RAW 264.7 macrophage activity. AS-P significantly reduced nitric oxide (NO) production by 50.95 % and 60.14 % at concentrations of 250 and 500 μg.mL-1, respectively, when co-incubated with lipopolysaccharide (LPS). Furthermore, AS-P induced a downregulation in the pro-inflammatory cytokine IL-6 and an upregulation of TNF and MCP-1 cytokines, indicating a dual modulatory effect. In vivo assays further demonstrated that the AS-P fraction was highly effective in reducing pain by 97 % during the inflammatory phase (15 to 30 min) at a dose of 30 mg.kg-1 following intraplantar injection of formalin solution. These findings highlight the potential of AS-P as a natural immunomodulatory compound, exhibiting dual regulation of cytokine production while exerting both anti-inflammatory and antinociceptive effects.

Mushroom-derived polysaccharides are known for their diverse biological activities, including immunomodulatory and anticancer properties. Breast cancer, the leading cause of cancer-related death among women worldwide, lacks targeted molecular therapies, underscoring the need for novel therapeutic agents. In this study, we extracted, purified, and characterized a novel heteropolysaccharide (HTP) - a rhamnogalactan - composed of galactose (72.6 %) and rhamnose (27.4 %) from the edible mushroom Lactarius quieticolor. Structural analysis revealed that HTP consists of a (1 → 6)-linked α-d-galactopyranose backbone, heavily substituted at O-2 by non-reducing rhamnose units, with a weight-average molar mass of 2.41 × 104 g.mol-1. In vitro assays using a triple-negative breast cancer (TNBC) cell model - the most aggressive subtype - showed a significant reduction in cell viability (50 % at 1200 μg·mL-1). Furthermore, an additive effect was observed when co-administered with the chemotherapy drug paclitaxel, reducing the cell viability to 42 %. These findings suggest potential therapeutic applications of the purified mushroom rhamnogalactan reported in this study against TNBC.

The mushroom Pleurotus ostreatus is widely produced in Brazil and its stipes are discarded before commercialization. In the present study, this agricultural fungal waste (mushroom stipes), was analyzed by preparing an aqueous extract and obtaining the polysaccharides by ethanol precipitation (POS-extract). The fraction presented 37 % of carbohydrates and small amounts of proteins and phenolic compounds. Rheological analyses showed a high viscosity and a gel-like behavior depending on temperature and concentration. The main polysaccharides present were (3-glucans and mannogalactans, which were confirmed by the enzymatic method, monosaccharide composition, and NMR. Methylation analyses showed a high content of 1,6-linkages for both polysaccharides. The POS-extract was tested on Caco-2 cells to observe cytotoxicity and cell migration ability. Incubation with different concentrations of POS-extract showed no significant cytotoxic effects (p >= 0.05) and promoted wound closure of the injured monolayer by 95 % after 72 h (at 300 mu g/mL). The preliminary results indicate a promising application of mushroom by-products, containing easily extractable bioactive compounds.

Ziziphus joazeiro ("juazeiro") is an endemic tree species of the Caatinga biome, found exclusively in Brazil. Its edible yellow fruits contain a sweet, mucilaginous pulp. Soluble dietary fibers (ZJ-SDF) were extracted from juá pulp using the enzymatic-gravimetric method, while the mucilage (ZJS) that surrounds the seeds was solubilized by water extraction. Three polysaccharides were purified and structurally characterized: a) an arabinogalactan, with Mw of 7.8 × 104 g/mol and composed of terminal, 6-, 3-, 4-, 3,4-, 3,6-, 4,6-, 3,4,6- and 2,3,6-O-linked Galp units, indicating a highly branched polysaccharide, that contains terminal, 3-, 5-, 2,5- and 3,5-O-linked Araf units as side chains; b) a rhamnogalacturonan (Mw = 9.0 × 105 g/mol) containing 2-O-, 4-O-, 2,4-di-O-linked Rhap units and 4-O-linked GalA units. Side chains attached to O-4 of Rhap units may be composed of arabinan, galactan, terminal Rhap, and terminal Galp/GalpA units; c) a linear (1 → 6)-α-D-glucan. This is the first report of complex polysaccharides from Ziziphus joazeiro fruits and the first identification of (1 → 6)-α-D-glucan in the genus Ziziphus. These findings highlight the structural diversity and complexity of polysaccharides from soluble dietary fibers and seed mucilage of this tropical edible fruit.

There is an incessant search for new therapies against cancer, able to unite effectiveness with specificity, leading to higher survival rates and lower deleterious effects. Herein, two fucogalactans (F-1 and F-2), isolated from Macrocybe titans, showed a (1 → 6)-linked α-D-Galp main chain partially substituted at O-2 by non-reducing end units of α-L-Fucp, with different Mw, being F-2 > 20 times higher than F-1. Both fucogalactans induced cell cycle arrest of MDA-MB-231 cells in G1 phase after 120 h of treatment. However, only F2 resulted in increased apoptosis and necrosis. For the MCF-7 cell line, no changes in the cell cycle and cell death were observed at 120 h. The non-tumoral cell line (VERO) was not affected. The results confirmed that polysaccharides with different Mw may have distinct effects and therefore this is an important feature to be considered on investigating anti-cancer treatments.

Mushroom-derived polysaccharides, particularly β-D-glucans, are valued for their biological activities and broad biotechnological applications. Although several conventional extraction methods have been established (cold-water, hot-water, and alkaline), the biomass that remains after exhaustive treatments is often discarded, despite its potential to retain valuable compounds. In this study, we evaluated whether residual laboratory materials from Pleurotus pulmonarius, Pholiota nameko, and Amanita muscaria still contain extractable polysaccharides by applying sequential autoclave extractions under aqueous, 5 % KOH, and 20 % KOH conditions. The extracts were precipitated with ethanol, quantified, and characterized by NMR, GC-MS, HPSEC-RI, and by total sugar and protein determinations. Significant yields were obtained, particularly with 5 % KOH, which produced up to 31.1 % in a single step, albeit with considerable protein content. Across extraction steps, a progressive decrease in molecular weight was observed, and monosaccharide analysis revealed that the extracts were composed predominantly of glucose. Structural characterization confirmed a predominance of β-D-glucans with (1→3)- and (1→6)-linkages, together with α-D-glucans with (1→3)- and (1→4)-linkages, as well as galactans and mannans. Overall, these findings demonstrate that autoclave extraction is a simple, efficient, and scalable approach to recover structurally diverse polysaccharides from mushroom residues, thereby enhancing resource utilization and opening new opportunities for fungal biomass valorization.

In situ forming hydrogels are promising for biomedical applications, especially in drug delivery. The precursor solution can be injected at the target site, where it undergoes a sol-gel transition to afford a hydrogel. In this sense, the most significant characteristic of these hydrogels is fast gelation behavior after injection. This study describes an all-polysaccharide, rapidly in situ-forming hydrogel composed of carboxymethyl chitosan (CMCHT) and hydroxyethyl cellulose functionalized with aldehyde groups (HEC-Ald). The HEC-Ald was synthesized through acetal functionalization, followed by acid deprotection. This innovative approach avoids cleavage of pyran rings, as is inherent in the periodate oxidation approach, which is the most common method currently employed for adding aldehyde groups to polysaccharides. The resulting hydrogel exhibited fast stress relaxation, self-healing properties, and pH sensitivity, which allowed it to control the release of an encapsulated model drug in response to the medium pH. Based on the collected data, the HEC-Ald/CMCHT hydrogels show promise as pH-sensitive drug carriers.

Ganoderma lucidum , widely used in traditional medicine, has several biological properties. Polysaccharides, mainly glucans, are known as one of its main bioactive compounds. Consequently, the achievement and chemical investigation of such molecules are of pharmaceutical interest. Herein, we obtained water-insoluble and watersoluble polysaccharides from G. lucidum by alkaline extraction. Fractionation process yielded three fractions (GLC-1, GLC-2, and GLC-3). All samples showed to be composed mainly of glucans. GLC-1 is a linear (1 -> 3)linked beta-glucan; GLC-2 is a mixture of three different linear polysaccharides: (1 -> 3)beta-glucan, (1 -> 3)alpha-glucan, and (1 -> 4)- alpha-mannan; while GLC-3 is a branched beta-glucan with a (1 -> 4)-linked main chain, which is branched at O-3 or O-6 by (1 -> 3)or (1 -> 6)-linked side chains. This research reports the variability of glucans in Ganoderma lucidum fruiting bodies and applicable methodologies to obtain such molecules. These polysaccharides can be further applied in biological studies aiming to investigate how their chemical differences may affect their biological properties.

Mushroom polysaccharides consist of a unique set of polymers that arrive intact in the human large intestine becoming available for fermentation by resident gut bacteria with potential benefits to the host. Here we have obtained four glucans from two mushrooms (Pholiota nameko and Pleurotus pulmonarius) under different extraction conditions and their fermentation profile by human gut bacteria in vitro was evaluated. These glucans were isolated and characterized as (1 -> 3),(1 -> 6)-beta-D-glucans varying in branching pattern and water-solubility. An aliquot of each (1 -> 3),(1 -> 6)-beta-D-glucan was subjected to controlled smith degradation process in order to obtain a linear (1 -> 3)-beta-D-glucan from each fraction. The four beta-D-glucans demonstrated different water solubilities and molar mass ranging from 2.2 x 10(5) g.mol(-1) to 1.9 x 10(6) g.mol(-1). In vitro fermentation of the glucans by human gut microbiota showed they induced different short chain fatty acid production (52.0-97.0 mM/50 mg carbohydrates), but an overall consistent high propionate amount (28.5-30.3 % of total short chain fatty acids produced). All glucans promoted Bacteroides uniformis, whereas Anaerostipes sp. and Bacteroides ovatus promotion was strongly driven by the beta-D-glucans solubility and/or branching pattern, highlighting the importance of beta-D-glucan discrete structures to their fermentation by the human gut microbiota.

Several beneficial properties have already been observed for mushrooms extracts containing polysaccharides, one of the main components present in these fungi. In order to obtain these extracts and clarify their composition, different extraction and characterization methods can be applied, contributing to understanding its biological effects. In this research, we evaluated three different crude fractions obtained through cold water, hot water, and alkaline solution, from the mushroom Lactarius quieticolor. Each sample was totally characterized by NMR (1H, 13C, HSQC-DEPT, COSY), GC-MS (monosaccharide composition and methylation analysis) and HPSEC (homogeneity and relative Mw). The results showed a mixture of different polysaccharides, including D-galactans and D-glucans in each fraction. All the extracts presented immunostimulant activity in murine RAW 264.7 macrophages, stimulating NO secretion and inducing morphological alterations that were typical of activated cells. These effects were more intense for alkaline fraction (EA), which have higher amounts of mannose and 3,6-di-O-substituted-Glcp units. The results obtained show that crude fractions can potentially stimulate immune cells, which indicates that the consumption of this edible mushroom can bring great benefits to health, being considered an excellent food due to its good biological properties.

Soursop fruits are widely used in the folk medicine to treat a variety of health conditions. Once the chemical structure of dietary fibers from fruits is closely related to its biological functions in the human body, we aimed to explore structural features and biological activity of dietary fibers from soursop. Polysaccharides that constitute the soluble and insoluble fibers were extracted and further analyzed using monosaccharide composition, methylation, molecular weight determination and 13C NMR data. Soursop soluble fibers (SWa fraction) were characterized as having type II arabinogalactan and a highly methyl esterified homogalacturonan, while non-cellulosic insoluble fibers (SSKa fraction) were mainly composed by a pectic arabinan, a xylan-xyloglucan complex and a glucuronoxylan. The oral pre-treatment with SWa and SSKa promoted antinociception in mice writhing test, reducing the number of pain-like behaviors (in 84.2 % and 46.9 %, respectively, at 10 mg/kg) and peritoneal leucocyte migration (55.4 % and 59.1 %, at 10 mg/kg), effects possibly associated with the pectins present in fruit pulp extractions. SWa also significantly inhibited the plasmatic extravasation of Evans blue dye in 39.6 % at 10 mg/kg. This paper describes for the first time the structural features of soursop dietary fibers that may be of biological significance in future.

C. brachyspora, a widespread dematiaceous fungus, was evaluated in this study to optimize the production of exopolysaccharides (CB-EPS). Optimization was performed using response surface methodology, and the best production yielded 75.05% of total sugar at pH 7.4, with 0.1% urea, after 197 h. The obtained CB-EPS showed typical signals of polysaccharides, which was confirmed by FT-IR and NMR. The HPSEC analysis indicated a polydisperse polymer, showing a non-uniform peak, with an average molar mass (Mw) of 24,470 g/mol. The major monosaccharide was glucose (63.9 Mol%), followed by mannose (19.7 Mol%), and galactose (16.4 Mol%). Methylation analysis encountered derivatives that indicated the presence of a β-d-glucan and a highly branched glucogalactomannan. CB-EPS was tested on murine macrophages to verify its immunoactivity, and the treated cells were able to produce TNF-α, IL-6, and IL-10. However, the cells did not produce superoxide anions or nitric oxide nor stimulated phagocytosis. The results demonstrated an indirect antimicrobial activity of macrophages by stimulating cytokines, showing another biotech applicability for the exopolysaccharides produced by C. brachyspora.

An optimized PLE method was applied to several truffle species using three different solvent mixtures to obtain bioactive enriched fractions. The pressurized water extracts contained mainly (1 → 3),(1 → 6)-β-D-glucans, chitins, and heteropolymers with galactose and mannose in their structures. The ethanol extracts included fatty acids and fungal sterols and others such as brassicasterol and stigmasterol, depending on the species. They also showed a different fatty acid lipid profile depending on the solvent utilized and species considered. Ethanol:water extracts showed interesting lipids and many phenolic compounds; however, no synergic extraction of compounds was noticed. Some of the truffle extracts were able to inhibit enzymes related to type 2 diabetes; pressurized water extracts mainly inhibited the α-amylase enzyme, while ethanolic extracts were more able to inhibit α-glucosidase. Tuber brumale var. moschatum and T. aestivum var. uncinatum extracts showed an IC50 of 29.22 mg/mL towards α-amylase and 7.93 mg/mL towards α-glucosidase. Thus, use of the PLE method allows o bioactive enriched fractions to be obtained from truffles with antidiabetic properties.