Identification and Quantification of Monosaccharides from Fungal Cell Walls and Exopolysaccharides by Gas Chromatography Coupled to Mass Spectrometry.

The fungal cell wall and secreted exopolysaccharides play an important role in the interactions between fungi and their environment. Despite their central role in fungal biology, ecology, and host-pathogen interactions, the composition of these polymers and their synthetic pathways are not well understood. The protocols presented in this article describe an approach to isolate fungal cell wall polysaccharides and to identify and quantify the monosaccharide composition of these polymers by gas chromatography-mass spectrometry (GC-MS). © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: O-methyl trimethylsilyl monosaccharide derivatives composition analysis by GC-MS Support Protocol: Fungal cell wall extraction.

Characterization and immunomodulatory effect of an alkali-extracted galactomannan from Morchella esculenta.

In our continuous exploration for bioactive polysaccharides, a novel polysaccharide FMP-2 was isolated and purified from the fruiting bodies of Morchella esculenta by alkali-assisted extraction. FMP-2 had an average molecular weight of 1.09 × 106 Da and contained mannose, glucuronic acid, glucose, galactose, and arabinose in a molar ratio of 4.10:0.22:1.00:5.75:0.44. The backbone of FMP-2 mainly consisted of 1,2-α-D-Galp, 1,6-α-D-Galp, and 1,4-α-D-Manp, with branches of 1,4,6-α-D-Manp and 1,2,6-α-D-Galp. FMP-2 can stimulate phagocytosis and promote the secretion of NO, ROS, and cytokines like IL-6, IL-1ß, and TNF-α in RAW264.7 cells ranging from 25 to 400 µg/mL. FMP-2 had great repairing effect on the immune injury of zebrafish induced by chloramphenicol. The phagocytosis ability of zebrafish macrophages and the proliferation of neutrophils can be greatly enhanced by polysaccharide FMP-2 with concentrations from 50 to 200 µg/mL. These findings suggest that FMP-2 might be used as a potential immunomodulator in the food and pharmaceutical industries.

Selenium-Containing Exopolysaccharides Isolated from the Culture Medium of Lentinula edodes: Structure and Biological Activity.

In continuation of our research on the influence of selenium incorporation on the biosynthesis, structure, and immunomodulatory and antioxidant activities of polysaccharides of fungal origin, we have isolated from a post-culture medium of Lentinula edodes a selenium (Se)-containing exopolysaccharide fraction composed mainly of a highly branched 1-6-α-mannoprotein of molecular weight 4.5 × 106 Da, with 15% protein component. The structure of this fraction resembled mannoproteins isolated from yeast and other mushroom cultures, but it was characterized by a significantly higher molecular weight. X-ray absorption fine structure spectral analysis in the near edge region (XANES) suggested that selenium in the Se-exopolysaccharide structure was present mainly at the IV oxidation state. The simulation analysis in the EXAFS region suggested the presence of two oxygen atoms in the region surrounding the selenium. On the grounds of our previous studies, we hypothesized that selenium-enriched exopolysaccharides would possess higher biological activity than the non-Se-enriched reference fraction. To perform structure-activity studies, we conducted the same tests of biological activity as for previously obtained mycelial Se-polyglucans. The Se-enriched exopolysaccharide fraction significantly enhanced cell viability when incubated with normal (human umbilical vein endothelial cells (HUVEC)) cells (but this effect was absent for malignant human cervical HeLa cells) and this fraction also protected the cells from oxidative stress conditions. The results of tests on the proliferation of human peripheral blood mononuclear cells suggested a selective immunosuppressive activity, like previously tested Se-polyglucans isolated from L. edodes mycelium. The Se-exopolysaccharide fraction, in concentrations of 10-100 µg/mL, inhibited human T lymphocyte proliferation induced by mitogens, without significant effects on B lymphocytes. As with previously obtained Se-polyglucans, in the currently tested Se-polymannans, the selenium content increased the biological activity. However, the activity of selenium exopolysaccharides in all tests was significantly lower than that of previously tested mycelial isolates, most likely due to a different mode of selenium binding and its higher degree of oxidation.

The chemical structure and immunomodulatory activity of an exopolysaccharide produced by Morchella esculenta under submerged fermentation.

The extracellular polysaccharide of Morchella esculenta cultivated under submerged fermentation was extracted. A single polysaccharide was purified through DEAE-Cellulose 52 and Sephadex G 100, and named as MEP 2a. The molecular weight of MEP 2a was determined by HPGPC and it is about 1391.5 kDa. MEP 2a is composed of mannose and glucose as the monosaccharide unit with a molar ratio of 8.15 : 1.07. The main polysaccharide chemical structure was analyzed by 1D and 2D NMR. Methylation and NMR analysis revealed that the backbone of MEP 2a consists of 1,3,4-linked-Manp, 1,2-linked-Manp and 1,6-linked-Glcp. 1D and 2D NMR results indicated that the main chain is based on →1)-ß-D-Glcp-(6→, →1)-α-D-Manp-(3,4→, →1)-α-D-Manp-(2→) and the branch chain is composed of α-D-Manp-(1→, →1)-ß-D-Glcp-(6→ and α-D-Glcp-(1→). MEP 2a promoted the phagocytosis function and secretion of NO, IL-1ß, IL-6 and TNF-α of macrophages. In the present study, the chemical structure and immunomodulatory ability of an extracellular polysaccharide of Morchella esculenta was investigated which guarantees further research studies and promising applications.

Structural characteristics, anti-proliferative and immunomodulatory activities of a purified polysaccharide from Lactarius volemus Fr.

Lactarius volemus Fr. is an edible mushroom widely consumed in China. Polysaccharide is an important nutritional component of L. volemus. This research aimed to isolate the polysaccharide from L. volemus and study its structure and bioactivities. A purified polysaccharide was identified and named as LVF-I whose primary structure was proposed considering the comprehensive results of monosaccharide composition, periodate oxidation-smith degradation, methylation analysis, FT-IR and 1D/2D NMR spectroscopy. Then the immunomodulation of LVF-I and its inhibition effect on H1299 and MCF-7 cells were investigated. Results showed that LVF-I (12,894 Da) contained fucose, mannose, glucose and galactose. It had a backbone consisting of →4)-α-D-Glcp-(1→, →6)-ß-D-Manp-(1→, →6)-α-D-Galp-(1 â†’ and →4)-ß-D-Manp-(1→. And its side chains were branched at C2 of →4)-ß-D-Manp-(1 â†’ by →6)-α-D-Galp-(1→, α-D-Glcp-(1→, α-D-Galp-(1 â†’ and α-L-Fucp-(1→. LVF-I (250-1000 µg/mL) could inhibit the proliferation of H1299 and MCF-7 cells, while enhance the proliferative response of splenocyte and the phagocytic ability of RAW264.7. Furthermore, LVF-I (250-1000 µg/mL) significantly induced the secretion of nitric oxide, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) by up-regulating their mRNA expression in macrophages. These results suggested that LVF-I had the potential to be developed as antitumor or immunomodulatory agents by inhibiting the proliferation of tumor cells and stimulating macrophages-mediated immune responses.

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.

Characteristics and antifatigue activity of graded polysaccharides from Ganoderma lucidum separated by cascade membrane technology.

In this paper, cascade membrane technology was utilized to classify polysaccharides from Ganoderma lucidum (GLPs). The properties and antifatigue activity of graded polysaccharides were identified and compared. GLPs were separated using cascade ultrafiltration membranes of 100 kDa, 10 kDa and 1 kDa in sequence. The molecular weights of polysaccharides in these GLP fractions were approximately 322.0 kDa, 18.8 kDa and 6.4 kDa, and all polysaccharides were in active ß-configurations. This showed that all graded GLPs could elongate swimming time, improve endurance and promote fatigue recovery, especially polysaccharides with molecular weights above 10 kDa. This demonstrated that GLPs could decrease the activities of SUN and CK and the levels of MDA and BLA. They also increased the level of Gly, accelerated fat transformation, and improved the activities of GPx, SOD and LDH in all treated mice. Accordingly, GLPs above 10 kDa might be potential agents with antifatigue activity.

The lipid lowering and antioxidative stress potential of polysaccharide from Auricularia auricula prepared by enzymatic method.

An efficient extraction method of Auricularia auricula polysaccharides (AAPs) by neutral protease was developed and optimized by response surface methodology. AAPs were graded by stepwise ethanol precipitation, the fraction with high recovery rate and strong radical scavenging rate were obtained, then its antioxidant and lipid lowering effect were studied using Caenorhabditis elegans as model organism. The extract yield and ABTS+ scavenging rates of AAPs could reach 14.90% and 86.0% at 50 °C, 75 mL/g of liquid-to-material ratio and pH 9.0. AAP3 obtained by 15% ethanol was a heteropolysaccharide comprised of mannose, glucose, glucuronic acid, xylose, galactose and glucosamine. AAP3 could significantly prolong the lifespan of C. elegans and enhance the activity of antioxidant enzymes including superoxide dismutase (SOD), catalases (CAT) at 0.25 mg/mL (p < 0.05). The qRT-PCR results showed that AAP3 could up regulate mRNA expression levels of daf-16 and skn-1 (>1.6 fold) at 0.25 mg/mL. Besides, AAP3 could significantly reduce the level of body fat and triglyceride in C. elegans (p < 0.05). These studies demonstrated that A. auricula polysaccharides prepared by neutral protease had a prominent protective effect to the damage induced by the intracellular free radical generating agents.

Submerged cultivation, characterization and in vitro antitumor activity of polysaccharides from Schizophyllum radiatum.

Production of polysaccharides by white-rot-fungi in submerged cultivation has several advantages due to process control. This work deals with the submerged cultivation, extraction and antitumor activity of polysaccharides from a wild strain of Schizophyllum radiatum isolated from a tropical forest of Colombia. The mushroom was cultivated in laboratory conditions, and classified by classical and molecular taxonomy. Submerged cultivation was performed in a bioreactor of 5 L using a ligninolytic residue as substrate. The fermentation conditions were 30 ± 1 °C, pH 4.5, 300 rpm and 1.5 vvm of air for 4 days. The yields were 16.8 g/L (w/v) of biomass, and after extraction, 0.6 g/L of water-soluble exopolysaccharide (SEPS) and 2.01 % (w/w) of water-soluble intrapolysaccharide (SIPS) were obtained. In each extract total carbohydrate, glucans and protein contents were determined. Also, nuclear magnetic resonance (NMR), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray diffractometry (XRD), high performance liquid chromatography with refraction index detection (HPLC-RI), high performance gel permeation chromatography (HPGPC) and Nuclear Magnetic Resonance (NMR) analysis were performed. Results indicated that SEPS and SIPS are heteropolysaccharides with amorphous structure and high molecular weights. Antitumor and immunostimulant activity was evaluated in different cancer cell lines. The results suggest these polysaccharides have direct and indirect antitumor activity activating immune cells such as macrophages. These findings enhance our knowledge about new sources of fungal metabolites that serve as adjuvant, cheaper and less harmful alternatives to cancer treatment.

Chemical structure and anti-inflammatory activity of a branched polysaccharide isolated from Phellinus baumii.

Phellinus baumii is used to treat inflammatory bowel disease (IBD) and gastroenteritis. In this study, a 46 kDa heteropolysaccharide SHPS-1 was isolated from fruiting bodies of P. baumii. SHPS-1 consisted of arabinose, mannose, glucose, and galactose at a molar ratio of 2.2:15.7:49.3:32.8. SHPS-1 had a backbone containing 1,3-linked ß-D-Glcp and 1,6-linked α-D-Galp residues, and Araf, Manp and Galp units were attached as oligosaccharidic side chains to the backbone at C-6 of some glucopyranoses. SHPS-1 decreased phosphorylation level of STAT-1 and expression levels of STAT-1 targeted genes such as iNOS and TNF-α in lipopolysaccharide-stimulated macrophage RAW 264.7 cells. Furthermore, SHPS-1 promoted the expression of IL-10 and macrophage mannose receptor CD 206, markers of tissue repairing macrophages. SHPS-1 alleviated ulcerative colitis in mice by decreasing pro-inflammatory genes and increasing anti-inflammatory and tissue repairing genes. Collectively, SHPS-1 polysaccharide from P. baumii had anti-inflammatory activity and can potentially treat IBD.

Fabrication and characterization of Rhizochitosan and its incorporation with platelet concentrates to promote wound healing.

Composite dressing composed of Rhizochitosan and Regenplex™ to promote wound healing were assessed. Rhizochitosan was fabricated by deacetylation of Rhizochitin, which obtained by simply depigmenting sporangium-free mycelial mattress produced from Rhizopus stolonifer F6. Physicochemical characterizations of Rhizochitosan demonstrated that it contained 13.5% chitosan with a water-absorption ability of 35-fold dry weight and exhibiting hydrogel nature after hydration. In a wound-healing study on SD rats with full-thickness injury, the composite dressing had a better healing effect than those for each individual components and control group and wound even healed as functional tissue instead of scar tissue. The underlying mechanism of the composite beneficial to wound remodeling is likely attributable to a more reduction level of matrix metalloproteinase (MMP)-9 expression in early stage and a higher MMP-2 expression level in a later stage of healing process. Conclusively, the composite dressing demonstrated to be highly beneficial to the healing of full-thickness injury wound.

Recent advances in polysaccharides from Lentinus edodes (Berk.): Isolation, structures and bioactivities.

Lentinus edodes, an important edible mushroom cultivated in East Asia for thousands of years, has been widely used as food and medicinal ingredient worldwide. Modern phytochemistry studies have demonstrated that L. edodes is very rich in bioactive polysaccharides, especially the ß-glucans. Over the past two decades, the isolation, chemical properties, and bioactivities of polysaccharides from fruiting bodies, mycelium and fermentation broth of L. edodes have been drawing much attention from scholars around the world. It has been demonstrated that L. edodes polysaccharides possess various remarkable biological activities, including anti-oxidant, anti-tumor, anti-aging, anti-inflammation, immunomodulatory, antiviral, and hepatoprotection effects. This review summarizes the recent development of polysaccharides from L. edodes including the isolation methods, structural features, bioactivities and mechanisms, and their structure-activity relationship, which can provide useful research underpinnings and update information for their further application as therapeutic agents and functional foods.

Polysaccharides and extracts from Agaricus brasiliensis Murill - A comprehensive review.

Edible mushrooms have been increasingly introduced into the human diet, which has driven research into their functional properties. Thus, Agaricus brasiliensis Murill or Agaricus blazei Murill (ABM) is a species native to the Brazilian biome, whose fruiting body has been used not only for dietary purposes, but also in the development of functional foods or as source of molecules of pharmacological interest. The bioactivity of ABM has been related to the presence of polysaccharides, although the contribution of other metabolites cannot be discharged. This work describes the polysaccharides isolation methodology and preparation of the extracts of ABM and their biological activities. Furthermore, it presents a general outline of its characterizations regarding composition, chemical structure and properties in solution. The ABM and its chemical constituents exhibit several biological activities that support their potential use for prevention or treatment of diseases with inflammatory background, such as cancer, diabetes and atherosclerosis. The mechanism of action of the extracts and polysaccharides from ABM is mainly related to a modulation of immune system response or reduction of inflammatory response. This review shows that the ABM has great potential in the pharmaceutical, biotechnological and food sectors that deserves additional research using standardized products.

Immunomodulatory effect of intracellular polysaccharide from mycelia of Agaricus bitorquis (QuéL.) Sacc. Chaidam by TLR4-mediated MyD88 dependent signaling pathway.

Agaricus bitorquis (QuéL.) Sacc. Chaidam is a valuable edible fungus in Qinghai-Tibet plateau and ABSP is a novel intracellular polysaccharide from its mycelia. GC and NMR analysis determined ABSP is galactoglucomannan-like polysaccharide that may have immunomodulatory effect. This study used RAW264.7 as model cell to determine immunomodulatory effect of ABSP. After ABSP treatment, viability and phagocytic ability promoted, and NO, ROS, TNF-α levels also raised which proved ABSP had immune regulation to RAW264.7. WB and qRT-PCR determined the key proteins and genes expression of TLR4, MyD88, TRAF-6 and NF-κB significantly increased while protein and gene expression of TRAM had no significant increase. Also, TNF-α level extremely decreased by adding inhibitors of TLR4 and MyD88 which confirmed ABSP could immunologically regulate RAW264.7 byTLR4-MyD88 dependent pathway. This study would provide theoretical basis for further study on ABSP and be helpful for development of beneficial functionally foods and exploitation of this resource.

A novel acid polysaccharide from Boletus edulis: extraction, characteristics and antitumor activities in vitro.

A novel cold-water-soluble polysaccharide (BEP), with a molecular weight of 6.0 × 106 Da, was isolated from Boletus edulis. BEP consists of galactose, glucose, xylose, mannose, glucuronic, and galacturonic acid in a ratio of 0.34:0.28:0.28:2.57:1.00:0.44. The IR results showed that BEP was an acid polysaccharide, containing α-type and ß-type glucoside bonds. MTT assay showed BEP could inhibit cell proliferation significantly. Morphological observation demonstrated that BEP-treated MDA-MB-231 and Ca761 cells exhibited typical apoptotic morphological features. Flow cytometry analysis revealed that BEP caused mitochondrial membrane potential collapse. Annexin V-FITC/PI staining indicated that BEP induced apoptosis of MDA-MB-231 and Ca761 cells through cell block in S phase and G0/G1 phase, respectively. Western blot results showed that BEP could increase the Bax/Bcl-2 ratios, promote the release of cytochrome C, and activate the expression of caspase-3 and caspase-9 in MDA-MB-231 and Ca761 cells. In conclusion, our results demonstrated that BEP could inhibit the proliferation of breast cancer cells and induce apoptosis through mitochondrial pathways.

Pink oyster mushroom Pleurotus flabellatus mycelium produced by an airlift bioreactor-the evidence of potent in vitro biological activities.

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.

Production, purification, characterization, and biological properties of Rhodosporidium paludigenum polysaccharide.

The yield of marine red yeast polysaccharide (MRYP) obtained from Rhodosporidium paludigenum was increased by optimizing fermentation conditions, and the pure polysaccharide was extracted by column chromatography. The molecular weight of pure MRYP and the ratio of mannose to glucose in components of MRYP were determined. Antioxidant and antibacterial abilities of MRYP were investigated in vitro and in vivo. The optimal fermentation parameters were as follows: Medium 4, pH = 6.72, temperature = 30.18°C, blades speed = 461.36 r/min; the optimized yield reached 4323.90 mg/L, which was 1.31 times the original yield. The sequence of factors that affected the MRYP yield was the blades speed>pH>temperature. The main components of MRYP were MYH-1 and MYH-2. The molecular weights of MYH-1 and MYH-2 were 246.92 kDa and 21.88 kDa, respectively; they accounted for 53.60% and 28.75% of total polysaccharide. In MYH-1 and MYH-2, the proportion of glucose and mannose accounted for 46.94%, 38.46%, and 67.10%, 7.17%, respectively. In vitro, the ability of scavenging DPPH•, •OH, and [Formula: see text] radical was 32.26%, 24.34%, and 22.09%; the minimum inhibitory concentration (MIC) of MRYP was 480 µg/mg. In vivo, MRYP improved the lambs' body weight, antioxidant enzyme activity, and the number of probiotics, but it reduced the feed/gain (F/G) ratio and the number of pathogenic bacteria in 60-days-old lambs.

Biologically active polysaccharide from edible mushrooms: A review.

Mushrooms are renewable natural gift for humankind, furnished with unique taste, flavor and medicinal properties. For the last few decades study of mushroom polysaccharides has become a matter of great interest to the researchers for their immunomodulating, antimicrobial, antioxidant, anticancer, and antitumor properties. Molecular mass, branching configuration, conformation of polysaccharides and chemical modification are the major factors influencing their biological activities. The mechanism of action of mushroom polysaccharides is to stimulate T-cells, B-cells, natural killer cells, and macrophage dependent immune responses via binding to receptors like the toll-like receptor-2, dectin-1. The present review offers summarized and significant information about the structural and biological properties of mushroom polysaccharides, and their potential for development of therapeutic materials.

Medicinal Properties and Bioactive Compounds from Wild Mushrooms Native to North America.

Mushrooms, the fruiting bodies of fungi, are known for a long time in different cultures around the world to possess medicinal properties and are used to treat various human diseases. Mushrooms that are parts of traditional medicine in Asia had been extensively studied and this has led to identification of their bioactive ingredients. North America, while home to one of the world's largest and diverse ecological systems, has not subjected its natural resources especially its diverse array of mushroom species for bioprospecting purposes: Are mushrooms native to North America a good source for drug discovery? In this review, we compile all the published studies up to September 2020 on the bioprospecting of North American mushrooms. Out of the 79 species that have been investigated for medicinal properties, 48 species (60%) have bioactivities that have not been previously reported. For a mere 16 selected species, 17 new bioactive compounds (10 small molecules, six polysaccharides and one protein) have already been isolated. The results from our literature search suggest that mushrooms native to North America are indeed a good source for drug discovery.

Comprehensive evaluation of alkali-extracted polysaccharides from Agrocybe cylindracea: Comparison on structural characterization.

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 → 6)-galactopyranosyl as main chain. JACP-50p and JACP-80r were characterized as ß-(1 → 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 → 6)-linked glucopyranosyl and ß-(1 → 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.