Comparison of the Secondary Metabolism of the Basidiomycetes Armillaria mellea and Desarmillaria ectypa.

During the course of our ongoing studies on the secondary metabolism of cultures of Basidiomycota, a new meroterpenoid named 10, 15-dihydroxydihydromelleolide (1) was isolated along with the known armillaridin (2) and arnamiol (3) from cultures of the rare saprotrophic species, Desarmillaria ectypa. These are the first secondary metabolites that were ever isolated from the latter species. A concurrently studied strain of the common pathogenic A. mellea yielded other melleolides, with 5'-O-methylmelledonal (4), melledonal C (5), 10 α-hydroxydihydromelleolide (6) and melledonal (7). The chemical structures were elucidated using 1D and 2D NMR spectroscopy and high-resolution electrospray ionization mass spectrometry (HR-ESI-MS). All compounds were studied for their antimicrobial and cytotoxic effects against a panel of microbes and mammalian cell lines, and the results are also reported.

Critical review of the phytochemical profiles and health-promoting effects of the edible mushroom Armillaria mellea.

Research on the nutritional and medicinal properties of wild edible mushrooms has witnessed a significant surge in recent years. Among these mushrooms, Armillaria mellea (AM) stands out due to its abundant biologically active components. The presence of biological compounds in AM, including carbohydrates, sterols, fatty acids, sesquiterpenes, non-hallucinogenic indole compounds and adenosine derivatives, has been demonstrated in previous studies. Notably, specific bioactive substances isolated from AM, such as armillarikin, have exhibited promising anticancer effects. In vitro studies have elucidated the mechanisms behind these effects, further emphasizing the potential of AM in cancer treatment. Consequently, the objective of this study is to provide a comprehensive overview of the phytochemical profiles of AM while thoroughly investigating its therapeutic benefits. Moreover, this research has uncovered novel and effective treatments, including the utilization of ultrasonic disruption extraction in food processing. These findings highlight the potential of AM as a functional food with possible medical applications. By exploring AM's phytochemical composition and therapeutic effects, this study aims to contribute to a deeper understanding of its potential as a valuable natural resource.

Study of anti-fatigue activity of polysaccharide from fruiting bodies of Armillaria gallica.

Fatigue is a common physiological response that is closely related to energy metabolism. Polysaccharides, as excellent dietary supplements, have been proven to have a variety of pharmacological activities. In this study, A 23.007 kDa polysaccharide from Armillaria gallica (AGP) was purified and performed structural characterization, including analysis of homogeneity, molecular weight and monosaccharide composition. Methylation analysis is used to analyze the glycosidic bond composition of AGP. The mouse model of acute fatigue was used to evaluate the anti-fatigue effect of AGP. AGP-treatment improved exercise endurance in mice and reduced fatigue symptoms caused by acute exercise. AGP regulated the levels of adenosine triphosphate, lactic acid, blood urea nitrogen and lactate dehydrogenase, muscle glycogen and liver glycogen of acute fatigue mice. AGP affected the composition of intestinal microbiota, the changes of some intestinal microorganisms are correlated with fatigue and oxidative stress indicators. Meanwhile, AGP reduced oxidative stress levels, increased antioxidant enzyme activity and regulated the AMP-dependent protein kinase/nuclear factor erythroid 2-related factor 2 signaling pathway. AGP exerted an anti-fatigue effect through modulation of oxidative stress, which is related to intestinal microbiota.

A Combination of Genome Mining with an OSMAC Approach Facilitates the Discovery of and Contributions to the Biosynthesis of Melleolides from the Basidiomycete Armillaria tabescens.

Genome mining revealed that the genomes of basidiomycetes may include a considerable number of biosynthetic gene clusters (BGCs), yet numerous clusters remain unidentified. Herein, we report a combination of genome mining with an OSMAC (one strain, many compounds) approach to characterize the spectrum of melleolides produced by Armillaria tabescens CPCC 401429. Using F1 fermentation medium, the metabolic pathway of the gene cluster mel was successfully upregulated. From the extracts of the wild-type strain, two new melleolides (1 and 2), along with five new orsellinic acid-derived lactams (10-14), were isolated, and their structures were elucidated by LC-HR-ESIMS/MS and 2D-NMR. Several melleolides exhibited moderate anti-carcinoma (A549, NCI-H520, and H1299) effects with IC50 values of 4.0-48.8 µM. RNA-sequencing based transcriptomic profiling broadened our knowledge of the genetic background, regulation, and mechanisms of melleolide biosynthesis. These results may promote downstream metabolic engineering studies of melleolides. Our study demonstrates the approach is effective for discovering new secondary metabolites from Armillaria sp. and will facilitate the mining of the unexploited biosynthetic potential in other basidiomycetes.

Study on antidepressant-like effect of protoilludane sesquiterpenoid aromatic esters from Armillaria Mellea.

Armillaria mellea, also known as Hazel mushroom, is a delicious food material and traditional herbal medicine in East Asia. Protoilludane sesquiterpenoid aromatic esters from A. mellea (PSAM) are the main active components with antibacterial and anticancer activities. This study explored the antidepressant-like activities of PSAM and its possible mechanisms of action using the open field test (OFT), tail suspension test (TST) and forced swimming test (FST) in mice for the first time. The results revealed that PSAM (1 mg/kg, i.p.) exhibited markedly antidepressant-like activity, which could be reversed by pretreatment with haloperidol (a non-selective D2 receptor antagonist), bicuculline (a competitive GABA antagonist), NMDA (an agonist at the glutamate site). Meanwhile, PSAM also effectively increased the hippocampus dopamine (DA) and γ-aminobutyric acid (GABA) and decreased the hippocampus glutamate (Glu) levels of mice, indicating that the antidepressant-like effect of PSAM might be mediated by the DAergic, GABAergic and Gluergic systems.

Protoilludane-type sesquiterpenoids from Armillaria sp. by co-culture with the endophytic fungus Epicoccumsp. associated with Gastrodia elata.

An investigation of a co-culture of the Armillaria sp. and endophytic fungus Epicoccum sp. YUD17002 associated with Gastrodia elata led to the isolation of eight new compounds, including five protoilludane-type sesquiterpenes (1-5) and three aryl esters (6-8), together with six known analogues (9-14). The assignments of their structures were conducted via extensive analyses of the spectroscopic data and comparison of experimental and calculatedelectronic circular dichroism(ECD)data. Notably, these new compounds were not present in the pure culture controls and were only detected in the co-cultures. Compound 4 is the first example of an ent-protoilludane sesquiterpenoid scaffold bearing a five-membered lactone. Compound 6 exhibited moderate in vitro cytotoxic activities against five human cancer cell lines (HL-60, A549, MCF-7, SMMC-7721, and SW480) with IC50 values ranging from 15.80 to 23.03 µM. Moreover, 6 showed weak acetylcholinesterase inhibitory activity (IC50 value of 23.85 µM).

Ultrasonic disruption extraction, characterization and bioactivities of polysaccharides from wild Armillaria mellea.

An efficient ultrasonic disruption extraction (UDE) of polysaccharides from Armillariamellea (AM) were optimized by response surface methodology (RSM). Under optimum conditions: ultrasonic power 915.00 W, temperature 69.27 °C and time 39.13 min, the crude polysaccharides (AMPs) yield was 19.5%. Two purified fractions AMPs-1-1 and AMPs-2-1 were obtained through anion-exchange and gel chromatography. AMPs-1-1 was a heteropolysaccharide with average molecular weights of 1.23 × 105 Da, and composed of Glc, Gal and GlcA with mole percentages of 89.06%, 9.59% and 1.34%, respectively, owning a backbone structure of (1→)-ß-d-Glcp, (1 â†’ 3,6)-α-d-Glcp and (1 â†’ 3)-ß-d-Glcp residues. AMPs-2-1 was a heteropolysaccharide with average molecular weights of 6.76 × 104 Da, and composed of Glc, Gal, GlcA and Man with mole percentages of 65.28%, 22.87%, 2.87% and 8.98%, containing a main backbone chain of (1 â†’ 3,6)-α-d-Glcp and (1 â†’ 6)-ß-d-Glcp residues. AMPs-2-1 possessed obviously antioxidant activities in terms of stronger scavenging activity against DPPH· and ABTS+⋅, higher FRAP and ORAC value than AMPs-1-1. AMPs-2-1 could promote splenocyte lymphocytes and RAW264.7 macrophages proliferation and enhanced the phagocytosis of macrophages, exhibited significant immunomodulatory activities. These results suggested that UDE is an effective extract technology, and AMPs-2-1 could be explored as potential natural antioxidants and immunomodulatory agents.

Induction of Autophagic Death of Human Hepatocellular Carcinoma Cells by Armillaridin from Armillaria mellea.

The honey mushroom, Armillaria mellea, is known to have medicinal qualities and has been used in recent years as a health food and dietary supplement worldwide. In Asia, it is commonly consumed as an herbal medicine, being a key component of the Chinese preparation "Tien-ma". Here, we examined the antitumor effects of armillaridin, a bioactive compound isolated from A. mellea, on human hepatocellular carcinoma (HCC) cells. Armillaridin inhibited the growth of human Huh7, HepG2, and HA22T HCC cells, and its cytotoxicity was confirmed by observations of its induction of mitochondrial transmembrane potential collapse. However, armillaridin treatment did not result in large numbers of cells with fragmented chromosomal DNA, suggesting that apoptosis was not responsible for these effects. We therefore tested for signs of autophagic cell death following armillaridin administration. Armillaridin induced LC3 aggregation in green fluorescent protein-LC3-overexpressing cells. Moreover, flow cytometry and immunoblotting revealed that it increased the number of acridine orange-positive cells and upregulated autophagy-related proteins, respectively. Furthermore, armillaridin cytotoxicity was suppressed by the autophagy inhibitor 3-methyladenine. In summary, our results indicated that armillaridin induces HCC cell death by autophagy, and demonstrated the potential of armillaridin as an antihepatoma agent.

Microbial Asymmetric Functionalization of ß-Cyclocitral-Derived Tetramethyl-Substituted γ-Lactone.

Searching for the new anticancer compounds we prepared three new ß-cyclocitral-derived hydroxyl-γ-lactones by microbial hydroxylation of tetramethyl-substituted bicyclic γ-lactone. The substrate was transformed by the enzymatic system of filamentous fungi. Three out of fifteen strains were selected as effective biocatalysts (Fusarium culmorum AM10, Armillaria mellea AM296, Trametes versicolor AM536). The hydroxylation processes were not only regioselective but also stereoselective. The hydroxylation products of each secondary carbon atom in the cyclohexane ring were obtained by the application of the selected fungal strains. The Fusarium culmorum AM10 introduced the hydroxy function at C-3 and C-4, Armillaria mellea AM296 incorporated the hydroxy function at C-3 and C-5 and Trametes versicolor AM536 transformed the substrate to the mixture of C-3, C-4 and C-5 hydroxylactones. The hydroxylactones obtained were enantiomericaly enriched (ee values in the range 17⁻99%). The in vitro antiproliferative activities of the functionalization products were also evaluated. Regardless of the hydroxy substituent location all tested lactones exhibited similar, significant activity towards selected cancer cell lines (IC50 in the range 22.8⁻33.9 µg/mL).

Melleolides from Honey Mushroom Inhibit 5-Lipoxygenase via Cys159.

5-Lipoxygenase (5-LO) initiates the biosynthesis of pro-inflammatory leukotrienes from arachidonic acid, which requires the nuclear membrane-bound 5-LO-activating protein (FLAP) for substrate transfer. Here, we identified human 5-LO as a molecular target of melleolides from honey mushroom (Armillaria mellea). Melleolides inhibit 5-LO via an α,ß-unsaturated aldehyde serving as Michael acceptor for surface cysteines at the substrate entrance that are revealed as molecular determinants for 5-LO activity. Experiments with 5-LO mutants, where select cysteines had been replaced by serine, indicated that the investigated melleolides suppress 5-LO product formation via two distinct modes of action: (1) by direct interference with 5-LO activity involving two or more of the cysteines 159, 300, 416, and 418, and (2) by preventing 5-LO/FLAP assemblies involving selectively Cys159 in 5-LO. Interestingly, replacement of Cys159 by serine prevented 5-LO/FLAP assemblies as well, implying Cys159 as determinant for 5-LO/FLAP complex formation at the nuclear membrane required for leukotriene biosynthesis.

Purification, structural elucidation, and anti-inflammatory activity of xylosyl galactofucan from Armillaria mellea.

A xylosyl 1,3-galactofucan (AMPS-III) was isolated and identified as a novel anti-inflammatory agent from an edible fungus, Armillaria mellea. The characteristics chemical structure of AMPS-III including the linkages of compositional monosaccharides and structure of the repeat unit were depicted and elucidated by proton, carbon and two-dimensional nuclear magnetic resonance techniques. AMPS-III was chemically proposed to have a partial 4-O-xylosylated 1,3-linked α-d-galactosyl-interlaced α-l-fucan composed of a pentadecasaccharide repeat unit with a molecular mass approximately 13 kDa. AMPS-III significantly suppressed the release of tumor necrosis factor-α (TNF-α) and cytokine monocyte chemotactic protein-1 (MCP-1) in RAW264.7 macrophages and EAhy926 following LPS and TNF-α induction. The results provide helpful evidences for application of AMPS-III as anti-inflammatory food supplements.

[Correlative analysis advance of chemical constituents of Polyporus umbellatus and Armillaria mellea].

Medicinal Polyporus umbellatus is the dry sclerotia of P. umbellatus, with the effect of diuresis; Armillaria mellea is a parasitic fungus which can infect plants up to 300 genera, with sedative, anticonvulsant and some other biological activities. As the medicinal value of P. umbellatus and A. mellea is increasingly wide concerned, the market quantity demanded of them is gradually increased and the demand outstrips the supply. The symbiotic A. mellea and P. umbellatus are both the medicinal and edible fungi with diverse activities, including hypoglycemic action, improve immunity and antitumor and so on. The growth of the sclerotia forming from the mycelium of P. umbellatus is related to the infection of the symbiotic A. mellea and their secondary products. In this study, by comparing the chemical constituents of the mycelium and sclerotia of P. umbellatus and A. mellea, we found that they all produced steroids and nitrogen-containing heterocycles. The sclerotia of P. umbellatus and A. mellea also produced triterpenes secondary metabolites. In addition, the mycelium and infected sclerotia of P. umbellatus mainly produced different steroids, and the sclerotia produced some other special secondary metabolites, such as long-chain fatty acids, ceramides, phenol and so on. By analyzing above all kinds of differences, speculated that these may be caused by the infection of the symbiotic A. mellea which mainly produced sesquiterpenes, diterpenes and other secondary metabolites. The contents and types of compounds of P. umbellatus and A. mellea are closely related to their symbiosis and reproduction, therefore, many symbiosis mechanisms should be found by utilizing more molecular biology technology to elucidate this complex symbiotic infection and provide scientific basis for improving the yield and quality of P. umbellatus and A. mellea.

Chemical, nutritive composition and a wide range of bioactive properties of honey mushroom Armillaria mellea (Vahl: Fr.) Kummer.

A. mellea fruiting bodies collected from nature were chemically characterized and shown to be rich in carbohydrates (81.25 g per 100 g dw), ash, fat and proteins (8.84 g per 100 g dw, 1.97 g per 100 g dw and 1.81 g per 100 g dw, respectively). Mannitol was the main free sugar while malic acid was the most abundant organic acid. δ-Tocopherol was the dominant form of tocopherols with 42.41 µg per 100 g dw. Polyunsaturated fatty acids were predominant, followed by saturated and monounsaturated fatty acids. A methanolic extract prepared from these samples was tested for antioxidant, quorum sensing and antimicrobial assays, as well as for its cytotoxicity effects. The extract showed antimicrobial activity against all tested microorganisms, including Candida albicans. Furthermore, when tested at sub-MIC concentration, it showed reduction of virulence factors and biofilm formation against Pseudomonas aeruginosa. The extract also exhibited antioxidant activity and did not show toxicity against tumor and non-tumor cells. Due to the observed bioactive properties and compounds of the honey mushroom and its well-balanced nutrients, this mushroom emerges as an interesting functional food and a source of nutraceuticals with applications in different diseases based on antioxidant and antimicrobial effects.

Bioassay-guided fractionation of ethyl acetate extract from Armillaria mellea attenuates inflammatory response in lipopolysaccharide (LPS) stimulated BV-2 microglia.

BACKGROUND: Armillaria mellea (A. mellea) is a traditional Chinese medicinal and edible mushroom, which is proved to possess a lot of biological activities, including anti-oxidation, immunopotentiation, anti-vertigo and anti-aging activities. However, little information is available in regard to its neuroprotection activity in inflammation-mediated neurodegenerative diseases. PURPOSE: We have found that A. mellea has an anti-inflammatory activity in LPS-induced RAW264.7 cells in our previous study. The objective of this study is to investigate the anti-neuroinflammatory mechanism of a bioassay-guided fractionation (Fr.2) and its active components/compounds. METHODS: Compounds were isolated by preparative high performance liquid chromatography (pre-HPLC) and their structures were established by mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopic analyses. The anti-neuroinflammatory effect of Fr.2 and each compounds were investigated in lipopolysaccharide (LPS)-stimulated murine microglia cell lineBV-2. RESULTS: We demonstrated that Fr.2 significantly decreased the production of inflammation mediator nitric oxide (NO) and inflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1beta (IL-1ß) in a dose-dependent manner (10, 30, 100µg/ml). In addition, Fr.2 markedly down-regulated the phosphorylation levels of nuclear factor kappa B p65 (NF-κB p65), inhibitory κB-α (IκB-α) and c-Jun N-terminal kinases (JNKs) pathways. Sevens compounds were isolated from Fr.2, among them, three compounds, 5-hydroxymethylfurfural (CP1), vanillic acid (CP4) and syringate (CP5) were reported for the first time in A. mellea. NO and inflammatory cytokines (TNF-α, IL-6, IL-1ß) secretion indicated that daidzein (CP6) and genistein (CP7) showed a more outstanding anti-inflammation potential at non-toxic concentrations (10, 30, 100µM) than the other five compounds. CONCLUSIONS: In conclusion, Fr.2 may have therapeutic potential for neurodegenerative diseases by inhibiting inflammatory mediators and suppress inflammation pathway in activated microglia. Daidzein and genistein may serve as the effective anti-inflammation compounds of Fr.2.

Novel Application of Peptidyl-Lys Metallopeptidase as a C-Terminal Processing Protease.

Adding fusion partners to proteins or peptides can aid or be a necessity to facilitate recombinant expression, folding, or purification. Independent of the reason it is desirable to remove the fusion partner to restore native functionality. Processing proteases catalyze the removal of fusion partners, however, most of these proteases have substrate specificity for the N-terminal of the scissile bond, leaving non-native termini if fusions are added to the C-terminal. The peptidyl-lys metallopeptidease of Armillaria mellea (Am-LysN) is unusual by having substrate specificity for the C-terminal side of the scissile peptide bond, allowing it to generate native C-termini. Am-LysN has strict specificity for lysine in P1', making all lysines of a protein or peptide a potential degradation site, however there are a number of amino acid side chains which lower hydrolysis significantly when located adjacent to the lysine. In this study we show that Am-LysN can be used as a processing protease to remove C-terminal extensions of peptides with no internal lysine to generate native Ctermini. Furthermore we show that removal of C-terminal extensions on peptides containing internal lysines can be achieved with little degradation of the product depending on the adjacent amino acids. These results demonstrate the utility of LysN allowing for novel ways to use fusion technology in the production of recombinant proteins.

Three New Sesquiterpene Aryl Esters from the Mycelium of Armillaria mellea.

Three new sesquiterpene aryl esters and eight known compounds were isolated from the EtOH extract of the mycelium of Armillaria mellea. The structures of new compounds were established by analysis of their spectroscopic data. Some of the isolates showed cytotoxicity to a variety of cancer cell lines, including MCF-7, H460, HT-29, and CEM.

Armillaria mellea: phenolic content, in vitro antioxidant and antihyperglycemic effects.

UNLABELLED: Armillaria mellea, known as honey mushroom, has been used both as food and medicine. AIM: In this work, the ethanolic and hydromethanolic extracts from Armillaria mellea fruiting bodies were investigated for their phenolic content, antioxidant and antihyperglycemic effects (in vitro studies). MATERIAL AND METHODS: The total phenolics were quantified using Folin-Ciocalteu assay. The antioxidant activity was evaluated by testing the free radical scavenging capacity, reducing power and ferrous ion chelating ability; in addition, the capacity to inhibit 15-lipoxygenase was also assessed. The antihyperglycemic activity was investigated by α-glucosidase assay. RESULTS: Total phenolic contents of 21.68 ± 0.06 and 5.70 ± 0.28 mg/g were determined in the hydromethanolic and ethanolic extracts, respectively. The hydromethanolic extract showed higher free radical scavenging and reducing abilities (EC50 = 452.6 ± 2.7, 140.57 ± 1.45 and 129.45 ± 0.98 µg/mL in DPPH, ABTS and reducing power assays, respectively). The ethanolic extract proved to be more efficient in the ferrous ion chelation, 15-lipoxygenase and α-glucosidase inhibition assays (EC55 = 67.93 ± 0.35, 290.93 ± 2.05 and 8.54 ± 0.06 µg/mL, respectively). CONCLUSIONS: Armillaria mellea extracts showed antioxidant and antihyperglycemic potential in in vitro models and therefore they are promising candidates for the development of dietary supplements and pharmaceutical products.

Chemical compositions and antioxidant activities of polysaccharides from the sporophores and cultured products of Armillaria mellea.

Armillaria mellea is a traditional Chinese medicinal and edible mushroom. Many cultured products of A. mellea have been used to develop commercial medicines in recent years. The chemical composition and activities of the major bioactive chemical components-polysaccharides-may be different because of differences in the raw materials used. Four polysaccharides (SP, CMP, CFBP and CFMP) were obtained from wild sporophores and cultured products (including mycelia, fermentation broth and fermentation mixture) of A. mellea. Their yields, carbohydrate contents, monosaccharide compositions, FT-IR spectra, NMR spectroscopy and antioxidant activities were investigated. All of the polysaccharides were composed of xylose, glucose and galactose without protein. Glucose was the dominant monosaccharide in SP, CMP and CFMP, whereas galactose was the dominant monosaccharide in CFBP. SP and CMP showed higher scavenging DPPH• and ABTS•+ activities and reducing power among four polysaccharides. The carbohydrate content and corresponding glucose percentage were positive influences on the antioxidant activities, whereas the corresponding xylose and galactose percentage were negative influences. A. mellea polysaccharides are potential natural antioxidants. Polysaccharides from cultured products, especially mycelia, are good substitutes for SP and are also potential sources for both dietary supplements and food industries.

Armillaridin, a Honey Medicinal Mushroom, Armillaria mellea (Higher Basidiomycetes) Component, Inhibits Differentiation and Activation of Human Macrophages.

Armillaridin (AM) is an aromatic ester compound isolated from honey medicinal mushroom, Armillaria mellea, which has anti-cancer potential. This study was designed to examine the effects of AM on differentiation and activation macrophages, the major ontogeny of innate immunity. Macrophages were derived from CD14+ monocytes which were sorted from human peripheral blood mononuclear cells. Cell viability was assessed by trypan blue exclusion test. Cells were stained with Liu's dye for observation of morphology. Expression of surface antigens was examined by flow cytometric analysis. Phagocytosis and generation of reactive oxygen species (ROS), as functional assays, were evaluated by counting engulfed yeasts and DCFH-DA reaction. The viability of macrophages was not significantly reduced by AM. AM at nontoxic concentrations markedly increased cytoplasmic vacuoles. The expression of surface CD14, CD16, CD36, and HLA-DR was suppressed. The phagocytosis function, but not ROS production, of macrophages was inhibited by AM. Armillaridin could inhibit the differentiation and activation of human macrophages. It may have potential to be developed as a biological response modifier for inflammatory diseases.

Antioxidant and antiedema properties of solid-state cultured honey mushroom, Armillaria mellea (higher Basidiomycetes), extracts and their polysaccharide and polyphenol contents.

Culinary-medicinal honey mushroom or Mi-Huan-Ku, Armillaria mellea (AM), is a popular ingredient in the traditional Chinese medicine for treating diseases of geriatric patients. This study aimed to examine the effect of cultured substrates on the mycelial growth of AM and evaluate its antioxidant and antiedema activities as well as its total polysaccharide and polyphenol contents. Results showed that AM grew best on the maize medium and worst on the potato medium. AM ethanol extract (AM-EtOH) showed stronger DPPH radical scavenging activity than AM aqueous extract (AM-H2O). However, they were weak in metal chelation and reducing power. AM-EtOH but not AM-H2O at 200 mg/kg showed antiedema activity in rats. The total ß-glucan content of AM-H2O and AM-EtOH was 21.95% and 3.50%, respectively. AM-EtOH showed higher phenol but lower flavonoid content than AM-H2O. These results indicate that maize is a good source of substrate for mass production of AM mycelia, and its potency of DPPH radical scavenging and antiedema activities was contributed mainly by the phenolic compounds, not the level of polysaccharide content.