Ganoderma Lucidum Mycelia Research Articles

A comparison of in vitro anticancerous activity and mechanism of ethanolic extracts from different Ganoderma genus

Five ethanolic extracts from the mycelia of Ganoderma lucidum, G. tsugae, G. oerstedii, G. subamboinense, and G. resinaceum were respectively studied on their anticancerous activities against leukemic HL-60 cell line in vitro. Results showed that all five extracts potently inhibited HL-60 proliferation. The extract from G. lucidum mycelia exerted the highest activity. Annexin V/PI bivariate flow cytometric analysis further revealed that the five extracts significantly induced early apoptosis in HL-60 cells. The results illustrate that not only G. lucidum but also other Ganoderma species can inhibit cancer cells, and their mechanisms are related to induction of apoptosis.

Optimization of nutrient medium for submerged cultivation of Ganoderma lucidum (Curt.: Fr.) P. Karst

The dependence of the amount of the grown vegetative mycelium of Ganoderma lucidum on the composition of the nutrient medium has been studied under conditions of submerged cultivation. The medium was optimized using full factorial and steepest ascent experimental designs. The addition of two carbon sources to the medium considerably improved the submerged growth of the fungus. An optimized medium provided for a high yield (20-20.95 g/l) of the morphologically homogeneous mycelium and shortened the cultivation period to 3-4 days.

In vitro and in vivo protective effects of proteoglycan isolated from mycelia of Ganoderma lucidum on carbon tetrachloride-induced liver injury

To investigate the possible mechanism of the protective effects of a bioactive fraction,Ganoderma lucidum proteoglycan (GLPG) isolated from Ganoderma lucidum mycelia, against carbon tetrachloride-induced liver injury. A liver injury model was induced by carbon tetrachloride. Cytotoxicity was measured by MTT assay. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined with an automatic multifunction-biochemical analyzer and the levels of superoxide dismutase (SOD)and TNF-alpha were determined following the instructions of SOD kit and TNF radioimmunoassay kit. Liver sections were stained with hematoxylin and eosin (H and E) for histological evaluation and examined under light microscope. We found that GLPG can alleviate the L-02 liver cells injury induced by carbon tetrachloride (CCl4) through the measurements of ALT and AST activities and the administration of GLPG to L-02 cells did not display any toxicity. Furthermore, histological analysis of mice liver injury induced by CCl4 with or without GLPG pretreatment indicated that GLPG can significantly suppress the toxicity induced by CCl4 in mice liver. We also found that GLPG reduced TNF-alpha level induced by CCl4 in the plasma of mice, whereas increased SOD activity in the rat serum. GLPG has hepatic protective activity against CCl4-induced injury both in vitro and in vivo. The possible anti-hepatotoxic mechanisms may be related to the suppression of TNF-alpha level and the free radical scavenging activity.

Ganoderma lucidumMycelium and Spore Extracts as Natural Adjuvants for Immunotherapy

Ganoderma lucidum (GL) is one of the most commonly used Chinese herbs in the oriental community, with more than 30% of pediatric cancer patients taking GL. The immunomodulating and anticancer effects exerted by GL extracts have been demonstrated by in vitro and in vivo studies. There was, however, no comparison between the immunomodulating effects of GL mycelium extract (GL-M) and spore extracts on human immune cells. Dendritic cells (DCs) are professional antigen-presenting cells and their role in DC-based tumor vaccine has been well defined. The possibility of GL as natural adjuvant for human DCs remains unknown. This study explored the differential effect of GL-M and GL spore extract (GL-S) on proliferation and Th1/Th2 cytokine mRNA expression of human peripheral blood mononuclear cells (PBMCs) and monocytes. Their effects on the phenotypic and functional maturation of human monocyte-derived DCs were also investigated. GL-M induced the proliferation of PBMCs and monocytes, whereas GL-S showed a mild suppressive effect. Both extracts could stimulate Th1 and Th2 cytokine mRNA expression, but GL-M was a relatively stronger Th1 stimulator. Different from GL-S, GL-M enhanced maturation of DCs in terms of upregulation of CD40, CD80, and CD86, and also reduced fluorescein isothiocyanate-dextran endocytosis. Interestingly, GLM- treated DCs only modestly enhanced lymphocyte proliferation in allogenic mixed lymphocyte culture with mild enhancement in Th development. These findings provide evidences that GL-M has immunomodulating effects on human immune cells and therefore can be used as a natural adjuvant for cancer immunotherapy with DCs.

Immunopotent Polysaccharides of Different Sources Selectively Activate Human Dendritic Cells.

Background: Purified polysaccharides extracted from plants and fungi have been shown to induce immune responses in-vivo and vitro over the past decade. Currently, most of these polysaccharides are found to be glucan but with different branch structure and sizes. Their relative potency and effect on human immune cells remains unknown. This study aims to compare their relative effect on human dendritic cell, the most potent antigen presenting cell.Materials & Methods: We selected 2 prototypes of purified polysaccharides extracted from: 1) Ganoderma lucidum (GL, Lingzhi, Reishi) mycelium, a widely used herb with long and branching β (1® 3), (1® 6) glucan structure (provided by Prof. Lin ZB, Beijing) and 2) Barley with shorter and different branching β (1® 3), (1® 4) structure (provided by Prof. Cheung VNK, NY). Their characteristics and chemical properties had been reported previously. Human peripheral blood mononuclear cells (PBMCs) proliferation was studied by XTT assay. Human dendritic cells (DCs) were derived from monocytes and maturation of DCs were determined by: a) immunophenotypic shift using flow cytometer; 2) dextran endocytosis assay and 3) mixed lymphocytes reaction. Cytokine secretions were determined by ELISA test. Comparisons between means were by nonparametric Student's t test (2-tailed).Results: We found that purified polysaccharides from GL but not barley could induce PBMCs proliferation and maturation of DCs. GL polysaccharides could enhance phenotypic and functional maturation of DCs with significant IL-12 and IL-10 production. DCs were relatively inert to Barley glucans stimulation. However, both polysaccharides did not polarize T cells into the direction of T helper 1, T helper 2 or regulatory T cells.Conclusions: Our study shown that purified polysaccharides extracted from plants and fungi have different effect on human DCs and their potency and effects are probably affected by their respective sources and structures.

Ganoderma lucidum mycelia enhance innate immunity by activating NF-κB

Ganoderma lucidum is a popular medicinal mushroom in China and Japan for its immunomodulatory and antitumor effects. The goal of this research is to investigate the effect of dried mycelia of Ganoderma lucidum produced by submerged cultivation on the enhancement of innate immune response. We found that Ganoderma lucidum mycelia (0.2–1.6 mg/ml) stimulated TNF-α and IL-6 production after 8 h treatment in human whole blood. IFN-γ release from human whole blood was also enhanced after 3 day-culture with Ganoderma lucidum mycelia (0.2–1.0 mg/ml). However, Ganoderma lucidum mycelia did not potentiate nitric oxide production in RAW264.7 cells. To better understand the possible immuno-enhancement mechanisms involved, we focused on nuclear factor (NF)-κB activation. Electrophoretic mobility shift assay revealed that the Ganoderma lucidum mycelia (1.6 mg/ml) activated κB DNA binding activity in RAW264.7 cells. These results provide supporting evidences for the immunomodulatory effect of Ganoderma lucidum mycelia.

Growth Characterization and Triterpenoid Derivatives Quantification of Ganoderma spp. Using HPLC

Mycelium of Ganoderma lucidum (W.Curt.:Fr.) Lloyd grew on the surface of molasses to form mycelial mats after about 12 days. When the mycelial mats became older, the color changed to a darker hue and wrinkled. It took about 9 days for the mycelium of G. tsugae Murrill to cover the whole surface of the molasses. Mycelial mats of G. tsugae were soft, and the old mycelia turned to yellow in color. Harvesting three pieces of mycelial mats of G. lucidum was enough to get 0.96 g of dried biomass. For G. tsugae, the mycelial mats were lighter. On wheat grains, the mycelium of G. lucidum needed 20 days to permeate throughout the grains. The mycelium of G. tsugae needed only 15 days. Both Ganoderma species produced primodia for approximately 65 days, and the mature basidiocarps were formed in about 100 days. The pH of extracts from both fungi became less acidic when harvested after longer periods. All the separation spots of the samples had an Rf value below 0.92. The TLC method detected the presence of triterpenoids in the extract. The different concentrations of thymol salt were used to obtain the standard curve through HPLC method. The values of triterpenoid concentrations, which refer to the peaks as P1, P2, P3, P4, P5, and P6 in the samples, were calculated based on this standard curve. P1, was obtained in every stage of incubation. P3 and P6 were detected only in the fruiting body, whereas P2, P3, P4 were the major triterpenoids of the mycelium. The highest triterpenoid production was found after 110 days; the fruiting body stage of G. tsugae is P5 with 1212.00 μg/mL. G. lucidum showed triterpenoid concentration and derivatives six times higher than G. tsugae.

Possible mode of action of antiherpetic activities of a proteoglycan isolated from the mycelia of Ganoderma lucidum in vitro

A bioactive fraction (GLPG) was extracted and purified from the mycelia of Ganoderma lucidum by EtOH precipitation and DEAE-cellulose column chromatography. GLPG was a proteoglycan and had a carbohydrate:protein ratio of 10.4:1. Its antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were investigated by the cytopathic effect (CPE) inhibition assay in cell culture. This kind of polysaccharide inhibited the development of the cytopathic effect in dose-dependent manner in HSV-infected cells, moreover did not show any cytotoxic effects on cells even when a concentration was as high as 2000 μg/ml. In order to study the possible mode of action of the antiviral activity of GLPG, cells were treated with GLPG before, during and after infection, and the viral titers in the supernatant of cell culture 48 h post-infection were tested by TCID 50 assay. The antiviral effects in pre-treated and treated during virus infection with GLPG were more remarkable than the treatment of post-infection. Although the precise mechanism has yet to be defined, our work suggested that GLPG inhibits viral replication by interfering with the early events of viral adsorption and entry into target cells. Thus, this proteoglycan seems to be a potential candidate for anti-HSV agents.

The inhibitory effect of the Chinese herb Ganoderma lucidum mycelium on gut immunoglobulin A responses to cholera toxin in mice

Five-week-old female C57BL/6J mice were fed AIN-93G diet containing 0, 0.5, 1, 3 and 5% (wt/wt) Ganoderma (G) lucidum mycelium for 4 wk. Mice were orally immunized with 5 μg cholera toxin (CT) on d 7 and d 21 of feeding period. The total IgA and specific anti-CT IgA level in luminal washes of small intestine, serum and fecal pellets as well as anti-CT IgG response in serum were determined by ELISA. The total IgA level generally was not affected by G. lucidum mycelium except in serum samples. By contrast, the anti-CT IgA antibody response was reduced in all the samples from mice fed diets containing G. lucidum mycelium except in the small intestine luminal washes of mice fed the diet containing 5% G. lucidum mycelium. The serum anti-CT IgG response was only decreased in mice fed 3% G. lucidum mycelium. Thus, G. lucidum mycelium reduced the mucosal specific IgA response of young adult mice orally immunized with CT.

Production of Ganoderma lucidum mycelium using cheese whey as an alternative substrate: response surface analysis and biokinetics

A novel approach to utilize cheese whey, cultivating mycelium of an edible mushroom Ganoderma lucidum using cheese whey as a substrate, was introduced. Response surface analysis (RSA) with central composite in cube design was successfully applied to determine the optimal conditions where the maximum mycelial production occurred, which was at pH 4.2 and 28.3 °C. The high extract ratio as well as high content of polysaccharide (i.e., 1.2 g/l) indicated that the whey could be an alternative substrate for the mycelial production. Soluble chemical oxygen demand (SCOD) removal ranged from 80.7 to 93.1% within the design boundary. Therefore, cultivation of G. lucidum mycelia using cheese whey can provide a unique solution to solve the dual problems of an alternative utilization of the whey and waste management. The substrate inhibition biokinetics at the optimal conditions were also evaluated using a method of fourth-order Runge–Kutta approximation. The nonlinear least-squares (NLLS) method with 95% confidence interval was used. The maximum microbial growth rate, μ max, and half saturation coefficient, K s, for lactose and SCOD were determined to be 2.28±0.11 and 2.27±0.15 per day, and 95.5±9.1 and 128.0±12.1 g/l, respectively. The microbial yield coefficient, Y, and microbial decay rate coefficient, k d, for lactose and SCOD were determined to be 0.49±0.03 and 0.39±0.03 g VSS/g of each substrate, and 0.05±0.01 and 0.05±0.01 per day, respectively. Inhibition coefficients were 37.6±2.9 and 49.3±3.3 g/l for lactose and SCOD, respectively.

Antiperoxidative, anti-inflammatory, and antimutagenic activities of ethanol extract of the mycelium of Ganoderma lucidum occurring in South India.

Free radical mediated genetic instability is widely thought to be a major etiological factor for initiation of carcinogenesis. Mushrooms represent a largely untapped source of powerful new pharmaceutical products. In the present study, we examined the antiperoxidative, anti-inflammatory, and antimutagenic activities of the ethanol extract of the mycelium of a medicinal mushroom, Ganoderma lucidum, occurring in south India. Antiperoxidative activity was evaluated using Fe(2+)-ascorbate-induced lipid peroxidation in rat liver homogenate and a phorbol ester (croton oil)-induced lipid peroxidation in mouse skin. Antiinflammatory activity was evaluated against carrageenan-induced acute and formalin-induced chronic inflammatory paw edema in mouse and phorbol ester-induced mouse skin inflammation. Antimutagenic activity was determined by the Ames mutagenicity assay using histidine mutant of Salmonella typhimurium strains TA 98, TA100, and TA102. Sodium azide (NaN(3)), N-methyl-N-nitro-N-nitrosoguanidine (MNNG), 4-nitro-o-phenylenediamine (NPD), and benzo[a]pyrene (B[a]P) were used as the mutagens. The extract showed significant inhibition of Fe(2+)-induced peroxidation of lipid in rat liver (IC(50) 510 +/- 22 microg/ml) and 37% inhibition of croton oil-induced peroxidation on the mouse skin at 20 mg/0.1 ml/skin. Carrageenan-induced acute and formalin-induced chronic inflammatory edema were inhibited by 56 and 60%, respectively, by the extract at 1,000 mg/kg body wt (i.p). The extract at a concentration of 5 mg/plate showed inhibition of mutagenicity elicited by direct acting mutagens, NaN(3) (55.5 and 75.7%) and MNNG (50.0 and 57.5%) for S. typhymurium strains TA100 and TA102, respectively. The extract at the same concentration also inhibited mutagenicity elicited by NPD (52.4 and 64.2%) and B[a]P (60.7 and 59.6%) for TA98 and TA100 strains, respectively. The B[a]P was activated in the presence of rat liver microsomal (S9) fraction. The results of our study revealed that ethanol extract of Ganoderma lucidum mycelium possessed significant antiperoxidative, antiinflammatory, and antimutagenic activities. The findings suggest a medicinal use for the ethanol extract of the mycelium of G. lucidum occurring in South India.

Fibrinolytic and Antithrombotic Protease from Ganoderma lucidum

A putative metalloprotease was purifed from mycelium of Ganoderma lucidum. The enzyme was purified by anion exchange, chromatofocusing, and gel filtration chromatography. The Mr was de- termined to be 52 000 by SDS-PAGE and 100 000 by gel filtration on a Sephadex G-150 column, indicat- ing that it is a dimer. The enzyme was inhibited by EDTA, 1,10-phenanthroline, and phosphoamidon. The presence of Zn2+ was detected by ICP mass spec- tral analysis as 1.1 mol of Zn2+ per mol of protease. This protease hydrolyzed A, and B, chains of human fibrinogen, but did not cleave thrombin, albumin, hemoglobin, and immunoglobulin under the same condition. It also showed an anticoagulant activity in human plasma. The enzyme delayed the activated partial thromboplastin time and thrombin time, but not the clotting induced by reptilase, indicating Gan- oderma protease is specific to thrombin. Ganoderma protease behaved as a competitive inhibitor of throm- bin-catalyzed fibrin formation, but no inhibition of thrombin was found with a small synthetic peptide. These observations indicate that Ganoderma protease could bind thrombin at an anion binding exosite dis- tinct from the active site and cause the delay of clot- ting time induced by thrombin.

Fibrinolytic and antithrombotic protease from Ganoderma lucidum

A putative metalloprotease was purified from mycelium of Ganoderma lucidum. The enzyme was purified by anion exchange, chromatofocusing, and gel filtration chromatography. The Mr was determined to be 52 000 by SDS-PAGE and 100 000 by gel filtration on a Sephadex G-150 column, indicating that it is a dimer. The enzyme was inhibited by EDTA, 1,10-phenanthroline, and phosphoamidon. The presence of Zn2+ was detected by ICP mass spectral analysis as 1.1 mol of Zn2+ per mol of protease. This protease hydrolyzed Aα and Bβ chains of human fibrinogen, but did not cleave thrombin, albumin, hemoglobin, and immunoglobulin under the same condition. It also showed an anticoagulant activity in human plasma. The enzyme delayed the activated partial thromboplastin time and thrombin time, but not the clotting induced by reptilase, indicating Ganoderma protease is specific to thrombin. Ganoderma protease behaved as a competitive inhibitor of thrombin-catalyzed fibrin formation, but no inhibition of thrombin was found with a small synthetic peptide. These observations indicate that Ganoderma protease could bind thrombin at an anion binding exosite distinct from the active site and cause the delay of clotting time induced by thrombin.

Growth and fruitbody formation of Ganoderma lucidum on media supplemented with vanadium, selenium and germanium

Responses of mycelia ofGanoderma lucidum to vanadium, selenium and germanium were examined over a wide range of concentrations (10–1, 120 μg/ml) in pure culture. Se and V were found to be highly toxic, but Ge was not toxic at the levels tested.Ganododerma lucidum cultivated on substrates of sawdust with V (30–80 μg/g) developed mature fruitbodies, but the bioaccumulation of V was quite low (2.5–7 μg/g in pileus, 12.5–21.5 μg/g in stipe and <1 μg/g in basidiospores). Se as Na2SeO4 labeled with75Se was effectively taken up from substrates and accumulated in fruitbodies (mainly in pileus), then depleted by discharge of basidiospores. Ge as GeCl4 labeled with77Ge was easily uptaken and translocated into fruitbodies.

Bistage control of pH for improving exopolysaccharide production from mycelia of Ganoderma lucidum in an air-lift fermentor

It was found that pH control definitely affects mycelial cell growth and exopolysaccharide (EPS) production of the mycelial cultivation of Ganoderma lucidum. Compared to the case of uncontrolled pH cultivation, a culture system whose pH was kept constant at 3 and 6 exhibited improved mycelial cell growth and EPS production, respectively. The bistage pH control technique, that is, shifting the pH from 3 to 6 at the initial phase of the exponential growth, is introduced to improve cell growth and EPS production. This technique can greatly increase EPS production to 20.1 g/ l from 4.1 g/ l in the case of uncontrolled pH cultivation, without adverse effects on cell growth as in the case of constant maintenance of a high pH. It was also proved that bistage pH control retained the desirable morphologies of the mycelia during cultivation and resulted in low viscosity and yield stress of the culture broth. It will be useful for the application of the culture process to mycelial growth in a large-scale fermentor.

A lectin from mycelia of the fungus Ganoderma lucidum

A lectin (GLL-M) was isolated from mycelia of Ganoderma lucidum using affinity chromatography on BSM-Toyopearl. GLL-M is a monomer in its native form with a M r of 18 000. Another lectin was also purified from fruiting bodies of the same fungus. The two lectins were partially compared with each other.

Molecular cloning of a cDNA and a gene encoding an immunomodulatory protein, Ling Zhi-8, from a fungus, Ganoderma lucidum

A large amount of the novel immunomodulatory protein Ling Zhi-8 (LZ-8) is synthesized in the mycelia of Ganoderma lucidum (Kino, K., Yamashita, A., Yamaoka, K., Watanabe, J., Tanaka, S., Ko, K., Shimizu, K., and Tsunoo, H. (1989) J. Biol. Chem. 264, 472-478). A cDNA and a gene for LZ-8 were isolated and characterized. The mixed oligonucleotide probes for LZ-8 cDNA were designed from the results of protein sequencing (Tanaka, S., Ko, K., Kino, K., Tsuchiya, K., Yamashita, A., Murasugi, A., Sakuma, S., and Tsunoo, H. (1989) J. Biol. Chem. 264, 16372-16377) and were used for screening the mycelial cDNA library. The nucleotide sequence of the cloned cDNA confirms the amino acid sequence of LZ-8 that was previously determined by protein sequencing. The clones containing the LZ-8 gene (lz-8) were obtained from the mycelial genomic DNA library using the cDNA probe. Two CCAAT-like sequences and one TATA box were found at the upstream region of the postulated transcription initiation site of lz-8. A small intron (61 nucleotides long) divided lz-8 into two exons at the 5'-untranslated region. The other characteristic sequences were also found around the postulated transcription initiation site and around the poly(A) additional site.

Ganoderic acid derivatives and ergosta-4,7,22-triene-3,6-dione from Ganoderma lucidum

Seven new triterpenoids, ganoderic acid T, ganoderic acid S, ganoderic acid R, ganoderic acid P, ganoderic acid Q, ganoderic acid 0, 7- O-methyl-ganoderic acid 0 and a known ergosterol derivative, ergosta-4,7,22-triene-3,6-dione were isolated from the cultured mycelium of Ganoderma lucidum. The structure of the first compound was determined using spectroscopic and X-ray analysis, and the structures of the other compounds were elucidated by spectroscopic data.

Structures and Antitumor Activities of the Polysaccharides Isolated from Fruiting Body and the Growing Culture of Mycelium of Ganoderma lucidum

Several β-D-glucans, appertaining to the same molecular species but having different degrees of branching, were isolated from water and alkali extracts of the fruiting body of Ganoderma lucidum (Reishi). The purified glucans that were mostly water-insoluble had a backbone of (1 →3)-linked D-glucose residues, attached mainly with single D-glucosyl units at 0-6 and also with a few short (l→4)-linked glucosyl units at 0-2 positions. However, their degrees of branching appeared to differ in the range of d.b. 1/3 ~ 1/23, depending on the extracted glucan fractions. In addition to the ^-glucans, the fruiting body contained water-soluble heteropolysaccharides, comprising D-glucose, D-galactose, D-mannose, L-(or D)-arabinose, D-xylose, and L-fucose. A branched (1 →3)-β-D-glucan was also isolated from the culture filtrate of G. lucidum grown in a glucose-yeast extract medium. The extracellular β-D-glucan was less soluble in water after purification, but soluble in dilute alkali. This glucan has essentially the same structure as that of hot-water extracted polysaccharide from the fruiting body. The repeating unit of the glucan contains a backbone chain of (1 →3)-linked D-glucose residues, five out of sixteen D-glucose residues being substituted at 0-6 positions with single D-glucosyl units and one D-glucose residue at 0-2 positions probably with a cellobiose unit. The hot-water extractable fruiting body glucan and the extracellular glucan of the culture of growing mycelium showed relatively high growth-inhibition activities against Sarcoma 180 solid tumor in mice, when administered by. successive intraperitoneal injections. When the moderately branched glucans were modified to D-glucan-polyols by periodate oxidation and borohydride reduction, they exhibited higher antitumor activities, confirming the previous conclusion that the attachment of polyol groups to the (1 →3)-lmked backbone significantly enhances its host-mediated antitumor effect.

Structures and antitumor activities of the polysaccharides isolated from fruiting body and the growing culture of mycelium of Ganoderma lucidum.

Several β-D-glucans, appertaining to the same molecular species but having different degrees of branching, were isolated from water and alkali extracts of the fruiting body of Ganoderma lucidum (Reishi). The purified glucans that were mostly water-insoluble had a backbone of (1→3)-linked D-glucose residues, attached mainly with single D-glucosyl units at O-6 and also with a few short (1→4)-linked glucosyl units at O-2 positions. However, their degrees of branching appeared to differ in the range of d.b. 1/3-1/23, depending on the extracted glucan fractions. In addition to the β-glucans, the fruiting body contained water-soluble heteropolysaccharides, comprising D-glucose, D-galactose, D-mannose, L-(or o)-arabinose, D-xylose, and L-fucose. A branched (1→3)-β-D-glucan was also isolated from the culture filtrate of G. lucidum grown in a glucose-yeast extract medium. The extracellular β-D-glucan was less soluble in water after purification, but soluble in dilute alkali. This glucan has essentially the same structure as that of hot-water extracted polysaccharide from the fruiting body. The repeating unit of the glucan contains a backbone chain of (1→3)-linked D-glucose residues, five out of sixteen D-glucose residues being substituted at O-6 positions with single D-glucosyl units and one D-glucose residue at O-2 positions probably with a cellobiose unit. The hot-water extractable fruiting body glucan and the extracellular glucan of the culture of growing mycelium showed relatively high growth-inhibition activities against Sarcoma 180 solid tumor in mice, when administered by- successive intraperitoneal injections. When the moderately branched glucans were modified to D-glucan-polyols by periodate oxidation and borohydride reduction, they exhibited higher antitumor activities, confirming the previous conclusion that the attachment of polyol groups to the (1→3)-linked backbone significantly enhances its host-mediated antitumor effect.