Antioxidant Activities of Hot Water Extracts from Mycelial Biomass of Different Combinations of Medicinal Agaricomycetes Mushrooms.

This study aimed to determine the free radical scavenging and antioxidant potential of hot water extracts prepared from different combinations and ratios of submerged cultivated mycelial biomass of medicinal mushrooms. Total phenolic compounds, flavonoid content, and antioxidant activity were evaluated for combined crude hot water extracts from medicinal higher Basidiomycetes mushrooms belonging to ten genera. The results demonstrate that almost all tested combinations were good sources of phenolic compounds and flavonoids, ranging between 16.42 and 18.83 gallic acid equivalents/g and 1.5 and 4.34 mg rutin equivalents/g, respectively. Moreover, free radical scavenging properties were evaluated with the DPPH and ABTS assays and metal chelating effects were investigated. All tested samples and/or extracts demonstrated significant free radical scavenging properties and antioxidant potential.

Chemical Composition and Antioxidant Properties of Different Combinations of Submerged Cultured Mycelia of Medicinal Mushrooms.

This research describes the investigation of submerged cultivated mycelial biomass and hot water extracts prepared from different combinations and ratios of medicinal mushroom (MM) dry powders, comprising various biologically active compounds/secondary metabolites. In particular, it was evaluated the proximate composition (moisture, ash, crude protein, fat, total carbohydrates, and total energy), γ-aminobutyric acid (GABA) and ergothioneine (ERG), amino acid content of mycelia of 16 higher Basidiomycetes MM species. The results obtained demonstrate that almost all tested combinations were found to be good sources of polysaccharides, with content varying in the ranges of 4.73 ± 1.33% and 58.46 ± 4.15%. Total protein contents varied in 1.97 ± 0.40% - 5.37 ± 0.40% range. ERG was detected in all tested samples, while GABA existed only in eight samples out of 15 and varied from 0.03 ± < 0.01 to 0.61 ± 0.03 mg/g, and from 0.16 ± 0.03 to 5.69 ± 0.41 mg/g respectively. Analyses of total phenolic and flavonoid contents demonstrate considerable content in all samples (15.53 ± 0.23 - 18.88 ± 0.34 mg gallic acid equivalents/g and 1.23 ± 0.04 - 4.34 ± 0.73 mg rutin equivalents/g respectively). In present research the complexity of samples/extracts were evaluated by multiple antioxidant assays to verify their antioxidant capacity. Determination of in vitro antioxidant activity was successfully carried out by several different methods such as 2,2-diphenyl-1-picrylhydrazyl scavenging activity, reducing power, chelating ability, hydroxyl radical scavenging activity, and 2,2'-azino-bis(3-ethylbenzothi-azoline-6-sulfonic acid scavenging activity. Therefore, all tested samples confirm the capable antioxidant activities of bioactive compounds extracted from MMs.

Antioxidant Activities of Selected Medicinal Mushroom Submerged Cultivated Mycelia.

Medicinal mushrooms fruiting bodies have been used as food or medicine for years but cultured mycelium is faster to grow and costs less. This research studied the antioxidant activities of three species (five strains) of medicinal mushroom mycelia (Cordyceps militaris, Ganoderma tsugae I and II, Trametes versicolor I and II). Two-stage extractions were performed: first the sample was extracted with 70% ethanol, and then the residue was extracted with 95°C hot water. Both ethanolic and hot water extracts showed effective concentration (EC50) values of 0.29-4.22 mg/mL, indicating that these extracts were remarkably effective in antioxidant activities. The ethanolic extracts displayed more effective reducing power, scavenging, and chelating ability (EC50 0.33-2.37 mg/mL) than hot water extracts (EC50 0.58-4.22 mg/g). Besides, ethanolic extracts contained higher total phenol content (75.49-144.99 GAE mg/g) than the hot water extracts (22.77-58.68 GAE mg/g). Furthermore, the ethanolic extracts contained flavonoids but not the hot water extract. Overall, these mycelia were highly effective in the antioxidant activities and might be potent antioxidants.

Effect of the King Oyster Culinary-Medicinal Mushroom Pleurotus eryngii (Agaricomycetes) Basidiocarps Powder to Ameliorate Memory and Learning Deficit in Ability in Aß-Induced Alzheimer's Disease C57BL/6J Mice Model.

One of the major causes of Alzheimer's disease (AD) is oxidative stress, which accelerates ß-amyloid peptide (AP) plaque and neurofibrillary tangle accumulation in the brain. Pleurotus eryngii is known to be rich in antioxidants, including ergothioneine, adenosine, and polyphenol, which can reduce oxidative stress-related aging. The aim of this study was to investigate the proximate and functional composition of P. eryngii, and evaluate the cognitive effects of low (LPE), medium (MPE), and high (HPE) P. eryngii dosages in an Aß-induced Alzheimer's disease C57BL/6J mouse model. Mice fed P. eryngii for six weeks showed no adverse effects on body weight gain, food intake efficiency, serum biochemical parameters, and liver and kidney histopathological features. The relative brain weight was significantly lower in Aß-injected mice (p < 0.05). Further, P. eryngii was shown to delay brain atrophy. Reference memory behavioral tasks showed that LPE, MPE, and HPE significantly decreased escape latency (49-85%) and distance (53-69%, p < 0.05). Probe and T-maze tasks showed that P. eryngii potently ameliorated memory deficit in mice. An AD pathology index analysis showed that P. eryngii significantly decreased levels of brain phosphorylated τ-protein, Aß plaque deposition, malondialdehyde, and protein carbonyl (p < 0.05). P. eryngii may therefore promote memory and learning capacity in an Aß-induced AD mouse model.

Effect of Ethanolic Extracts from Taiwanofungus camphoratus and T. salmoneus (Agaricomycetes) Mycelia on Osteoporosis Recovery.

Osteoporosis is common in postmenopausal women and elderly people. In this study, the ovariectomized mice were used as an in vivo test to evaluate the effects of 70% ethanolic extracts of Taiwanofungus camphoratus and T. salmoneus (Polyporales, Agaricomycetes) on postmenopausal osteoporosis. Ovariectomized mice had significantly higher body weight and histopathological alterations of the liver were found to have diffused fatty infiltrated vesicles. The bone parameters of the femur were determined by microcomputed tomography. In addition, the relative weight of the uterus is significantly lower and atrophy of the uterine glands was found in histopathological alterations. The results of trabecular bone parameters showed that feeding high doses of T. camphoratus mycelia ethanolic extract to ovariectomized mice had the ability to delay bone loss. The bone density of trabecular bone and cortical bone were also significantly higher than those of ovariectomized mice, indicating that the ethanolic extract of T. camphoratus has the potential to delay the occurrence of osteoporosis.

Morphological Characteristics, Molecular Identification and Antioxidant Activities of Phallus atrovolvatus (Agaricomycetes) Isolated from Thailand.

Phallus atrovolvatus is a wild edible mushroom found in Thailand. Three strains of Ph. atrovolvatus (DOAP-1, DOAP-2, and DOAP-3) were collected from forests in Central Thailand. Some requirements for mycelial growth were obtained in different media. Potato dextrose agar was determined as the best medium to support mycelial growth (83.50 mm after incubation for 7 days). Molecular phylogenetic analyses based on the sequence of internal transcribed spacers (ITS1 and ITS4) confirmed DOAP-1 species status within Phallaceae as Ph. atrovolvatus with high levels of similarity at 99.34%. Antioxidant properties of hot water extract from the fruiting body of three isolates (CMP-1, CMP-2, and CMP-3) were also evaluated. Highest free radical scavenging ability was found in CMP-1 (94.94% at 2.0 mg/mL) whereas crude mushroom extracts exhibited very strong ferrous-ion chelating effects of 99.16% at 10 mg/ mL. Results indicating that all CMP isolates from Ph. atrovolvatus possess excellent antioxidant properties from natural sources.

Composition of Mycelia and Basidiomata of the Culinary-Medicinal Golden Oyster Mushroom, Pleurotus citrinopileatus (Agaricomycetes) by Nuclear Magnetic Resonance Spectroscopy.

Nuclear magnetic resonance (NMR) spectroscopy has been used to obtain the profile of soluble metabolites on fungus. To enhance the ergothioneine content in Pleurotus citrinopileatus mycelium, amino acid precursors were used for submerged fermentation. The study aimed to analyze the metabolites of high-ergothioneine and regular P. citrinopileatus mycelia (HEPM and RPM) and P. citrinopileatus basidiomata (PCB) using NMR spectroscopy. Principal component analysis (PCA) was applied to differentiate and to describe the differences among three sets of metabolites. The results showed that 55 water-soluble metabolites of PCB, HEPM, and RPM were identified and classified into five groups including amino acids, carbohydrates, organic acids, purines and pyrimidines, and others. Contents of total soluble metabolites were in descending order as follows: PCB (135 mg/g) > HEPM (58 mg/g) > RPM (44 mg/g). The score plot and loading plot separated using PCA showed that the first two components were responsible for R2X = 97.1% and the variance [R2X(1) = 74% for principal component PC1 and R2X(2) = 23.1% for PC2]. A metabolic pathway map of P. citrinopileatus mycelium was established and the differences in the metabolites of RPM and HEPM with precursors added were illuminated.

Extraction of Ergothioneine from Pleurotus eryngii and P. citrinopileatus (Agaricomycetes) and Preparation of Its Product.

Ergothioneine is an effective antioxidant and is abundant in species of genus Pleurotus. This research focused on developing an ergothioneine extract from P. eryngii and P. citrinopileatus under optimal extraction conditions. The hot-water and 70% ethanol extractions yielded more ergothioneine than microwave, ultrasonic, and autoclaving extractions. Hot-water extraction with optimal conditions-125 rpm at 75°C for 5 minutes-produced P. eryngii and P. citrinopileatus extracts with 0.86 and 3.73 mg ergothioneine/g dry weight, respectively. In addition to 50% of added maltodextrin, spray-dried products from mushrooms contained 23.07-16.58% carbohydrates and 18.32-21.40% protein. The microstructure of spray-dried products showed shrunken spheres with mean particle diameters of 15.82-19.94 µm. After in vitro simulated gastric and intestinal digestion, the spray-dried P. eryngii and P. citrinopileatus products contained 88% and 91% residual ergothioneine, respectively. Overall, the spray-dried products could be used as antioxidative supplements.

Apoptotic Effect of Taiwanofungus salmoneus (Agaricomycetes) Mycelia and Solid-State Fermented Products on Cancer Cells.

Because of the high incidence of and high mortality rates associated with cancer, effective therapy for cancer is currently an imperative issue. Taiwanofungus salmoneus, a medicinal mushroom indigenous to Taiwan, has been demonstrated to exhibit various physiological activities and harmless effects. The bioactive compounds and anticancer abilities of T. salmoneus mycelia and fermented products were evaluated in this study. Total phenols, flavonoids, and ergothioneine were detected in mycelia and fermented products, and the amount of ergothioneine was evaluated in buckwheat (56.25-118.64 µg/g extract) and oats (53.60-138.70 µg/g extract) after fermentation. Results showed that T. salmoneus-fermented buckwheat and oats exhibited better apoptotic effects on Sk-Hep-1 and Caco-2 cells than sorafenib and celecoxib via receptor-linked apoptosis and mitochondria-mediated apoptosis. In addition, T. salmoneus-fermented buckwheat may have potential for replacing anticancer drugs or for consumption as a dietary supplement to retard cancer development.

Anti-Inflammation and Lipogenic Inhibition of Taiwanofungus salmonea Mycelium and Grifola frondosa Fruiting Body.

A long-term inflammatory response can cause the development of nonalcoholic steatohepatitis (NASH), which is mainly caused by abnormal metabolism of lipids in the liver. Taiwanofungus salmonea and Grifola frondosa, which are valuable medicinal mushrooms in Taiwan and exhibit many beneficial bioactivities, are used to evaluate effects on immune regulation in Raw 264.7 cells and on NASH-related responses in Hep G2 cells. According to the results, the production of proinflammatory cytokines was effectively decreased, whereas the production of interleukin-10 was increased. Lipid accumulation and reactive oxygen species production also declined, and free glycerol was released. The expression of acetyl-CoA carboxylase, fatty acid synthase, and sterol regulatory element-binding protein 1c was decreased. These results indicate that T. salmonea and G. frondosa are capable of reducing inflammatory responses and ameliorating NASH, indicating their potential as hepatoprotective dietary supplements.

Quality of Bread Supplemented with Antrodiasalmonea-Fermented Grains.

Fermented grains of buckwheat, oat, embryo rice and wheat, which were prepared by solid-state fermentation with Antrodia salmonea, and the mycelium was used to substitute 7% of wheat flour to make bread. No difference in proximate composition, texture profile and contents of non-volatile taste components was observed among bread samples. White bread and bread supplemented with mycelium and fermented grains looked different. Bread supplemented with fermented grains had similar thermal properties, which differed from those of white bread and bread supplemented with mycelium. Bread supplemented with fermented grains contained substantial mass fractions (on dry mass basis) of adenosine (0.92-1.96 µg/g), ergosterol (24.53-30.12 µg/g), ergothioneine (2.16-3.18 µg/g) and γ-aminobutyric acid (2.20-2.45 µg/g). In addition, bread supplemented with mycelium contained lovastatin (0.43 µg/g). White bread and bread supplemented with fermented grains had similar sensory results. Overall, fermented grains could be incorporated into bread to provide beneficial effects.

Nonvolatile Taste Components and Antioxidant Properties of Fruiting Body and Mycelium with High Ergothioneine Content from the Culinary-Medicinal Golden Oyster Mushroom Pleurotus citrinopileatus (Agaricomycetes).

Pleurotus citrinopileatus mycelium was prepared with high ergothioneine (Hi-Ergo) content and its proximate composition, nonvolatile taste components, and antioxidant properties were studied. The ergothioneine contents of fruiting bodies and Hi-Ergo and regular mycelia were 3.89, 14.57, and 0.37 mg/g dry weight, respectively. Hi-Ergo mycelium contained more dietary fiber, soluble polysaccharides, and ash but less carbohydrates, reducing sugar, fiber, and fat than regular mycelium. However, Hi-Ergo mycelium contained the smallest amounts of total sugars and polyols (47.43 mg/g dry weight). In addition, Hi-Ergo mycelium showed the most intense umami taste. On the basis of the half-maximal effective concentration values obtained, the 70% ethanolic extract from Hi-Ergo mycelium showed the most effective antioxidant activity, reducing power, and scavenging ability, whereas the fruiting body showed the most effective antioxidant activity, chelating ability, and Trolox-equivalent antioxidant capacity. Overall, Hi-Ergo mycelium could be beneficially used as a food-flavoring material or as a nutritional supplement.

Anti-Inflammation Properties of Fruiting Bodies and Submerged Cultured Mycelia of Culinary-Medicinal Higher Basidiomycetes Mushrooms.

This research shows the phenolic composition and anti-inflammation properties of fruiting bodies and mycelia of 15 strains of 12 species of higher Basidiomycetes medicinal mushrooms. In this research, 15 extracts were prepared and their effects on inflammation-related mediators in RAW 264.7 cells were evaluated. In the extracts, amounts of total phenols ranged from 8.47 to 70.32 gallic acid equivalents mg/g and amounts of flavonoids ranged from 0.13 to 15.21 rutin equivalents mg/g. The production of nitric oxide, tumor necrosis factor-α, and interleukin-6 was decreased at different levels by these extracts, whereas the production of interleukin-10 was increased by 6 of the extracts. Overall, Cordyceps militaris fruiting bodies, Grifola frondosa fruiting bodies, and Ophiocordyceps sinensis mycelia might be used to ameliorate inflammatory responses.

Quantification of Water-Soluble Metabolites in Medicinal Mushrooms Using Proton NMR Spectroscopy.

The water-soluble metabolites in 5 mushrooms were identified and quantified using proton nuclear magnetic resonance (NMR) spectroscopy and software for targeted metabolite detection and quantification. In total, 35 compounds were found in Agaricus brasiliensis, 25 in Taiwanofungus camphoratus, 23 in Ganoderma lucidum (Taiwan) and Lentinus edodes, and 16 in G. lucidum (China). Total amounts of all identified metabolites in A. brasiliensis, T. camphoratus, G. lucidum, G. lucidum (China), and L. edodes were 149,950.51, 12,834.18, 9,549.09, 2,788.41, and 111,726.51 mg/kg dry weight, respectively. These metabolites were categorized into 4 groups: free amino acids and derivatives, carbohydrates, carboxylic acids, and nucleosides. Carbohydrates were the most abundant metabolites among all 4 groups, with mannitol having the highest concentration among all analyzed metabolites (848-94,104 mg/kg dry weight). Principal components analysis (PCA) showed obvious distinction among the metabolites of the 5 different kinds of mushrooms analyzed in this study. Thus PCA could provide an optional analytical way of identifying and recognizing the compositions of flavor products. Furthermore, the results of this study demonstrate that NMRbased metabolomics is a powerful tool for differentiating between various medicinal mushrooms.

A steroid like phytochemical Antcin M is an anti-aging reagent that eliminates hyperglycemia-accelerated premature senescence in dermal fibroblasts by direct activation of Nrf2 and SIRT-1.

The present study revealed the anti-aging properties of antcin M (ANM) and elucidated the molecular mechanism underlying the effects. We found that exposure of human normal dermal fibroblasts (HNDFs) to high-glucose (HG, 30 mM) for 3 days, accelerated G0/G1 phase arrest and senescence. Indeed, co-treatment with ANM (10 µM) significantly attenuated HG-induced growth arrest and promoted cell proliferation. Further molecular analysis revealed that ANM blocked the HG-induced reduction in G1-S transition regulatory proteins such as cyclin D, cyclin E, CDK4, CDK6, CDK2 and protein retinoblastoma (pRb). In addition, treatment with ANM eliminated HG-induced reactive oxygen species (ROS) through the induction of anti-oxidant genes, HO-1 and NQO-1 via transcriptional activation of Nrf2. Moreover, treatment with ANM abolished HG-induced SIPS as evidenced by reduced senescence-associated ß-galactosidase (SA-ß-gal) activity. This effect was further confirmed by reduction in senescence-associated marker proteins including, p21CIP1, p16INK4A, and p53/FoxO1 acetylation. Also, the HG-induced decline in aging-related marker protein SMP30 was rescued by ANM. Furthermore, treatment with ANM increased SIRT-1 expression, and prevented SIRT-1 depletion. This protection was consistent with inhibition of SIRT-1 phosphorylation at Ser47 followed by blocking its upstream kinases, p38 MAPK and JNK/SAPK. Further analysis revealed that ANM partially protected HG-induced senescence in SIRT-1 silenced cells. A similar effect was also observed in Nrf2 silenced cells. However, a complete loss of protection was observed in both Nrf2 and SIRT-1 knockdown cells suggesting that both induction of Nrf2-mediated anti-oxidant defense and SIRT-1-mediated deacetylation activity contribute to the anti-aging properties of ANM in vitro. Result of in vivo studies shows that ANM-treated C. elegens exhibits an increased survival rate during HG-induced oxidative stress insult. Furthermore, ANM significantly extended the life span of C. elegans. Taken together, our results suggest the potential application of ANM in age-related diseases or as a preventive reagent against aging process.

Antimicrobial and antitumor activities of chitosan from shiitake stipes, compared to commercial chitosan from crab shells.

Chitosan was prepared by alkaline N-deacetylation of chitin obtained from shiitake stipes and crab shells and its antimicrobial and antitumor activities were studied. Chitosan from shiitake stipes and crab shells exhibited excellent antimicrobial activities against eight species of Gram positive and negative pathogenic bacteria with inhibition zones of 11.4-26.8mm at 0.5mg/ml. Among chitosan samples, shiitake chitosan C120 was the most effective with inhibition zones of 16.4-26.8mm at 0.5mg/ml. In addition, shiitake and crab chitosan showed a moderate anti-proliferative effect on IMR 32 and Hep G2 cells. At 5mg/ml, the viability of IMR 32 cells incubated with chitosan was 68.8-85.0% whereas that of Hep G2 cells with chitosan was 60.4-82.9%. Overall, shiitake chitosan showed slightly better antimicrobial and antitumor activities than crab chitosan. Based on the results obtained, shiitake and crab chitosan were strong antimicrobial agents and moderate antitumor agents.

Submerged Cultivation of Mycelium with High Ergothioneine Content from the Culinary-Medicinal Golden Oyster Mushroom, Pleurotus citrinopileatus (Higher Basidiomycetes).

The optimization of submerged culture of the culinary-medicinal golden oyster mushroom, Pleurotus citrinopileatus, was studied using a one-factor-at-a-time, two-stage stimulation and central composite rotatable design to produce mycelia with high ergothioneine content. The optimal culture conditions for mycelia harvested at day 22 were a temperature of 25°C, an inoculation ratio of 5%, 2% glucose, 0.5% yeast extract, and adjustment of the initial pH value to 10. The biomass and ergothioneine content were 8.28 g/L and 10.65 mg/g dry weight (dw), respectively. The addition of an amino acid precursor increased the ergothioneine content of mycelia; cysteine was the most effective. In addition, the results obtained from central composite rotatable design showed that the recommended combination for cysteine, histidine, and methionine was 8, 4, and 0.5 mmol/L, respectively. The predicted ergothioneine content was 13.90 mg/g dw, whereas the experimental maximal ergothioneine content was 14.57 mg/g dw. With the addition of complex precursors and under optimal culture conditions, mycelia harvested at days 16-20 had higher ergothioneine content. Accordingly, the information obtained could be used to produce mycelia with high ergothioneine content.

Apoptotic Effect of Extract from Medicinal Mushroom from Taiwan Taiwanofungus salmoneus (Higher Basidiomycetes) Mycelium Combined with or without Cisplatin on Hepatocellular Carcinoma Cells.

Hepatocellular carcinoma is a cancer of high mortality; therefore, the effective therapy on this cancer is an imperative issue. Recently, anticancer agent combined with natural products has been demonstrated to increase apoptosis of various cancer cells effectively. Accordingly, we investigated the apoptotic effect and possible mechanism of the ethanol extract from Taiwanofungus salmoneus (=Antrodia salmonea) mycelium (TsE) alone or in combination with cisplatin in SK-Hep-1 cells. In this study, the proliferation of SK-Hep-1 cells could be inhibited at various concentrations of TsE for 24 h whereas TsE combined with cisplatin would inhibit the cell proliferation more notably. Moreover, the DNA damage and the interruption of cell cycle of SK-Hep-1 cells would be effectively raised after incubation with TsE combined with cisplatin for 24 h. The apoptosis of cells was dramatically induced, and the expression of caspases 3, 8, and 9, apoptosis-related protein, were significantly upregulated. Therefore, we proposed that the TsE combined with cisplatin inhibited cell proliferation by elevating sub-G1 phase, inducing DNA damage, activating caspases 3, 8, and 9 activities, and triggering cells apoptosis. These results reveal that TsE could be a potential adjuvant chemotherapeutic agent.

Antiproliferative Activities of Hot Water Extracts from Culinary-Medicinal Mushrooms, Ganoderma tsugae and Agrocybe cylindracea (Higher Basidiomycetes) on Cancer Cells.

Using anticancer agents to progress chemotherapy to inhibit the proliferation of cancer cells is an effective means. Two medicinal mushrooms, Ganoderma tsugae and Agrocybe cylindracea, exhibited various physiological effects, and the antiproliferation effect on HL-60, Hep 3B, and C6 cells was studied. The viability of different cancer cells was decreased significantly by hot water extracts from different forms of G. tsugae and A. cylindracea. The hot water extracts from the fruit body, mycelium, and filtrate of A. cylindracea were less effective in inhibiting the antiproliferation of C6, Hep 3B, and HL-60 cells than were those from G. tsugae, as evidenced by their IC50 values. The IC50 values of G. tsugae on C6, Hep 3B, and HL-60 cells were 1.13, 2.73, and 2.60 mg/mL, respectively, whereas those of baby G. tsugae were 1.87, 2.63, and 3.12 mg/mL, respectively. In addition, the filtrates of G. tsugae on C6 and Hep 3B cells were 2.81 and 2.80 mg/mL, respectively. The morphological transformation of 3 cancer cells was observed clearly, and the possible mechanism would be necrosis, apoptosis, or differentiation. Owing to the noticeable effect on antiproliferation of hot water extracts, especially those from G. tsugae, the extract could be of great potential to be used as an alternative cancer therapy.

Effect of UV-B Irradiation on Physiologically Active Substance Content and Antioxidant Properties of the Medicinal Caterpillar Fungus Cordyceps militaris (Ascomycetes).

Ultraviolet-B (UV-B) light irradiation is a well-known technique for converting vitamin D2 from ergosterol in mushroom fruit bodies. Mushrooms are a natural and nonanimal food source of vitamin D2. We studied the effect of UV-B light irradiation on the amount of vitamin D2 and physiologically active substances in Cordyceps militaris and their antioxidant properties. After UV-B irradiation for 2 hours, the vitamin D2 content of freshly harvested C. militaris fruiting bodies, mycelia, whole submerged culture (WSC), and homogenized submerged culture (HSC) increased from 0 to 0.03 to 0.22 to 1.11 mg/g, but the ergosterol content was reduced from 1.36 to 2.50 to 1.24 to 2.06 mg/g, respectively. After UV-B irradiation, the amount of adenosine, cordycepin, and ergothioneine of fruiting bodies dramatically increased 32-128%, but the polysaccharide content slightly decreased 36%. The reverse trends were observed in mycelia, WSC, and HSC. UV-B irradiation could reduce the effective concentrations at 50% of fruiting bodies for ethanolic and hot water extracts in reducing power, scavenging, and chelating abilities, whereas mycelia, WSC, and HSC of ethanolic extracts increased effective concentrations at 50% in reducing power, scavenging, and chelating abilities. UV-B irradiation slightly increased flavonoid content (10-56%) and slightly affected total phenol content.