Antifungal Activity of Medicinal Mushrooms and Optimization of Submerged Culture Conditions for Schizophyllum commune (Agaricomycetes).

The main goal of the present study was the exploration of the antifungal properties of Agaricomycetes mushrooms. Among twenty-three tested mushrooms against A. niger, B. cinerea, F. oxysporum, and G. bidwellii, Schizophyllum commune demonstrated highest inhibition rates and showed 35.7%, 6.5%, 50.4%, and 66.0% of growth inhibition, respectively. To reveal culture conditions enhancing the antifungal potential of Sch. commune, several carbon (lignocellulosic substrates among them) and nitrogen sources and their optimal concentrations were investigated. Presence of 6% mandarin juice production waste (MJPW) and 6% of peptone in nutrient medium promoted antifungal activity of selected mushroom. It was determined that, extracts obtained in the presence of MJPW effectively inhibited the grow of pathogenic fungi. Moreover, the content of phenolic compounds in the extracts obtained from Sch. commune grown on MJPW was several times higher (0.87 ± 0.05 GAE/g to 2.38 ± 0.08 GAE/g) than the extracts obtained from the mushroom grown on the synthetic (glycerol contained) nutrient medium (0.21 ± 0.03 GAE/g to 0.88 ± 0.05 GAE/g). Flavonoid contents in the extracts from Sch. commune varied from 0.58 ± 0.03 to 27.2 ± 0.8 mg QE/g. Identification of phenolic compounds composition in water and ethanol extracts were provided by mass spectrometry analysis. Extracts demonstrate considerable free radical scavenging activities and the IC50 values were generally low for the extracts, ranging from 1.9 mg/ml to 6.7 mg/ml. All the samples displayed a positive correlation between their concentration (0.05-15.0 mg/ml) and DPPH radical scavenging activity. This investigation revealed that Sch. commune mushroom has great potential to be used as a source of antifungal and antioxidant substances.

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.

Evaluation of Lignin-Modifying Enzyme Activity of Trametes spp. (Agaricomycetes) Isolated from Georgian Forests with an Emphasis on T. multicolor Biosynthetic Potential.

In this study, a wide diversity in lignin-modifying enzyme (LME) secretion by 11 Trametes spp. strains isolated from the forests of Georgia was revealed in their submerged cultivation in both synthetic and lignocellulose-based media. Among them, T. multicolor BCC 511 was distinguished by simultaneous production of laccase, manganese peroxidase (MnP), and lignin peroxidase (LiP) in the presence of high carbon and nitrogen concentrations. Mannitol at the concentration of 15 g/L provided an accumulation of 23.7 U/mL laccase and 0.56 U/mL MnP. Significant modulation of LME activity by lignocellulosic substrates, metals, aromatic compounds, and their concentrations was established. Mandarin peels manifold increased the fungus laccase and LiP activities, while the ethanol production residue and banana peels activated manganese-oxidizing and Phenol Red-oxidizing manganese peroxidases, respectively. The addition of 2 mM of copper sulfate to the control medium induced the laccase production 28-fold and did not significantly affect the MnP and LiP activities. Fe2+ at a concentration of 0.1 mM enhanced the fungus volumetric and specific laccase activities almost 8-fold; at a concentration of 0.25-0.5 mM, there was a 2-fold increase in the MnP activity. Mn2+ appeared to be an effective inducer of the Mn-oxidizing MnP, increasing specific activity of the enzyme 14-fold. Supplementation of the copper-containing medium with 1 mM veratryl alcohol or guaiacol favored laccase and MnP production. The high yields of laccase (110 U/mL), MnP (0.62 U/mL), and LiP (0.71 U/mL) obtained in a laboratory fermenter make T. multicolor 511 useful for industrial and environmental applications.

Physiological Peculiarities of Lignin-Modifying Enzyme Production by the White-Rot Basidiomycete Coriolopsis gallica Strain BCC 142.

Sixteen white-rot Basidiomycota isolates were screened for production of lignin-modifying enzymes (LME) in glycerol- and mandarin peel-containing media. In the synthetic medium, Cerrena unicolor strains were the only high laccase (Lac) (3.2-9.4 U/mL) and manganese peroxidase (MnP) (0.56-1.64 U/mL) producers while one isolate Coriolopsis gallica was the only lignin peroxidase (LiP) (0.07 U/mL) producer. Addition of mandarin peels to the synthetic medium promoted Lac production either due to an increase in fungal biomass (Funalia trogii, Trametes hirsuta, and T. versicolor) or enhancement of enzyme production (C. unicolor, Merulius tremellosus, Phlebia radiata, Trametes ochracea). Mandarin peels favored enhanced MnP and LiP secretion by the majority of the tested fungi. The ability of LiP activity production by C. gallica, C. unicolor, F. trogii, T. ochracea, and T. zonatus in the medium containing mandarin-peels was reported for the first time. Several factors, such as supplementation of the nutrient medium with a variety of lignocellulosic materials, nitrogen source or surfactant (Tween 80, Triton X-100) significantly influenced production of LME by a novel strain of C. gallica. Moreover, C. gallica was found to be a promising LME producer with a potential for an easy scale up cultivation in a bioreactor and high enzyme yields (Lac-9.4 U/mL, MnP-0.31 U/mL, LiP-0.45 U/mL).

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.

Trinitrotoluene and mandarin peels selectively affect lignin-modifying enzyme production in white-rot basidiomycetes.

Five white-rot basidiomycetes (WRB) species have been evaluated for their potential to tolerate and to degrade 0.2 mM 2, 4, 6-trinitrotoluene (TNT) as well as to produce laccase and manganese peroxidase (MnP) in presence of this xenobiotic. The tested fungal strains produced laccase in both glycerol and mandarin peels-containing media, whereas in the glycerol-containing medium only Cerrena unicolor strains and Trametes versicolor BCC 775 secreted MnP. Replacement of glycerol by milled mandarin peels 3- to 45-fold increased laccase activity, promoted C. unicolor strains and T. versicolor MnP secretion and induced this enzyme production by Fomes fomentarius BCC 38 and Funalia trogii BCC 146. Differential response of the WRB strains to the TNT addition was observed. In particular, laccase activity of C. unicolor increased 2- to 3-fold in both media whereas no stimulation of the laccase production was revealed in cultivation of F. fomentarius. TNT practically did not affect the MnP activity. Two strains of C. unicolor followed by T. versicolor producing laccase and MnP almost completely removed 0.2 mM TNT from the synthetic medium. Increase of TNT concentration from 0 to 0.4 mM in the mandarin peels-based medium and from 0 to 0.3 mM in the glycerol-containing medium stimulated C. unicolor BCC 300 laccase production from 92.4 to 240.7 U/ml and from 17.1 to 48.6 U/ml, respectively. This strain has been resistant to the TNT high concentration and has ability to remove 85 % of initial 0.3 mM TNT content during 6 days of the submerged cultivation.

Chemical Composition and Medicinal Value of Fruiting Bodies and Submerged Cultured Mycelia of Caterpillar Medicinal Fungus Cordyceps militaris CBS-132098 (Ascomycetes).

In this paper, we report the results of a proximate analysis (i.e., moisture, ash, protein, fat, carbohydrates, and energy); a bioactive compounds analysis (i.e., cordycepin and ergothioneine); fatty and amino acid analysis; and analyses of vitamin content, macro- and microelement composition of fruiting body (FB), and mycelial biomass (MB) of medicinal caterpillar fungus Cordyceps militaris strain CBS-132098. These results demonstrate that the FB and MB of C. militaris are good sources of proteins: 59.8% protein content in the FB and 39.5% in the MB. The MB was distinguished by its carbohydrate content (39.6%), which was higher than that of the FB (29.1% carbohydrate). In the FB of C. militaris, the total amino acid content was 57.39 mg/g and in the MB it was 24.98 mg/g. The quantification of the identified fatty acids indicated that palmitic acid, oleic acid, linoleic acid, and linolenic acid were the major fatty acids. The micro- and macroelement compositions were studied. The highest results were calcium (797 mg/kg FB; 11 mg/kg MB); potassium (15,938 mg/kg FB 12,183 mg/kg MB); magnesium (4,227 mg/kg FB; 3,414 mg/kg MB); sodium (171 mg/kg FB; 1,567 mg/kg MB); phosphorus (7,196 mg/kg FB; 14,293 mg/kg MB); and sulfur (5,088 mg/kg FB; 2,558 mg/kg MB). The vitamin composition was studied, and the most abundant vitamins were vitamin A, vitamin B3, and vitamin E. The bioactive components were cordycepin, cordycepic acid (D-mannitol), and ergothioneine. There were differences in cordycepin and ergothioneine contents between the FB and the MB. The cordycepin concentration was 0.11% in the FB and 0.182% in the MB, the cordycepic acid was 4.7 mg/100g in the FB and 5.2 mg/100 g in the MB, and the ergothioneine content was 782.37 mg/kg in the FB and 130.65 mg/kg in the MB. The nutritional values of the FB and the MB of C. militaris detected indicate its potential use in well-balanced diets and sources of bioactive compounds.

Chemical Composition and Medicinal Value of the New Ganoderma tsugae var. jannieae CBS-120304 Medicinal Higher Basidiomycete Mushroom.

In this research, the chemical composition and anticancer and antioxidant activity of the new medicinal mushroom Ganoderma tsugae var. jannieae CBS-120304 were evaluated. The chemical composition assay includes amounts of total carbohydrates and proteins, amino acids, fatty acids, micro- and macroelements, and vitamins. The investigated medicinal mushroom seemed to be a rich source of nutritional components. Mycelium accumulated more than 2-fold more total protein compared with the fruiting body and reached 37% and 16% of dry weight, respectively. Carbohydrate content in the fruiting body seemed to be conspicuously higher than in the mycelium (50% of dry weight) and reached 80% of dry weight. Quantification of the identified fatty acids indicated that, in general, palmitic acid, oleic acid, and linoleic acid were the major fatty acids. Toxic elements, such as silver, arsenic, cadmium, and mercury, were found only in trace amounts in mycelium and were not detected in the fruiting body. Furthermore, the 1,1-diphenyl-2-picrylhydrazyl free radical scavenging assay was used to evaluate antioxidant activity. The highest radical scavenging activity was 9.0 mg/mL (65.9%) by ethanol extract. In addition, mycelial extracts were tested to inhibit MCF7 breast cancer cells. Ganoderma tsugae var. jannieae ethyl acetate extract (GTEAE) extract showed high potential by inhibiting reporter activity by more than 70%. Results demonstrated that GTEAE had a strong effect on inhibitory protein κΒα level in the higher concentration used (200 gg/mL), which could be compared with the effect of parthenolide. Furthermore, GTEAE demonstrated strong inhibition of IκΒα phosphorylation.

Shiitake Medicinal Mushroom, Lentinus edodes (Higher Basidiomycetes) Productivity and Lignocellulolytic Enzyme Profiles during Wheat Straw and Tree Leaf Bioconversion.

Two commercial strains of Lentinus edodes have been comparatively evaluated for their productivity and lignocellulolytic enzyme profiles in mushroom cultivation using wheat straw or tree leaves as the growth substrates. Both substrates are profitable for recycling into shiitake fruit bodies. L. edodes 3715 gave the lowest yield of mushroom during tree leaves bioconversion with the biological efficiency (BE) 74.8% while the L. edodes 3721 BE achieved 83.4%. Cultivation of shiitake on wheat straw, especially in the presence of additional nitrogen source, increased the L. edodes 3721 BE to 92-95.3% owing to the high hydrolases activity and favorable conditions. Despite the quantitative variations, each strain of L. edodes had a similar pattern for secreting enzymes into the wheat straw and tree leaves. The mushrooms laccase and MnP activities were high during substrate colonization and declined rapidly during primordia appearance and fruit body development. While oxidase activity decreased, during the same period cellulases and xylanase activity raised sharply. Both cellulase and xylanase activity peaked at the mature fruit body stage. When mushrooms again shifted to the vegetative growth, oxidase activity gradually increased, whereas the hydrolases activity dropped rapidly. The MnP, CMCase, and FP activities of L. edodes 3721 during cultivation on wheat straw were higher than those during mushroom growth on tree leaves whereas the laccase activity was rather higher in fermentation of tree leaves. Enrichment of wheat straw with an additional nitrogen source rather favored to laccase, MnP, and FPA secretion during the vegetative stage of the L. edodes 3721 growth.

Chemical composition and nutritional and medicinal value of fruit bodies and submerged cultured mycelia of culinary-medicinal higher Basidiomycetes mushrooms.

This research gives the results of a proximate analysis (moisture, ash, crude protein, fat, total carbohydrates, and total energy); a bioactive compounds analysis (γ-aminobutyric acid [GABA], ergothioneine, lovastatin, and cordycepin); fatty acid and amino acid analysis; and an analysis of macro- and microelement content of fruit bodies and mycelia of 15 higher Basidiomycetes medicinal mushroom strains belonging to 12 species. The results obtained demonstrate that almost all investigated mushrooms were found to be good sources of proteins and carbohydrates, with content varying in the ranges of 8.6-42.5% and 42.9-83.6%, respectively. Different species exhibited distinct free amino acid profiles. The total amino acid content was highest in Ophiocordyceps sinensis (MB) (23.84 mg/g) and Cordyceps militaris (FB) (23.69 mg/g). The quantification of the identified fatty acids indicated that, in general, palmitic acid, oleic acid, stearic acid, and linoleic acid were the major fatty acids. The micro- and macroelement compositions were studied, and the highest results were (as milligrams per kilogram) 224-7307 for calcium, 1668-38564 for potassium, 1091-11676 for phosphorus, and 5-97 for zinc. Bioactive components were lovastatin, GABA, and ergothioneine, which are commonly found in most mushrooms. C. militaris (FB), Pleurotus ostreatus (FB), and Coprinus comatus (FB) were most abundant and contained a high amount of GABA (756.30 µg/g, 1304.99 µg/g, 1092.45 µg/g, respectively) and ergothioneine (409.88 µg/g, 2443.53 µg/g, 764.35 µg/g, respectively). The highest lovastatin content was observed in Hericium erinaceus (FB) (14.38 µg/g) and Ganoderma lucidum (FB) (11.54 µg/g). In contrast to C. militaris (FB), cordycepin was not detected in O. sinensis (MB). The fruit body biomass of C. militaris cordycepin content reached 1.743 mg/g dry weight. The nutritional values of the mushroom species studied here could potentially be used in well-balanced diets and as sources of bioactive compounds.

Comparative study of contents of several bioactive components in fruiting bodies and mycelia of culinary-medicinal mushrooms.

Mushrooms (including fruiting bodies and mycelia) contain several bioactive components such as lovastatin, γ-aminobutyric acid (GABA), and ergothioneine. This article reports the results of 49 samples, including 9 fruiting bodies, 39 mycelia, and 1 vegetative cell, of 35 species of culinary-medicinal mushrooms from 18 genera: Agaricus, Agrocybe, Coprinus, Cordyceps, Cyathus, Daedalia, Flammulina, Fomes, Ganoderma, Grifola, Laetiporus, Lentinus, Morchella, Ophiocordyceps, Pleurotus, Trametes, Tremella, and Verpa. The results show that Cyathus striatus strain 978 contained the highest amount of lovastatin (995.66 mg/kg) in mycelia. Among fruiting bodies, 6 samples contained a high amount of GABA (274.86-822.45 mg/kg), whereas among mycelia, contents of GABA in 27 samples ranged from 215.36 to 2811.85 mg/kg. Among mycelia, Pleurotus cornucopiae strain 1101 contained the highest amount of ergothioneine (3482.09 mg/kg). Overall, these 3 bioactive components were commonly found in most mushrooms, and the results obtained might be related to their beneficial effects.

The Shaggy Inc Cap medicinal mushroom, Coprinus comatus (O.F.Mull.: Fr.) Pers. (Agaricomycetideae) substances interfere with H2O2 induction of the NF-kappaB pathway through inhibition of Ikappaalpha phosphorylation in MCF7 breast cancer cells.

Breast cancer is the most commonly diagnosed cancer among women. Currently, there is no effective therapy for malignant estrogen-independent breast cancer. In our study, we used hydrogen peroxide, a well-known strong oxidative reagent capable of activating the nuclear factor kappa B (NF-kappaB) transcription factor. The IC50 value of the culinary-medicinal Shaggy Inc Cap mushroom Coprinus comatus culture liquid crude extract on MCF7 cell viability was found to be as low as 76 microg/mL, and the IC50 value of C. comatus ethyl acetate extract was only 32 microg/ mL. Our results also showed that both extracts significantly affected IkappaBalpha phosphorylation in a dose-dependent manner. The effect of ethyl acetate extract was comparable to the effect of curcumin, a known NF-kappaB pathway inhibitor, and seemed to be the most active inhibitor of H2O2-dependent IkappaBalpha phosphorylation. In addition, the data obtained showed that only ethyl acetate extract inhibited the activity of IKK complex, at close to 90% as compared to the control of the untreated sample. These results suggest that C. comatus contains potent compounds capable of inhibiting NF-kappaB function and also possibly acts as an antitumor agent.

Free-radical scavenging activity of submerged mycelium extracts from higher basidiomycetes mushrooms.

Twenty-four Basidiomycetes strains were evaluated to determine their free-radical scavenging capacity in submerged cultivation. The scavenging capacity of the extracts varied from 1 to 85% depending on the mushroom species, solvent used, and concentration. A calculation of EC(50) of extracts from several wood-rotting Basidiomycetes showed high scavenging abilities at low effective concentration.