Clitocybe Maxima Research Articles

Exploring the Potential of Fungal Biomass for Bisphenol A Removal in Aquatic Environments

Bisphenol A is a plastic component, which shows endocrine activity that is detrimental to humans and aquatic ecosystems. The elimination of BPA from the environment is one of the solutions for BPA contaminant management. Adsorption is a cost-effective, easy-to-use method generating low harmful byproducts; nevertheless, contaminant sorbent treatment is a challenge that still needs to be addressed. Fungal fruiting bodies biomass is rarely studied sorbent but is promising due to its high polysaccharide content and availability. Our preliminary studies showed BPA sorption (100 mg/L) by 50 cultivated and wild fungi. The cultivated species: Clitocybe maxima (82%), Pholiota nameko (77%), and Pleurotus columbinus (74%), and wild fungi Cantharellus cibarius (75%) and Lactarius deliciosus (72%) were the most efficient. The biomass was able to sorb BPA over a broad range of temperature and pH levels, with an optimum at 20 °C and pH 7. Although saturation of sorbents was rapid, the regeneration process using ethanol was effective and allowed to recover up to 75% of sorbents’ initial efficiency. A single use of 1 g of sorbent would allow the treatment of 8.86 to 10.1 m3 of wastewater effluent, 16.5 to 18.7 m3 of surface water, and 411 to 469 m3 of drinking water, assuming the concentrations of BPA reported in the literature.

Effect of oxidative torrefaction on the pyrolysis of Clitocybe maxima stipe: Pyrolysis behaviour, and products' properties

In this study, oxidative torrefaction (OT) was used as the pretreatment process for the pyrolysis of Clitocybe maxima stipe (CMS) which is one kind of typical mushroom wastes, in order to evaluate the influence of OT on pyrolysis behaviour of torrefied CMS and products’ properties. The experimental results demonstrated that OT enhanced the degree of aromatization, and improved the thermal stability of the torrefied CMS compared with inert torrefaction (IT). The biochar (T9-P700) obtained by OT (9% oxygen) coupled pyrolysis (700 °C) had the improved pore structure with the highest specific surface area of 661.61 m2/g, while the obtained liquid products were less acidic with an increased formation of N-containing compounds (up to 19.15%) and phenolic compounds (8.08%). Furthermore, T9-P700 had a higher adsorption efficiency and adsorption capacity (147.31 mg/g) for Pb(Ⅱ). C-O, CO, -COOH, and -OH on the obtained biochar participated in the adsorption process of Pb(Ⅱ), attributing to the adsorption mechanism interpreted as physical adsorption, precipitation, and functional group complexation. Higher oxygen concentrations increased the severity of torrefaction, which facilitated a better pore structure in the CMS biochar generated during the subsequent pyrolysis process and the generation of liquid products with better qualities. Therefore, OT coupled pyrolysis may be an effective way to improve the quality of pyrolysis products.

Multi-targeting cytotoxic drug leads from mushrooms

Due to genetic and epigenetic mechanisms, cancer have become a resistant disease and there is a need for new molecules having multiple targeting action that promotes apoptosis. Phyto-molecules having multiple targeting anticancer activity are in high demand and there is less documentation or information available on these metabolites. It is evident that mushrooms are became the store houses of new anticancer molecules and mushrooms like Agaricus blazei, Antrodia camphorate, Albatrellus confluens, Bolteus badius, Cordyceps militaris, Clitocybe maxima, Funalia trogii, Grifola frondosa, and Inocybe umbrinella, are some of the medicinal mushrooms reported for their cytotoxic activity. Cytotoxic molecules like lentinan, grifolin, illudin-S, psilocybin, ganoderic acid, theanine, and hispolon, have been isolated from various mushroom species. However, studies have been limited only to in vitro cytotoxic mechanisms and very few trials have been conducted to prove the clinical efficacy of these drug leads. Hence, the current review focuses on new anticancer metabolites isolated from various mushrooms having multiple targeting mechanisms in cancer. However, an extensive research is needed to define the biosynthesis and clinical mechanism of these multiple acting metabolites. This review provides a platform for researchers new anticancer drugs and to bring out potent multiple acting anticancer newer drugs.

Andioxidant and structural analysis of polysaccharide extracted from Clitocybe maxima Stipe using ultrasonication: simulation and validation

Andioxidant and structural analysis of polysaccharide extracted from Clitocybe maxima Stipe using ultrasonication: simulation and validation

The effect of different substrates on the growth of six cultivated mushroom species and composition of macro and trace elements in their fruiting bodies

There is an ongoing interest in the production of mushrooms as food, and with their consumption on the rise, there is a need to establish different safety measures. In the present study, six mushroom species (Agrocybe cylindracea, Clitocybe maxima, Flammulina velutipes, Ganoderma lucidum, Lentinula edodes and Pleurotus eryngii) were cultivated on two commonly used substrates (A—based on alder and beech sawdust; B—based on oak sawdust and flax shives). The aims were to determine their growth and the accumulation of 70 elements in their fruiting bodies, 33 of which were detected in all analysed samples and were used for statistical evaluation. Cultivation of C. maxima, G. lucidum and L. edodes resulted in higher yield and mineral content when substrate A was used, cultivation of A. cylindracea and F. velutipes yielded better results on substrate B, P. eryngii cultivation outcomes were similar for both substrates. L. edodes was found to have a high affinity to accumulate Cd, C. maxima can bioconcentrate Al and Ni, Ganoderma lucidum—Pb, while F. velutipes—Hg. The study indicated that the chemical composition of substrates could affect both the yield and the level of various toxic and nutritional elements.

Optimization of Submerged Cultivation Conditions for Production of Big Cup Culinary-Medicinal Mushroom Clitocybe maxima (Agaricomycetes) Biomass with Significant Antioxidative and Antihyperlipidemic Activities.

In this study, a large Clitocybe maxima mycelium biomass was obtained by submerged cultivation under optimal conditions. Three test samples from lyophilized mycelia, including hot water extract (CW) and elutes from solvents with different polarity (CA and CB), were combined and used to explore antioxidant and antihyper-lipidemic activities in vitro and in vivo. The CA group showed the highest DPPH free radical scavenging activity and iron-reducing capability at concentrations of 6.0% and 3.0% (w/v), respectively. Further, the CA group showed the highest glutathione peroxidase and superoxide dismutase activities at a dose of 0.25 mg/kg body weight (CA-0.25 group) in all hyperlipidemic hamsters tested. Serum lipid levels (apart from high-density lipoprotein cholesterol levels) of hamsters in the CA-0.25 group were lower than those of hamsters in the negative control group in antihy-perlipidemic tests. Therefore, we believe that extracts from C. maxima mycelia are rich reservoirs of antioxidant and antihyperlipidemic activities.

Mycoextraction by Clitocybe maxima combined with metal immobilization by biochar and activated carbon in an aged soil

To develop an eco-friendly and efficient route to remediate soil highly polluted with heavy metals, the idea of mycoextraction combined with metal immobilization by carbonaceous sorbents (biochar and activated carbon) was investigated in this study. Results showed that the application of carbonaceous amendments decreased acid soluble Cd and Cu by 5.13–14.06% and 26.86–49.58%, respectively, whereas the reducible and oxidizable fractions increased significantly as the amount of carbonaceous amendments added increased. The biological activities (microbial biomass, soil enzyme activities) for treatments with carbonaceous sorbents were higher than those of samples without carbonaceous amendments. Clitocybe maxima (C. maxima) simultaneously increased soil enzyme activities and the total number of microbes. Biochar and activated carbon both showed a positive effect on C. maxima growth and metal accumulation. The mycoextraction efficiency of Cd and Cu in treatments with carbonaceous amendments enhanced by 25.64–153.85% and 15.18–107.22%, respectively, in response to that in non-treated soil, which showed positive correlation to the augment of biochar and activated carbon in soil. Therefore, this work suggested the effectiveness of mycoextraction by C. maxima combined the application of biochar and activated carbon in immobilising heavy metal in contaminated soil.

Bioremediation of soils co-contaminated with heavy metals and 2,4,5-trichlorophenol by fruiting body of Clitocybe maxima

Pot experiments were performed to investigate the single effect of 2,4,5-trichlorophenol (TCP) or heavy metals (Cu, Cd, Cu+Cd) and the combined effects of metals-TCP on the growth of Clitocybe maxima together with the accumulation of heavy metals as well as dissipation of TCP. Results showed a negative effect of contaminations on fruiting time and biomass of the mushroom. TCP decreased significantly in soils accounting for 70.66–96.24% of the initial extractable concentration in planted soil and 66.47–91.42% in unplanted soil, which showed that the dissipation of TCP was enhanced with mushroom planting. Higher biological activities (bacterial counts, soil respiration and laccase activity) were detected in planted soils relative to unplanted controls, and the enhanced dissipation of TCP in planted soils might be derived from the increased biological activities. The metals accumulation in mushroom increased with the augment of metal load, and the proportion of acetic acid (HOAc) extractable metal in soils with C. maxima was larger than that in unplanted soils, which may be an explanation of metal uptake by C. maxima. These results suggested that the presence of C. maxima was effective in promoting the bioremediation of soil contaminated with heavy metals and TCP.

Interaction of copper and 2,4,5-trichlorophenol on bioremediation potential and biochemical properties in co-contaminated soil incubated with Clitocybe maxima

The bioremediation of soil co-contaminated with heavy metal and organic pollutants has attracted considerable attention in recent years.

Antitumor and Immunomodulating Activities of Exopolysaccharide Produced by Big Cup Culinary- Medicinal Mushroom Clitocybe maxima (Higher Basidiomycetes) in Liquid Submerged Culture.

Water-soluble polysaccharides extracted from mushrooms have been found to have some physiological effects. In this study, exopolysaccharides (EPSs) were extracted by alcohol precipitation from cultivated broth of the mushroom Clitocybe maxima. EPSs with molecular weights of 10(4) and 10(5) Da were obtained by ultrafiltration; they are referred to as EPA and EPB, respectively. The major components of these EPSs were glucose, galactose, mannose, rhamnose, and arabinose. ICR mice with artificially induced metastatic pulmonary tumors were fed a daily diet containing EPA or EPB at doses of 8, 20, or 50 mg/kg. Results showed that the proliferation of pulmonary sarcoma lesions was lower in the groups fed EPS. In addition, the numbers of total T cells, CD4+ cells, CD8+ cells, and macrophages significantly increased in EPS-fed mice compared with the negative control group. The antitumor and immunomodulating effects observed in the EPB-fed groups were higher than those of EPA-fed groups. These results demonstrate the ability of EPSs of C. maxima to inhibit tumor cells while enhancing immune response.

Effect of Extraction Methods on Polysaccharide of <em>Clitocybe maxima</em> Stipe

Clitocybe maxima (Gartn. ex Mey. Fr.) Quel. is a favorable edible fungi species. The proportion of its stipe is about 45% of entire fruit biomass, which is a low value defined byproduct. To increase its value added utilization, three extraction methods (as hot water, microwave-assisted and complex-enzyme-hydrolysis-assist) were conducted. The extraction effect on the polysaccharide of Clitocybe maxima stipe was compared and the processing conditions in extraction were optimized. The content of polysaccharide was determined with Phenol Sulfuric Acid Method. With three methods, the extraction yield and the polysaccharide amount were different greatly, which were 5.86 and 40.23% for hot water extraction, 9.4 and 52.62% for microware-assist extraction, 10.26 and 53.13% for complex-enzyme-hydrolysis extraction, respectively. Results indicated that the complex-enzyme-hydrolysis-assist extraction was the optimal approach used for extracting polysaccharide from Clitocybe maxima stipe. It showed with the advantages in high efficiency, energy and running time saving.

The novel role of fungal intracellular laccase: used to screen hybrids between Hypsizigus marmoreus and Clitocybe maxima by protoplasmic fusion

Laccase has been proved important in decolorization of Remazol Brilliant Blue R (RBBR), oxidation of 2, 2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, lignin degradation and fruiting-body formation. The decolorization of RBBR by laccase was firstly used to screen protoplast fusants. Fusants were obtained by protoplast fusion between the strains of Hypsizigus marmoreus and Clitocybe maxima, and two fusants (IM1 and IIIM5) were screened on PDA medium containing RBBR. These fusants were significant higher in laccase activity than H. marmoreus, nearly 413 and 395 times, respectively. Their hyphal growth rates were also remarkable higher than H. marmoreus, nearly 1.5 and 1.4 times, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed these fusants contained the laccase, and the molecular mass of the laccase was consistent with the laccase of C. maxima, nearly 62 kDa. The pileus color of the IM1 and IIIM5 also showed partial recombined characteristics comparing to the parental strains, while biological efficiency ratios were prominent higher than that of H. marmoreus, up to 14.58 and 10.87 %, respectively. Randomly amplified polymorphic DNA bands of fusants not only were similar to parental bands, but presented new non-parental bands. Using the Unweighted pair-group method together with mathematic averages method to gain a dendrogram, in which the fusants showed intra-cluster variations. Significantly, H. marmoreus was the dominant parent, while C. maxima were distant from the fusants. The differences among IM1, IIIM5 and H. marmoreus, and the similarities among IM1, IIIM5 and C. maxima indicated IM1 and IIIM5 were somatic hybrids of H. marmoreus and C. maxima. Accordingly, it is feasible to use laccase to screen fusants of H. marmoreus and C. maxima.

Chemical composition of five wild edible mushrooms collected from Southwest China and their antihyperglycemic and antioxidant activity

Evaluation of the chemical composition and antihyperglycemic and antioxidant activity of five wild edible mushrooms (Clitocybe maxima, Catathelasma ventricosum, Stropharia rugoso-annulata, Craterellus cornucopioides and Laccaria amethystea) from Southwest China. The chemical composition assay includes proximate analysis (moisture, ash, crude protein, crude fat, total carbohydrates and total energy), bioactive compounds analysis (total phenolic, flavonoid, ascorbic acid, ergosterol, tocopherol), fatty acid analysis, amino acid analysis, phenolic compounds analysis and mineral analysis of these mushrooms. Furthermore, assays of α-glucosidase inhibitory and α-amylase inhibitory activity were used for evaluating antihyperglycemic activity of the mushrooms, and assays of reducing power, chelating effect on ferrous ions, scavenging effect on hydroxyl free radicals and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were used for evaluating antioxidant activity of the mushrooms. Based on the results, ethanolic and aqueous extract of these mushroom all showed antihyperglycemic and antioxidant potential. In particular, the aqueous extract of C. ventricosum revealed the highest α-glucosidase inhibitory activity (EC50 value 2.74μg/mL), DPPH radical scavenging activity (EC50 value 2.86mg/mL) and reducing power (EC50 value 0.96mg/mL), while the aqueous extract of L. amethystea showed the highest α-amylase inhibitory activity (EC50 value 4.37μg/mL) and metal chelating activity (EC50 value 2.13mg/mL).

Purification and characterization of a novel laccase from the edible mushroom Clitocybe maxima

A novel laccase from the edible mushroom Clitocybe maxima was purified and characterized. The purification protocol involved ammonium sulfate saturation, ion-exchange chromatography on DEAE-cellulose, SP-Sepharose, and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. It was a monomeric protein with a molecular mass of 62 kDa as estimated by SDS-PAGE. Its N-terminal amino acid sequence was DIGPVTPLAI, which exhibited partial sequence homology to those of published mushroom laccases. Its optimum pH was 3.0 and its optimum temperature was 60 °C. It manifested degrading activity towards a variety of phenolic compounds. The most sensitive substrate for the enzyme was 2,7′-azinobis[3-ethylbenzothiazolone-6-sulfonic acid]diammonium salt (ABTS), with a K m of 61.7 μM at pH 3.0 and 37 °C. The ranking of degradation activity toward phenolic substrates was ABTS > hydroquinone > N,N-dimethyl-1,4-phenylenediamine > pyrogallol > catechol > 2-methylcatechol. The laccase showed antiproliferative activity against Hep G2 and MCF-7 tumor cells with IC 50 of 12.3 μM and 3.0 μM, respectively. It also lowered the activity of human immunodeficiency virus-1 (HIV-1) reverse transcriptase with an IC 50 of 14.4 μM.

Flavour components and antioxidant properties of several cultivated mushrooms

Three mushrooms, Clitocybe maxima, Pleurotus ferulae and Pleurotus ostreatus grey strain were used to study their flavour components and antioxidant properties. The volatile flavour components found comprised of six eight-membered carbon compounds and two aromatic compounds. The content total of soluble sugars and polyols was 125–270 mg/g. The content of monosodium glutamate-like components was 1.76–8.89 mg/g. The contents of flavour 5′-nucleotides ranged from 1.89 to 7.59 mg/g. Based on the results obtained, three mushrooms possessed highly intense umami taste. Ethanolic extracts were more effective in the inhibition of conjugated diene and scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals, whereas hot water extracts were more effective in the scavenging ability of hydroxyl radicals. EC 50 values were less than 14 and 30 mg/ml for ethanolic and hot water extracts, respectively, indicating that the three mushrooms were relatively effective as they exhibited antioxidant properties, despite having scavenging abilities for hydroxyl radicals. Phenols were the major antioxidant components and the total contents were 5.10–11.1 mg gallic acid equivalents/g.

Isolation of a new ribonuclease from fruiting bodies of the silver plate mushroom Clitocybe maxima

A ribonuclease, with an N-terminal sequence exhibiting some homology to ribonuclease from Pleurotus ostreatus (Family Pleurotaceae), has been purified from fruiting bodies of the silver plate mushroom Clitocybe maxima (Family Tricholomataceae). However, there is little resemblance between the N-terminal sequences of ribonucleases from various Pleurotus species, and a lesser extent of resemblance between ribonucleases from C. maxima and Pleurotus tuber-regium. No structural relationship exists between ribonuclease from C. maxima, and those from Volvariella volvacea, Lentinus edodes and Irpex lacteus. The purification protocol involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-Sepharose, and fast protein liquid chromatography on Superdex 75. The ribonuclease was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-Sepharose. It exhibited a molecular mass of 17.5 kDa in both gel filtration and sodium dodecyl sulfate–polyacrylamide gel electrophoresis. It manifested roughly the same ribonucleolytic potency toward poly A and poly G followed by poly U. Its activity toward poly C was, by comparison, meager. The temperature and pH required for its optimal activity were, respectively, 70 °C and 6.5–7.0.