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Four new meroterpenoids, Clavilactone M-P, possessing novel aminoglycoside moiety (1–4) and a 10-membered carbocycle fused with an α,β-epoxy-γ-lactone, were isolated from Clitocybe clavipes, a basidiomycete. Their structures with absolute configurations were determined by extensive analysis of their spectroscopic data, and the ECD method. All the isolated compounds (1–4) were evaluated for their antitumor activity against three human cancer cell lines using the MTT assay. Compound 1 and 2 exhibited a significant suppression of cell viability in the Hela (IC50 = 22.8 and 19.7 μM) cell line.

The purpose of this study was to determine Fe, Cd, Cr, Se, P, Cu, Mn, Zn, Al, Ca, Mg, and K contents of some edible ( Chlorophyllum rhacodes , Clavariadelphus truncatus , Clitocybe nebularis , Hydnum repandum , Hygrophorus pudorinus , Infundibulicybe gibba , Lactarius deliciosus , L. piperatus , L. salmonicolor , Macrolepiota mastoidea , Russula grata , Suillus granulatus , and Tricholoma imbricatum ), inedible ( Amanita pantherina , Geastrum triplex , Gloeophyllum sepiarium , Hypholoma fasciculare , Phellinus vorax , Pholiota limonella , Russula anthracina , and Tapinella atrotomentosa ), and poisonous mushroom species ( Amanita pantherina and Hypholoma fasciculare ) collected from Ilgaz Mountain National Park (Western Black Sea, Turkey). The element contents of the mushrooms were determined to be 18.0–1239.1, 0.2–4.6, 0.1–3.4, 0.2–3.2, 1.0–8.9, 3.3–59.9, 3.7–220.4, 21.3–154.1, 6.4–754.3, 15.8–17,473.0, 413.0–5943.0, and 2803.0–24,490.0 mg·kg -1 , respectively. In addition to metal contents, the daily intakes of metal (DIM) and Health Risk Index (HRI) values of edible mushrooms were also calculated. Both DIM and HRI values of mushroom species except L. salmanicolor , M. mastoidea , and R. grata were within the legal limits. However, it was determined that the Fe content of L. salmanicolor and M. mastoidea and Cd content of R. grata were above the legal limits.

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.

Changes in the functional properties and structures of Clitocybe squamulosa protein isolate (CSPI) in the process of freeze–thaw (F–T) cycles were explored. Remarkable alterations and the reduced content of protein ordered structure were revealed through structural analysis of CSPI after F–T treatments. The surface hydrophobicity and free sulfhydryl content of CSPI first increased and then decreased. However, after the F–T treatments, the carbonyl content of CSPI continued to increase. Similarly, the water holding capacity (WHC), oil holding capacity (OHC), and solubility of CSPI all declined as the number of F–T cycles increased. The foaming properties and emulsifying properties of CSPI were significantly improved and reached maximum values after three F–T cycles. CSPI undergoing two F–T cycles showed the highest digestibility, maximum polypeptide content, and highest DPPH and ·OH-radical-scavenging activities. The ·OH-radical-scavenging activities and reducing power of the gastrointestinally digested CSPI had the highest value after one F–T cycle. Therefore, it has been demonstrated that F–T treatments could be a residue-free and cost-effective tool for improving mushroom protein functional properties.

The research was carried out between 2021 and 2023 in zones A‒D, 400, 600, 1000, and 1500 m away from the magnesite plant in Lubeník (central Slovakia), which are polluted by magnesite immissions. Due to the fall of magnesite dust, the Organic horizons became neutral to weakly acidic, the mineral layers of soils in zones A and B strongly to weakly alkaline, and neutral in zones C and D. Ca, Mg, and Na in the Organic horizon and 0–5 cm soil layers decreased from zone A toward zone D. Mn and K increased from zone A to zone D. Cd generally increased from zone A to zone C. A total of 107 species of macromycetes from the phyla Ascomycota (15) and Basidiomycota (92) were detected. The absolute best accumulator of Ca and Mg was Hymenochaete rubiginosa and Ca, Mg, and Mn Geastrum rufescens . The best K accumulator was Inosperma erubescens together with Russula cyanoxantha , and the absolute best Na accumulator was Clitocybe phyllophila . Cd was best accumulated by Hymenochaete rubiginosa and Inosperma erubescens . PCA showed that soil properties depended on pH values and C, N, K, and Na content. The mentioned variables were significantly correlated with PC1 (share 46%), while the contents of Ca, Mg, Mn, and Cd were correlated with PC2 (share 35%). Ca and Cd were the most important environmental variables accumulated in macromycete fruiting bodies. PC1 represents a significant axis for the prediction of Ca content (40.2%) and PC2 for the prediction of Cd content in macromycetes (19%). Environmental pollution by Mg emissions significantly contributed to soil degradation and the composition of macromycete communities. The knowledge gained can help in the implementation of corrective measures in areas affected by alkaline immissions.

Basidiomycete mushrooms are a rich source of unique substances, including lectins, that could potentially be useful in biotechnology or biomedical applications. Lectins are a group of carbohydrate-binding proteins with diverse biological activities and functions. Here, we demonstrate the presence of a number of lectins in the basidiomycete mushroom Clitocybe nebularis. Glucose-, galactose-, sucrose-, lactose-, and Sepharose-binding lectins were isolated from fruiting bodies using affinity chromatography on Sepharose-immobilized sugars or on Sepharose. The lectins were characterized biochemically and their binding specificities examined by agglutination and agglutination inhibition assays. In addition, insecticidal and anti-nutritional properties of the lectins were studied against a model organism, fruit fly (Drosophila melanogaster), and Colorado potato beetle (Leptinotarsa decemlineata). Of the several basidiomycete mushrooms screened, C. nebularis extract showed the most potent insecticidal activity. Sucrose-binding lectin showed the strongest activity against D. melanogaster, followed by lactose- and galactose-binding lectins. Feeding bioassays with Colorado potato beetle revealed that C. nebularis extract exhibited high anti-nutritional activity against the insect; and of those tested, only lactose-binding lectin, named CNL showed the effect. Mushroom C. nebularis is shown to be rich in a variety of lectins with versatile biological activities, including insecticidal and anti-nutritional effects. C. nebularis lectins could thus have potential for use as natural insecticides.[PUBLICATION ABSTRACT]

Wild edible mushrooms are very popular for both their flavors and nutritional values. However, some mushroom species can be harmful to human health as they accumulate some elements in excessive amounts. The aim of this study was to determine the concentrations of Al, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, P, Se, and Zn of some wild edible ( Agaricus arvensis , A. bitorquis , A. sylvaticus , Amanita vaginata , Armillaria mellea , Clavariadelphus pistillaris , Clitocybe nebularis , Clitopilus prunulus , Hygrophorus marzuolus , H. russula , Lactarius volemus , Lycoperdon molle , and Macrolepiota mastoidea ) and non-edible mushroom species ( A. citrina , Auricularia mesenterica , Chanterellus melanoxeros , Chondrostereum purpureum , Clathrus ruber , L. controversus , L. helvus , and L. zonarius ) collected from Belgrad forest (Istanbul, Turkey). Daily intakes of element (DIE) and health risk index (HRI) values of the edible mushroom species were also calculated. The concentrations of the elements in question were determined to be in the ranges of 9.7–556.8, 2.5–2226.7, 0.06–2.52, 0.03–13.17, 3.74–100.19, 13.3–507.4, 2635.0–28614.0, 493.0–2412.0, 6.97–3150.73, 0.29–13.26, 0.38–3.67, and 9.1–293.8 mg/kg, respectively. The Cd concentration of H. russula (DIE: 1.08, HRI: 1.08), Cr concentration of C. nebularis (DIE: 5.64, HRI: 1.88), and the Cu concentration of M. mastoidea (DIE: 42.94, HRI: 1.07) were above the reference values. The results showed that the long-term consumption of H. russula , C. nebularis , and M. mastoidea collected from Belgrad forest can have a negative impact on human health. Therefore, it was concluded that the element concentrations of edible wild mushrooms in this region should be examined periodically.

We studied neuroprotective, antioxidative, antimicrobial and cytotoxic activity of acetone extracts of Clitocybe geotropa and Clitocybe nebularis . The ability of extracts to inhibit acetylcholinesterase was tested in order to examine neuroprotective potential of selected mushroom species. DPPH, superoxide anion and reducing power assays were used in assessing the antioxidative potential of studied mushrooms. Total phenolics content was estimated as pyrocatechol equivalent. Antimicrobial activity was evaluated through microdilution method. MTT assay was utilized in determining the cytotoxic activity of extracts. The results demonstrated that studied extracts express relatively strong neuroprotective, antioxidative, antimicrobial and moderate cytotoxic activity. The results of acetylcholinesterase inhibition for Clitocybe geotropa were in range 46.95–14.82% and for Clitocybe nebularis 36.73–12.49%. DPPH assay’s IC50 for Clitocybe geotropa were 246.58 μg/mL and for Clitocybe nebularis 46.49 μg/mL. IC50 in superoxide anion assay was 151.36 μg/mL for Clitocybe geotropa and 35.69 μg/mL for Clitocybe nebularis . In reducing power assay the absorbance for Clitocybe geotropa was in range 0.0708–0.0132 and for Clitocybe nebularis 0.0324–0.0141. Total phenolics content of Clitocybe geotropa was 95.71 µg PE/mg and of Clitocybe nebularis 93.94 µg PE/mg. MIC values for Clitocybe geotropa were in range 0.78–25 mg/mL and for Clitocybe nebularis 3.12–25 mg/mL. Cytotoxic activity of extracts was moderate, IC50 values for Clitocybe geotropa ranged from 124.2 to 155.3 μg/mL and for Clitocybe nebularis from 134.03 to 199.03 μg/mL. Both mushroom species are edible and can be referred to as functional food.

Five new meroterpenoids, clavipols A–B (1–2) with a 12-membered ether ring and clavilactones G–I (3–5) having a 10-membered carbocycle connected to a hydroquinone and an α,β-epoxy/unsaturated lactone, were obtained from the fruiting bodies of the basidiomycete Clitocybe clavipes. Their structures were determined by comprehensive analysis of their spectroscopic data, and the absolute configuration of 1 was established by quantum chemical calculations of electronic circular dichroism (ECD). All the isolated compounds (1–5) were tested for their cytotoxic activity against three human tumor cell lines (Hela, SGC-7901, and SHG-44) in vitro after treatment for 48 h. Compound 4 exhibited moderate cytotoxic activity against Hela and SGC-7901 tumor cell lines, with IC50 values of 23.5 and 14.5 µM, respectively.

Lectins have been increasingly utilized as carriers for targeted drug delivery based on their specific binding to glycans located on mammalian cells. This study employed two lectins, B subunit of bacterial Shiga holotoxin (Stx1B) and fungal Clitocybe nebularis lectin (CNL), for surface display on the lactic acid bacterium Lactococcus lactis. The specific adhesion of these engineered, lectin-displaying L. lactis to cancer cells was evaluated. The expression and surface display of both lectins on L. lactis were demonstrated by western blotting and flow cytometry, respectively. MTS assays revealed that recombinant Stx1B had no effect on Caco-2 cell viability at concentrations of ≤25 µg/mL, whereas CNL was non-toxic even at relatively high concentrations of ≤250 µg/mL. Stx1B bound to Caco-2, HT-29 and HeLa cells after 1 h of incubation. CNL bound to Caco-2 cells and recognized several glycoproteins in HT-29 and Caco-2 cell homogenates of which a 70 kDa protein predominated. Confocal microscopy revealed adhesion of Stx1B-displaying L. lactis to HeLa, Caco-2, and, to a lesser extent, HT-29 cells; CNL-displaying L. lactis showed a relatively similar level of adherence to HT-29 and Caco-2 cells. Thus, lectin-displaying L. lactis might serve as a carrier in targeted drug delivery when coupled to a therapeutic moiety.