Discovery of novel secondary metabolites from the basidiomycete Lentinus cf. sajor-caju and their inhibitory effects on Staphylococcus aureus biofilms.

Three novel derivatives of microporenic acid, microporenic acids H-J, were identified from submerged cultures of a Lentinus species obtained from a basidiome collected during a field trip in the tropical rainforest in Western Kenya. Their structures were elucidated via HR-ESIMS spectra and 1D/2D NMR spectroscopic analyses, as well as by comparison with known derivatives. Applying biofilm assays based on crystal violet staining and confocal microscopy, two of these compounds, microporenic acids H and I, demonstrated the ability to inhibit biofilm formation of the opportunistic pathogen Staphylococcus aureus. Thereby, they were effective in a concentration range that did not affect planktonic growth. Additionally, microporenic acid I enhanced the anti-biofilm activity of the antibiotics vancomycin and gentamicin when used in combination. This opens up possibilities for the use of these compounds in combination therapy to prevent the formation of S. aureus biofilms.

Lentinuses A-B, two alkaloids from the marine-derived fungus Lentinus sajor-caju with potent anti-pulmonary fibrosis activity.

By adding natural amino acids into the medium as sole nitrogen source, twenty-four compounds, including two new alkaloids lentinuses A-B (1-2) with a rare oxazinone core in marine natural products, one new natural product 3-acetamido-4-phenylfurazan (3), 9ß-ergosterol (22) were firstly discovered from a marine fungus, and twenty known compounds (4-21, 23-24) were isolated from the marine-derived fungus Lentinus sajor-caju. The chemical structures of all these compounds were elucidated by HRMS, NMR spectroscopy and X-ray diffraction. Compounds 1-24 were evaluated for their inhibitory activity against TGF-ß1-induced collagen accumulation in human fetal lung fibroblasts (HFL1). Compounds 2, 3, 12, 22, and 23 showed potent activity against TGF-ß1-induced collagen accumulation and low toxicity to HFL1 cells. The binding mode of lentinus B (2) with TGF-ß1 receptor was then performed by using Schrödinger software, and the result showed that lentinus B possesses a strong binding force such as hydrogen bonding and hydrophobic interactions to the protein, which may provide a theoretical basis to design more potent anti-fibrotic drugs in the future.

A Study of the Therapeutic Potential of Lentinus squarrosulus (Agaricomycetes) from Gabon.

Lentinus squarrosulus is a mushroom widely used in Gabon for its culinary and medicinal properties. The bioactive potential of mushrooms might be attributable to the presence of several pharmaceutically important mycocompounds that need to be ascertained scientifically. A study of the therapeutic potential of L. squarrosulus, the species of lignicolous fungus exploited in Gabon, was carried out on the basis of a chemical screening performed on three extracts in order to highlight different important chemical groups. This chemical screening was followed by a study of the fungus's antioxidant activity and prediction of its additional pharmacological activities. Chemical screening revealed that three extracts (aqueous, hydroethanolic, and ethanolic) of L. squarrosulus were almost free of tannins and were poor in total flavonoids and moderately rich in reducing sugars. Aqueous and ethanolic extracts were rich in total polyphenols, whereas aqueous and hydroethanolic extracts were rich in alkaloids. The aqueous extract was rich in saponosides and the hydroethanolic extract was rich in coumarins. The dosage of phenolic compounds confirmed the fungus's richness in total polyphenols, especially for aqueous and ethanolic extracts, its poverty in flavonoids and absence of tannins in ethanolic and hydroethanolic extracts. Regarding antioxidant activities, the results obtained for diphenyl picryl hydrazyl trapping tests showed that the different extracts tested had antioxidant activity ranging from low to moderate (0.12 ≤ antioxidant activity index [IAA] ≤ 0.6); the greatest activity was obtained with ethanolic extract (IAA = 0.6). Hence, we conclude that L. squarrosulus extracts can be used as easily accessible sources of natural antioxidants for potential preventative therapies.

Chemosensitizing activity of peptide from Lentinus squarrosulus (Mont.) on cisplatin-induced apoptosis in human lung cancer cells.

The limitations of cisplatin, a standard chemotherapy for lung cancer, have been documented with serious adverse effects and drug resistance. To address the need for novel therapy, this study firstly reveals the potential of peptide from Lentinus squarrosulus (Mont.) as a chemotherapeutic adjuvant for cisplatin treatment. The purified peptide from L. squarrosulus aqueous extracts was obtained after eluting with 0.4 M NaCl through FPLC equipped with anion exchange column. Preincubation for 24 h with 5 µg/mL of the peptide at prior to treatment with 5 µM cisplatin significantly diminished %cell viability in various human lung cancer cells but not in human dermal papilla and proximal renal cells. Flow cytometry indicated the augmentation of cisplatin-induced apoptosis in lung cancer cells pretreated with peptide from L. squarrosulus. Preculture with the peptide dramatically inhibited colony formation in lung cancer cells derived after cisplatin treatment. Strong suppression on integrin-mediated survival was evidenced with the diminution of integrins (ß1, ß3, ß5, α5, αV) and down-stream signals (p-FAK/FAK, p-Src/Src, p-Akt/Akt) consequence with alteration of p53, Bax, Blc-2 and Mcl-1 in cisplatin-treated lung cancer cells preincubated with peptide from L. squarrosulus. These results support the development of L. squarrosulus peptide as a novel combined chemotherapy with cisplatin for lung cancer treatment.

Molecular characterization, gas chromatography mass spectrometry analysis, phytochemical screening and insecticidal activities of ethanol extract of Lentinus squarrosulus against Aedes aegypti (Linnaeus).

Mosquito-transmitted diseases like zika, dengue, chikungunya, and yellow fever are known to affect human health worldwide. Numerous synthetic insecticides have been used as vector control for these diseases, but there is the challenge of environmental toxicity and vector resistance. This study investigated the medicinal and insecticidal potential of Lentinus squarrosulus against Aedes aegypti. The fruiting bodies were identified morphologically as well as using internal transcribed spacer (ITS) sequences for its molecular characterization. Genomic deoxyribonucleic acid (DNA) yield was confirmed with NanoDrop Spectrophotometer ND-1000 and amplified with ITSl and ITS4 primers. The amplicons were sequenced and the National Center for Biotechnology Information (NCBI) database identified the nucleotides. Its ethanol extract was subjected to phytochemical screening and gas chromatography mass spectrometry (GC-MS) analysis and tested against the pupa and fourth instar larva of Aedes aegypti with percentage mortality monitored. The Macrofungus was identified morphologically and confirmed with molecular characterization as Lentinus squarrosulus (LS). The gene sequence was deposited in GenBank (Accession number MK629662.1). GC-MS analysis showed that its ethanol extract has 25 bioactive compounds with 9,12-Octadecadienoic acid, ethyl ester having the highest percentage of 43.32% as well as methyl-2-oxo-1-pyrrolidine acetate and 17-octadecynoic acid having the lowest percentage (0.09%). The macrofungus contained varied concentrations of phytochemicals including phenols (159 mg/g GAE), tannins (1.6 mg/g TAE), and flavonoids (31.4 mg/g QE). The ethanol extract had significant potent effects on Aedes aegypti larva and pupa which could be due to the occurrence and abundance of 9,12-octadecadienoic acid in LS. The LC50 of the extract for larvicidal and pupicidal activities were 2.95 mg/mL and 3.55 mg/mL, respectively, while its LC90 were 6.31 mg/mL and 5.75 mg/mL respectively. Lentinus squarrosulus had insecticidal effects against the Aedes aegypti larva and pupa and possessed great potential as a source of alternative medicine and eco-friendly insecticides.

Characterization of the Nutritional Composition of a Biotechnologically Produced Oyster Mushroom and its Physiological Effects in Obese Zucker Rats.

SCOPE: Sustainable protein sources are needed to meet the increasing protein demands of a continuously growing world population. This study is focused on the biotechnological production of a protein rich oyster mushroom (Pleurotus sajor-caju; PSC) by valorization of an agricultural side stream and the evaluation of the physiological effects of PSC in a rat model of metabolic syndrome. METHODS AND RESULTS: PSC is produced via submerged cultivation in a 150 L bioreactor that utilizes isomaltulose molasses as its sole carbon source, and is further analyzed for its nutritional composition. A feeding trial is performed using Zucker rats which are fed a 5% PSC supplemented diet, for 4 weeks. Biochemical analyses reveal a significant reduction of the liver lipid concentrations and liver inflammation in the PSC fed obese rats in comparison to the obese rats from the control group. Hepatic qPCR analyses, differential transcript profiling, and enzyme activity measurements reveal a number of altered pathways that may be responsible for these anti-steatotic and anti-inflammatory effects of the mushroom. CONCLUSION: Bioconversion of a low quality agricultural side stream to an improved protein source is performed by submerged cultured PSC, and the obtained mycelium shows strong anti-steatotic and anti-inflammatory effects.

Rapid quality control of medicine and food dual purpose plant polysaccharides by matrix assisted laser desorption/ionization mass spectrometry.

With their multiple biological activities and health benefit effects, polysaccharides from medicine and food dual purpose plants (MFDPPPs) have been extensively applied in many fields, including in medical treatments, stock farming, and cosmetics. However, to date, quality issues of MFDPPPs and technologies for the analysis of polysaccharides have posed challenges to chemists. Reported herein is a rapid and high-throughput quality control method for analyzing MFDPPPs, based on matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). For the analysis of illegally added and doped substances, ferroferric oxide nanoparticles were employed as the MALDI matrix to avoid small molecule interference. Qualitatively, high sensitivity was obtained for both illegal drugs and glucose. Quantitatively, the best linear response (R2 > 0.99) was attained in the concentration range from 0.005 to 1 mg mL-1 for glucose. For the analysis of polysaccharides, 2,5-dihydroxybenzoic acid/N-methylaniline was employed as the MALDI matrix to increase the detection sensitivity and mass range coverage. Furthermore, the established method was successfully applied to the analysis of supplements from Astragalus polysaccharides and Lentinan real samples, showing its potential in quality control for MFDPPPs.

Toxicity and Nutritional Assessment of Extracts of Medicinal Tiger Sawgill Mushroom, Lentinus tigrinus (Agaricomycetes), a Newly Domesticated in Iran.

Information on the biosafety of tiger sawgill mushroom, Lentinus tigrinus, is limited and controversial. In the present study, the toxicity of a native L. tigrinus strain was evaluated in both cell cultures and mice. In addition to proximate analysis, the amino acid composition and the substrate-dependent uptake of elements were also evaluated. The mushroom (dry weight) had 7.53 ± 0.11% ash, 4.23 ± 0.23% lipid, 13.4 ± 0.04% fiber, 74.84 ± 0.38% total carbohydrate, and 4.16 ± 0.08 (mg GAEs/g) total phenol. Lysine was found at the highest amount among the L-amino acids determined in the L. tigrinus soluble protein. Comparison of the elemental profile of L. tigrinus with that of the substrate demonstrated a great capacity for absorption of K, P, S, and Mg elements, while the Na uptake was low. Neither the substrate nor the mushroom contained toxic elements beyond the standards of the joint FAO/WHO. The viability of normal human and mouse-derived cells was not influenced by the extract up to 250 µg/mL, where 70% of cancerous PC3 and MCF-7 cells were killed. Selectivity index analysis suggested that the L. tigrinus extract was selective only against MCF-7 cells. The extract also did not affect mice treated orally or via i.p. injection, while i.v. injection caused some mortality in mice with an LD50 of 47.19 mg/mL. In conclusion, L. tigrinus may be considered a source of macronutrients and micronutrients with a selective anticancer activity, while it is much less likely to have detrimental effects on humans at low serving levels.

A Review on Phytochemistry and Pharmacology of Medicinal as well as Poisonous Mushrooms.

Mushrooms have been used as traditional medicine from last few decades. Mushrooms as higher Basidiomycetes contain secondary metabolites in fruit bodies, cultured mycelium, and cultured broth. Medicinal mushrooms possess medicinal properties such as anti-tumor, immunomodulating, antioxidant, cardiovascular, anti-hypercholesterolemic, anti-viral, anti-bacterial, anti-parasitic, antifungal, detoxification, hepatoprotective, and anti-diabetic effects. Phase-I, II, and III clinical trials were studied on various biologically active compounds isolated from medicinal mushrooms and are used adequately to treat various diseases including cancer. The present review focuses on various edible, medicinal and poisonous species of mushrooms belong to genera; Auricularia, Cantherallus, Ganoderma, Pleurotus, Lentinus, Trametes (Coriolus), Tremella and Amanita along with their chemical composition, biologically active compounds isolated and their pharmacological potential.

Lentinoid and Polyporoid Fungi, Two Generic Conglomerates Containing Important Medicinal Mushrooms in Molecular Perspective.

Polyporoid and lentinoid fungi contain the important producers of substances having immunomodulatory, antitumoral, antiviral, and antihyperlipidemic effects. The discovery of several phylogenetic lines within the lentinoid-polyporoid continuum will help with target metabolomic analysis of species still not studied in pharmacological respects. The purpose of the present work was to increase a resolution in the lentinoid-polyporoid phylogenetic zone by means of selection of both the main representatives of Lentinus-related genera and poorly known/intermediate taxa such as Lentinus suavissimus, Neofavolus spp., and the resupinate part of Polyporus (genera Perenniporia and Pachykytospora) in the context of the basic structure of the Polyporales tree. The molecular phylogeny of highlighting all the polyporoid and lentinoid nodes was reconstructed using nLSU ITS rDNA and TEF datasets. The data obtained from ITS, TEF, and LSU coincide in support of core Polyporaceae of 10 clades corresponded to the generic level and 7 of these (Cerioporus, Cladomeris, Favolus, Lentinus, Neofavolus, Picipes, and Polyporus s.str.) contain generic units characterized by polyporoid or lentinoid morphotypes. The other 2 clades containing lentinoid taxa are outside the core Polyporaceae, namely Panus (Meruliaceae, Polyporales) and Neolentinus (Gloeophyllaceae, Gloeophyllales). A new genus, Picipes, is described and 25 new combinations are proposed.

Prenylhydroquinone-Derived Secondary Metabolites from Cultures of the Basidiomycete Lentinus similis BCC 52578.

Two new prenylhydroquinone-derived compounds, Ientinospirol (1) and 1-(2,5-dihydroxyphenyl)-4-hydroxy-3-methyl-l-butanone (2), were isolated from cultures of the basidiomycete Lentinus similis BCC 52578, together with the known compounds panepoxydone (3), panepoxydione (4), isopanepoxydone (5), 2,2-dimethyl-6-hydroxy-2H-chromene (6), and (3R,4S)-3,4-dihydroxy-2,2-dimethyl-6-methoxychroman (7). Compounds 3 and 4 exhibited cytotoxicity against all cell-lines tested, while the other compounds were inactive.

Nutritional Composition of Three Domesticated Culinary-Medicinal Mushrooms: Oudemansiella sudmusida, Lentinus squarrosulus, and Tremella aurantialba.

The nutritional composition of three recently domesticated culinary-medicinal mushroom species (Oudemansiella sudmusida, Lentinus squarrosulus, and Tremella aurantialba) was evaluated for contents of protein, fiber, fat, total sugar content, amino acid, carbohydrate, and nucleotide components. The data indicated that fruiting bodies of these three mushroom species contained abundant nutritional substances. The protein contents of L. squarrosulus and O. submucida were 26.32% and 14.70%, which could be comparable to other commercially cultivated species. T. aurantialba contained 74.11% of carbohydrate, of which soluble polysaccharide was 40.55%. Oudemansiella sudmusida contained 15.95% of arabitol as the highest sugar alcohol in three mushrooms. These mushrooms also possessed distinct taste by their flavor component composition. Among them, L. squarrosulus contained 10.68% and 9.25% of monosodium glutamate-like and sweet amino acids, which were higher than the other two mushrooms. However, the nucleotide amounts of the three mushrooms were all lower than those of other commercially cultivated mushrooms. Among them, L. squarrosulus contained the highest amount of flavor nucleotides, which was 1.01‰. Results revealed that these three mushroom species are potentially suitable resources for commercial cultivation and healthy food.

Taxonomy and compositional analysis of two new for science medicinal mushroom taxa from India.

Taxonomy and compositional evaluation of two new medicinal and edible varieties, Lentinus squarrosulus var. mangiferae and Pleurotus pulmonarius var. indicus, have been detailed in the present paper. Two new varieties were taxonomically identified by morphological, microscopical and ITS-region sequencing. Composition of proteins, carbohydrates, fat, ash content, minerals, fibers, ascorbic acid, phenolics, ß-carotene, and lycopene and alkaloids was analyzed for these varieties.

HS/GC-MS analyzed chemical composition of the aroma of fruiting bodies of two species of genus Lentinus (Higher Basidiomycetes).

The chemical composition of the aroma of fresh fruiting bodies of the cultivated mushroom Lentinus boryanus is described here and compared with medicinal shiitake mushroom L. edodes. Volatile compounds were analyzed through headspace sampling coupled with gas chromatography-mass spectrometry. The mushrooms under study were grown on different substrates based on barley straw, sugarcane bagasse, oak wood sawdust, and beech leaf litter. It was determined that L. boryanus as well as L. edodes contain an abundant amount of a volatile compound identified as 3-octanone with a sweet fruity aroma. On the other hand, only L. boryanus produced 3-octanol a characteristic aroma of cod liver oil. In total, 10 aromatic compounds were identified, some of which were obtained exclusively in one species or substrate.

Proximate composition and functionality of the culinary-medicinal tiger sawgill mushroom, Lentinus tigrinus (higher Basidiomycetes), from the Philippines.

The proximate composition and functionality of Lentinus tigrinus were evaluated to establish and popularize this mushroom as functional food source. The evaluation of functionality focused on the antibacterial and hypoglycemic activities of the mushroom extracts. An acute single oral dose toxicity test in mice was used for its biosafety analysis. The pileus contained higher amounts of protein (25.9%), fat (2.1%), and ash (7.4%) and a higher energetic value (142.1 kcal/100 g) than the corresponding stipe, whereas the stipe contained higher amounts of total carbohydrates (67.7%), which consist of dietary fiber (63.0%) and reducing sugar (4.7%), than the pileus. Biosafety analysis confirmed that L. tigrinus is an edible mushroom species; it was found to be toxicologically safe in imprinting control region mice. The administration of lyophilized hot water extract of the fruiting body (both 100 and 250 mg/ kg doses) to diabetic mice significantly lowered the glucose level by 26.9% in the third week, which was significantly comparable to the results of the antidiabetic agent glibenclamide, which was used as a positive control. In vitro antibacterial assay showed that the ethanolic extract of the fruiting body and the immobilized secondary mycelia had high antibacterial activities against Staphylococcus aureus but not on Escherichia coli. Combining its useful nutrients and significant biological properties, L. tigrinus can be considered a natural source of safe nutraceuticals.

Cytotoxic and immunomodulatory effects of polyhydroxyoctane isolated from Lentinus polychrous mycelia.

A polyhydroxyoctane, 6-methylheptane-1,2,3,4,5-pentaol (MHP), was first isolated from mycelia of the Thai edible mushroom Lentinus polychrous. MHP was evaluated for its cytotoxic and immunomodulatory effects in vitro. MHP was slightly cytotoxic to murine splenocytes but not to RAW264.7 cells or peripheral blood mononuclear cells. MHP decreased nitric oxide and intracellular O2 (-) production from lipopolysaccharide- and phorbol-12-myristate-13-acetate-activated RAW264.7 cells at levels of 78.98 ± 4.72 and 78.48 ± 2.41 % of controls, respectively. The mRNA expression of pro-inflammatory mediators, including iNOS, TNF-α, IL-1ß, IL-6, COX-1 and COX-2, were significantly suppressed by MHP. In addition, MHP significantly increased the proliferation of phytohemagglutinin and pokeweed mitogen-induced splenocytes. These results indicate that MHP is able to modulate inflammatory responses and the proliferation of both T- and B-lymphocyte cells, suggesting that MHP may be a good natural immunomodulator.

Mycelial biomass and biochemical properties of proteases produced by Lentinus citrinus DPUA 1535 (Higher Basidiomycetes) in submerged cultivation.

The cultivation of Lentinus citrinus for mycelial biomass and protease production under different carbon and nitrogen sources was studied in submerged cultivation. The nutritional source concentration for protease production was evaluated using a full factorial design. For mycelial biomass maltose (4.94 mg/mL) and beef extract (5.45 mg/mL), carbon and nitrogen sources presented the best results, respectively. The maximum protease activity was 73.33 U/mL with fructose (30.0 g/L) and beef extract (10.0 g/L). Proteases showed maximum activity at 40°C and pH 7.0, which exhibited high stability at experimental conditions. The final part of this work was devoted to estimating the main thermodynamic parameters of the irreversible enzyme inactivation (ΔH* = 17.86 kJ/mol, ΔG* =102.09 kJ/mol, ΔS* = -260.76 J/mol×K) through residual activity tests carried out at 25-70°C, by making use of Arrhenius and Eyring plots.

Secondary metabolite production by the basidiomycete, Lentinus strigellus, under different culture conditions.

The basidiomycete Lentinus strigellus was cultivated in three different culture media and the secondary metabolites produced under different culture conditions were isolated and identified. When cultivated in a liquid medium with peptone, L. strigellus afforded the benzopyrans, 2,2-dimethyl-6-methoxychroman-4-one, 4-hydroxy-2,2-dimethyl-6-methoxychromane and (3R,4S)-3,4-dihydroxy-2,2-dimethyl-6-methoxychromane. The indole alkaloid echinuline and the anthraquinone fiscione, both unprecedented for the genus Lentinus, were isolated from the mycelium of the fungus. When cultured in Czapek medium enriched with potato broth, the fungus afforded the same benzopyrans except (3S,4S)-3,4-dihydroxy-2,2-dimethyl-6-methoxychromane. Panepoxydone and isopanepoxydone were also isolated when the microorganism was grown in Czapek medium.

Antioxidant and antityrosinase activities of various extracts from the fruiting bodies of Lentinus lepideus.

Lentinus lepideus is an edible mushroom currently available in Korea. The acetone, methanol and hot water extracts were prepared and assayed for their antioxidant and antityrosinase inhibitory activities. The hot water extract showed the strongest ß-carotene-linoleic acid inhibition compared to the other extracts. At 8 mg/mL, the methanolic extract showed a high reducing power of 1.21. The acetone and methanol extracts were more effective in scavenging DPPH radicals than the hot water extract. The strongest chelating effect was obtained from the methanolic extract. Xanthine oxidase and tyrosinase inhibitory activities of the acetonic, methanol and hot water extracts increased with increasing concentration. Gallic acid, chlorogenic acid, vanillin, naringin, naringenin, formononetin, and biochanin-A were detected in the acetonitrile and hydrochloric acid (5:1) solvent extract. This study suggests that fruiting bodies of L. lepideus can potentially be used as a readily accessible source of natural antioxidants.

In vitro activity of hypnophilin from Lentinus strigosus: a potential prototype for Chagas disease and leishmaniasis chemotherapy.

Hypnophilin and panepoxydone, terpenoids isolated from Lentinus strigosus, have significant inhibitory activity on Trypanosoma cruzi trypanothione reductase (TR). Although they have similar TR inhibitory activity at 10 µg/mL (40.3 µM and 47.6 µM for hypnophilin and panepoxydone, respectively; ~100%), hypnophilin has a slightly greater inhibitory activity (~71%) on T. cruzi amastigote (AMA) growth in vitro as well as on in vitro phytohemagglutinin (PHA)-induced peripheral blood mononuclear (PBMC) proliferation (~70%) compared to panepoxydone (69% AMA inhibition and 91% PBMC inhibition). Hypnophilin and panepoxydone at 1.25 µg/mL had 67% inhibitory activity onLeishmania (Leishmania) amazonensis amastigote-like (AMA-like) growth in vitro. The panepoxydone activity was accompanied by a significant inhibitory effect on PHA-induced PBMC proliferation, suggesting a cytotoxic action. Moreover, incubation of human PBMC with panepoxydone reduced the percentage of CD16(+) and CD14(+) cells and down-regulated CD19(+), CD4(+) and CD8(+) cells, while hypnophilin did not alter any of the phenotypes analyzed. These data indicate that hypnophilin may be considered to be a prototype for the design of drugs for the chemotherapy of diseases caused by Trypanosomatidae.