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A lectin from an ascomycete mushroom, Melastiza chateri: no synthesis of the lectin in mycelial isolate.

Using an affinity adsorbent prepared from L-fucose and starch, a lectin was isolated from fruit bodies of an ascomycete mushroom, Melastiza chateri. The lectin was found to cross-react with antiserum against Aleuria aurantia lectin (AAL), that had been obtained from another ascomycete mushroom. The N-terminal amino acid sequence was analyzed, and among 20 residues 12 were the same as AAL. The molecular mass of the lectin estimated by SDS-PAGE was approximately 40 kDa, which is larger than that of AAL. Mycelial isolate was obtained from M. chateri by germinating ascospores, and identified by analyzing restriction fragment length polymorphisms (RFLP) of DNA. The isolate from M. chateri did not synthesize the lectin, although the isolate from A. aurantia had been known to synthesize AAL as much as the fruit body.

Amino Acid Sequence↗

Mushroom tyrosinase inhibitory activity of esculetin isolated from seeds of Euphorbia lathyris L.

A tyrosinase inhibitor was isolated from the seeds of Euphorbia lathyris L. by bioassay-guided fractionation and purification, using silica gel column chromatography. It was identified as esculetin by comparing its physical properties and spectral data with those of an authentic sample. The IC50 value of esculetin in the mushroom tyrosinase activity test was 43 microM. The kinetic study indicates that esculetin exhibited competitive inhibition against the oxidation of 3-(3,4-dihydroxyphenyl)-alanine by mushroom tyrosinase. The structure-activity relationships among five esculetin analogs suggest that hydroxyl groups at the C6 and C7 positions of the coumarin skeleton played an important role in the expression of tyrosinase inhibitory activity.

Agaricales↗

Cloning and characterization of farnesyl diphosphate synthase from the rubber-producing mushroom Lactarius chrysorrheus.

Farnesyl diphosphate is involved in rubber biosynthesis as an initiating substrate for both polyprenol and mushroom rubber. So far, we have isolated the cDNA of a farnesyl diphosphate synthase (FPS) for the first time from a rare rubber-producing mushroom, Lactarius chrysorrheus, by the degenerate RT-PCR technique based on sequence information of FPS genes from fungi and yeasts. The open reading frame was clarified to encode a protein of 381 amino acid residues with a calculated molecular weight of 42.9 kDa. The deduced amino acid sequence of L. chrysorrheus FPS showed about 50% identity with those of other fungi and yeasts as well as plants. We expressed the cDNA of L. chrysorrheus FPS in Escherichia coli as a glutathione-S-transferase (GST)-fusion protein. The purified obtained protein showed FPS activity in which geranyl diphosphate (GPP) served as primary substrate, with a 2.4-fold higher k(cat)/K(m) value for GPP than for dimethylallyl diphosphate (DMAPP).

Agaricales↗

A phytochemical in the edible Tamogi-take mushroom (Pleurotus cornucopiae), D-mannitol, inhibits ACE activity and lowers the blood pressure of spontaneously hypertensive rats.

D-Mannitol, one of the main phytochemicals of the edible Tamogi-take mushroom (Pleurotus cornucopiae), was found to inhibit an angiotensin I converting enzyme (ACE). The antihypertensive effect of D-mannitol and a hot water extract of Tamogi-take mushroom was demonstrated in spontaneously hypertensive rats (SHR) by oral administration.

Angiotensin-Converting Enzyme Inhibitors↗

Medicinal mushrooms and cancer therapy: translating a traditional practice into Western medicine.

Modern medical practice relies heavily on the use of highly purified pharmaceutical compounds whose purity can be easily assessed and whose pharmaceutical activity and toxicity show clear structure-function relationships. In contrast, many herbal medicines contain mixtures of natural compounds that have not undergone detailed chemical analyses and whose mechanism of action is not known. Traditional folk medicine and ethno-pharmacology coupled to bioprospecting have been an important source of many anticancer agents as well as other medicines. With the current decline in the number of new molecular entities from the pharmaceutical industry, novel anticancer agents are being sought from traditional medicine. As the example of medicinal mushrooms demonstrates, however, translating traditional Eastern practices into acceptable evidence-based Western therapies is difficult. Different manufacturing standards, criteria of purity, and under-powered clinical trials make assessment of efficacy and toxicity by Western standards of clinical evidence difficult. Purified bioactive compounds derived from medicinal mushrooms are a potentially important new source of anticancer agents; their assimilation into Western drug discovery programs and clinical trials also provides a framework for the study and use of other traditional medicines.

Agaricales↗

Mushroom (Agaricus bisporus).

We have devised an easy and effective genetic transformation method for the preeminent edible mushroom, Agaricus bisporus. Our method exploits the T-DNA transfer mechanism in Agrobacterium tumefaciens and relies on the reproductive fruiting body as the recipient tissue. The use of fruiting body explants, particularly the gill, provided high-frequency transformation, overcoming the inefficacy of Agrobacterium-based methods targeting fungal spores or vegetative mycelium. The protocol entails incubation of A. tumefaciens for 3 h with acetosyringone, a signaling molecule that launches the gene transfer mechanism, co-cultivation of the induced bacterium and gill explants for 3 d, and selection for transformants based on an inherited resistance to the antibiotic hygromycin. Between 7 and 28 d on the selection medium, upwards of 95% of the gill explants generate hygromycin-resistant colonies. About 75% of the mushroom transformants show a single-copy of the hygromycin-resistant gene integrated at random sites in the genome.

Agaricus↗

Contents of metals in some wild mushrooms: its impact in human health.

The concentrations of 7 metals (lead, cadmium, manganese, copper, nickel, silver, and chromium) were determined in 32 different species of wild mushrooms. The mushroom samples, which have been using for food and some medical purposes, were collected from Konya, an Inner Anatolian region of Turkey. The highest metal concentrations were determined as 39 mg/kg Pb and 3.72 mg/kg Cd in Trichaptum abietinum, 467 mg/kg Mn in Panaeolus sphinctrinus, 326 mg/kg Cu in Trametes versicolor, 69.4 mg/kg Ni in Helvella spadicea, 6.97 mg/kg Ag in Agaricus campestris, and 84.5 mg/kg Cr in Phellinus igniarius. The maximum contents are 1.52, 2.22, and 60.2 mg/kg in Pleurotus eryngii (for Pb), Amanita vaginata (for Cd), and Helvella leucomelana (for Cu), respectively. These results were compared according to the WHO/FAO standard.

Agaricales↗

Preliminary study on the determination of selenium compounds in some selenium-accumulating mushrooms.

Using various chromatographic techniques (size exclusion, anion exchange, and cation exchange) combined with several detectors (neutron activation analysis and atomic fluorescence spectrometry), an attempt was made to characterize selenium compounds in some edible, selenium-accumulating mushrooms (Albatrellus pes-caprae and Boletus edulis). The mushrooms contained mostly low-molecular-weight (6 kDa) selenium compounds. After proteolysis, only a small fraction of the extractable selenium could be identified as selenite (3.0-9.2%, Albatrellus pes-caprae), selenocystine (minor, Albatrellus pes-caprae; 7.5%, Boletus edulis), or selenomethionine (1.0%, Boletus edulis), leaving the form of the bulk still to be elucidated.

Agaricales↗

Proteins as active compounds involved in insecticidal activity of mushroom fruitbodies.

Many mushrooms are toxic to insects. To identify the chemicals involved in insecticidal activity, the toxicity of 14 species has been studied for water solubility, thermolability, and dialysis. The data strongly suggest that proteins are responsible for most of the insecticidal activity of mushroom fruitbodies and may be a source of genes available for plant protection against insects. Among proteins, lectins and hemolysins were good insecticide candidates because the toxicities were not affected by protease.

Agaricales↗

Successful treatment of hypersensitivity pneumonitis caused by Grifola frondosa (Maitake) mushroom using a HFA-BDP extra-fine aerosol.

We successfully treated a patient with occupational hypersensitivity pneumonitis (HP) caused by Grifola frondosa (Maitake) mushroom spore with an extra-fine aerosol corticosteroid; beclomethasone dipropionate (BDP) dissolved in hydrofluoroalkane-134a (HFA). A 49-year-old woman developed respiratory symptoms 3 months after beginning work on a mushroom farm. She was diagnosed as HP based on radiological and serological findings. Oral prednisolone therapy improved her HP and she returned to the same farm. Her HP relapsed after 5 months, and daily 400 microg of HFA-BDP was administered with gradual improvement. An extra-fine particle inhaled corticosteroid might reach appropriate alveoli to be effective therapy for mild HP.

Administration, Inhalation↗

A sialic acid-specific lectin from the mushroom Paecilomyces Japonica that exhibits hemagglutination activity and cytotoxicity.

The mushroom Paecilomyces japonica, grown on the silkworm larvae, has been used in Asia as a nutraceutical, tea, and Chinese medicine. In the present study, a sialic acid-specific lectin has been purified from the mushroom P. japonica using affinity chromatography on a fetuin-agarose column. Electrophoretical analyses indicated that this lectin, designated P. japonica agglutinin (PJA), is an acidic protein with a molecular mass of 16 kDa, and has no intermolecular disulfide bonds. PJA induced hemagglutination activity in human ABO, mouse, rat, and rabbit erythrocytes. This activity was inhibited by sialic acid and sialoglycoproteins, but not by any other carbohydrates. PJA was stable at pH 4.0-8.0, and at temperatures below 55 degrees C. The activity of PJA was independent of EDTA and divalent cations. In addition, PJA exerts cytotoxic effects on the following cancer cell lines: human stomach cancer SNU-1, human pancreas cancer AsPc-1, and human breast cancer MDA-MB-231.

Animals↗

Fiber number in the mushroom bodies of adult Drosophila melanogaster depends on age, sex and experience.

The mushroom bodies are two characteristically shaped structures of the insect central brain. In Drosophila melanogaster they contain more fibers in females than in males. Within the first week of adult life the total number of fibers increases by about 15% and decreases again in flies older than 3-4 weeks. The number of mushroom body fibers is significantly reduced in flies kept under social isolation or deprived of their antennal input, but not in flies subjected to visual deprivation.

Aging↗

Drosophila mushroom body mutants are deficient in olfactory learning.

Two Drosophila mutants are described in which the connections between the input to and the output from the mushroom bodies is largely interrupted. In all forms of the flies (larva, imago, male, female) showing the structural defect, olfactory conditioning is impaired. Learning is completely abolished when electroshock is used as reinforcement and partially suppressed in reward learning with sucrose. No influence of the mushroom body defect on the perception of the conditioning stimuli or on spontaneous olfactory behavior is observed. The defect seems not to impair learning of color discrimination tasks or operant learning involving visual cues.

Animals↗

Generation of oxygen radicals from iron complex of orellanine, a mushroom nephrotoxin; preliminary ESR and spin-trapping studies.

Orellanine, [2,2'-bipyridine]-3,3',4,4'-tetrol-1,1'-dioxide, is the toxin responsible for the lethal nephrotoxicity of some Cortinarius mushrooms. Our present ESR and spin-trapping studies of the redox properties of the system of non-illuminated orellanine, ferrous iron and dioxygen contribute to understanding the molecular mechanism of its toxicity. UV-visible spectrophotometry, cyclic voltammetry and ESR in frozen medium showed the formation of a wine-red tris complex, Fe(III)Or3. This ferric complex is easily reducible (Ep = -565 mV vs Ag/AgCl/3M KCl at pH 7), involving a one-electron reversible process. Spin-trapping using DMPO is employed to detect the generation of superoxide anion and hydroxyl radicals. The instantaneous one-electron oxidation of ferrous ions in the presence of the toxin under air is concomitant with dioxygen consumption as supported by dioxygen consumption. GSH involves the toxin and ferrous ions under air in a redox cycling process resulting in the production of glutathionyl and oxygen free radicals, observed for the first time with an iron complex of a mushroom toxin. In most cases, EDTA is not able to prevent the Fe(III)Or3 and radical formation. The ortho-dihydroxylated groups borne by the di-N-oxidized bipyridine structure and not the bipyridine structure itself, are responsible for the formation of a stable ferric complex at pH 7, as they are for the generation of an apparently stable ortho-semiquinone anion radical. These one-electron mechanisms may play a major role in some of the known toxic effects of orellanine.

2,2'-Dipyridyl↗

Quantitative determination of the free amino acids and their derivatives in the common edible mushroom, Agaricus bisporus.

Nitrogen in the ethanol extract of the fruiting bodies of the common edible mushroom, Agaricus bisporus, accounted for about 60% of the total nitrogen of the fruiting bodies, and about 70% of the ethanol-extractable nitrogen was attributable to the free amino acids and their derivatives. Identification and quantitative determination of the free amino acids and their derivatives contained in this fraction were performed after group separation. Of the 44 ninhydrin-positive compounds detected, alanine, glutamic acid, proline and arginine were the most predominant protein amino acids occurring in the free form, and ornithine, N-(gamma-L-glutamyl)-4-hydroxyaniline and gamma-aminobutyric acid were the most plentiful non-protein amino acids. An analytical method of the aromatic compounds characteristic of the mushroom is also described.

Amino Acids↗

Dietary mushrooms reduce blood pressure in spontaneously hypertensive rats (SHR).

The blood pressure of spontaneously hypertensive rats (SHR) were significantly reduced by Maitake feeding for 8 weeks period beginning at a time when the animals were 10 weeks of age with well-established high blood pressure. There was no difference in the plasma total and free cholesterol, triglyceride and phospholipid levels between the Maitake fed animals and the control. On the other hand, Shiitake mushroom did not reduce the blood pressure, but significantly lower the plasma free cholesterol, triglyceride and phospholipid in compared with the control. The results suggest that dietary Maitake mushroom reduce the blood pressure.

Animals↗

Isolation of plasma cholesterol-lowering components from ningyotake (Polyporus confluens) mushroom.

The present study was undertaken to isolate component(s) which contributes to the hypocholesterolemic action of Ningyotake (Polyporus confluens) mushroom. The mushroom powder was extracted with 80% ethanol, and the extract and residue were fractionated into five fractions according to the solubility to solvents. When each fraction was added to a diet containing 1% cholesterol and 0.25% sodium cholate and fed to rats, the plasma cholesterol level was significantly decreased only by ethyl acetate-soluble fraction. Therefore, ethyl acetate-soluble fraction was further fractionated by silica gel column chromatography. Two major compounds, which comprised 45.0% and 28.5% of the ethyl acetate-soluble fraction, were obtained in a pure form by the chromatography, and the compounds were identified as grifolin (2-trans, trans-farnesyl-5-methylresorcinol) and neogrifolin (4-trans, trans-farnesyl-5-methylresorcinol), respectively. The addition of grifolin and neogrifolin to the high cholesterol diet was found to lower plasma cholesterol level significantly.

Animals↗