Bodies Of Mushrooms Research Articles

A new broad-spectrum protease inhibitor from the entomopathogenic bacterium Photorhabdus luminescens.

A new protease inhibitor was purified to apparent homogeneity from a culture medium of Photorhabdus luminescens by ammonium sulfate precipitation and preparative isoelectric focusing followed by affinity chromatography. Ph. luminescens, a bacterium symbiotically associated with the insect-parasitic nematode Heterorhabditis bacteriophora, exists in two morphologically distinguishable phases (primary and secondary). It appears that only the secondary-phase bacterium produces this protease inhibitor. The protease inhibitor has an M:(r) of approximately 12000 as determined by SDS-PAGE. Its activity is stable over a pH range of 3.5-11 and at temperatures below 50 degrees C. The N-terminal 16 amino acids of the protease inhibitor were determined as STGIVTFKND(X)GEDIV and have a very high sequence homology with the N-terminal region of an endogenous inhibitor (IA-1) from the fruiting bodies of an edible mushroom, Pleurotus ostreatus. The purified protease inhibitor inactivated the homologous protease with an almost 1:1 stoichiometry. It also inhibited proteases from a related insect-nematode-symbiotic bacterium, Xenorhabdus nematophila. Interestingly, when present at a molar ratio of 5 to 1, this new protease inhibitor completely inactivated the activity of both trypsin and elastase. The activity of proteinase A and cathepsin G was partially inhibited by this bacterial protease inhibitor, but it had no effect on chymotrypsin, subtilisin, thermolysin and cathepsins B and D. The newly isolated protease inhibitor from the secondary-phase bacteria and its specific inhibition of its own protease provides an explanation as to why previous investigators failed to detect the presence of protease activity in the secondary-phase bacteria. The functional implications of the protease inhibitor are also discussed in relation to the physiology of nematode-symbiotic bacteria.

Concentrations of mercury, cadmium, lead and copper in fruiting bodies of edible mushrooms in an emission area of a copper smelter and a mercury smelter

Four metals were determined by AAS techniques in 56 samples of 23 wild mushroom species collected in a heavily polluted area in eastern Slovakia in 1997 and 1998. The area has been contaminated from historical polymetallic ores mining and smelting and by emissions from a mercury smelter between 1969 and 1993 and from a copper smelter since 1951. No significant differences in metal concentrations ( P<0.05) were found in four species when comparing the periods 1992–1993 and 1997–1998. Considerable contamination of most species was observed mainly for mercury and cadmium. The highest levels of mercury, up to 50 mg kg −1 dry matter, were found in Boletus reticulatus, Lycoperdon perlatum and Marasmius oreades, and of cadmium up to 20 mg kg −1 dry matter in Xerocomus chrysenteron and Lycoperdon perlatum. The latter species also had extremely high lead and copper concentrations in hundreds of milligrams per kilogram dry matter. Concentrations of mercury and copper in caps of four Boletaceae species were significantly ( P<0.05) higher than those in stipes.

Flammulin: A novel ribosome-inactivating protein from fruiting bodies of the winter mushroom &lt;i&gt;Flammulina velutipes&lt;/i&gt;

Flammulin: A novel ribosome-inactivating protein from fruiting bodies of the winter mushroom &lt;i&gt;Flammulina velutipes&lt;/i&gt;

Three new species of anamorphic yeasts phenotypically and phylogenetically related to Candida mesenterica: the description of Candida fungicola sp. nov., Candida sagamina sp. nov., and Candida fukazawae sp. nov. isolated from fruit bodies of mushrooms

Four strains of anamorphic yeasts isolated from fruit bodies of mushrooms collected in Japan were found to represent three new species of the genusCandida. These species resembleCandida mesenterica in characteristics commonly employed in the classification of yeasts. On the basis of DNA-DNA reassociation, however, they were clearly distinguished fromC. mesenterica and from one another. Three new species,Candida fungicola, Candida sagamina, andCandida fukazawae, are proposed for these yeasts. The analysis of SSU rDNA sequences suggested that these three species were closely related to each other and toC. mesenterica andC. suecica.

Isolation of a New Ribonuclease from Fresh Fruiting Bodies of the Straw Mushroom

Although ribonucleases have been isolated from numerous organisms in the animal and plant kingdoms, only a few have been reported from mushrooms. In this investigation, a novel ribonuclease, with an N-terminal amino acid sequence distinctly different from those of the previously published mushroom ribonucleases, was isolated from the fruiting bodies of the straw mushroom. It exhibited a molecular weight of 42.5 kDa in sodium dodecyl sulfate–polyacrylamide gel electrophoresis which was larger than those of ribonucleases from other organisms. The isolation procedure entailed extraction with aqueous buffer, (NH4)2SO4 precipitation, ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-Sepharose CL6B, and gel filtration on Superdex 75 using fast protein liquid chromatography. Straw mushroom ribonuclease was relatively specific for polyU and hydrolyzed yeast tRNA with a pH optimum between 6.5 and 7.5.

Hypocholesterolemic and antiatherogenic effect of oyster mushroom (Pleurotus ostreatus) in rabbits.

The addition of 10% dried fruiting bodies of oyster mushroom (Pleurotus ostreatus) to the diet containing 1% of cholesterol reduced serum cholesterol levels by 65% and cholesterol content in liver, heart, long extensor muscle and aorta of male rabbits (Chinchilla) by 60, 47, 25 and 80%, respectively. The decrease in total serum cholesterol was affected primarily (by 70%) by reduced cholesterol content in very-low-density lipoproteins (VLDL) while the contribution of high density lipoproteins (HDL) cholesterol increased by a factor of 3. Oyster mushroom diet reduced by 60-70% the content of conjugated dienes in plasma, erythrocytes and liver, however, it did not affect significantly the activities of antioxidant enzymes. Oyster mushroom diet reduced significantly the incidence of atherosclerotic plaques as estimated by sudanophilia (absence of positive-reaction in 3 of 5 animals) as well as plaque size (26% vs. 2% of the area with positive reaction in control and oyster mushroom-treated animals, respectively). While all animals on control diet showed atherogenic changes in aorta, oyster mushroom diet prevented the development of these changes in three animals. Fatty streaks and fibromatous plaques were found in the remaining two animals from this group. Oyster mushroom prevented the formation of atheroma plaques (found in three cases from the control diet group) and reduced the incidence of segmental injury of coronary arteria and of focal fibrosis of myocardium. Oyster mushroom caused lower incidence of foam cells in all types of lesions.

The uptake of cadmium and zinc by mycelia and their accumulation in mycelia and fruiting bodies of edible mushrooms

Cd and Zn concentrations were determined by optical emission spectroscopy in various parts of the fruiting body and the mycelium of two wild mushrooms, Agaricus macrosporus and Agaricus silvicola, and in cultivated Stropharia rugosoannulata. Cd was distributed in a characteristic manner within the fruiting body of all three species. The Cd content of the cap was a function of its radius as well as its height. Concentrations of Cd and the chemically related Zn in the investigated mushroom segments were strongly correlated, whereas Al, Cu and Ag correlated poorly with Cd. To our knowledge, this is the first study of Cd contents in wild mycelia. We found similar concentrations of Cd and Zn in isolated mycelia and stems of the corresponding fruiting bodies. In addition, substrates were analysed to study soil-specific effects on Cd accumulation. The extent of Cd and Zn transfer from soil to mushroom was species-specific and influenced by the availability of these two heavy metals, as well as the age of the mushroom. Interestingly, the typical Cd and Zn distributions described here were not affected by the extent of accumulation, indicating that uptake and distribution of Cd and Zn are actually two separate mechanisms.

The inhibitory properties and primary structure of a novel serine proteinase inhibitor from the fruiting body of the basidiomycete, Lentinus edodes.

A novel proteinase inhibitor, Lentinus proteinase inhibitor, has been purified from the fruiting bodies of the edible mushroom, Lentinus edodes, by buffer extraction and affinity chromatography on immobilized anhydrotrypsin. The protein simultaneously inhibits bovine beta-trypsin and alpha-chymotrypsin at independent sites, with apparent dissociation constants of 3.5 x 10(-10) M and 4 x 10(-8) M, respectively. The purified protein is eluted as two well-separated peaks on reversed-phase HPLC, one of which is inhibitory-active and the other inactive, and they are interconvertible under folding/unfolding conditions. Among the mammalian and microbial serine proteinases examined, including human enzymes of blood coagulation and fibrinolysis, activated factor XI was inhibited by the Lentinus proteinase inhibitor. Chemical modification studies suggest involvement of one or more arginine residues in the inhibition of trypsin. The complete primary structure composed of 142 amino acids with an acetylated N-terminus was determined by protein analysis. The theoretical molecular mass (15999.2) from the sequence is close to the experimental value of 15999.61 +/- 0.61 determined by mass spectrometry. Although there are no apparently homologous proteinase inhibitors in the protein database, there is a rather striking similarity to the propeptide segment of a microbial serine proteinase, as well as to the N-terminal region of the mature enzyme.

14-Hydroxy-12-oxo-10E,13E,15E-octadecatrienoic acid, a new fatty acid from a Vietnamese mushroom,Cantharellus friesiiQuel.

The new 14-hydroxy-12-oxo-10E,13E,15E-octadecatrienoic acid (1) was isolated from the fruit bodies of a Vietnamese edible mushroom, Cantharellus friesii Quel. Purification was done by liquid chromatography and gel-permeation chromatography. The structure was settled by mass spectrometric and NMR investigations.

Medicinal Properties of Substances Occurring in Higher Basidiomycetes Mushrooms: Current Perspectives (Review)

This review highlights some of the recently isolated and identified substances of higher Basidiomycetes mushroom origin that express promising antitumor, immune modulating, cardiovascular and antihyper-cholesterolemia, antiviral, antibacterial, antiparasitic, hepatoprotective, and antidiabetic effects. Medicinal mushrooms have a long history of use in folk medicine. Mushrooms useful against cancers of the stomach, esophagus, lungs, etc., are known in China, Russia, Japan, and Korea, as well as the United States and Canada. There are approximately two hundred species of mushrooms that have been found to markedly inhibit the growth of different kinds of tumors. However, most of the mushroom origin antitumor substances have not been clearly defined. Several antitumor polysaccharides, such as hetero-β-glucans and their protein complexes (e.g., xyloglucans, and acidic β-glucan containing uronic acid), as well as dietary fibers, lectins, and terpenoids, have been isolated from medicinal mushrooms. In Japan, Russia, China, and the United States, several different polysaccharide antitumor agents have been developed from the fruiting body, mycelia, and culture medium of various medicinal mushrooms (Lentinus edodes, Ganoderma lucidum, Schizophyllum commune, Trametes versicolor, Inonotus obliquus, and Flammulina velutipes). Both cellular components and secondary metabolites of a large number of mushrooms have been shown to effect the immune system of the host and therefore could be used to treat a variety of disease states. The information presented in this review is helpful in exploring and understanding the rich traditions of medicinal mushrooms in Eastern and Western cultures and medicine.

A fibrinolytic metalloprotease from the fruiting bodies of an edible mushroom, Armillariella mellea.

A fibrinolytic metalloprotease has been purified from the fruiting bodies of the edible honey mushroom (Armillariella mellea). The enzyme has a molecular weight of 18538.1508, as measured by MALDI-TOF mass spectrometry and includes Zn2+ ion as found by ICP/MS. The N-terminal amino acid sequence, XXYNGXTXSRQTTLV, do not match any known protein or open reading frame. It hydrolyzes fibrinogen as well as fibrin, but does not show any proteolytic activity for other blood proteins such as thrombin, human albumin, bovine albumin, human IgG, hemoglobin, or urokinase. This protease hydrolyzes both A alpha and B beta subunits of human fibrinogen with equal efficiency. The enzyme activity was strongly inhibited by EDTA and 1,10-phenanthroline, indicating that the enzyme is a metalloprotease. No inhibition was found with PMSF, E-64, pepstatin, and 2-mercaptoethanol. The activity of the purified enzyme was slightly increased by Mg2+, Zn2+, and Co2+, but the enzyme was totally inhibited by Hg2+. It has broad substrate specificity for synthetic peptides, and a pH optimum at 7, suggested that the purified enzyme was a neutral protease. It was thermally stable up to 60 degrees C and the maximum fibrinolytic activity was at 55 degrees C.

Therapeutic Effects of Substances Occurring in Higher Basidiomycetes Mushrooms: A Modern Perspective

This review highlights some of the recently isolated and identified substances of higher Basidiomycetes mushrooms origin that express promising antitumor, immune modulating, cardiovascular and hypercholesterolemia, antiviral, antibacterial, and antiparasitic effects. Medicinal mushrooms have a long history of use in folk medicine. In particular, mushrooms useful against cancers of the stomach, esophagus, lungs, etc. are known in China, Russia, Japan, Korea, as well as the U.S.A. and Canada. There are about 200 species of mushrooms that have been found to markedly inhibit the growth of different kinds of tumors. Searching for new antitumor and other medicinal substances from mushrooms and to study the medicinal value of these mushrooms have become a matter of great significance. However, most of the mushroom origin antitumor substances have not been clearly defined. Several antitumor polysaccharides such as hetero-beta-glucans and their protein complexes (e.g., xyloglucans and acidic beta-glucan-containing uronic acid), as well as dietary fibers, lectins, and terpenoids have been isolated from medicinal mushrooms. In Japan, Russia, China, and the U.S.A. several different polysaccharide antitumor agents have been developed from the fruiting body, mycelia, and culture medium of various medicinal mushrooms (Lentinus edodes, Ganoderma lucidum, Schizophyllum commune, Trametes versicolor, Inonotus obliquus, and Flammulina velutipes). Both cellular components and secondary metabolites of a large number of mushrooms have been shown to effect the immune system of the host and therefore could be used to treat a variety of disease states.

Selective oviposition by adult females and larval growth of Dacne picta Crotch (Coleoptera : Erotylidae) on different growing stages of the shiitake mushroom, Lentinula edodes

Dacne picta is one of the most important pests of cultivated shiitake mushrooms (Lentinula edodes) in Japan. To clarify the preferential oviposition of adults among different growing stages of mushrooms, survival and development of offspring, a census was taken in a cultivated field of the mushroom. Fruiting bodies of mushrooms were classified into three stages : growing, mature and decaying stages. A large number of adults and eggs were observed in and on the growing and mature stages of mushrooms, but few were observed on the decaying stage. Adult beetles, thus, disappeared from the mushrooms when they started to decay. An inoculation experiment of adult beetles on mushrooms revealed that the mortality of first stadium larvae was much higher in the decaying stage of the mushrooms. Rearing experiments on powdered mushrooms suggested that chemical inhibitors and/or nutritional deficiency was an important cause for the higher mortalities of the 1st stadium larvae. However, the hardening of decaying mushrooms may have prevented hatched larvae from actually consuming them. Thus, the mushroom stage chosen by ovipositing adults is closely related to the mortality of hatched larvae.

Plasma and Hepatic Cholesterol Levels and Fecal Neutral Sterol Excretion Are Altered in Hamsters Fed Straw Mushroom Diets

The effect of the fruiting body and mycelium of Volvariella volvacea (straw mushroom) on the concentrations of plasma lipids, liver cholesterol, fecal neutral sterol and bile acid excretions was investigated in male Golden Syrian hamsters. The hamsters were fed a purified hypercholesterolemic diet (0.1% cholesterol, 10% fat) for 4 wk to elevate plasma lipid concentrations. Twelve hamsters with elevated plasma total cholesterol were randomly assigned to each treatment group: control (5% cellulose), mushroom fruiting body (5%) and mushroom mycelium (5%). After 4 wk of mushroom diet consumption, the plasma total cholesterol, HDL cholesterol, and combined VLDL + LDL cholesterol concentrations (mmol/L) were significantly lower than control in the group fed the fruiting body-diet (40, 38 and 43%, respectively) (P < 0.05). The liver cholesterol levels were significantly lower in both the mushroom fruiting body– and the mycelium-fed groups (28 and 21% in terms of concentration; 39 and 30% in terms of total content, respectively) (P < 0.05) than that in the control group. Fecal neutral sterol excretion in the mushroom fruiting body– and mycelium-fed groups was significantly higher (81 and 74%, respectively) (P < 0.05) than that in the control group. Although no significant differences (P > 0.05) in the excretion of fecal bile acids were observed among groups fed the mushroom diets and the control diet, the mushroom fruiting body diet–fed hamsters apparently had less bacterial degradation of cholic acid as indicated by a significantly greater proportion (P < 0.05) of fecal cholic acid than in controls. They also had a significantly lower proportion of fecal deoxycholic acid (P < 0.05). This study suggests that the fruiting body of the straw mushroom lowers elevated plasma cholesterol in hypercholesterolemic hamsters, whereas the mycelium does not.

Characterization of ribonucleases from culture medium of Lentinus edodes.

Lentinus edodes (shiitake) cultivated in potato dextrose medium produced five RNases in the culture filtrate. The two major RNases (RNase Le37 and RNase Le45) were highly purified and their molecular masses, base specificities, N-terminal amino acid sequences, and amino acid compositions were analyzed and compared to RNase Le2 isolated from the fruit bodies of the same mushroom. RNase Le37 and RNase Le45 are base non-specific and adenylic acid preferential RNases like RNase Le2 and their N-terminal sequences are very similar to RNase Le2, but they are glycoproteins and their amino acid compositions are significantly different from that of RNase Le2. In addition to these enzymes, a guanylic acid-specific RNase with a molecular mass 13 kDa was partially purified. Since RNase Le2, which has very similar N-terminal sequence to RNase Le37 and RNase Le45, was not excreted from the mycelia, the analysis of the structures of these two excreted RNase may shade a light on the mechanism of excretion of RNases in this organism.

Studies on Metabolites of Mycoparasitic Fungi, 6. 15‐Hydroxyacorenone: New Acorane‐Type Sesquiterpene from the Culture Broth of the Mycoparasitic Fungus Trichoderma harzianum

AbstractA new acorane‐type sesquiterpene, 15‐hydroxyacorenone, has been isolated from the culture broth of Trichoderma harzianum, which in turn was isolated from the fruiting body of a medicinal mushroom, Ganoderma lucidum (Fr.) Karst. (the oriental crude drug “Lin‐Chi”). The structure of 15‐hydroxy‐acorenone was determined by spectroscopic methods to be (1S,4S,5S)‐8‐hydroxymethyl‐1‐isopropyl‐4‐methylspiro[4.5]‐dec‐8‐en‐7‐one (1).

Changes in In vitro Digestibility of Wheat Straw During Fungal Growth and after Harvest of Oyster Mushrooms (Pleurotus spp.) on Laboratory and Industrial Scale

Zadrazil, F. 1997. Changes in in vitro digestibility of wheat straw during fungal growth and after harvest of Oyster mushrooms (Pleurotus spp.) on laboratory and industrial scale. J. Appl. Anim. Res., 11: 37–48. The in vitro digestibilities of wheat straw during fungal growth and spent substrate left after the harvest of fruit bodies of Oyster mushrooms, (Pleurotus spp.) cultivated under laboratory and industrial conditions, were determined. Some strains of Pleurotus increased the in vitro digestibility of cereal straw under laboratory conditions by 63%. In an industrial production unit with a capacity of 6000kg mushroom/day, the mean in vitro digestibility of spent substrate after the harvest of mushrooms was 49.3±3.0% as compared to 42.8% of untreated straw. The yield of mushrooms was 170 kg per 1000 kg of original substrate. The quality of spent substrate from “low cost production” of mushrooms varied more in chemical composition and digestibility than substrates from industrial production.

Concentrations of mercury, copper, cadmium and lead in fruiting bodies of edible mushrooms in the vicinity of a mercury smelter and a copper smelter

Concentrations of mercury, copper, cadmium and lead in fruiting bodies of edible mushrooms in the vicinity of a mercury smelter and a copper smelter

Cadmium removal from water environment by a fungus Volveriella volvacea

Biosorption technique was used for removal of cadmium under different conditions from water environment using a biosorbent, Volveriella volvaceas, locally growing fruit bodies of mushroom. Effects of different parameters like pH, sorbent concentration, ionic strength on the removal efficiency of cadmium by V. volvacea were carried out in continuation with adsorption kinetics and equilibrium isotherm experiments. From the kinetics studies it was found that nearly 95% of the total cadmium removal was achieved from cadmium spiked distilled water within first 15 minutes. Isotherm data was best fitted to linearised Langmuir equation and the sorption capacity was found to be varying from 9.13 to 9.33 mg/g for different sizes of sorbent. The uptake of cadmium(II) is a function of pH of the solution and increases with the increasing pH. Increasing ionic strength and the presence of soluble complexing agents such as ethylene diamine tetraacetic acid (EDTA) decrease the sorption of cadmium (II). The presence of other diavalent cations like calcium and magnesium impedes the uptake of cadmium (II). The presence of chloride ion has no significant effect on cadmium (II) removal. The spent biosorbent can effectively be regenerated with acid and can then be reused.

Insecticidal properties of mushroom and toadstool carpophores

In order to find compounds with insecticidal or antifeedant properties from mushrooms and toadstools, a wide screening was undertaken using the non-mycophagous Drosophila melanogaster as a model insect. Powdered fruit bodies of edible and poisonous mushrooms were incorporated with the Drosophila's rearing medium, and their development was observed. Among the 175 different species of fungi tested, 79 were found to inhibit insect development, hence making the isolation of new compounds look hopeful.