Antiviral activity of Basidiomycete mycelia against influenza type A(serotype H1N1) and herpes simplex virus type 2 in cell culture

In this study, we investigated the in vitro antiviral activity of the mycelia of higher mushrooms against influenza virus type A(serotype H1N1) and herpes simplex virus type 2(HSV-2), strain BH. All 10 investigated mushroom species inhibited the reproduction of influenza virus strain A/FM/1/47(H1N1) in MDCK cells reducing the infectious titer by 2.0–6.0 lg ID50. Four species, Pleurotus ostreatus, Fomes fomentarius, Auriporia aurea, and Trametes versicolor, were also determined to be effective against HSV-2 strain BH in RK-13 cells, with similar levels of inhibition as for influenza. For some of the investigated mushroom species—Pleurotus eryngii, Lyophyllum shimeji, and Flammulina velutipes—this is the first report of an anti-influenza effect. This study also reports the first data on the medicinal properties of A. aurea, including anti-influenza and antiherpetic activities. T. versicolor 353 mycelium was found to have a high therapeutic index(324.67), and may be a promising material for the pharmaceutical industry as an anti-influenza and antiherpetic agent with low toxicity. Mycelia with antiviral activity were obtained in our investigation by bioconversion of agricultural wastes(amaranth flour after CO2 extraction), which would reduce the cost of the final product and solve some ecological problems.

A New Ceramide from the Basidiomycetes Armillaria mellea

A new phytosphingosine-type ceramide, armillaramide 1, has been isolated from the fruiting bodies of Basidiomycetes Armillaria mellea (Vahl ex Fr.) Quel. Its structure was established as (2S, 3S. 4R)-2-N-(palmitoyl)-phytosphingosine by spectroscopic and chemical methods.

Six novel steroids from culture of basidiomycete Polyporus ellisii

Investigation of the culture of basidiomycete Polyporus ellisii led to the isolation of a novel compound 3β,9α,15α-trihydroxy-(22E,24R)-10(5→4)-abeo-ergosta-6,8(14),22-trien-5-one(1)with a new 5/7/6/5 ring system of ergosterol skeleton.In addition,five new steroids,5β,6β-epoxy-3β,7α,9α-trihydroxy-(22E,24R)-ergosta-8(14),22-dien-15-one(2),5β,6β-epoxy-3β,7α-dihydroxy-(22E,24R)-ergosta-8(14),22-dien-15-one(3),5α,6α-epoxy-3β,9α,15α-trihydroxy-(22E,24R)-ergosta-8(14),22-dien-7-one(4),15α-acetoxy-(22E,24R)-ergosta-4,6,8(14),22-tetraen-3-one(5),15β-methoxy-(22E,24R)-ergosta-4,6,8(14),22-tetraen-3-one(6),along with four known ergosterols(7-10),were obtained.All structures were elucidated based on 1D and 2D NMR spectral data.New compounds were evaluated for cytotoxicity against five human cancer cell lines,only compound 4 was found to exhibit a favorable cytotoxicity profile toward all tested tumor cell lines.

A New p-Terphenyl Derivative from Basidiomycetes Boletopsis grisea

A new p-terphenyl derivative 1 was isolated from the fruiting bodies of Boletopsis grisea. Its structure was established as 2. 3-diacetoxy-4 ', 4 ', 5, 6-tetrahydroxy-p-terphenyl by spectroscopic and chemical means.

Lepida Acid A from Basidiomycetes Russula lepida

A new compound, lepida acid A 1, has been isolated from the fruiting bodies of Basidiomycetes Russula lepida. Its structure was elucidated by spectral methods.

Hirsutane Sesquiterpenes from Cultures of the Basidiomycete Marasmiellus sp.BCC 22389

Two new hirsutane sesquiterpenes,marasmiellins A(1)and B(2),were isolated from cultures of the basidiomycete Marasmiellus sp.BCC 22389.The structures were elucidated on the basis of NMR spectroscopic and mass spectrometry data.The absolute configuration of marasmiellin B was determined by application of the modified Mosher’s method.

Rufoolivacin B,a novel polyketide pigment from the fruiting bodies of the fungus Cortinarius rufo-olivaceus(basidiomycetes)

A novel polyketide pigment （1） with the 4＇,10-coupled linkage between 1-naphthalenol and 1,4-anthraquinone, named rufoolivacin B together with the known analog rufoolivacin （2）, has been isolated from the fruiting bodies of the Chinese toadstool Cortinarius rufo-olivaceus （basidiomycetes）. Their structures were characterized by means of analysis of spectroscopic methods, including 2D-NMR experiments and HR-ESI-MS.

The Ability of Edible Mushrooms to Act as Biocatalysts: Preparation of Chiral Alcohols Using Basidiomycete Strains

To examine the potential ability of edible mushrooms to act as biocatalysts, 19 basidiomycete strains were screened. Modified media (PG, O, and PGO medium) for liquid cultivation of these basidiomycete strains were designed and tested. Wet cells (>10 g) of 4 basidiomycete strains (Pleurotus salmoneostramineus H7, P. salmoneostramineus H13, Ganoderma lucidum NBRC31863, Flammulina velutipes NBRC31862) were harvested from PGO medium for 7 days. The stereoselective reduction of α-keto esters using the 4 strains was tested. It was found that each of these strains had a reducing activity toward 6 aliphatic α-keto esters. In the presence of L-alanine as an additive, the reduction of ethyl 2-oxobutanoate and ethyl 2-oxopentanoete by P. salmoneostramineus H7 produced the corresponding alcohol with a high conversion ratio and with excellent enantiomeric excess (>99% e.e. (R)). Furthermore, ethyl pyruvate, ethyl 2-oxobutanoate, and ethyl 2-oxopentanoate were predominantly reduced to the corresponding (R)-hydroxy ester (>99% e.e.) by G. lucidum. Thus, we found that these edible mushrooms have great potential to be used as biocatalysts for the stereoselective reduction of carbonyl compounds.

Assessment of Factors Influencing Trace Element Content of the Basidiomycetes in the European Part of Russia

The article presents concentrations of 29 trace elements in different groups of basidiomycetes: saprotrophic, parasitic and symbiotic. The potential of using the concentration factor (Kc) is shown for the assessment of trace element accumulation by the fungi. The variations of trace element concentrations in different ecological and geochemical environment are presented. The data obtained in this research made it possible to mark out the group of trace elements that are mainly accumulated by fungi. This information helped to assess the correlation between the trace element concentrations in the mycothalluses of fungi and their mobile forms in the soil. The expected patterns of the trace element accumulation in fungi are described.

Seven New Sesquiterpenes from Cultures of the Basidiomycete Conocybe siliginea

Seven new tremulane-type sesquiterpenes （1 7） have been isolated from cultures of the basidiomycete Cono- cybe siliginea. Their structures and relative configurations were elucidated by extensive spectroscopic data （HSQC, HMBC, 1H-tH COSY, ROESY） and MS analysis.

Tetra-,penta-,and hexa-nor-lanostane triterpenes from the medicinal fungus Ganoderma australe

Chemical investigation on the medicinal fungus Ganoderma australe led to the identification of ten new nor-lanostane triterpenes,namely two hexa-nor ones,ganoaustratetraenones A(1)and B(2),five penta-nor ones,ganoaustraldehydes A-E(3-7),and three tetra-nor ones ganoaustrenoic acids A-C(8-10).The chemical structures along with the absolute configurations were determined by extensive spectroscopic analysis of 1D&2D NMR and HRESIMS data.The postulated biosynthesis pathways of these compounds were proposed.Ganoaustraldehydes A(3)and B(4)showed moderate inhibition against nitric oxide production in RAW264.7 macrophage cells with the respec-tive IC_(50) values of 32.5,34.2μM(the IC_(50) of positive control pyrrolidine dithiocarbamate was 20.0μM).

Structures and cytotoxicities of three new sesquiterpenes from cultures of Armillaria sp.

Three new sesquiterpene aryl esters,named 10-dehydroxy-melleoliede B(1),1-O-formyl-10-dehydroxy-melleoliede B(2)and 10-oxo-melleoliede B(3)together with six known ones(4-9),were isolated from the cultures of Armillaria sp.The structures of the new compounds were elucidated based on the extensive spectroscopic methods.Compounds 1,2,and 5-9 exhibited moderate cytotoxicities.

Proteins from <i>Punctularia atropurpurascens</i>with Biotechnological Applications

Basidiomycetes are able to biodegrade waste and xenobiotic molecules through the production of extracellular enzymes. For example, white-rot fungi produce lignin-degrading enzymes which are capable of efficiently decolorizing dye solutions. Many mushrooms also produce lectins, a group of proteins which bind specifically to the carbohydrates in glycoconjugates. Several fungal lectins target their specificities towards oligosaccharides present in mammalian glycoproteins, thus constituting excellent ligands for the preparation of affinity adsorbents useful in isolation and characterization of these glycoproteins. In this study we isolated and characterized two different proteins, a lectin and a laccase, present in extracts from Punctularia atropurpurascens. The lectin isolated from the mycelium extract, was immobilized on activated-Sepharose and used to evaluate the interaction with three glycoproteins. The adsorbent was able to efficiently adsorb and elute bovine lactoferrin, constituting a promising tool for the purification of this glycoprotein. In vitro experiments revealed that the lectin also exhibited antimicrobial activity against Aspergillus niger. Laccase activity was detected in the extracellular extract from P. atropurpurascens. This enzyme, in both soluble and immobilized forms, was able to degrade Remazol Brilliant Blue R and Acid Blue 25 dyes. The biological activities found in this fungus demonstrate its potential for various biotechnological applications.

Optimization of the Production Process of <i>Enzymatic Activity</i>of Lentinula edodes (Berk.) Pegler in Holocelulases

Issues such as fossil fuels and oil supplies have stimulated the search for renewable alternatives such as biofuels. Agricultural crop residues represent an abundant renewable resource for the future of bioethanol. For it to be a viable alternative, the second-generation ethanol which ought to provide a net energy gain, environmental benefits, should be economically viable, and also be produced in large quantities without reducing food supplies. The current difficulty of lignocellulosic biofuel production is the hydrolysis of biomass into sugar. This is a work in which the white-rot Lentinula edodes fungus secretes substantial amounts of hydrolytic enzymes and is useful for degradation of lignocellulosic biomass which have not been described yet. The objective of this investigation was to evaluate the pH effect (5, 6 and 7), agitation (0, 100 rpm and 200 rpm) and also the cultivation time (6, 9 and 12 days). The culture medium was supplemented with agro-industrial residue and the EF 52 strain of the fungus Lentinula edodes was used as a processing agent. A factorial design 22 repeating the central point was performed. Submerged cultivation was conducted in a synthetic medium and was incubated at 25°C. The total protein content was determined as well as the activity of xylanase and cellulase (endoglucanase, exoglucanase and β-glucosidase). By Pareto diagram, the agitation and pH variables were significant for enzymatic activities. The highest enzyme expression occurred at pH values between 5.0 and 6.0 and above 100 rpm agitation. The exoglucanase was the enzyme which showed the highest activity in terms of cellulases, despite the cultivation time. Regarding the production of other enzymes and proteins, the most significant cultivation time was 12 days.

Production of Manganese Peroxidase by Trametes villosa on Unexpensive Substrate and Its Application in the Removal of Lignin from Agricultural Wastes

Manganese peroxidase (MnP) is a ligninolytic enzyme that is involved in the removal of lignin from the cell wall of plants. This removal facilitates the access of hydrolytic enzymes to the carbohydrate polymers that are hydrolyzed to simple sugars, which allows the subsequent fermentation to obtain bioproducts, such as ethanol. In this work, response surface methodology (RSM) was employed to optimize the culture conditions on unexpensive substrate for MnP secretion by Trametes villosa. Three independent variables were evaluated (i.e., temperature, moisture content and pH). The crude extract containing MnP was used in the delignification experiment and it caused a reduction in lignin content for all residues tested: 35.05 ± 1.45 (%) for the sugar cane bagasse;63.11 ± 0.06 (%) for the sisal fiber and 39.61 ± 0.39 (%) for the coconut shell, under the reaction conditions tested after 4 hours of fermentation. The preliminary results exhibited the potential application of this enzyme in the removal of lignin from plant residues. However, the conditions should be evaluated and optimized for each residue type.

Vibralactone Biogenesis-Associated Analogues from Submerged Cultures of the Fungus Boreostereum vibrans

A scale-up fermentation of the fungus Boreostereum vibrans facilitated the isolation of six new vibralactone biogenesis-associated analogues,namely vibralactamide A(1),vibralactone T(2),13-O-lactyl vibralactone(3),10-O-acetyl vibralactone G(4),(11R,12R)-and(11S,12R)-vibradiol(5,6).Their structures were established via extensive spectroscopic analyses,specific optical rotation comparison,and Snatzke’s method.The biosynthetic pathway for vibralactamide A was postulated.The absolute configuration of vibralactone B was revised by single crystal X-ray diffraction analysis.This work puts the divergent vibralactone biosynthesis pathway one step further and expands the structural diversity of vibralactone-associated compounds.

Potential Pathogens among Fungi Identified as Nonsporulating Molds from Blood Cultures

Twenty-eight molds were isolated from clinical blood cultures and were unidentifiable by cellular morphology using conventional microscopy. Using the utility of amplification and direct sequencing of internal transcribed spacer region of ribosomal RNA gene, 93% of these fungi were identified. Seventy-one percents of the molds were found to be associated with plants or soil with no or few published cases of human disease. These include species of basidiomycetes and ascomycetes such as Botryosphaeria dothidea, Phomopsis flavodonflavus, Inonotus pachyphloeus, Earlilella scabrosa, Calocybe indica, Athelia pellicularis, Tinctoporellus epimiltinus, Trametes lactinea, Coprinellus aureogranulat and Xylaria feejeensis. Some of the nonsporulating molds were identified as pathogen or potential pathogens in immunocompetent or immunocompromised hosts. These include Schizophylum commune and hyphomycetes such as Cladosporium cladosporoides, Aspergillus niger and Fusarium equiseti. Basidiomycetes and hyphomycetes identified in the current study are ubiquitous in the environment and are almost similar to the species of molds reported from cutaneous and respiratory samples suggesting that the fungi may represent contaminants rather than true fangaemia. Results of this study emphasize the need of an effort to minimise blood culture contamination and support the recommendation to incorporate clinical, radiologic findings and positive blood culture for molds in the diagnosis and management of invasive mycosis.

Biological Active Components of Selected Medical Fungi

According to official data, about 2000 mushrooms belong to the category of medical mushrooms, while over 600 have already confirmed medical properties. The aim and task of this work were to collect, analyze and process scientific and expert data of biologically active components of selected Basidiomycetes fungi: Letinula edodes, Ganoderma Lucidum, Grifola frondosa, Trametes versicolor and Inonotus obliquus. Areas of fungi therapy and the search for new immunomodulatory agents are far from being restricted to these species alone, however, these five may serve as typical representatives of widespread medicinal fungi used in both traditional medicine and modern biomedical research. Their biologically active components have different pharmacological effects, and beta-glucan polysaccharides, which are recognized as immunomodulators, are of particular importance. Many of the fungal beta-glucans tested have switched to pharmaceuticals such as Lentinan, Sonifilan, Krestin and GanoPoly, which speaks to their pharmacological and research potential. Citing the results of scientific advances in the last two decades, the results of preclinical tests and the results of clinical studies can confirm that supplementation with medical fungi can increase treatment success or mitigate the negative side effects of different therapies. A long-term weakened immune system is a risk factor for malignancies, so it can be concluded that disease prevention is beneficial for each individual and deserves the same attention paid to treating the disease.

Production of Enzymes and Biotransformation of Orange Waste by Oyster Mushroom, <i>Pleurotus pulmonarius</i>(Fr.) Quél.

The wood-decay fungi are able to bioconvert a wide variety of lignocellulosic residues due to the secretion of extracellular enzymes. The use of agricultural wastes as substrate for mushroom cultivation or enzymes production can help to solve environmental problems caused by inadequate discharge in the nature. The production of hydrolytic and oxidative enzymes by Pleurotus pulmonarius developed in solid state system using orange waste as substrate was evaluated in this work. Among the hydrolytic enzymes, pectinase was the main enzyme produced by the fungus, presenting the highest enzymatic activity of 9.4 U/mL after 35 days of cultivation. Considering the oxidative enzymes, laccase was the main enzyme produced with maximal activity of 12.2 U/mL obtained after 20 days of cultivation. Low enzyme levels of manganese peroxidase, β-glucosidase and β-xy-losidase were detected with activity peaks at the end of the cultivation. The enzymatic levels of amylase, carboxymethyl cellulase and xylanase were similar and less than 1.5 U/mL. No aryl-alcohol oxidase activity was detected. NDF, ADF and cellulose values increased during 45 days of cultivation. There was no lignin degradation during the study period and the fungus culture in orange solid waste caused protein enrichment in the substrate. Our results demonstrate that P. pulmonarius was an efficient producer of two important industrial enzymes, pectinase and laccase in a cheap solid state system using orange waste as substrate.

A Wood Preservative Based on Commercial Silica Nanodispersions and Boric Acid against Fungal Decay through Laboratory and Field Tests

The paper is based on the development of a wood preservative without metal salts to be used in use classes 3 and 4 (EN 335), eco-friendly and harmless to humans and animals. Boric acid was used as a biocide, due to its effectiveness against fungi and insects. It is also known to be easily leached from wood exposed to weather action. Colloidal silica was therefore added in the formulations to guarantee the fixation of boric acid to wood. The different formulations were tested for the protective efficacy against decay fungi through laboratory tests (EN 113) and field trials (EN 252). The results were promising, especially those concerning boron fixation and efficacy against decay fungi through laboratory tests, where some formulations and retentions gave a durability class 1 (very durable) according to EN 350-1. The fourth evaluation, after 50 months of field trials showed only a slight difference between the treated samples and controls.