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Amauroderma rugosum (AR) is a basidiomycete in the Ganodermataceae family that has been used traditionally to prevent epileptic attacks and constant crying in babies. However, AR has not been widely studied scientifically. In this review, we summarize the phytochemical components and pharmacological properties of AR that have been reported in the literature. Chemical analyses have revealed that the components of AR include sterols, flavonoids, fatty acids and esters, aromatic acids and esters, phenols, polysaccharides, and triterpenes. Pharmacological properties of AR include antioxidant, anti-inflammatory, neuroprotective, anti-cancer, anti-hyperlipidemic, anti-epileptic, and antibacterial effects. These findings suggest that AR and its bioactive ingredients have potential therapeutic applications, particularly for age-related diseases.

Canine splenic hemangiosarcoma (HSA) is an aggressive tumour of vascular endothelium that carries a grave prognosis following standard of care treatment with surgery and doxorubicin. A previous pilot study revealed potential anti-tumour activity of I'm-Yunity polysaccharopeptide (PSP) for canine HSA. The aim of this prospective study was to assess patient outcome when treated with PSP alone or in combination with doxorubicin post-splenectomy compared to patients treated with surgery and doxorubicin that received a placebo in place of PSP. Dogs undergoing splenectomy for splenic HSA were eligible. Following splenectomy, owners were offered treatment with PSP alone or adjuvant doxorubicin chemotherapy (unblinded). Patients with owners that selected to proceed with doxorubicin chemotherapy were blindly randomized to receive placebo or PSP. Dogs were evaluated weekly for 15 weeks, then scheduled for monthly visits until death. One hundred and one dogs were included in the final analysis: 51 PSP alone, 25 doxorubicin/placebo, and 25 combination PSP/doxorubicin. On multivariate analysis, dogs treated with single agent PSP, female dogs, decreased haematocrit at diagnosis, and stage III disease were negatively significantly associated with outcome; however, an interaction between treatment group and sex was documented. Gender-specific outcomes revealed no significant difference in survival between treatment groups for male dogs, but female dogs treated with PSP alone had significantly reduced survival compared to females receiving doxorubicin/placebo (HR 0.21; p = .004). Anaemia (HR 5.28; p .001) and stage III disease (HR 2.9; p = .014) remained negatively associated with survival when controlling for sex and treatment group. The addition of PSP to doxorubicin post-splenectomy did not improve survival in dogs with splenic HSA.

The structure and complete nuclear magnetic resonance (NMR) assignments of orbifomitellic acid, a novel lanostane triterpene isolated from the fruiting bodies of a Gabonese Ganoderma orbiforme (Polyporaceae), are reported. Within the vast catalogue of lanostanes documented from Ganoderma spp., orbifomitellic acid is the first disclosing a -COOH group at C-4.

Bisphenol A (BPA) is one of the toxic chemicals, which is widely used for manufacturing epoxy, polyester resin, and polycarbonates. These materials are extensively used in manufacturing of reusable bottles, baby bottles, dental sealants, various medical devices, and so forth. Moreover, canned and packaged foods are sources of bisphenol A, which is unknowingly consumed by many people worldwide. Its endocrine disrupting and teratogenic properties impose potential risk to the wildlife and human health. BPA has been linked to reproductive, metabolic, and immunity disorders in humans. Regardless of BPA ban in reusable and baby bottles, annually, 15 billion pounds of BPA still being produced. BPA pollution and its cleanup are major challenges. Therefore, it is essential to develop a suitable strategy to bioremediate BPA. The Trametes versicolor (TV) laccase-based nanoemulsion calcium alginate bead organogel was able to transform 94% of BPA within 2 h of treatment. Organogel showed 60% of BPA removal from actual industrial wastewater in packed bed batch reactor and 67% of BPA removal in continuous flow packed bed reactor. The biological oxygen demand (BOD) of treated industrial effluent was 14 mg/L, which is very much less than untreated effluent's BOD, which was 48 mg/L. The chemical oxygen demand of industrial effluent was 1240 mg/ml, and treated effluent was 248 mg/L, respectively. Hence, application of nanoemulsion-based organogel in packed bed reactor found to be a potential candidate for the bioremediation of industrial effluent containing BPA. PRACTITIONER POINTS: The TV laccase-based nanoemulsion calcium alginate bead organogel was able to transform 94% of BPA. Organogel showed 67% of BPA removal from industrial wastewater in continuous flow packed bed reactor. The nanoemulsion-based organogel in packed bed reactor found to be potential candidate for the bioremediation of industrial effluent containing BPA.

International audience; Lignin is the second most abundant polymer after cellulose in lignocellulosic biomass. Its aromatic composition and recalcitrant nature make its valorization a major challenge for obtaining low molecular weight aromatics compounds with high value-added from the enzymatic depolymerization of industrial lignins. The oxidation reaction of lignin polymer using laccases alone remains inefficient. Therefore, researches are focused on the use of a laccase-mediator system (LMS) to facilitate enzymatic depolymerization. Until today, the LMS system was studied using water-soluble lignin only (commercial lignins, modified lignins, or lignin model compounds). This work reports a study of three LMS systems to depolymerize the three major industrial lignins (organosolv lignin, Kraft lignin, and sodium lignosulfonate). We show that an enzymatic depolymerization of these lignins can be achieved by LMS using laccase from Trametes versicolor, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt as mediator and a cosolvent (25% of 1,4-dioxane) to enhance the solubilization of lignins.

Poly(2‐oxazoline)s (POxs) with 2,2′‐iminodiacetate (IDA) end groups were investigated as inhibitors for laccase. The polymers with the IDA end groups are reversible, competitive inhibitors for this enzyme. The IC50 values were found to be in a range of 1–3 mm. Compared with IDA alone, the activity was increased by a factor of more than 30; thus indicating that attaching a polymer chain to an inhibitor can already improve the activity of the former. The enzyme activity drops to practically zero upon increasing the concentration of the most active telechelic inhibitor, IDA‐PEtOx30‐IDA (PEtOx: poly(2‐ethyl‐2‐oxazoline)), from 5 to 8 mm. This unusual behavior was investigated by means of dynamic light scattering, which showed specific aggregation above 5 mm. Furthermore, the laccase could be stabilized in the presence of POx‐IDA, upon addition at a concentration of 20 mm and higher. Whereas laccase becomes completely inactive at room temperature after one week, the stabilized laccase is fully active for at least a month in aqueous solution., Bulking up: Poly(2‐oxazoline)s (POx) with iminodiacetate (IDA) end groups competitively inhibit the enzyme laccase. A telechelic IDA‐POx‐IDA derivative shifts the inhibition mechanism at higher concentrations to become a dead‐end inhibitor. Simultaneously, the polymers stabilize the enzyme to preserve its activity, even for 28 days at room temperature.

Novel liquid supports for enzyme immobilization and reuse based on aqueous biphasic systems (ABS) constituted by cholinium-based ionic liquids (ILs) and polymers for the degradation of dyes are here proposed. The biocatalytic reaction for dye decolorization using laccase occured in the biphasic medium, with the enzyme being "supported" in the IL-rich phase and the dye and degradation products being enriched in the polymer-rich phase. An initial screening of the laccase activity in aqueous solutions of ABS constituents, namely cholinium dihydrogen citrate ([Ch][DHC]), cholinium dihydrogen phosphate ([Ch][DHP]), cholinium acetate ([Ch][Acet]), polypropylene glycol 400 (PPG 400), polyethylene glycol 400 (PEG 400) and K2 HPO4 was carried out. Compared to the buffered control, a relative laccase activity of up to 170%, 257%, and 530% was observed with PEG 400, [Ch][DHP], and [Ch][DHC], respectively. These ABS constituents were then investigated for the in situ enzymatic biodegradation of the Remazol Brilliant Blue R (RBBR) dye. At the optimized conditions, the ABS constituted by PPG 400 at 46 wt% and [Ch][DHC] at 16 wt% leads to the complete degradation of the RBBR dye, further maintaining the enzyme activity. This ABS also allows an easy immobilization, recovery, and reuse of the biocatalyst for six consecutive reaction cycles, achieving a degradation yield of the dye of 96% in the last cycle. In summary, if properly designed, high enzymatic activities and reaction yields are obtained with ABS as liquid supports, while simultaneously overcoming the safety and environmental concerns of conventional organic solvents used in liquid-liquid heterogeneous reactions, thus representing more sustainable biocatalytic processes.

Sclerotial powder of a cultivated species of the Tiger Milk Mushroom, Lignosus cameronensis was analysed for its nutritional components and compared against species of the same genus, Lignosus rhinocerus and Lignosus tigris. All three species have been used by indigenous tribes in Peninsular Malaysia as medicinal mushrooms. Content of carbohydrate, fibre, mineral, amino acid, palatable index, fat, ash and moisture were determined. L. cameronensis sclerotial material consists of carbohydrate (79.7%), protein (12.4%) and dietary fibre (5.4%) with low fat (1.7%) and no free sugar. It has the highest content of total carbohydrate (791 g kg-1 ), energy value (3,700 kcal kg-1 ) and calcium (0.85 g kg-1 ). The crude protein content (123 g kg-1 ) is comparable to that of L. rhinocerus with its main amino acids consisting of glutamic acid, aspartic acid and leucine. The umami index is determined to be 0.27. The total essential amino acid (45 g kg-1 ) is comparable to that of L. tigris. The main mineral is potassium (1.51 g kg-1 ) and the Na/K ratio was

Fungal mycelia can eliminate almost all cocultured cyanobacterial cells within a short time. However, molecular mechanisms of algicidal fungi are poorly understood. In this study, a time‐course transcriptomic analysis of algicidal fungus Bjerkandera adusta T1 was applied to investigate gene expression and regulation. A total of 132, 300, 422, and 823 differentially expressed genes (DEGs) were identified at 6, 12, 24, and 48 hr, respectively. Most DEGs exhibited high endopeptidase activity, cellulose catabolic process, and transmembrane transporter activity by using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Many decomposition genes encoding endopeptidases were induced a little later in B. adusta T1 when compared with previously investigated algicidal fungus Trametes versicolor F21a. Besides, the accumulated expression of Polysaccharide lyases8 (PL8) gene with peptidoglycan and alginate decomposition abilities was greatly delayed in B. adusta T1 relative to T. versicolor F21a. It was implied that endopeptidases and enzymes of PL8 might be responsible for the strong algicidal ability of B. adusta T1 as well as T. versicolor F21a., Algicidal process of fungus Bjerkandera adusta T1 was investigated by a time‐course transcriptomic analysis. The results showed that most differentially expressed genes exhibited high endopeptidase activity, cellulose catabolic process, and transmembrane transporter activity. Comparison of gene expression between B. adusta T1 and Trametes versicolor F21a during the algicidal process suggested that endopeptidases together with enzymes of the polysaccharide lyases family 8 might be responsible for the strong algicidal activity of B. adusta T1 as well as T. versicolor F21a.

Micro total analysis system (μTAS) is expected to be applied in various fields. In particular, since electrochemical measurement is inexpensive and easy, electrochemical measurement can be integrated with a microchannel. However, electrochemical detection sensitivity in a microchannel is lowered because the diffusion of the detection target is limited. In an ordinary electrochemical detection system, using a stirrer in a beaker can overcome limited diffusion. We previously proposed a new detection system that combines a microliquid solution agitation technology using surface acoustic waves (SAWs) with the μTAS. The SAWs function as microstirrers, thus making electrochemical detection possible by overcoming limited diffusion of the sample. However, when the solution is stirred by the SAWs, the temperature of the solution increases to 70°C due to vibrational energy. This leads to enzyme inactivation and impaired electrochemical response. Therefore, in this study, we used a hyperthermophile-derived enzyme. Temperature and electrochemical characteristics of the detection system using SAWs and a multi-copper oxidase (MCO) derived from the hyperthermophilic archaea Pyrobaculum aerophilum were studied. Laccase, which is an MCO derived from the thermophilic fungus Trametes versicolor, was also studied. We also characterized the enzyme-electrochemical reaction using SAWs by comparing the magnitude of the reduction current obtained using the two enzymes with different heat resistances. We observed an increase in the electrochemical response with the SAWs, without impaired enzyme activity. Thus, the use of a thermostable enzyme is suitable for the development of a biosensor that uses SAWs for agitation.

Laccases are multi copper oxidases that can oxidize both phenolic and nonphenolic lignin related compounds. Consequently, there has been continuous demand for laccases for the oxidative degradation of phenolic dyes in effluents. In view of this, the present work was focused on laccase production by solid substrate fermentation using a newly isolated fungus Perenniporia tephropora-L168. To intensify the laccase production, the process parameters pH, nitrogen, inducer, and substrate: water ratio were optimized by using statistical model. A set of optimal conditions noted were pH 3, nitrogen 0.001 g/L; inducer 0.5% and substrate: water ratio (1:10), which yielded laccase 1,160 U/g. The crude laccase exhibited noteworthy potential to degrade a triaryl-methane dye especially Malachite green. Also, during bioremediation studies, the statistical process optimization could achieve 81% decolourization within 180 min. The laccase treatment brought chemical transformation in malachite green as evident from UV-Visible spectra, FTIR, HPLC while toxicity against bacteria and fungi was also reduced. During phytotoxicity study, effect of treated and untreated dye on germination of seed was analyzed. Interestingly, the germination index for Vigna aconitifolia and Vigna radiata was increased by two and fourfold, respectively. Overall, this work demonstrates optimized production of laccase using Perenniporia tephropora-L168 and its efficient bioremediation potential for triaryl-methane dye.

Poria cocos is an edible mushroom known as “Fuling” in Chinese, which belongs to the fungus family of Polyporaceae. Poria cocos has been used as a Chinese traditional medicine for > 2000 years. Indications for using it include promoting urination, to invigorate the spleen function, and to calm the mind. The bioactive components in Poria cocos include polysaccharides, triterpenoids, fatty acids, sterols, enzymes, etc. Poria cocos polysaccharide (PCP) accounts for 84% by weight among all constituents in the dried sclerotium. Biochemical and pharmacological studies reveal that PCP is the major bioactive component in Poria cocos and has a wide range of biological activities including antitumor, immunomodulation, anti-inflammation, anti-oxidation, anti-aging, anti-hepatitis, anti-diabetes, and anti-hemorrhagic fever effects. As a result, “Poria cocos polysaccharide oral solution” was developed and sold as an over-the-counter health supplement since 1970s. In 2015, “Polysaccharidum of Poria cocos oral solution” was approved as a drug by Chinese Food and Drug Administration (SFDA) for treating multiple types of cancers, hepatitis, and other diseases alone or during chemo- or radiation therapy for cancer patients. In this article, biochemical, preclinical and clinical studies of Poria cocos polysaccharide from 74 independent studies during the past 46 years (1970–2018) based on PubMed, VIP (Chongqing VIP Chinese Scientific Journals Database), CNKI (China National Knowledge Infrastructure), and Wanfang database searches are summarized. The structure, pharmacological effects, clinical efficacy, immune regulatory molecular mechanisms, and toxicity of PCP are deliberated to provide the data basis for PCP to serve as a clinically used antitumor drug.

PD was supported by a grant of the Netherlands Scientific Organization NWO 824.15.023 to RPdV. The Academy of Finland grant no. 308284 to MRM is acknowledged. Part of the research was performed at the Environmental Molecular Sciences Laboratory (EMSL), a national scientific user facility sponsored by the Department of Energy's Office of Biological and Environmental Research, located at the Pacific Northwest National Laboratory in Richland, WA, USA. The work conducted by the U.S. Department of Energy Joint Genome Institute (JGI), was supported by the Office of Science of the U.S. Department of Energy under Contract No. DE‐AC02‐05CH11231. CSL was supported by the CatchBio program. White‐rot fungi, such as Dichomitus squalens, degrade all wood components and inhabit mixed‐wood forests containing both soft‐ and hardwood species. In this study, we evaluated how D. squalens responded to the compositional differences in softwood [guaiacyl (G) lignin and higher mannan content] and hardwood [syringyl/guaiacyl (S/G) lignin and higher xylan content] using semi‐natural solid cultures. Spruce (softwood) and birch (hardwood) sticks were degraded by D. squalens as measured by oxidation of the lignins using 2D‐NMR. The fungal response as measured by transcriptomics, proteomics and enzyme activities showed a partial tailoring to wood composition. Mannanolytic transcripts and proteins were more abundant in spruce cultures, while a proportionally higher xylanolytic activity was detected in birch cultures. Both wood types induced manganese peroxidases to a much higher level than laccases, but higher transcript and protein levels of the manganese peroxidases were observed on the G‐lignin rich spruce. Overall, the molecular responses demonstrated a stronger adaptation to the spruce rather than birch composition, possibly because D. squalens is mainly found degrading softwoods in nature, which supports the ability of the solid wood cultures to reflect the natural environment. Publisher PDF

The ability to obtain carbon and energy is a major requirement to exist in any environment. For several ascomycete fungi, (post-)genomic analyses have shown that species that occupy a large variety of habitats possess a diverse enzymatic machinery, while species with a specific habitat have a more focused enzyme repertoire that is well-adapted to the prevailing substrate. White-rot basidiomycete fungi also live in a specific habitat, as they are found exclusively in wood. In this study, we evaluated how well the enzymatic machinery of the white-rot fungus Dichomitus squalens is tailored to degrade its natural wood substrate. The transcriptome and exoproteome of D. squalens were analyzed after cultivation on two natural substrates, aspen and spruce wood, and two non-woody substrates, wheat bran and cotton seed hulls. D. squalens produced ligninolytic enzymes mainly at the early time point of the wood cultures, indicating the need to degrade lignin to get access to wood polysaccharides. Surprisingly, the response of the fungus to the non-woody polysaccharides was nearly as good a match to the substrate composition as observed for the wood polysaccharides. This indicates that D. squalens has preserved its ability to efficiently degrade plant biomass types not present in its natural habitat.

Here we present the results of the exploration of laccase multigene families (MGFs) in basidiomycetous fungi from different taxonomic groups using a next generation sequencing (NGS) technology. In our study, multiple laccase genes were identified in all of the investigated fungi (13 species) from Polyporaceae, Phanerochaetaceae, Meruliaceae, Pleurotaceae, Physalacriaceae, and Peniophoraceae families. It was shown that phylogenetic positioning of the newly identified sequences exhibit patterns of clusterization with respect to enzyme properties. This can be a potentially useful tool for selecting naturally existing laccases with different physicochemical characteristics relevant to different biotechnological applications. Moreover, the method developed in this study can be used in the screening of environmental samples and fast characterization of laccase MGFs in newly identified fungal species.

The lignocellulolytic platform of the wood-decaying organism Dichomitus squalens is important for production of biodegradable elements; however, the system has not yet been fully characterized. In this study, using statistical target optimization, we analysed substrate selectivity based on a variety of D. squalens metabolic pathways using combined omics tools. As compared with the alkali-lignin (AL) programme, the rice straw (RS) programme has the advantage of multilayered signalling to regulate cellulolytic-related genes or to connect their pathways. The spontaneous instability of the AL programme was accelerated by harsh starvation as compared with that of the RS programme. Therefore, the AL programme converged on cellular maintenance much easier and more rapidly. However, regardless of external substrate/concentration type, the compensatory pattern of the major targets (especially peroxidases and growth regulators) was similar, functioning to maintain cellular homeostasis. Interestingly, ligninolytic-mediated targets under non-kaleidoscopic conditions were induced by a substrate-input-control, and especially this mechanism had an important effect on the early stages of the biodegradation process. This optimized target analysis could be used to understand lignocellulolytic network and to improve downstream efficiency.

The present work investigated the effects of environmental factors on the growth of fungal colonies of the white-rot basidiomycetes Physisporinus vitreus using a lattice-free discrete modeling approach called the fungal growth model (FGM), in which hyphae and nutrients are considered as discrete structures. A discrete modeling approach enables the underlying mechanistic rule concerning the basic architecture and dynamics of fungal networks to be studied on the scale of a single colony. By comparing simulations of the FGM with laboratory experiments of fungal colonies growing on malt extract agar we show that the combined effects of water activity, temperature and pH on the radial growth rate of fungal mycelia on the macroscopic scale may be explained by a power law for the costs of hyphal maintenance and expansion on the microscopic scale. Information about the response of the fungal mycelium at the micro- scopic level to environmental conditions is essential for simulating its behavior in complex structure substrates such as wood, where the effect of the fungus on the wood (i.e. the degradation of the cell wall) changes the local environmental condition (e.g. the permeability of the substrate and therefore the water activity in a colonized wood cell lumen). Using a combination of diffusion and moisture processes with the FGM may increase our understanding of the colonization strategy of P. vitreus and help to optimize its growth behavior for biotechnological applications such as bioincising.

Ganoderma lucidum is known as a medicinal mushroom used in traditional medicine. In our study, the cytotoxic activities of 17 compounds (1-17) isolated from the fruiting bodies of G. lucidum were investigated. Among them, ergosta-7,22-diene-2β,3α,9α-triol (EGDT) induced apoptosis in HL-60 human premyelocytic leukemia cells. EGDT activated the apoptotic process, including DNA fragmentation and caspase-3 activity. In immunoblotting analysis, treatment with EGDT resulted in the cleavage of procaspase-3 and poly(ADP-ribose) polymerase (PARP) into active forms. In the in vivo study, the administration (i.p.) of EGDT to Lewis lung carcinoma (LLC)-inoculated mice evidenced a significant inhibition of tumor growth. These results indicate that EGDT was one of the apoptotic constituents of G. lucidum, and might be an antitumor agent.

A hierarchical set of assays were used to evaluate the interspecific competitive ability of the bioincising fungus Physiporinus vitreus (response species). The competitiveness and growth mode of P. vitreus against a range of blue stain fungi and Trichoderma species (challenge species) was investigated in dual culture tests (a) and a spatially heterogeneous system of tessellation agar (b) on different media. In addition, the robustness of the bioincising process against biotic influences was determined by controlled interaction tests with different inocula of the challenge species (colony-forming units: 10(2), 10(4), 10(6) ml(-1)) on Norway spruce (Picea abies Karst.) heartwood. The results of the dual culture and tessellation agar tests revealed a significant (P ≤ 0.001) antagonistic potential (lethal effect) of the T. atroviride strain (15603.1), whereas no or only a slight effect of the blue stain fungi on P. vitreus was detectable. The interaction tests on Norway spruce heartwood confirmed that an increase in inoculum size of the challenge fungi correlated with an increased negative effect (Spearman's rho coefficient (ρ) = -0.521). Among the challenge species, T-15603.1 revealed the most negative effect on the homogeneous development of P. vitreus on wood and thus on the outcome of the bioincising process. To improve the uniformity of colonization and to reduce the rate of contamination, the incubation of wood in a sealed system (bioreactor) after sterilization is suggested.

Violins produced by Antonio Stradivari during the late 17th and early 18th centuries are reputed to have superior tonal qualities. Dendrochronological studies show that Stradivari used Norway spruce that had grown mostly during the Maunder Minimum, a period of reduced solar activity when relatively low temperatures caused trees to lay down wood with narrow annual rings, resulting in a high modulus of elasticity and low density. The main objective was to determine whether wood can be processed using selected decay fungi so that it becomes acoustically similar to the wood of trees that have grown in a cold climate (i.e. reduced density and unchanged modulus of elasticity). This was investigated by incubating resonance wood specimens of Norway spruce (Picea abies) and sycamore (Acer pseudoplatanus) with fungal species that can reduce wood density, but lack the ability to degrade the compound middle lamellae, at least in the earlier stages of decay. Microscopic assessment of the incubated specimens and measurement of five physical properties (density, modulus of elasticity, speed of sound, radiation ratio, and the damping factor) using resonance frequency revealed that in the wood of both species there was a reduction in density, accompanied by relatively little change in the speed of sound. Thus, radiation ratio was increased from 'poor' to 'good', on a par with 'superior' resonance wood grown in a cold climate.

Three new compounds, 2,4-dimethoxy-6-methylbenzene-1,3-diol (1), salmoquinone (2), and 3-(4-hydroxyphenyl)-4-isobutyl-1H-pyrrole-2,5-dione (3), together with six known compounds, 2-methoxy-6-methyl-p-benzoquinone (4), 2,3-dimethoxy-5-methyl-p-benzoquinone (5), 2-hydroxy-5-methoxy-3-methyl-p-benzoquinone (6), eburcoic acid (7), fomefficinic acid C (8), and a pyrroledione (9), were isolated from the mycelium of Antrodia salmonea. The structures of these compounds were elucidated by spectrometric analyses including IR, NMR, and MS. Among these compounds, 4 and 5 exhibited cytotoxicity against KB, HepG2, and H2058 cell lines.

In the course of our investigation on biologically active substances from inedible mushrooms in Japan, Germany, and Vietnam, we studied the chemical constituents of 22 species belonging to five families: Scutigeraceae, Polyporaceae, Xylariaceae, Thelephoraceae, and Paxillaceae. Various types of chemical substances were purified and characterized based on the modern spectroscopic methods and also on chemical reactions. These metabolites have shown a broad activity in many biological systems, such as antimicrobial, nematicidal, inhibition of NO production, anti-human immunodeficiency virus, tumor necrosis factor-α, and antioxidant activities. These isolated metabolites did not only show interesting activities, but also are employed as chemical markers supported for chemosystematics of these families. This review paper deals with the chemical constituents of 22 species, their biological activities, and also a discussion on chemosystematics. © 2006 The Japan Chemical Journal Forum and Wiley Periodicals, Inc. Chem Rec 6: 79–99; 2006: Published online in Wiley InterScience (www.interscience.wiley.com) DOI 10.1002/tcr.20074

1. Fungivorous insects are generally viewed as polyphagous, largely because most fungal fruiting bodies constitute an unpredictable resource. To examine the validity of this hypothesis, and degree of phylogenetic relatedness between the preferred hosts of the insects, host selection in the insect fauna of bracket fungi was studied, using data obtained both from the field and the literature. 2. More than half (53%) of the insect species breeding in them appeared to be monophagous. 3. Modern phylogenies explained the host selection patterns better than older classifications, since non-monophagous species of beetles frequently used hosts that are closely related to each other. 4. The hypothesis that polyphagous species use more heavily decayed fruiting bodies than monophagous species was verified for insects breeding in Fomes fomentarius. The results indicate that the chemical composition of the fungi influences host selection. 5. It is suggested that fruiting bodies of bracket fungi differ from most other fungi in that their occurrence is more predictable. Therefore, the primary colonising fungivores generally attack only one host species, or a few hosts that are closely phylogenetically related. Polyphagous species generally colonise fruiting bodies after they have reached a certain stage of decay, thus escaping their chemical defence.

ummary Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used for the discrimination among wood decay fungal species of the genera Fomitopsis, Rhodofomes, Fomes and Ganoderma (Polyporaceae, Basidiomycota). For all taxa studied, distance levels as high as 400 were observed and intraspecies variability and reproducibility accounted for distance levels up to 150, indicating the applicability of this method for the identification of filamentous basidiomycete fungi. The method was found to work on in vitro mycelia.

Among antifungal products of natural origin, Pycnoporellus fulgens (Fr) Donk, is active against pathogenic yeasts. Our study was carried out to observe the ultrastructural modifications produced by P. fulgens crude extract on Candida glabrata. Yeasts were submitted to different concentrations (50, 100, 200, 400 micrograms ml-1) for 72 h under constant stirring at 35 degrees C. Transmission electron microscopy revealed that the extract acted on the cell envelope (cell wall and plasmalemma). Cell divisions were also affected by thickening of the septum (50 micrograms ml-1) and a deficiency in the daughter cell wall texture. The extent of the antifungal effect clearly depended on the extract concentration.

In the search for antiparasite compounds from the Colombian flora, an active compound against Leishmania (Viannia) panamensis amastigotes was isolated from the fungi Pycnoporus sanguineus. The structural elucid-ation was achieved with spectroscopic methods ((1)H and (13)C NMR and MS). This compound was identified as ergosterol 5,8-endoperoxide.

Lucidenic acid A (1) and a new lactone, lucidenolactone (2) were isolated from the fruiting bodies of Ganoderma lucidum. Their structures were elucidated by spectroscopic method and single-crystal X-ray analysis. Lucidenolactone (2) showed significant antiplatelet aggregation activity. The NMR spectral data of lucidenic acid A were also reassigned.

The ligninolytic basidiomycetes Pleurotus eryngii, Pleurotus ostreatus, Pleurotus pulmonarius and Pleurotus sajor-caju did not exhibit detectable levels of manganese peroxidase (MP) when grown in liquid media with ammonium tartrate as N source. However, after examination of cells grown on different organic N-based media, high MP activity was obtained in peptone medium, up to nearly 3 U/ml in cultures of P. eryngii. Moreover, Mn2+ supplementation was not used to produce MP, since all Mn2+ concentrations assayed (1–4000 μM) inhibited production of this enzyme in liquid medium. Two MP isoenzymes were purified to homogeneity from shaken or stationary cultures of P. eryngii grown in peptone medium. The purification process (which included chromatography on Biorad Q-cartridge, Sephacryl S-200 and Mono-Q) attained 56% activity yield with a purification factor of 25. The isoenzymes differed in pI (3.75 and 3.65), N-terminal sequence and some catalytic properties. They were in some aspects (e.g., molecular mass of 43 kDa) similar to Phanerochaete chrysosporium MP but exhibited some distinct characteristics, including Mn2+ -independent peroxidase activities against 2,6-dime-thoxyphenol and veratryl alcohol, and higher resistance to H2O2. Recent studies have shown that MP are ubiquitous enzymes in ligninolytic fungi, but the results obtained suggest that differences in catalytic properties probably exist between different Mn2+ -oxidizing peroxidases produced by these fungi.

Summary The effect of fungal and yeast glucan on different immune functions in mice was examined and compared. The simultaneous administration of glucan and a sensitizing dose of DNFB on the different sites significantly stimulated delayed-type hypersensitivity (DTH) response only when using fungal glucan. Both glucans tested, when administered before sensitization, significantly increased DTH response, but with a significantly higher level at the beginning of the investigation (on day 7) when using fungal glucan. The increase in phagocytic activity by the blood leucocytes started in the 1st week after fungal-glucan treatment, and in the 2nd week after yeast-glucan treatment, and took longer after administration of fungal glucan. The values of the phagocytic-activity index were significantly influenced only after fungal-glucan injection. The results of the study indicate that fungal glucan isolated from Pleurotus ostreatus could be a prospective immunomodulating substance.

Generic variation within and among one Finnish and three Swedish populations of Fomitopsis pinicola (Schwarts: Fr.) Karst. were studied by amplifying DNA from hap-loid isolates originating from single spore cultures using two arbitrary primers. Analysis offspring from single fruit bodies revealed only three pairs of codominant alleles among 42 variable genetic markers, the remaining 38 segregated independently. Genetic similarity was measured in terms of Euclidean distance. Individuals in the Finnish population tended to form a distinct cluster in the principal component analysis. Variation within and among populations/regions was partitioned by Analysis of Molecular Variance — AMOVA. Within population variation accounted for 91.6% of the total genetic variation. The remaining 7.68% was accounted for by variation between the Finnish population and each of the three Swedish ones. Variation among the Swedish populations accounted for only 0.72% of the total variation. Wright's Fst was 0.17 for all four populations and 0.13 for the three Swedish populations. These relatively low values indicate that there is gene flow among all populations or that they are derived from a common ancestral population. The observed pattern of genetic variation is probably the result of effective spore dispersal and the continuous distribution of this common early successional species.

Summary. The antifungal activity of 38 Polyporaceae sensu lato against yeasts and dermatophytes was tested in vitro by the agar dilution method. Strains were typed organisms and clinical isolates. In this first report Pycnoporellus fulgens (Fr. Donk was found to be the most active species against pathological fungi and showed broad-spectrum antifungal activity against yeasts (Candida albicans, Candida glabrata) and dermatophytes (Trichophyton mentagrophytes, Trichophyton rubrum. Microsporum canis. Microsporum gypseum. Epidermophyton floccosum). Other Polyporaceae species also showed antifungal activity, but only against dermatophytes. Zusammenfassung. Alkoholische Rohextrakte von 38 Polyporaceae-Arten wurden in vitro mittels Agardilutiontest auf ihre antimyzetische Wirkung gegen Hefen und Dermatophyten gepruft. Die Teststamme waren einige Sammlungsstamme, in der Mehrzahl aber klinische Isolate. In dieser Studie erwies sich der Extrakt von Pycnoporellus fulgens (Fr.) Donk als antimyzetisch aktivste Spezies gegen pathogene Hefen (Candida albicans, C. glabrata) sowie gegen Dermatophyten (Trichophyton mentagrophytes, T. rubrum, Microsporum canis, M. gypseum, Epidermophyion floccosum). Andere Polyporaceae zeigten ebenfalls antimyzetische Aktivitat, jedoch nur gegen Dermatophyten.

Polysaccharide fractions TG-1 and TG-2 were isolated from the fruit bodies of Trametes gibbosa (Pers.: Fr.) Fr.(Polyporaceae). Both fractions are glucans with trace amounts of other sugars. The polysaccharide fractions administered to rats intravenously, strongly inhibited vessel permeability in the rat pleural exudate model, induced by carragenan, but did not influence its density. Both polysaccharide fractions decreased considerably the total protein level in the pleural effusion. Fractions TG-1 and TG-2 antagonized effectively the inflammation mediator complex induced by carragenan and they neutralized the pharmacological effects with respect to small vessel permeability. At the same time they increased the number of neutrophils and eosinophils and decreased the number of lymphocytes in the peripheral blood. The pharmacological activity of polysaccharide fractions TG-1 and TG-2 confirms their function as potential factors responsible for the stabilization of a wall vessel, especially in pathological conditions leading to endothelial damage.

Morphological and cultural studies of Polyporus maculatissimus and P. portentosus are presented. Neolentiporus gen. nov. is proposed to accommodate P. maculatissimus, a species with a striking macromorphological similarity with the north temperate P. squamosus. The new genus is characterized by medium to large stipitate fruitbodies with a poroid hymenophore, circular to flabellate pilei with a scaly surface and an excentric or lateral stipe that sometimes is reduced to a short, robust umbo. The hyphal system is dimitic with clamped, irregularly thick-walled generative hyphae that do not react with cresyl-blue, and terminal, unbranched, thick-walled skeletal hyphae that are strongly metachromatic in cresyl-blue. Spores are cylindric, hyaline, thin-walled, inamyloid and binucleate. The sexuality is bipolar, the nuclear behavior is astatocoeno-cytic and the associated wood-rot is brown. The new genus is regarded as the poroid counterpart of the agaricoid Neolentinus. Polyporus portenrosus is included in Laetiporus on the basis of its yellowish fruitbodies, its soft, punky context, its hyphal system composed of simple septate generative hyphae and binding hyphae, its holocoenocytic nuclear behavior and its associated brown wood-rot. A new code symbol, i.e. ‘9s’ is proposed to codify the presence, in cultures, of simple-septate generative hyphae with irregularly thickened walls.

Four conserved disulfide bonds and N-linked and O-linked glycans of extracellular fungal peroxidases have been identified from studies of a lignin and a manganese peroxidase from Trametes versicolor, and from Coprinus cinereus peroxidase (CIP) and recombinant C. cinereus peroxidase (rCIP) expressed in Aspergillus oryzae. The eight cysteine residues are linked 1-3, 2-7, 4-5 and 6-8, and are located differently from the four conserved disulfide bridges present in the homologous plant peroxidases. CIP and rCIP were identical in their, glycosylation pattern, although the extent of glycan chain heterogeneity depended on the fermentation batch. CIP and rCIP have one N-linked glycan composed only of GlcNAc and Man at residue Asn142, and two O-linked glycans near the C-terminus. The major glycoform consists of single Man residues at Thr331 and at Ser338. T. versicolor lignin isoperoxidase TvLP10 contains a single N-linked glycan composed of (GlcNAc)(2)Man(5) bound to Asn103, whereas (GlcNAc)(2)Man(3) was found in T. versicolor manganese isoperoxidase TvMP2 at the same position. In addition, mass spectrometry of the C-terminal peptide of TvMP2 indicated the presence of five Man residues in O-linked glycans. No phosphate was found in these fungal peroxidases. (Less)

Four new sterols, 5α, 6α-epoxy-(22E, 24R)-ergosta-8(14), 22-diene-3β, 7β-diol (1), (22E, 24R)-ergosta-8, 22-diene-3β, 5α, 6β, 7α-tetrol (2), (22E, 24R)-ergosta-7, 9(11), 22-triene-3β, 5α, 6β-triol (3) and 3β, 5α, 6β-trihydroxy-(22E, 24R)-ergost-22-en-7-one (4), have been isolated from the fruit bodies of Grifola frondosa (FR.) S. F. GRAY (Polyporaceae)together with fourteen known ones (5-18), of which two (5 and 6) are reported for the first time from a natural source. The structures of these compounds were elucidated on the basis of spectral data.

The production in a 5-1 fermenter of the extracellular enzymes laccase and aryl-alcohol oxidase by the fungus Pleurotus eryngii was studied. The latter enzyme has been purified 50-fold by Sephacryl S-200 and Mono Q chromatography. Purified aryl-alcohol oxidase is a unique flavoprotein with 15% carbohydrate content, a molecular mass of 72.6 kDa (SDS/PAGE) and a pI of 3.9. The enzyme presents wide specificity, showing activity on benzyl, cinnamyl, naphthyl and aliphatic unsaturated alcohols. Neither activity nor inhibition of veratryl alcohol oxidation was found with saturated alcohols, but competitive inhibition was produced by aromatic compounds which were not aryl-alcohol oxidase substrates, such as phenol or 3-phenyl-1-propanol. From these results, it was apparent that a double bond conjugated with a primary alcohol is necessary for substrate recognition by aryl-alcohol oxidase, and that activity is increased by the presence of additional conjugated double bonds and electron donor groups. Both affinity and maximal velocity during enzymic oxidation of methoxybenzyl alcohols were affected in a similar way by ring substituents, increasing from benzyl alcohol (Km = 0.84 mM, Vmax = 52 U/mg) to 4-methoxybenzyl alcohol (Km = 0.04 mM, Vmax = 208 U/mg). Aryl-alcohol oxidase presents also a low oxidase activity with aromatic aldehydes, but the highest activity was found in the presence of electron-withdrawing groups.

The antitumorigenic effect of cryptoporic acid E (CPA-E), a dimeric drimane sesquiterpenoid isolated from the fungus Cryptoporus volvatus, on colon carcinogenesis was investigated. Female F344 rats given an intrarectal instillation of 2 mg of N-methyl-N-nitrosourea 3 times weekly in weeks 1 and 2 were fed diet containing 0.2% CPA-E from week 3. Female ICR mice given 15 weekly intraperitoneal injections of 10 mg of 1,2-dimethylhydrazine/kg body weight during weeks 1 to 15 were fed diet containing 0.06% CPA-E from week 1. The experiment was terminated at week 35 for rats and at week 25 for mice. The incidence and the number of tumors per animal were reduced in CPA-E-fed animals compared to the controls: 31% vs. 75% (P less than 0.05) and 0.4 +/- 0.2 (SEM) vs. 0.9 +/- 0.2 (0.1 greater than P greater than 0.05) in rats, and 31% vs. 63% (0.1 greater than P greater than 0.05) and 0.4 +/- 0.2 vs. 2.4 +/- 0.8 (P less than 0.05) in mice (16 animals in each group). Intrarectal deoxycholic acid-induced colonic mucosal ornithine decarboxylase activity was significantly lowered in CPA-E-fed animals compared to controls. This shows an antipromoting activity of CPA-E against colon carcinogenesis. Thus, it was concluded that CPA-E inhibits colon cancer development in both rats and mice treated with 2 different colon carcinogens.

A 42-year-old female shiitake grower was investigated to clarify the etiology of skin lesions which developed during the planting of shiitake hyphae into bed logs. She complained of repeated eczematous skin lesions during the planting season, from March to July, for 10 years. She handled 7,000 pieces of small conic blocks made of beech, with shiitake hyphae attached to their surface, per day, and 300,000 pieces altogether per season. She was positive on patch testing with extracts of shiitake hyphae. In contrast, female shiitake growers with skin lesions associated with work other than planting, and without skin lesions, were negative on patch testing to the hyphae. Moderate allergenicity was observed to extracts of shiitake hyphae in a guinea pig maximization test. These findings indicated the etiology of skin lesions in shiitake growers to be allergic contact dermatitis induced by shiitake hyphae.

Shiitake (Lentinus edodes) is a mushroom which is eaten in Chinese and Japanese meals and is nowadays the second most commonly produced edible mushroom in the world. Shiitake dermatitis was first described by Nakamura in 1977. This disease presents with very characteristic skin manifestations. From April 1974 to April 1991, I have observed 51 patients with shiitake dermatitis. The following description reviews the clinical manifestations, laboratory investigations and sources of shiitake dermatitis.

Polyporus umbellatus is a widely used diuretic herbal medicine. In this study, a high-performance liquid chromatography coupled with atmospheric pressure chemical ionization-mass spectrometric detection (HPLC-APCI-MS) method was developed for qualitative and quantitative analysis of steroids, as well as for the quality control of Polyporus umbellatus. The selectivity, reproducibility and sensitivity were compared with HPLC with photodiode array detection and evaporative light scattering detection (ELSD). Selective ion monitoring in positive mode was used for qualitative and quantitative analysis of eight major components and beta-ecdysterone was used as the internal standard. Limits of detection and quantification fell in the ranges 7-21 and 18-63 ng/mL for the eight analytes with an injection of 10 microL samples, and all calibration curves showed good linear regression (r(2) > 0.9919) within the test range. The quantitative results demonstrated that samples from different localities showed different qualities. Advantages, in comparison with conventional HPLC-diode array detection and HPLC-ELSD, are that reliable identification of target compounds could be achieved by accurate mass measurements along with characteristic retention time, and the great enhancement in selectivity and sensitivity allows identification and quantification of low levels of constituents in complex Polyporus umbellatus matrixes.

Ganoderma applanatum polysaccharide (GAPS) was extracted and purified from Polyporaceae Ganoderma applanatum (Pers.) Pat. It was found that in vitro GAPS (65.2–250 μg/mL) increased the spleen cell proliferation response to ConA; GAPS (500 μg/mL) enhanced IL-2 production of spleen cells stimulated with ConA; GAPS (125–500 μg/mL) augmented the IFNγ production of spleen cells stimulated with ConA. Administering GAPS (20 mg/kg/day s.c.) to normal mice for 10 consecutive days enhanced spleen cell proliferation responses to ConA and the IL-2 and IFNγ production of spleen cells stimulated with 5 μg/mL ConA. It also increased spleen cell primary antibody responses to sheep red blood cells. Administering GAPS (20 mg/kg/day, s.c.) to sarcoma 180 transplanted mice for 15 consecutive days restored the NK activity and the IL-2 and IFNγ production of the spleen cells which were suppressed by the tumour.

Reinvestigation of the pigments of Pycnoporus sanguineus (Polyporaceae) yielded 4 new pigments of the 2-amino-phenoxazin-3-one type and the new phenoxazine ether pycnosanguin (9) besides cinnabarine (1). Unsubstituted phenoxazin-3-one was also shown among the pigments. Isolation and structure elucidation was performed after methylation of the crude acetone extract. The structures were established by spectral analysis and chemical degradations or interconversions. Untersuchung der Pigmente von Pycnoporus sanguineus - Pycnosanguin und neue Phenoxazin-3-one Erneute Untersuchung der Pigmente von Pycnoporus sanguineus (Polyporaceae) fuhrte neben Cinnabarin (1) zur Isolierung von 4 neuen Pigmenten vom 2-Amino-phenoxazin-3-on-Typ und dem neuen Phenoxazinether Pycnosanguin (9). Unsubstituiertes Phenoxazin-3-on befand sich ebenfalls unter den isolierten Verbindungen. - Die Isolierung erfolgte nach Methylierung des rohen Aceton-Extraktes. Die Strukturen wurden durch Spektroskopie sowie chemische Umwandlungen und Abbau aufgeklart.

New triterpene glucosides, oligoporins A (1), B (2), and C (3), were isolated from the methanolic extract of the fruiting bodies of Oligoporus tephroleucus (Polyporaceae). Their structures were established by spectroscopic methods. These compounds significantly exhibited protective effect to plasmid DNA damage by hydroxyl radical (*OH) generated from the Fenton reaction with hydrogen peroxide and ferrous.

Five new lanostane-type triterpenes, named fomefficinic acid A-E (1-5), were isolated from the dried sclerotium of Fomes officinalis, respectively. Their structures were established as 24-methylene-3-oxo-lanost-8-en-21-oic acid (1), 3alpha,15alpha-dihydroxy-24-methylene-lanosta-7,9(11)-dien-21-oic acid (2), 3alpha,15alpha-dihydroxy-24-methylene-lanost-8-en-21-oic acid (3), 15alpha-hydroxy-3-oxo-24-methylenelanost-8-en-21-oic acid (4), 15alpha-acetoxy-3-oxo-24-methylenelanosta-7,9(11)-dien-21-oic acid (5), by spectral analysis and chemical methods as well as comparison with known compounds.

1. Introduction Out of the four copper atoms per molecule of lacc- ase two copper atoms, Type 3, have an undetermined redox state, are not detectable by EPR and are non- paramagnetic. However, these oxidases can accept four electron equivalents [ 1 ] . Redox titrations have shown [ 1,2] that a 330 nm chromophore is not associated with Type 1 or Type 2 Cu2+ but with a unit accepting two electrons. During reduction of this chromophore no new EPR signals are formed. It has been suggested that this chromophore is associated with Type 3 copper, which may be present in the divalent state as a binuclear complex [3]. In an alternative proposal, it is not the copper that accepts the two electrons, but a disulfide bridge closely associated with a copper pair [4]. The aim of this study was to present additional evidence about the state of copper in relation to sulfhydryl groups and disulfide bridges in order to choose a model for the Type 3 copper. We studied fungal lactase A and B from

Of the 93 polypores described by Montagne, either alone or with other mycologists, 32 are accepted, 53 are synonyms, 3 names are invalid, 2 species are of unknown affinities, 2 types have not been found while 1 name is rejected. The following three combinations are proposed: Phylloporia capucina (Mont.) Ryv. comb, nov., Coltricia fragilissima (Mont.) Ryv. comb, nov., and Gloeophyllum mexicanum (Mont.) Ryv. comb. nov.

154 species of Ahyllophorales, mostly Corticiaceae and Polyporaceae are reported from Thailand, 116 out of which are new to the country. Aleurodiscus cremicolor Hjortst. & Ryv., Byssocorticium naviculare Hjortst. & Ryv., Grammothele ochraceus Ryv., Hyphoderma tuberculare Hjortst. & Ryv., Boletopsis atrata Ryv., Ceriporia subreticulala Ryv. and Oxyporus subulatus Ryv. are described as new. A key to all known Byssocorticium species is given.