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As indústrias têxteis e de alimentos são responsáveis pela liberação de efluentes corados ao meio ambiente, que podem ser tóxicos ou cancerígenos . A biodegradação por diferentes microrganismos constitui uma alternativa menos agressiva ao meio ambiente e economicamente viável, envolvendo enzimas ligninolíticas como as lacases. Este trabalho objetivou a comparação entre o Pleurotus ostreatus e o Pycnoporus spp na descoloração de efluentes e corantes têxteis e determinar suas atividades de lacase. Os experimentos foram realizados em meios líquidos de Vogel suplementados com glucose, extrato de levedura e efluente corado, incubados a 28°C e 180 rpm de agitação até se observar a descoloração. Os resultados demonstraram uma descoloração de 87,97% do Remazol Brilliant Orange pelo Pleurotus ostreatus contra 35,60% pelo Pycnoporus, 63,88% do Remazol Preto Intenso contra 43,93%, em relação ao efluente obteve-se 68,53% de descoloração contra 70,69%, respectivamente. Indicando uma melhor performance dos tratamentos com o Pleurotus ostreatus.

C4 grasses are common species in rangelands around the world and represent an attractive option for second-generation biofuel production. Although they display high polysaccharide content and reach great levels of biomass accumulation, there is a major technical issue to be addressed before they can be used for bioethanol industrial production: lignin removal. Concerning this, Pycnoporus and Ganoderma fungal genera have been highlighted due to their ability to hydrolyze lignocellulose in biological pretreatments. Our goals here were to evaluate the pretreatment efficiency using the secretome of species from Pycnoporus and Ganoderma spp. harvested from a glucose-free inductive medium (using a C4 grass) and to identify the fungal enzymatic activities responsible for the lignin degradation and glucose release. Our results show that P. sanguineus secretome exhibits a higher activity of lignocellulolytic enzymes such as cellulases, xylanases, laccases, and manganese peroxidases compared with that from G. resinaceum. Interestingly, zymograms in the presence of 2 M glucose suggest that a β-glucosidase isoform from P. sanguineus could be glucose tolerant. The proteomic approach carried out allowed the identification of 73 and 180 different proteins in G. resinaceum and P. sanguineus secretomes, respectively, which were functionally classified in five main categories and a miscellaneous group. These results open new avenues for future experimental work that lead to a deeper comprehension and a greater application of the mechanisms underlying lignocellulosic biomass degradation.

The biological treatment efficiency of dye wastewater using activated sludge (AS) is largely limited to the chromaticity and ecotoxicity of dyestuff. To alleviate this limitation, eleven industrial-grade disperse dyes were obtained from a fiber-dyeing factory, and for the first time, we studied the decolorization and detoxification effects of using the

Marore Island (Province of North Sulawesi) is one of the outermost island in Indonesia. The island is located at the borderline within the Philippines. Till date, there is no publication about islands macrofungi diversity from this island. In fact, the diversity of macrofungi has the potency to support the development of global bioeconomic in Indonesia. The purpose of this study is to make an inventory list of macrofungi on Marore Island in order to enhance the potential in the future. A total was of 22 mushrooms from 13 genera were obtained from this study which were divided into two phylums, namely Basidiomycota and Ascomycota. The identification results confirmed the species obtained: Ganoderma sp.1, Ganoderma sp.2, Ganoderma sp.3, Ganoderma sp.4, Ganoderma sp.5, Ganoderma sp.6, Trametes sp.1, Trametes sp.2, Trametes sp.3, Pycnoporus sp.1, Pycnoporus sp.2, Polyporus sp., Daedalea sp., Lentinus sp., Marasmius sp.1, Marasmius sp.2, Schizophyllum sp., Panellus sp., Tricholoma sp., Phallus sp., Dacryopinax spathularia, and Daldinia sp.

Woody biomass represents an important source of carbon on earth, and its global recycling is highly dependent on Agaricomycetes fungi. White-rot Basidiomycetes are a very important group in this regard, as they possess a large and diverse enzymatic repertoire for biomass decomposition. Among these enzymes, the recently discovered lytic polysaccharide monooxygenases (LPMOs) have revolutionized biomass processing with their novel oxidative mechanism of action. The strikingly high representation of LPMOs in fungal genomes raises the question of their functional versatility. In this work, we studied an AA9 LPMO from the white-rot basidiomycete Pycnoporus sanguineus, PsAA9A. Successfully produced as a recombinant secreted protein in Pichia pastoris, PsAA9A was found to be a C1-specific LPMO active on cellulosic substrates, generating native and oxidized cello-oligosaccharides in the presence of an external electron donor. PsAA9A boosted cellulolytic activity of glysoside hydrolases from families GH1, GH5, and GH6.This study serves as a starting point towards understanding the functional versatility and biotechnological potential of this enzymatic family, highly represented in wood decay fungi, in Pycnoporus genus. KEY POINTS: • PsAA9A is the first AA9 from P. sanguineus to be characterized. • PsAA9A has activity on cellulose, producing C1-oxidized cello-oligosaccharides. • Boosting activity with GH1, GH5, and GH6 was proven.

Rose dregs were used for laccase production by the strain Pycnoporus sp. W-9 under liquid fermentation, and the obtained laccase was used for phenol degradation. The conditions for laccase production were optimized by Box-Behnken design, and the phenol degradation conditions using the crude laccase were optimized by central composite design. The optimal conditions for laccase production by the strain W-9 were as follows: rose dregs (40 g/L), KCl (0.144 g/L), KH2PO4 (1 g/L), K2HPO4 (0.699 g/L), NaCl (0.2 g/L), MgSO4·7H2O (0.3 g/L), CuSO4·5H2O (0.306 g/L), Tween 80 (0.2 g/L), and CaCO3 (1 g/L), at a pH of 6.0, incubated at 30 °C and 200 rpm for 7 d. The corresponding laccase yield reached 17.4 U/mL, which was approximately 1.8 times the original production. The optimal conditions for phenol removal by the crude laccase were as follows: laccase (12.2 U/mL), 0.1 mol/L citric acid-disodium hydrogen phosphate buffer (pH 3.5), phenol (100 μg/mL), and Triton X-100 (284 μg/mL), incubated at 45 °C for 2 h. The corresponding phenol degradation rate reached 86.6%. These results should be useful for utilization of rose dregs and bioremediation of soil and wastewater.

Carboxylic acid reductases (CARs) are attracting burgeoning attention as biocatalysts for organic synthesis of aldehydes and their follow-up products from economic carboxylic acid precursors. The CAR enzyme class as a whole, however, is still poorly understood. To date, relatively few CAR sequences have been reported, especially from fungal sources. Here, we sought to increase the diversity of the CAR enzyme class. Six new CAR sequences from the white-rot fungus Pycnoporus cinnabarinus were identified from genome-wide mining. Genome and gene clustering analysis suggests that these PcCAR enzymes play different natural roles in Basidiomycete systems, compared to their type II Ascomycete counterparts. The cDNA sequences of all six Pccar genes were deduced and analysis of their corresponding amino acid sequence showed that they encode for proteins of similar properties that possess a conserved modular functional tri-domain arrangement. Phylogenetic analyses showed that all PcCAR enzymes cluster together with the other type IV CARs. One candidate, PcCAR4, was cloned and over-expressed recombinantly in Escherichia coli. Subsequent biotransformation-based screening with a panel of structurally-diverse carboxylic acid substrates suggest that PcCAR4 possessed a more pronounced substrate specificity compared to previously reported CARs, preferring to reduce sterically-rigid carboxylic acids such as benzoic acid. These findings thus present a new functionally-distinct member of the CAR enzyme class.

The genus Pycnoporus is a member of Phylum Basidiomycota. This fungus is one of polypore genera that is easily recognized from the orange color of the fruiting body and it likely has high morphological features. Currently, there are 207 number fungal specimens kept in Herbarium Bogoriense (BO) under the genus Pycnoporus, and they were identified as P. sa-nguineus (L.) Murrill, P. cinnabarinus (Jacq.) P. Karst., and Pycnoporus sp. To ensure that those species name of the Pycnoporus collections is correct, thus this study was carried out. Re-identification process on the fungal collections was done based on macro-micromorphological features of all Pycnoporus specimens. This study revealed that species Pycnoporus collections stored in BO represent 2 species: P.sanguineus (L.) Murrill and P. puniceus (Fr.) Ryvarden. In addition, some specimens under the name P. cinnabarinus should be validated as P. puniceus. This information will be very useful for fungal taxonomists, non-taxonomists, and people who work on Pycnoporus species from Indonesia.

Mushroom strains of Polyporales from the genera Coriolus, Trametes, Pycnoporus, Ganoderma, and Formitella were explored in terms of mycelial growth characteristics for the application of mushroom mycelia as alternative sources of materials replacing fossil fuel-based materials. Among the 64 strains of Polyporales, G. lucidum LBS5496GL was selected as the best candidate because it showed fast mycelial growth with high mycelial strength in both the sawdust-based solid medium and the potato dextrose liquid plate medium. Some of the Polyporales in this study have shown good mycelial growth, however, they mostly formed mycelial mat of weak physical strength. The higher physical strength of mycelial mat by G. lucidum LBS5496GL was attributed to its thick hyphae with the diameter of 13 µm as revealed by scanning electron microscopic analysis whereas the hyphae of others exhibited less than 2 µm. Glycerol and skim milk supported the best mycelial growth of LBS5496GL as a carbon and a nitrogen source, respectively.

RESUMEN Pycnoporus es un género representativo de los homobasidiomicetos saprófitos que tiene un potencial lignocelulitíco: Los metabolitos secundarios de este hongo son importantes en la búsqueda de sustancias antivirales, antioxidantes, antifúngicas, y antibacterianas. El objetivo de este trabajo fue la producción de los pigmentos producidos por Pycnoporus sanguineus y su actividad contra hongos y bacterias fitopatógenas. El mayor rendimiento del pigmento (8.8%) se obtuvo en caldo papa, donde se produjo 1.1631 g de pigmento. Con los extractos crudos obtenidos, se determinó la CMI en 1.5 mg mL-1 para Erwinia amylovora y Pectobacterium carotovorum, CMB de 3 mg mL-1 y 5 mg mL-1 para E. amylovora y P. carotovorum, respectivamente. ABSTRACT Pycnoporus is a representative genus of saprophytic homobasidiomycetes that has lignocellulitic potential: The secondary metabolites of this fungus are important in the search for antiviral, antioxidant, antifungal, and antibacterial substances. The objective of this work was the production of pigments produced by Pycnoporus sanguineus and their activity against fungi and phytopathogenic bacteria. The highest pigment yield (8.8%) was obtained in potato broth, where 1.1631 g of pigment were produced. With the crude extracts obtained, the MIC was determined at 1.5 mg mL-1 for Erwinia amylovora and Pectobacterium carotovorum, CMB of 3 mg mL-1 and 5 mg mL-1 for E. amylovora and P. carotovorum, respectively., {"references":["Cruz-Muñoz, R. 2012. Producción de extractos de Pycnoporus sanguineus con actividad antimicrobiana en hongos y bacterias fitopatógenas. Tesis de Maestría. UPIBI-IPN, México, DF. pp: 64.","Kirk, P.M., Cannon, P.F., David, J.C. & Stalpers, J.A. 2001. Ainsworth & Bisby's Dictionary of the Fungi. 9th Edition. CABI Publishing.","Rosa, L. E., M. K. M. Gomes, J. C. Cristina, M. Capelari, R.C. Augusto, and Z. C. Leomar. 2003. Screening of Brazilian basidiomycetes for antimicrobial activity. Mem. Inst. Owaldo Cruz 98: 967-974.","Sommer, N.F., Fortalage, J., Edwuards, D.C. 2002.Postharvest Diseases of selected commodities. En Kader,A.A.(Ed)., Postharvest Tecnology of Horticultural Crops (pp.223-227).University of California.","Thomson. 2007. Diccionario de especialidades agroquímicas patrocinado por BASF The chemical company (versión electrónica).","Wilson, C.L., Wisniewski, M.E., 1994.Biological Control of Postharvest Diseases.Theory and Practice.CRC Press, Boca Raton, USA."]}

The degradation of 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) by Pycnoporus sanguineus was investigated in order to explore the impact of the heavy metal Cu2+ on BDE-47 decomposition and the subsequent formation of metabolites, as well as to further elucidate the degradation mechanism of BDE-47. An increase in degradation rate from 18.63% to 49.76% in the first four days and its stabilization at (51.26 ± 0.08)% in the following days of BDE-47 incubation were observed. The presence of Cu2+ at 1 and 2 mg/L was found to promote the degradation rate to 56.41% and 60.79%, respectively, whereas higher level of Cu2+ (≥ 5 mg/L) inhibited the removal of BDE-47. The similar concentration effects of Cu2+ was also found on contents of fungal protein and amounts of metabolites. Both intracellular and extracellular enzymes played certain roles in BDE-47 transportation with the best degradation rate at 27.90% and 27.67% on the fourth and third day, individually. During the degradation of BDE-47, four types of hydroxylated polybrominated diphenyl ethers (OH-PBDEs), i.e., 6′-OH-BDE-47, 5′-OH-BDE-47, 4′-OH-BDE-17, 2′-OH-BDE-28, and two bromophenols, i.e., 2,4-DBP and 4-BP were detected and considered as degradation products. These metabolites were further removed by P. sanguineus at rates of 22.42%, 23.01%, 27.04%, 27.96%, 64.21%, and 40.62%, respectively.

Some plant species colonize mining-contaminated soils. They are adapted to harsh growth conditions and show high native phytoremediation abilities. For environmental cleanup, these species can be improved by genetic manipulation. Laccases are ligninolytic enzymes that oxidise a broad range of substrates by a radical-catalysed reaction mechanism using oxygen as the electron acceptor. Fungi of the genus Pycnoporus produce laccases with important biotechnological, industrial and environmental applications. Here we describe a successful attempt of Agrobacterium rhizogenes mediated stable transformation of Nicotiana glauca, a species naturally resistant to metal-contaminated soils. The coding region of a Pycnoporus sanguineus laccase gene was fused with the Nicotiana plumbaginifolia calreticulin apoplast targeting signal and driven by the constitutive cauliflower mosaic virus (CaMV) 35S promoter (2X35S). The obtained transgenic N. glauca callus cells secreted a recombinant fungal laccase into the growth medium and were capable to degrade an anthraquinone dye, Remazol Brilliant Blue R. Under phenol and cadmium stresses, the transgenic calli not only maintained their growth capacity but their relative growth rate was greater compared to callus transformed with empty vector. This work shows that stable expression of a fungal laccase in a plant species resistant to heavy metals presents a successful strategy that potentially can be used to combat combined organic and inorganic contamination. A fungal laccase was heterologously expressed in a heavy metal-tolerant plant species. The transgenic callus was capable of growth in medium supplemented with organic and inorganic contaminants.

Background 5-Hydroxymethylfurfural (HMF), a major residual component of a lignocellulosic bio-refinery process, can be transformed into fundamental building blocks for green chemistry via oxidation. While chemical methods are well established, interest is also being directed into the enzymatic oxidation of HMF into the bio-plastic precursor 2,5-furandicarboxylic acid (FDCA). Results We demonstrate that three glyoxal oxidases (PciGLOX) isoenzymes from the Basidiomycete fungus Pycnoporus cinnabarinus were able to oxidize HMF, with PciGLOX2 and PciGLOX3 being the most efficient. The major reaction product obtained with the three isoenzymes was 5-hydroxymethyl-2-furancarboxylic (HMFCA), a precursor in polyesters and pharmaceuticals production, and very little subsequent conversion of this compound was observed. However, small concentrations of FDCA, a substitute for terephthalic acid in the production of polyesters, were also obtained. The oxidation of HMF was significantly boosted in the presence of catalase for PciGLOX2, leading to 70% HMFCA yield. The highest conversion percentages were observed on 2,5-furandicarboxaldehyde (DFF), a minor product from the reaction of PciGLOX on HMF. To bypass HMFCA accumulation and exploit the efficiency of PciGLOX in oxidizing DFF and 5-formyl-2-furan carboxylic acid (FFCA) towards FDCA production, HMF was oxidized in a cascade reaction with an aryl alcohol oxidase (UmaAAO). After 2 h of reaction, UmaAAO completely oxidized HMF to DFF and further to FFCA, with FDCA only being detected when PciGLOX3 was added to the reaction. The maximum yield of 16% FDCA was obtained 24 h after the addition of PciGLOX3 in the presence of catalase. Conclusions At least two conversion pathways for HMF oxidation can be considered for PciGLOX; however, the highest selectivity was seen towards the production of the valuable polyester precursor HMFCA. The three isoenzymes showed differences in their catalytic efficiencies and substrate specificities when reacted with HMF derivatives. Electronic supplementary material The online version of this article (10.1186/s40694-019-0067-8) contains supplementary material, which is available to authorized users.

C4 grasses are common species in rangelands around the world and represent an attractive option for second-generation biofuels production. Although they display a high polysaccharide content and reach great levels of biomass accumulation, there is a major technical issue to be solved before they can be considered as biofuels feedstock: lignin removal. Concerning this, Pycnoporus and Ganoderma fungal genera have been highlighted due to their ability to hydrolyze lignocellulose. The goals here were to evaluate the pretreatment efficiency using P. sanguineus and G. applanatum secretomes harvested from a glucose-free inductive medium and to identify the fungal enzymatic activities responsible for the lignin degradation and glucose release. The findings show that P. sanguineus secretome exhibits a higher activity of lignocellulolytic enzymes compared to the one from G. applanatum. Interestingly, zymograms in presence of glucose suggest that a β-glucosidase isoform from P. sanguineus could be glucose-tolerant. The proteomic approach carried out allowed to identify 73 and 180 different proteins for G. applanatum and P. sanguineus secretomes, respectively, which were functionally classified in five main categories, and a miscellaneous group. Many uncharacterized proteins were found in both secretomes, reflecting that greater research is still needed for a better comprehension of lignocellulose degradation.

The purpose of this study was to investigate the effect of diet supplementations on biochemical, hematological, and redox metabolism parameters in streptozotocin-induced diabetic rats. Healthy male Wistar rats and streptozotocin-induced diabetic rats were provided diets supplemented with 20% of Pinus sp. sawdust or Pycnoporus sanguineus mycelium for 4 weeks. Diabetic rats treated with both Pinus sp. sawdust- and P. sanguineus mycelium-supplemented diets presented a significant decrease in non-HDL cholesterol of 38.43% and 33.53% and triglycerides of 70.03% and 69.81%, respectively, compared to diabetic control. As far as we know, this is the first report of a significant decrease in serum lipids attributed to these supplementations. Even though with the alterations in hematological and redox metabolism parameters related to these diet treatments, our data suggest that Pinus sp. sawdust and Pycnoporus sanguineus mycelium could be a useful a diet supplement to control diabetic dyslipidemia in animals. PRACTICAL APPLICATIONS: Pinus sp. sawdust is a residue from the wood industry that can be reused as a substrate to cultivate Pycnoporus sanguineus mycelium. Both species have specific and rich natural compounds. The results of the present study surprisingly showed that diets supplemented with the isolated substrate (Pinus sp. sawdust) and Pycnoporus sanguineus mycelium significantly decrease non-HDL cholesterol and triglycerides in induced diabetic animal model. These new natural approaches can be interesting to develop a nutraceutical to treat dyslipidemia.

14 p.-8 fig.-1 tab., White-rot (WR) fungi are pivotal decomposers of dead organic matter in forest ecosystems and typically use a large array of hydrolytic and oxidative enzymes to deconstruct lignocellulose. However, the extent of lignin and cellulose degradation may vary between species and wood type. Here we combined comparative genomics,transcriptomics and secretome proteomics to identify conserved enzymatic signatures at the onset of wood decaying activity within the Basidiomycota genus Pycnoporus. We observed strong conservation in the genome structures and the repertoires of protein coding genes across the four Pycnoporus species described to date,despite the species having distinct geographic distributions. We further analyzed the early response of P. cinnabarinus, P. coccineus and P. sanguineus to diverse (ligno)-cellulosic substrates. We identified a conserved set of enzymes mobilized by the three species for breaking down cellulose, hemicellulose and pectin.The co-occurrence in the exo-proteomes of H2O2 producing enzymes with H2O2 consuming enzymes was a common feature of the three species, although each enzymatic partner displayed independent transcriptional regulation. Finally, cellobiose dehydrogenase-coding genes were systematically co-regulated with at least one AA9 LPMO gene, indicative of enzymatic synergy in vivo. This study highlights a conserved core white-rot fungal enzymatic mechanism behind the wood decaying process., This work was supported by The French National Agency for Research (ANR-14-CE06-0020). The work conducted by the U.S. Department of Energy Joint Genome Institute, a DOE Office of Science User Facility, is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231. The work by CSIC was supported by the Ministerio de Ciencias, Innovación y Universidades de España (BIO2017-86559-R).

Biorecovery is emerging as a promising approach to retrieve gold from various sources, while its efficiency is usually restricted by the limited functional groups on natural microbial biomass surface. This study aims to intensify Pycnoporus sanguineus boosted sorption-reduction coupled gold biorecovery process via microbial surface modification. Results showed that grafting polyallylamine hydrochloride onto P. sanguineus biomass surface increased amino group content on microbial biomass surface from 1.29 to 2.81 mmol/g. When applying modified biomass to gold biorecovery with initial gold concentrations of 1.0, 2.0 and 3.0 mM, biosorption equilibrium time shortened to the 12.5%, 37.5% and 41.7% of those obtained with pristine biomass, and sorption rate constants correspondingly increased to 11.2, 3.1 and 3.7 folds as well. Maximum sorption capacity increased 30% and the affinity between biomass and gold enhanced heavily after microbial surface modification. Meanwhile, microbial surface modification favored gold reduction and gold nanoparticles (AuNPs) formation. The change of microbial biomass morphology from smooth surface with some branched structure to layered stacking structure with many pores and the increase of amino group content on microbial biomass surface were the main impetus for the gold bioreocovery process intensification.

Background and Aims: The production of edible fungi is affected by bacterial, fungal and viral diseases, which very often cause large losses. In the productionof mushrooms of the genus Pleurotus, the fungi of Trichoderma spp. represent a serious problem of contamination and although there are some chemical compoundsthat control the infection, they are not entirely safe for human consumption. As a consequence, alternatives are being searched for through biotechnology,such as the one presented in this paper.Methods: Strains of fungi of the genus Trichoderma were isolated from the substrate where Pleurotus ostreatus was being cultivated. These were identifiedmorphologically and molecularly, followed by tests to inhibit the growth of Trichoderma strains in both agar and wheat straw, using a cetonic extract of thedehydrated fruiting body of Pycnoporus sp.Key results: Two strains of Trichoderma (T. pleuroti and T. atrobrunneum, belonging to the clade of T. harzianum) were isolated from infected substrate obtainedin production modules of Pleurotus ostreatus located in Tlaquitenango and Cuernavaca, in the state of Morelos, Mexico. The effect of a cetonic extract of thefruiting body of Pycnoporus sp. on the mycelial growth of the isolated strains of Trichoderma was also evaluated, observing decrease in mycelial growth rate inPetri dish up to 72% and on lignocellulosic substrate both mycelial growth and sporulation were delayed up to 10 days.Conclusions: The extract of Pycnoporus sp. could be an alternative to control the infection by Trichoderma spp. in mushroom cultures of the genus Pleurotus.Key words: disease control, edible mushrooms, lignocellulosic substrate.

Bioremediation is a strategy to mitigate environmental impacts of hazardous pollutants from anthropogenic sources. Natural byproducts, including agroindustrial wastes (AW) can be used to induce enzyme biosynthesis, leading up to enhancement of pollutants degradation process. Therefore, this study aimed to evaluate the use of cupuacu, Theobroma grandiflorum AW as Pycnoporus sanguineus Laccase (Lac) inducer in order to promote 17-α-ethinylestradiol (EE2) bioremediation. The macro and micro-nutrients levels of cupuacu AWs were evaluated in order to establish further correlations with enzymatic biosynthesis induction. The fungus was cultivated for 7 days in temperature of 28 ± 2 °C and agitation of 150 rpm. For bioremediation, Lac enzymatic extract was added to EE2 solution (10 µg mL−1) and the percentage of removal was evaluated by HPLC after 1–24 hr of reaction. At optimized conditions, the enzyme extract production was remarkably enhanced by adding only 1% (w/v) of cupuacu AW. Lac activity reached ...

Wood offers natural advantages tremendously such as their elasticity, flexibility and resistance against compression as well as force impact. Therefore, wood material is astonishingly better material than concrete for railway sleepers. In spite of wood material’s advantages, the material is susceptible to attack by organisms. This paper observes the service life of wood sleepers in Java, Indonesia. Wood sleepers were observed visually from the ground railway system, Banten Province includes wood species identification macroscopically and organism’s attack. Results reveal that natural wood resistance of the sleepers are relatively high, then sleepers were predicted to be last long. Sleepers wood species are teak ( Tectona grandis ), balau ( Hopea sp.), merbau ( Intsia bijuga ), and ulin ( Eusideroxylon zwageri ). Sleepers were also found deteriorated by fungus during five months instalment. Decaying fungi observed are Dacryopinax sp. , D. spathularia, Lentinus sp ., Panus sp ., Pycnoporus sp. , P. sanguineus, Polyporus spp., Schizophyllum commune , and Trametes sp.

Since fossil fuel resources are limited there is a necessity to produce alternative types of fuel that are renewable and eco-friendly. Basidiomycetes are potential sources of enzymes that can be used for biofuel production. The current study aimed to isolate basidiomycetes from Sri Lanka, screen them for lignocellulose degrading enzymes, namely cellulase, xylanase, laccase, Mn peroxidase and lignin peroxidase and study the effect of potential inducers of laccase production. Among the eighteen basidiomycetes isolated, Pycnoporus sp. produced the highest cellulase activity (0.23 FPU/ml) whereas Phlebiopsis sp. produced the highest xylanase activity (5.4 U/ml). Earliella scabrosa produced the highest laccase (91.2 U/l) and Mn peroxidase (17.5 U/l) activities. Lignin peroxidase activity was not detected from the isolates. Effect of alkali lignin, Cu2+ and rice bran, three potential inducers, on laccase production by E. scabrosa, Pycnoporus sp. and Trametes hirsuta (M40) was studied. Results indicated that alkali lignin (2 g/l) significantly increased laccase production from Pycnoporus sp. and T. hirsuta (M40) while Cu2+ increased laccase production from E. scabrosa and T. hirsuta at 200 μM. Use of rice bran (10 g/l) resulted in higher laccase production from E. scabrosa and Pycnoporus sp. High laccase activity (79600 U/l) was obtained from E. scabrosa by using 50 g/l of rice bran and by extending the incubation period to 18 days. The study concluded that some of the basidiomycetes isolated can produce significant lignocellulose degrading enzyme activities.

Fungi are an important source of traditional medicine in Asian and Western countries. Globally, cancer is one of the leading causes of death with 22 million cases predicted by 2030. The cytotoxicity of 13 South African macrofungal species against HeLa, HT-29, MCF-7, MIA PaCa-2 and PC-3 cancer cell lines was measured using the 3-(4,5-dimethyl-2-thiazolul)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, and the IC 50 values determined for the most cytotoxic macrofungal species. Fomitopsis lilacinogilva had IC 50 values of 42.2, 55.1, 63, 48.8 and 79.8 μg/mL against the HeLa, HT-29, MCF-7, MIA PaCa-2 and PC-3 cancer cell lines, respectively. Gymnopilus junonius had IC 50 values of 55.1 and 52.5 μg/mL against the HeLa and MIA PaCa-2 cancer cell lines, respectively. Pycnoporus sanguineus had IC 50 values of 24.2, 48.1, 32.7, F. lilacinogilva , G. junonius and P. sanguineus was confirmed using Hoechst and phalloidin staining. Morphological changes included condensed chromatin, membrane blebbing, loss of cytoskeletal arrangement and rounding up of cells, as well as multi- and micronuclei. Macrofungal research in South Africa remains a novel field, and this study provides interesting data on the cytotoxicity of South African macrofungal species against some of the most common cancers affecting humans worldwide.

Mono and dual cultures of four white-rot basidiomycete species were evaluated for cellulase and xylanase activity under submerged fermentation conditions. Co-cultivation of Pycnoporus coccineus or Trametes hirsuta with Schizophyllum commune displayed antagonistic interactions resulting in the decrease of endoglucanase and total cellulase activities. In contrast, increases in cellulase and xylanase activity were revealed through the compatible interactions of Irpex lacteus with S. commune. Co-cultivation conditions were optimized for maximum enzyme production by I. lacteus and S. commune, the best producers of cellulase/xylanase and β-glucosidase, respectively. An optimized medium for the target enzyme production by the mixed culture was established in a laboratory fermenter yielding 7U/mL total cellulase, 142U/mL endoglucanase, 104U/mL xylanase, and 5.2U/mL β-glucosidase. The dual culture approach resulted in an enzymatic mixture with 11% improved lignocellulose saccharification potential compared to enzymes from a monoculture of I. lacteus.

White rot fungi have been proved to be a promising option for the removal of heavy metals, understanding their toxic response to heavy metals is conducive to developing and popularizing fungi-based remediation technologies so as to lessen the hazard of heavy metals. In this study, Cr(VI)-induced oxidative stress and apoptosis in Pycnoporus sanguineus, a species of white rot fungi were investigated. The results suggested that high level of Cr(VI) promoted the formation of ROS, including H 2 O 2 , O 2 •− and ·OH. With the increment of Cr(VI) concentration, the SOD and CAT activity along with GSH content increased within the first 24 h, but decreased afterward, companied with a significant enhancement of MDA content. Cr(VI)-induced oxidative damage further caused and aggravated apoptosis in P. sanguineus, especially at Cr(VI) concentrations above 20 mg/L. Cr(VI)-induced apoptosis was involved with mitochondrial dysfunction including mitochondrial depolarization, the enhancement of mitochondrial permeability and release of cytochrome c. The early and late apoptosis hallmarks, such as metacaspase activation, phosphatidylserine (PS) externalization, DNA fragmentation and the nuclear condensation and fragmentation were observed. Moreover, we also found disturbances of ion homeostasis, which was featured by K + effluxes and overload of cytoplasmic and mitochondrial Ca 2+ .Based on these results, we suggest that Cr(VI) induced oxidative stress and apoptosis in white rot fungi, P. sanguineus .

Functionalization of thin films with organic ligands has been the subject of intense research due to their potential application as heterogeneous molecular nanosystems. In this work, self-assembled monolayers of thiols (16-mercaptohexadecanoic acid and 11-mercaptoundecanol) were used to bind laccase (from Pycnoporus sanguineus CS43) to nanostructured gold thin films obtained by DC sputtering. Sputtering power, sputtering pressure and substrate temperature were optimized to enhance the activity of the immobilized biomolecules. Scanning electron microscopy, confocal microscopy, X-ray diffraction and UV-vis spectroscopy were used to characterize the SAM-functionalized gold substrates. Our results demonstrate that the highest immobilized enzyme activity values can be achieved on substrates of surface roughness ˜200 nm and Au particle size of about 14 nm. The outstanding quality of the as-prepared substrates makes them particularly attractive as bionanosensors.

Inflammatory bowel disease (IBD) is a chronic autoimmune disease associated with various risk factors. Pycnoporus sanguineus (L.) Murrill is a saprotrophic fungus used worldwide for its industrial and medical purposes. Here, polysaccharide from P. sanguineus (PPS) was explored for its antiinflammatory potential in a murine colitis model of IBD induced by dextran sulfate sodium (DSS). PPS ameliorated the colitis as manifested by the lowered disease activity index (DAI), prolonged colon, and reduced serum lipopolysaccharide (LPS). PPS recovered the histological lesion by upregulating the expressions of Zonula occludens-1 (ZO-1), E-cadherin, and proliferating cell nuclear antigen (PCNA). PPS inhibited the helper T cells (Th)-mediated immune response by decreasing the proportions of Th cells (including Th2 cells, Th17 cells, and regulatory T cells), which was accompanied with reductions on myeloperoxidase (MPO) activity and releases of several interleukins and chemokines within the colon. Moreover, PPS exhibited an evident inhibition on autophagy, in which the ratio of light chain 3 (LC3) II/I was declined, while the expression of p62 and Beclin-1 was increased. The present study highlighted important clinical implications for the treatment application of PPS against IBD, which relies on the regulation of Th cells repertoire and autophagy suppression to restore epithelium barrier.

Pycnoporus sanguineus, an edible mushroom, produces antimicrobial and antitumor bioactive compounds and pH- and thermo- stable laccases that have multiple potential biotechnological applications. Here we reported the complete genome of the species Pycnoporus sanguineus ACCC 51,180 by using the combination of Illumina HiSeq X Ten and the PacBio sequencing technology. The represented genome is 36.6 Mb composed of 59 scaffolds with 12,086 functionally annotated protein-coding genes. The genome of Pycnoporus sanguineus encodes at least 19 biosynthetic gene clusters for secondary metabolites, including a terpene cluster for biosynthesis of the antitumor clavaric acid. Seven laccases were identified, while 22 genes were found to be involved in the kynurenine pathway in which the intermediate metabolite 3-hydroxyanthranilic acid were catalyzed by laccases into cinnabarinic acid. This study represented the third genome of the genus Pycnoporus, and wound facilitate the exploration of useful sources from Pycnoporus sanguineus for future industrial applications.

The influence of Cr(VI) on the degradation of tetrabromobisphenol A (TBBPA) by a typical species of white rot fungi, Pycnoporus sanguineus , was investigated in this study. The results showed that P. sanguineus together with its intracellular and extracellular enzyme could effectively degrade TBBPA. The degradation efficiency of TBBPA by both P. sanguineus and its enzymes decreased significantly when Cr(VI) concentration increased from 0 to 40 mg/L. The subsequent analysis about cellular distribution of TBBPA showed that the extracellular amount of TBBPA increased with the increment of Cr(VI) concentration, but the content of TBBPA inside fungal cells exhibited an opposite variation tendency. The inhibition of TBBPA degradation by P. sanguineus was partly attributed to the increase of cell membrane permeability and the decrease of cell membrane fluidity caused by Cr(VI). In addition, the decline of H + -ATPase and Mg 2+ -ATPase activities was also an important factor contributing to the suppression of TBBPA degradation in the system containing concomitant Cr(VI). Moreover, the activities of two typical extracellular lignin-degrading enzymes of P. sanguineus , MnP and Lac, were found to descend with ascended Cr(VI) level. Cr(VI) could also obviously suppress the gene expression of four intracellular enzymes implicated in TBBPA degradation, including two cytochrome P450s, glutathione S-transferases and pentachlorophenol 4-monooxygenase, which resulted in a decline of TBBPA degradation efficiency by fungal cells and intracellular enzyme in the presence of Cr(VI). Overall, this study provides new insights into the characteristics and mechanisms involved in TBBPA biodegradation by white rot fungi in an environment where heavy metals co-exist.

International audience; Production of second-generation bioethanol uses lignocellulose from agricultural by-products such as sugarcane bagasse (SCB). A lignocellulose pre-treatment is required to degrade lignin, ensuring further efficient sacchar-ification. Two experimental designs were set up to define culture conditions of Pycnoporus sanguineus in meso-cosms to increase laccase activities and thus delignification. The first experimental design tested the effect of phenolic complementation (via coffee pulp) and the use of urea as a simple nitrogen source and the second defined more precisely the percentages of coffee pulp and urea to enhance delignification. The responses measured were: lignocellulolytic activities, laccase isoform profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the chemical transformation of the substrate using solid-state NMR of 13C. Adding 10% of coffee pulp increased laccase activities and fungal biomass (32.5% and 16% respectively), enhanced two constitutive isoforms (R f 0.23 and 0.27), induced a new isoform (R f 0.19) and led to a decrease in total aromatics. However, higher concentrations of coffee pulp (25%) decreased laccase and cellulase activities but no decrease in aromaticity was observed, potentially due to the toxic effect of phenols from coffee pulp. Moreover, laccase production was still inhibited even for lower concentrations of urea (0-5%). Our findings revealed that an agricultural by-product like coffee pulp can enhance laccase activity-though to a threshold-and that urea limited this process, indicating that other N-sources should be tested for the biological delignification of SCB.

Emerging contaminants like hormones can interfere ecological life health. Both liquid cultivation and direct enzymatic process have been proved effective for hormonal removal. This study aimed to assess direct removal of 17 β -estradiol (E2) from solid medium poultry litter (PL) by cultivating laccase-producing fungi toward clean treatment of PL. Before E2 removal test, laccase production was optimized by addition of citric acid and lignocellulosic biomasses into PL, which turned out to dramatically enhance laccase production to varying degrees. The highest laccase activity of approximately 16 U/g medium was produced by Pycnoporus sp. SYBC-L3 on PL mixed with sweat sorghum bagasse at a 2 to 3 (g/g) ratio. An extra dose of E2 spiked in the PL/biomass mixture (final concentration of 10 μg/g) was used to test its effect on laccase production and subsequent E2 removal. Over 80% E2 was removed from the above cultivation system, and E2 removal appeared to be positively correlated with laccase activity. Our results might present an alternative clean strategy for livestock manure treatment before land application.

A unique, non-leaching and durable novel anti-biocide polyacrylamide-starch coating (pAam-g-S) has successfully been synthesized. In this study, a grafted cationic-functionalized polymeric coating was designed and cross-linked with native starch extracted from Dioscorea hispida sp. yam to afford multifunctional anti-fungal and anti-termite coating. Leaching of the disinfectant into the environment was eliminated because no toxic chemicals were used. The copolymerization was successfully carried out to produce various ratios of pAam-g-S coating using modification of starch-gel. Results obtained from Fourier Transform Infrared-Attenuated Total Reflectance (FTIR-ATR) confirmed the monomer pAam was grafted onto the starch backbone as shown by the cross-linked peak at 1642 cm−1. Supported by differential scanning calorimetry (DSC), the highest transition glass temperature was observed at 154 °C. The coating was designed to continuously decontaminate against pathogenic fungus and termite in addition to afford preliminary anti-biocides properties. Facile anti-fungal and anti-termite evaluations were conducted via total weight loss of wood (%) studies using white rot fungus, Pycnoporus. sanguineus and termite Coptotermes curvignathus respectively. In order to verify the coating performance, gel fractions, morphologies and heat resistant properties were also investigated.

The genome of the white rot fungus Pycnoporus cinnabarinus includes a large number of genes encoding enzymes implicated in lignin degradation. Among these, three genes are predicted to encode glyoxal oxidase, an enzyme previously isolated from Phanerochaete chrysosporium . The glyoxal oxidase of P. chrysosporium is physiologically coupled to lignin-oxidizing peroxidases via generation of extracellular H 2 O 2 and utilizes an array of aldehydes and α-hydroxycarbonyls as the substrates. Two of the predicted glyoxal oxidases of P. cinnabarinus , GLOX1 ( Pci GLOX1) and GLOX2 ( Pci GLOX2), were heterologously produced in Aspergillus niger strain D15#26 ( pyrG negative) and purified using immobilized metal ion affinity chromatography, yielding 59 and 5 mg of protein for Pci GLOX1 and Pci GLOX2, respectively. Both proteins were approximately 60 kDa in size and N-glycosylated. The optimum temperature for the activity of these enzymes was 50°C, and the optimum pH was 6. The enzymes retained most of their activity after incubation at 50°C for 4 h. The highest relative activity and the highest catalytic efficiency of both enzymes occurred with glyoxylic acid as the substrate. The two P. cinnabarinus enzymes generally exhibited similar substrate preferences, but Pci GLOX2 showed a broader substrate specificity and was significantly more active on 3-phenylpropionaldehyde. IMPORTANCE This study addresses the poorly understood role of how fungal peroxidases obtain an in situ supply of hydrogen peroxide to enable them to oxidize a variety of organic and inorganic compounds. This cooperative activity is intrinsic in the living organism to control the amount of toxic H 2 O 2 in its environment, thus providing a feed-on-demand scenario, and can be used biotechnologically to supply a cheap source of peroxide for the peroxidase reaction. The secretion of multiple glyoxal oxidases by filamentous fungi as part of a lignocellulolytic mechanism suggests a controlled system, especially as these enzymes utilize fungal metabolites as the substrates. Two glyoxal oxidases have been isolated and characterized to date, and the differentiation of the substrate specificity of the two enzymes produced by Pycnoporus cinnabarinus illustrates the alternative mechanisms existing in a single fungus, together with the utilization of these enzymes to prepare platform chemicals for industry.

A laccase from the basidiomycete Pycnoporus sanguineus strain RVAN5 was evaluated for its ability to decolorize synthetic dyes and denim bleaching. Dye color reduction and denim bleaching were monitored at different dye concentrations and incubation times. Dye decolorization by Pycnoporus sanguineus fungal crude extract (FCE) ranged from 80 to 96% within 2-4 h at 25-65 °C. Comparable results were obtained when violuric acid (VA) was added as mediator to the FCE, however, the number of decolorized dyes increased significantly. Dye decolorization rates with VA varied of initial and final optical density (595 nm) values of 2.5-3.0 and 0.2-0.02, respectively. P. sanguineus FCE had no substantial effect on denim bleaching when used alone, notwithstanding, the mixture of FCE with VA (10 mM) showed significant denim color reduction values and considerably higher than those obtained with a bleaching enzyme from a commercial formulation; CIElab values obtained with FCE/VA mixture were of ΔL = 6.4, versus a ΔL 1.4 value obtained with an enzyme from commercial formulation.

Laccase has been emerged as a valuable green biocatalyst in cleaner production, which requires vast amount of laccase at low production cost and with good properties. The current work evaluated a wild strain Pycnoporus sp. SYBC-L3 as an efficient microbial cell factory for laccase production. High-cell density cultivation (HCDC) of L3 was developed in a 5-L bioreactor by adjusting the ratio of inoculum to medium, achieving over 4-fold of both laccase activity (240 U/mL) and fungal biomass (20 g/L) than those obtained under routine procedures. Laccase productivity was further assessed at pilot-scale in a 500-L and a 5-ton bioreactor, where the highest activity of around 60 U/mL and 80 U/mL were produced, respectively. Cost percentage analysis for the 5-ton reactor demonstrated that labor was the highest portion (50%) in total cost for industrial production in this case. Using a one-step purification scheme, laccase was purified to homogeneity from the culture broth with yield of 44% and 97% purity. Circular dichroism (CD) analyses suggested that the secondary structure of laccase was mainly composed of β-sheet that played a vital role in maintaining laccase activity, and also revealed the potential mechanisms for the changes in laccase activities under varying conditions. Powdered laccase was efficiently prepared from cell-free culture broth using spray drying technique with above 74% activity retained. This study provides important information for an efficient and scalable production of laccase.

A new poroid wood-inhabiting fungal genus, Leifiporia, is proposed, based on morphological and molecular evidence, which is typified by L. rhizomorpha sp. nov. The genus is characterized by an annual growth habit, resupinate basidiocarps with white to cream pore surface, a dimitic hyphal system with generative hyphae bearing clamp connections and branching mostly at right angles, skeletal hyphae present in the subiculum only and distinctly thinner than generative hyphae, IKI–, CB–, and ellipsoid, hyaline, thin-walled, smooth, IKI–, CB– basidiospores. Sequences of ITS and LSU nrRNA gene regions of the studied samples were generated, and phylogenetic analyses were performed with maximum likelihood, maximum parsimony and Bayesian inference methods. The phylogenetic analysis based on molecular data of ITS + nLSU sequences showed that Leifiporia belonged to the core polyporoid clade and was closely related to Diplomitoporus overholtsii and Lopharia cinerascens, and then grouped with Pycnoporus and Trametes. Further investigation was obtained for more representative taxa in the Polyporaceae based on nLSU sequences, in which the results demonstrated that the genus Leifiporia formed a monophyletic lineage with a strong support (100 % BS, 100 % BP, 1.00 BPP). Both morphological and molecular evidence confirmed the placement of the new genus in the core polyporoid clade. In addition, a new combination, Leifiporia eucalypti, is proposed based on examination of its type material and phylogeny.