Pycnoporus Sanguineus Research Articles

Production, Purification, Characterization, and Application of Halotolerant and Thermostable Endoglucanase Isolated from Pycnoporus sanguineus

Production, Purification, Characterization, and Application of Halotolerant and Thermostable Endoglucanase Isolated from Pycnoporus sanguineus

Detection of Acetaminophen in Groundwater by Laccase-Based Amperometric Biosensors Using MoS2 Modified Carbon Paper Electrodes.

The use of enzyme-based biosensors for the detection and quantification of analytes of interest such as contaminants of emerging concern, including over-the-counter medication, provides an attractive alternative compared to more established techniques. However, their direct application to real environmental matrices is still under investigation due to the various drawbacks in their implementation. Here, we report the development of bioelectrodes using laccase enzymes immobilized onto carbon paper electrodes modified with nanostructured molybdenum disulfide (MoS2). The laccase enzymes were two isoforms (LacI and LacII) produced and purified from the fungus Pycnoporus sanguineus CS43 that is native to Mexico. A commercial purified enzyme from the fungus Trametes versicolor (TvL) was also evaluated to compare their performance. The developed bioelectrodes were used in the biosensing of acetaminophen, a drug widely used to relieve fever and pain, and of which there is recent concern about its effect on the environment after its final disposal. The use of MoS2 as a transducer modifier was evaluated, and it was found that the best detection was achieved using a concentration of 1 mg/mL. Moreover, it was found that the laccase with the best biosensing efficiency was LacII, which achieved an LOD of 0.2 µM and a sensitivity of 0.108 µA/µM cm2 in the buffer matrix. Moreover, the performance of the bioelectrodes in a composite groundwater sample from Northeast Mexico was analyzed, achieving an LOD of 0.5 µM and a sensitivity of 0.015 µA/µM cm2. The LOD values found are among the lowest reported for biosensors based on the use of oxidoreductase enzymes, while the sensitivity is the highest currently reported.

Production, characterization, and application of a new chymotrypsin-like protease from Pycnoporus sanguineus

Proteases are the most commercialized enzymes and are widely used in industrial processes, especially in the dairy industry. Cheese production is increasing disproportionately to the availability of calf rennet, the main coagulant used in this process. In addition to the cost of rennet, there is also a religious and dietary objection to the use of calf proteases. An alternative is the production of microbial enzymes via solid state fermentation (SSF), which uses agroindustrial residues as a source of nutrients and substrate. To obtain proteases with milk-clotting activity, Pycnoporus sanguineus was produced via SSF using agroindustrial residue wheat bran as the culture medium. Caseinolytic activity and maximum enzyme production were obtained after 96 h of cultivation. The protease was identified as a chymotrypsin-like serine protease after testing it against specific substrates and inhibitors. The protease from P. sanguineus had a specific activity of 741.3 U mg−1 and was able to coagulate reconstituted skim milk and whole milk, with and without the addition of calcium. Coagulation was affected by time, temperature, and calcium and enzyme concentrations. Therefore, we describe herein for the first time a protease obtained from P. sanguineus, a chymotrypsin-like serine protease with milk-clotting activity that has potential applications in the dairy industry.

Cardioprotective effect of crude polysaccharide fermented by Trametes Sanguinea Lyoyd on doxorubicin-induced myocardial injury mice

Doxorubicin (DOX) is a broad-spectrum anti-tumor drug, but its clinical application is greatly limited because of the cardiotoxicity. Thus, exploration of effective therapies against DOX-induced cardiotoxicity is necessary. The aim of this study is to investigate the effects and possible mechanisms of Trametes Sanguinea Lyoyd fermented crude polysaccharide (TSLFACP) against DOX-induced cardiotoxicity. We investigated the protective effects of TSLFACP on myocardial injury and its possible mechanisms using two in vitro cells of DOX-treated cardiomyocytes H9C2 and embryonic myocardial cell line CCC-HEH-2 and a in vivo mouse model of DOX-induced myocardial injury. We found that TSLFACP could reverse DOX-induced toxicity in H9C2 and CCC-HEH-2 cells. Similarly, we found that when pretreatment with TSLFACP (200 mg/kg, i.g.) daily for 6 days, DOX-induced myocardial damage was attenuated, including the decrease in serum myocardial injury index, and the amelioration in cardiac histopathological morphology. Additionally, immunohistochemistry and western blotting were used to identify the underlying and possible signal pathways. We found that TSLFACP attenuated the expression of LC3-II, Beclin-1 and PRAP induced by DOX. In conclusion, our results demonstrated that TSLFACP could protect against DOX-induced cardiotoxicity by inhibiting autophagy and apoptosis.

Modification of Wood by Tannin-Furfuryl Alcohol Resins–Effect on Dimensional Stability, Mechanical Properties and Decay Durability

Furfurylation is a well-known wood modification technology. This paper studied the effect of tannin addition on the wood furfurylation. Three kinds of dicarboxylic acids, adipic acid, succinic acid, and tartaric acid, as well as glyoxal as a comparing agent, were used to catalyse the polymerisation of furanic or tannin-furanic solutions during wood modification. Impregnation of furanic or tannin-furanic solution at a certain concentration into the wood followed with curing at 103°C for a specific duration was performed for the wood modification. Different properties of the modified woods like dimensional stability, resistance of treatment to leaching, mechanical properties, decay durability against white-rot (Coriolus versicolor, Pycnoporus sanguineus) and brown-rot (Coniophora puteana) as well as their chemical and anatomical characteristics were evaluated. Results revealed that the partial substitution of FA by the tannins improved the fixation of the chemicals impregnated in wood. Further, dimensional stability, leaching resistance, Brinell hardness, modulus of elasticity/modulus of rupture, and decay durability properties of the furfurylated wood were also improved in the presence of tannins. Scanning electron microscopy revealed the deposition of the polymer in the wood lumen cells and in the wood cell walls.

Oxalic acid degradation in wood-rotting fungi. Searching for a new source of oxalate oxidase

Oxalate oxidase (EC 1.2.3.4) is an oxalate-decomposing enzyme predominantly found in plants but also described in basidiomycete fungi. In this study, we investigated 23 fungi to determine their capability of oxalic acid degradation. After analyzing their secretomes for the products of the oxalic acid-degrading enzyme activity, three groups were distinguished among the fungi studied. The first group comprised nine fungi classified as oxalate oxidase producers, as their secretome pattern revealed an increase in the hydrogen peroxide concentration, no formic acid, and a reduction in the oxalic acid content. The second group of fungi comprised eight fungi described as oxalate decarboxylase producers characterized by an increase in the formic acid level associated with a decrease in the oxalate content in their secretomes. In the secretomes of the third group of six fungi, no increase in formic acid or hydrogen peroxide contents was observed but a decline in the oxalate level was found. The intracellular activity of OXO in the mycelia of Schizophyllum commune, Trametes hirsuta, Gloeophyllum trabeum, Abortiporus biennis, Cerrena unicolor, Ceriosporopsis mediosetigera, Trametes sanguinea, Ceriporiopsis subvermispora, and Laetiporus sulphureus was confirmed by a spectrophotometric assay.

Transforming second-generation ethanol production waste into sustainable enzymatic cosmetic inputs

Agro-industrial residues were used as a successful, cheap and renewable substrate, capable of increasing activity of ligninolytic enzymes for further use in melanin degradation assays to develop inputs for the treatment of skin hyperpigmentation. The analysis of melanin degradation was performed in a quantitative and qualitative way. The enzymatic crude broth extract, rich in laccases , produced by Marasmiellus palmivorus VE111 in medium formulated with lignocellulosic residues of Araucaria angustifolia subjected to hydrolysis and alcoholic fermentation , degraded almost 33% of the melanin content and enzymatic crude broth extract produced by Pycnoporus sanguineus PR_32 degraded 13%, in just 8 h, using vanillin as a mediator. Proton 2D NMR spectra show melanin changes after laccase treatment. This work has provided an insight for the development of ecofriendly, sustainable and innovative cosmetics to deal with skin hyperpigmentation. • By-products from 2nd generation ethanol production act as inducer to laccases activity. • Vanillin can be used as a safe redox mediator for laccase activity in cosmetics. • Laccase from Marasmiellus palmivorus VE111 can degrade melanin. • HSQC 2D NMR data shows changes in melanin after enzymatic treatment.

Community structure of polypores (Basidiomycota) in a restored Brazilian Forest

Polypores (Basidiomycota) are of great importance in several forest areas since they are fundamental for wood decomposition and nutrient recycling, which is essential for the functioning of ecosystems. This study assessed the polypore community structure in a restoration area and its use as a parameter to monitor restoration. Our study was carried out in Parque Florestal São Marcelo Private Natural Heritage Reserve (RPPN), a protected area of 240 ha, formed in 2002 from reforestation with native species, 13 years after the restoration measures. The polypore community in the area was characterized according to the richness, abundance, and functionality. Results were compared with data from two natural reference areas near the study site and in the same forest type, i.e., riparian forest, but with different land use history. One hundred and eighteen specimens belonging to 31 species were collected; 6.45% were abundant (Funalia rigida and Pycnoporus sanguineus), 12.90% common, 32.26% occasional, and 48.39% rare. Four functional groups based on the species' relative frequency on each substrate were found. Higher frequency of polypores was observed in substrates of smaller diameter (dead branches). The restored area showed a similar richness to the preserved area used as reference. In addition, the polypore community showed a structure similar to well-established areas. This result indicates that the RPPN restoration project was successful and that the polypore community structure can be suitable for evaluating and monitoring regions restored over time.

Pigment production by a newly isolated strain Pycnoporus sanguineus SYBC-L7 in solid-state fermentation.

Natural pigments are playing important roles in our daily lives. They not only make products colorful but also provide various health benefits for humans. In addition, Pycnoporus genus, listed as food- and cosmetic-grade microorganism, is one of the promising organisms for developing natural pigments. In this study, a new fungal strain with high efficiency in producing intense orange pigments was isolated and identified as Pycnoporus sanguineus SYBC-L7. Different agro-industrial wastes were applied to evaluate the growth and pigment production of strain SYBC-L7. SYBC-L7 can grow rapidly and effectively produce pigments using wood chips as substrate in solid-state fermentation (SSF). Culture conditions were also optimized for value-added pigments production and the optimum production conditions were glucose as carbon source, ammonium tartrate as nitrogen source, initial pH 6.0, and relative humidity of 65%. Pigment components, cinnabarinic acid, tramesanguin, and 2-amino-9-formylphenoxazone-1-carbonic acid were confirmed by liquid chromatography-mass spectrometry. Meanwhile, an agar plate diffusion assay was performed to evaluate the antimicrobial activity of the pigment. These pigments showed more significant inhibition of Gram-positive than Gram-negative bacteria. The results showed that Pycnoporus sanguineus SYBC-L7 was able to cost-effectively produce intense natural orange pigments with antibacterial activity in SSF, which is the basis of their large-scale production and application.

Antibacterial activity of fruiting body extracts from Pycnoporus sanguineus mushroom

Antibacterial activity of fruiting body extracts from Pycnoporus sanguineus mushroom

Efficient biodegradation of straw and persistent organic pollutants by a novel strategy using recombinant Trichoderma reesei

Efficient biodegradation of lignocellulosic biomass needs a battery of enzymes targeting cellulose, hemicellulose, and lignin. In this study, recombinant Trichoderma reesei ZJ-09 with Pycnoporus sanguineus laccase gene was used to degrade rice straw by in situ production of laccase, xylanase, and cellulases under solid-state fermentation (SSF). Effects of parameters on key enzymes (cellulase, xylanase, and laccase) in biodegradation during SSF were investigated. Under the optimized SSF conditions, the FPA, xylanase activity, and laccase activity reached 110.47 FPU/g, 5787.59 IU/g, and 24.45 IU/g, respectively, on day 12. The obtained recombinant T. reesei SSF system achieved efficient degradation of rice straw with the final mass loss up to 51.16% which was 1.4-fold higher than the host strain. Further, bioconversion of rice straw into a novel laccase-enriched koji for persistent organic pollutants bioremediation (LKPB) was conducted by the optimized SSF system. LKPB was found to degrade persistent organic pollutants (POPs) effectively without the addition of mediators. 4-h removal rates of three POPs mediated by LKPB (87.21% for 2,4,5-trichlorophenol, 92.45% for nonylphenol, and 90.73% for oxytetracycline) were comparable to those achieved by laccase-co-mediator system. The newly established recombinant T. reesei SSF system could be potential to effectively degrade lignocellulosic wastes as well as organic pollutants.Graphical abstract

Optimization and Implementation of Fed-Batch Strategy to Produce Ligninolytic Enzyme from the White-Rot Basidiomycete Pycnoporus sanguineus in Bubble Column Reactor

The current work evaluates the production of ligninolytic enzyme optimization via response surface methodology using different inducers: acid cellulignin (CA); MnSO4 (Mn2+); CuSO4·5H2O (Cu2+); veratryl (3, 4-dimethoxybenzyl); alcohol (VA); Tween 80% (T80); and the carbon-to-nitrogen ratio (C/N). A further goal was implementing a fed-batch strategy to produce ligninolytic enzyme extracts from P. sanguineus 2512 using a bubble column reactor (BCR). The best optimized experimental condition in the shake flasks was a 7.5 C/N ratio, 0.025 g/L Cu2+, 1.5 mM Mn2+, 3.0 mM VA and 0.025 mM T80, resulting in 64,580, 9.10 and 80.72 U/L for Laccase (Lac), Manganese (MnP) and Lignin peroxidase (LiP) activities, respectively. In the BCR, three feedings were performed at 24 h intervals on the 6th, 7th and 8th days with a significant increase in Lac (99,600 U/L) and MnP (47.53 U/L) activities on the 8th day and a reduction on the 9th day of cultivation. The LiP activity peak was achieved on the 5th day (416 U/L) of cultivation, decreasing thereafter. Enzyme cocktails concentrated in hollow fiber in the third cultivation batch showed contents of 4 × 105 U/L, 220 U/L and 2.5 g/L for Lac, MnP and total proteins, respectively. The enzymatic cocktail with the highest LiP activity (1200 U/L) was obtained in the first batch. The results showed that the optimization of the biosynthesis of the ligninolytic enzymes provided satisfactory improvement in terms of Lac and MnP production per run.

Mycochemical analysis, anti-inflammatory and cytotoxic activities of Pycnoporus sanguineus (L.) Murrill, a medicinal mushroom from Gabon

A mycochemical study followed by an evaluation of anti-inflammatory and cytotoxic activities of Pycnoporus sanguineus, a species of lignicole mushroom exploited in traditional medicine in Gabon, was carried out on four extracts. Bioactive compounds were extracted by successive macerations with water, hydro-ethanol (50-50), ethanol solvents and sonication was used for dichloromethane-methanol (50-50) extraction. Agilent LC-MS system was used for the molecular profile of the different extracts and fractions. The fractionation of the extracts was made by flash chromatography and the purification using preparative chromatography. Structures of compounds responsible for the pharmacological activities were determinated by NMR. Cytotoxicity essay was carried out on PNT2 cells and the anticancer activities on A549 and PC3 cell lines. Two molecules have been isolated from this fungus; namely m/z 301.04 [M-H] + corresponding to cinnabarinic acid and m/z 287.06 [M-H]+ for which the NMR data were insufficient to confirm the molecular identification. Anti-inflammatory activity of the aqueous extract was stronger (IC50 = 197.82 μg/ml) than niflumic acid (IC50 = 809.27 μg/ml) but weaker than diclofenac (IC50 = 15.28 μg/ml). However, the hydro-ethanolic and ethanolic extracts of P. sanguineus, have a weaker anti-inflammatory activity than niflumic acid and diclofenac. About cytotoxicity, the crude extracts and fractions tested have anticancer and cytotoxic activities on A549, PC3 and PNT2 lines. However, these activities are more pronounced in the healthy line PNT2 than in the cancerous lines A549 and PC3.

Habitat Distribution of Fungal Species Collected at Lung NGOC Hoangnature Reserve

Through the survey of 4 different habitats in Lung Ngoc Hoang Nature reserve in Vietnam, totally 63 macro fungi have been found with 313 individuals. On forest habitat, 49 species with 116 individuals were obtained. In which, Pycnoporus sanguineus and Chaetocalathus columellifer had the highest number of individuals with 9 individuals, accounting for 7.76% of the total number of individuals collected in the forest. On the riverside habitat, 47 species with 89 individuals were obtained; The roadside habitat found 71 individuals with 42 species, in these two habitats Pycnoporus sanguineus was the most dominant species. On the agricultural habitat, there are 24 species with 37 individuals out of the total number of individuals collected. In this habitat, species Coprinus disseminates, Chaetocalathus columellifer and Lenzites palisoti were the most dominant with 3 individuals found, accounting for 8.11% of the total population here.

SUBSTRATO COM RESÍDUOS LIGNOCELULÓSICOS PARA CULTIVO DE Pycnoporus sanguineus

ABSTRACT Basidiomycete fungi that decompose wood produce substances with promising biological activity for the alternative control of plant diseases. The production of these substances can change according to the climatic conditions and the substrate used for fungal cultivation. The objective of this study was to develop a substrate with sawdust from Eucalyptus sp. and to verify its influence on biomass and cinnabarin production by Pycnoporus sanguineus. Sawdust was used in two particle sizes: less than 500 microns (G1) and between 500–841 microns (G2). Four isolates of P. sanguineus were plated on Petri dishes containing potato broth and agar media added with 0%, 1%, 5%, 10%, and 15% sawdust for each particle size. The largest final diameter of the colony and speed of mycelial growth were observed in the substrate with G1 particle size, with the Ps14 isolate showing the highest averages. For these variables, the sawdust concentration did not influence G1 granulometry and provided the highest values in G2 granulometry. Fresh mycelium mass and cinnabarin production showed the highest values in G2, with the isolated Ps13 and Ps08 showing the highest averages, whereas in G1, Ps14 had the best performance for the analyzed variables. These results indicate that sawdust from Eucalyptus sp., at concentrations of 10% and 15%, is an alternative for the in vitro cultivation of P. sanguineus, and that particle size influences the growth speed, fresh mass production, and cinnabarin content.

Isolation and characterization of wood-decomposing basidiomycetes from the Andean Forest in Boyacá, Colombia.

This study explores the biotechnological potential of lignocellulolytic fungi collected in an oak forest. Fungal collections were obtained from natural reserves located in Boyacá-Colombia, ranging from 2700 to 3000m.a.s.l. Twenty-three strains were isolated on malt agar, molecular characterization was performed, and ligninolytic and cellulolytic enzymatic activities were screened. Several white-rot fungi of biotechnological importance were identified as follows: Trametes sp., Trametes versicolor, Trametes villosa, Pycnoporus sanguineus, Bjerkandera adjusta, Lentinula boryana, Panus conchatus, Antrodia neotropica, Brunneoporus malicola, Laetiporus gilbertsonii, Stereum sp., Ganoderma sp., and Dichomitus sp. The strains T. versicolor 0554 and 0583, T. villosa 0562, and B. adusta 0556 showed the highest response in the qualitative enzymatic assays. These strains were used to determine their ability to decolorate the dyes aniline blue and Congo red, and it was found that T. villosa 0562 reached a level of decolorization close to 90% after 48h of submerged culture. The fungal strains obtained here could offer alternatives to develop a process to accomplish sustainable development objectives.

ANTIFUNGAL ACTIVITY OF ESSENTIAL OILS FROM NATIVE TREE SPECIES IN SOUTHERN BRAZIL

The objective of this exploratory work was to analyze the yield and chemical composition of essential oils (EO) from Blepharocalyx salicifolius [BS] and from two pools of EO obtained from Nectandra megapotamica [NM1 and NM2], as well as to evaluate their antifungal properties against the fungi Pycnoporus sanguineus (white-rot) and Gloeophyllum trabeum (brown-rot). The EO were obtained by hydrodistillation and its components were identified by gas chromatography coupled to mass spectrometry (GC-MS) and quantified with flame ionization detection (GC-FID). For the evaluation of antifungal activity, the mycelial growth index (MGI) and the mycelial growth inhibition (GI) of BS were determined at concentrations of 1, 2 and 5 µL.mL-1, and 0.25; 0.5 and 1 µL.mL-1 for MN1 and NM2. The yield of EOs was 2.29% (BS), 0.14% (NM1) and 0.17% (NM2). The major components of BS EO were eucalyptol (63.44%) and β-caryophyllene (13.68%), from NM1 were bicyclogermacrene (16.14%) and α-pinene (15.93%), and for NM2, α-pinene (19.04%) and sabinene (16.66%). EO NM1 (1μL.mL-1) showed the best result for antifungal activity, inhibiting about 57.55 ± 0.68 % of P. sanguineus growth and 58.82 ± 0.78 % in the test with G. trabeum.

Antiprotozoal Activity of Mycelial Extracts of Several Medicinal Agaricomycetes Mushrooms against Giardia duodenalis.

Despite knowledge on the therapeutic properties of fungal bio-compounds, few studies have been reported on their anti-parasitic activities. The anti-parasitic activity (APA) of mycelial extracts from seven medicinal agaricomycetous mushrooms (Polyporus lipsiensis, Ganoderma applanatum, Pleurotus ostreatus, P. flabellatus, Oudemansiella canarii, Lentinula edodes, and Pycnoporus sanguineus) against Giardia duodenalis, and identification of chemical compounds produced by mycelium P. lipsiensis mycelium, have been reported. The extracts of mycelia and fermented culture broths of tested mushroom species were evaluated against G. duodenalis by biological assays. P. lipsiensis showed the highest APA. The chemical analysis of mycelial extract of P. lipsiensis by gas chromatography-mass spectrometry (GC-MS) identified 73 molecules, including steroids, terpenes, and lipids. According to literature data, among these molecules, 11 possess APA. The present study revealed the diversity of compounds with anti-protozoal potential produced by mycelia of Agaricomycetes mushrooms, particularly P. lipsiensis against G. duodenalis.

Medicinal fungi used by rural communities in Northeastern Brazil

The use of fungi for diversified purposes is part of the history of mankind. We sought to report what is known about fungal species used for medicinal purposes by rural populations in Northeastern Brazil. To this end, semi-structured forms were applied to 176 informants from four rural communities located in the southern region of Piauí. The fungi were deposited in the Graziela Barroso Herbarium (TEPB) of the Federal University of Piauí. The use value (UV) and relative importance (RI) of each species were calculated and similarity was estimated with the Sorensen index. Medicinal use was identified for four basidiomycete species belonging to the families Agaricaceae [Podaxis pistillaris (L.) Fr.], Hymenochaetaceae (Fomitiporia sp.) and Polyporaceae [Hexagonia hydnoides (Sw.) M. Fidalgo and Pycnoporus sanguineus (L.) Murrill]. Indicated for intestinal colic in infants, P. sanguineus was the species that presented the greatest current and potential UV and greater cultural importance. Studies on the chemical and pharmacological composition of the mentioned species were found, indicating their antibacterial, antiparasitic, antiviral, antifungal and antioxidant potential. Knowledge about these fungi has been transmitted from generation to generation, with greater similarity among individuals of the female sex and adult age. The results showed that there is ethnomycological knowledge in rural communities still to be explored in the semi-arid region of Brazil and highlighted the importance of studies about the ethnological knowledge of medicinal fungi.

Habitat characteristics and utilization of edible wild mushrooms by local communities in the protected forest in Pinrang Regency, Indonesia

This study aims to determine the habitat characteristics and utilization of edible wild mushrooms by local communities in protected forest areas in Pinrang Regency, Indonesia. The study was conducted using observation, survey, interview, and questionnaire methods. The characteristics of the wild mushroom habitat are climate type B, with average rainfall is 2,780.2 mm/year, a temperature of 24-32°C, and relative humidity of 59-82%. In general, a wild mushroom grows on dead wood, especially candlenut (Aleurites moluccana) and mango (Mangifera indica), soil, and litter. There were 18 types of wild mushrooms found in the protected forest areas in Pinrang Regency, namely Termitomyces clypeate, Pleurotus ostreatus, Pycnoporus sanguineus, Tyromyces chioneus, Trametes hirsute, Schizophyllum commune, Lepiota clypeolaria, Lepiota brunneoincarnata, Auricularia auricular, Psavinea, squarrosulus, Leucocoprinus sp., Coprinellus micaceus, Ganoderma lucidum, Oudemansiella mucida. There were 5 types which include edible wild mushrooms that can be consumed by the community as a source of food and medicine, namely Termitomyces clypeatus, Pleurotus ostreatus, Schizophyllum commune, Auricularia auricular, and Pluteus cervinus, and 13 species include non-edible wild mushrooms and some of them are known as poisonous mushrooms.