Different wavelengths of LED irradiation promote secondary metabolite production in Pycnoporus sanguineus for antioxidant and immunomodulatory applications.

Pycnoporus sanguineus is a fungus of the phylum Basidiomycota that has many applications in traditional medicine, modern pharmaceuticals, and agricultural industries. Light plays an essential role in the metabolism, growth, and development of fungi. This study evaluated the mycelial growth and antioxidant and anti-inflammatory activities in P. sanguineus fermentation broth (PFB) cultured under different wavelengths of LED irradiation or in the dark. Compared to the dark cultures, the dry weight of mycelia in red- and yellow-light cultures decreased by 37 and 35% and the yields of pigments increased by 30.92 ± 2.18 mg and 31.75 ± 3.06 mg, respectively. Compared with the dark culture, the DPPH free radical scavenging ability, ABTS+ free radical scavenging capacity, and reducing power of yellow-light cultures increased significantly, and their total phenolic content peaked at 180.0 ± 8.34 µg/mL. However, the reducing power in blue-light cultures was significantly reduced, though the total phenol content did not vary with that of dark cultures. In LPS- and IFN-γ-stimulated RAW 264.7 cells, nitrite release was significantly reduced in the red and yellow light-irradiated PFB compared with the dark culture. In the dark, yellow-, and green-light cultures, TNF-α production in the inflamed RAW 264.7 cells was inhibited by 62, 46, and 14%, respectively. With red-, blue-, and white-light irradiation, TNF-α production was significantly enhanced. Based on these results, we propose that by adjusting the wavelength of the light source during culture, one can effectively modulate the growth, development, and metabolism of P. sanguineus.

Deciphering ligninolytic enzymes in the secretome of Pycnoporus sp. and their potential in degradation of 2-chlorophenol.

Chlorophenols and their derivatives are persistent environmental pollutants, posing a threat to terrestrial and aquatic life. The biological approach for eliminating toxic contaminants is an effective, sustainable, and environmental friendly method. In this study, the crude enzymes present in the secretome of white-rot fungus (Pycnoporus sp.) were explored for the degradation of 2-chlorophenol. The activity of ligninolytic enzymes in the secretome was analyzed and characterized for their kinetics and thermodynamic properties. Laccase and manganese peroxidase were prevalent ligninolytic enzymes and exhibited temperature stability in the range of 50-65 °C and pH 4-5, respectively. The kinetic parameters Michaelis constant (Km) and turnover number (Kcat) for Lac were 42.54 µM and 45 s-1 for 2,2'-azino-bis (3-ethylben- zothiazoline-6-sulfonic acid), and 93.56 µM and 48 s-1 towards 2,6-dimethoxyphenol whereas Km and Kcat for MnP were 2039 µM and 294 s-1 for guaiacol as substrate. Treatment with the crude enzymes laccase and manganese peroxidase results in the reduction of 2-chlorophenol concentration, confirmed by UV-visible absorption spectra and high-performance liquid chromatography analysis. The detoxification of 2-chlorophenol into less toxic forms was confirmed by the plate toxicity assay. This study demonstrated that crude enzymes produced by Pycnoporus sp. could potentially minimize the toxicity of phenolic compounds in a sustainable way.

Effect of hexavalent chromium on the biodegradation of tetrabromobisphenol A (TBBPA) by Pycnoporus sanguineus.

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 Mg2+-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.

Degradation of 2,2',4,4'-tetrabromodiphenyl ether by Pycnoporus sanguineus in the presence of copper ions.

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 ±â€¯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.

Production of lactobionic acid from lactose using the cellobiose dehydrogenase-3-HAA-laccase system from Pycnoporus sp. SYBC-L10.

The aim of this study was to produce lactobionic acid from lactose by a new Pycnoporus sp. SYBC-L10 strain. Recently, studies on enzymatic production of lactobionic acid mostly focus on cellobiose dehydrogenase from Sclerotium rolfsii CBS 191·62 and laccase from Trametes pubescens MB 89 oxidize lactose to lactobionic acid with redox mediators. In this study, we converted lactose to lactobionic acid by shaking flask fermentation without exogenous mediator in the reaction mixture. In this bioconversion process, lactose is efficiently converted into lactobionic acid with a specific productivity of up to 3·1 g l-1  h-1 and 96% yield. 3-Hydroxyanthranilic acid added externally to the reaction mixture can obviously accelerate the conversion of lactose to lactobionic acid. The results showed that 3-hydroxyanthranilic acid produced by the fungus itself is an important influencing factor in this bioconversion process. This study presents the first attempt to efficiently produce lactobionic acid by white-rot fungi, suggesting definite potential for Pycnoporus to produce lactobionic acid. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobionic acid has been applied to a wide range of applications in pharmaceutical, food, nanotechnology and chemical industries. Here, an attempt was done to produce lactobionic acid from lactose using the cellobiose dehydrogenase-3-HAA-laccase system in a fermentation system. After a survey of other methods to produce lactobionic acid by cellobiose dehydrogenase, this study explores a new and significant perspective for the production of lactobionic acid.

Lambertellin from Pycnoporus sanguineus MUCL 51321 and its anti-inflammatory effect via modulation of MAPK and NF-κB signaling pathways.

Lambertellin (1) and ergosta-5,7,22-trien-3-ol (2) were isolated from the solid rice fermentation of the plant pathogenic fungus Pycnoporus sanguineus MUCL 51321. Their structures were elucidated using comprehensive spectroscopic methods. The isolated compounds were tested on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Lambertellin (1) exhibited promising inhibitory activity against nitric oxide (NO) production with IC50 value of 3.19 µM, and it significantly inhibited the expression of inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2). Lambertellin (1) also decreased the expression of pro-inflammatory cytokines IL-6 and IL-1ß. The study of the mechanistic pathways revealed that lambertellin (1) exerts its anti-inflammatory effect in LPS-stimulated RAW 264.7 macrophage cells by modulating the activation of the mitogen activated protein kinase (MAPK) and nuclear factor κB (NF-κB) signaling pathways. Therefore, lambertellin (1) could be a promising lead compound for the development of new anti-inflammatory drugs.

Simultaneous removal of ciprofloxacin, norfloxacin, sulfamethoxazole by co-producing oxidative enzymes system of Phanerochaete chrysosporium and Pycnoporus sanguineus.

Pycnoporus sanguineus could remove 98.5% ciprofloxacin (CIP), 96.4% norfloxacin (NOR), 100% sulfamethoxazole (SMX), and 100% their mixture through biotransformation within 2 d, while Phanerochaete chrysosporium could only remove 64.5% CIP, 73.2% NOR, and 63.3% SMX through biosorption and biotransformation within 8 d, respectively. The efficiencies of antibiotic bioremoval under co-culture were more than that under the pure culture of P. chrysosporium but less than that under the pure culture of P. sanguineus. However, only 2% CIP and 3% NOR under co-culture were detected in the mycelia. In vitro enzymatic degradation and in vivo cytochrome P450 inhibition experiments revealed that laccase and cytochrome P450 could play roles in the removal of above all antibiotics, while manganese peroxidase could only play role in SMX removal. Transformation products of CIP and NOR under the pure culture of P. chrysosporium could be assigned to three different reaction pathways: (i) defluorination or dehydration, (ii) decarboxylation, and (iii) oxidation of the piperazinyl substituent. Additionally, other pathways, (iv) monohydroxylation, and (v) demethylation or deethylation at position N1 also occurred under the co-culture and pure culture of P. sanguineus. Antibacterial activity of antibiotics could be eliminated after treatments with pure and co-culture of P. chrysosporium and P. sanguineus. The cytotoxicity of the metabolites of SMX and NOR under co-culture was lower than that under the pure culture of P. sanguineus, indicating co-culture is a more environmentally friendly strategy to eliminate SMX and NOR.

Oil palm fiber biodegradation: physico-chemical and structural relationships.

MAIN CONCLUSION: X-ray microtomography results revealed that delignification process damaged the oil palm fibers, which correlated well with reduction of lignin components and increase of the phenolic content. Biodegradation investigation of natural fibers normally focuses on physico-chemical analysis, with less emphasis on physical aspect like fiber structures affect from microbial activity. In this work, the performance of Pycnoporus sanguineus to delignify oil palm empty fruit bunch fibers through solid-state fermentation utilizing various ratio of POME sludge was reported. In addition to tensile testing, physico-chemical and X-ray microtomography (µ-CT) analyses on the oil palm fibers were conducted to determine the effectiveness of the degradation process. The best ratio of fiber to fungi (60:40) was chosen based on the highest lignin loss and total phenolic content values and further investigation was performed to obtain fermentation kinetics data of both laccase and manganese peroxidase. µ-CT results revealed that delignification process damaged the pre-treated and untreated fibers structure, as evident from volume reduction after degradation process. This is correlated with reduction of lignin component and increase of the phenolic content, as well as lower stress-strain curves of the pre-treated fibers compared to the untreated ones (from tensile testing). It is suggested that P. sanguineus preferred to consume the outer layer of the fiber, before it penetrates through the cellular structure of the inner fiber.

Sorption-reduction coupled gold recovery process boosted by Pycnoporus sanguineus biomass: Uptake pattern and performance enhancement via biomass surface modification.

Biorecovery is emerging as a promising process to retrieve gold from secondary resources. The present study aimed to explore the uptake pattern of Pycnoporus sanguineus biomass for gold, identify the effective functional groups in gold recovery process, and thus further intensify the process via microbial surface modification. Results showed that P. sanguineus biomass could effectively recover gold with the formation of highly crystal AuNPs without any exogeneous electron donor. Under the conditions of various initial gold concentrations (1.0, 2.0, and 3.0 mM), biomass dosage of 2.0 g/L, solution pH value of 4.0, and incubation temperature of 30°C, the uptake equilibrium established after 4, 8, and 12 h, respectively. The uptake process could be well described by pseudo-second order kinetics model (R2  = 0.9988) and Langmuir isotherm model (R2  = 0.9958). The maximum uptake capacity of P. sanguineus reached as high as 358.69 mg/g. Further analysis indicated that amino, carboxyl and hydroxyl groups positively contributed to the uptake process. Among them, amino group significantly favored the uptake of gold during recovery process. When P. sanguineus biomass was modified by introduction of amino group, the gold uptake process was successfully intensified by shortening the uptake period and enhancing the uptake capacity. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1314-1322, 2017.

Influence of co-existed tetrabromobisphenol A (TBBPA) and hexavalent chromium on the cellular characteristics of Pycnoporus sanguineus during their removal and reduction.

Simultaneous TBBPA removal and Cr(VI) reduction by Pycnoporus sanguineus together with the effect of these co-existed pollutants on the fungal cellular characteristics were investigated in this study, aiming at illuminating the mechanism involved in the interactions between contaminants and microbial cells. The results revealed that Cr(VI) reduction and TBBPA removal declined from 92.5%, 75.4-30.6%, 44.8% when Cr(VI) concentration increased from 5 to 40mg/L, respectively. The removal efficiencies for Cr(VI) and TBBPA reached 61.4% and 94% separately under the optimum concentration of TBBPA at 10mg/L. Subsequent analyses indicated that the negative effect of Cr(VI) of high concentrations on Cr(VI) reduction and TBBPA removal was mainly attributed to the inhibition of fungal growth, intracellular proteins synthesis, cell viability and ATP enzyme activity. Compared with the moderate impact of TBBPA, the cell membrane of P. sanguineus was impaired severely and the surface morphology and intracellular structure changed dramatically in the presence of high concentration of Cr(VI) (above 10mg/L). This study also suggested that high level of TBBPA (15 and 20mg/L) promoted the synthesis of intracellular proteins and improved ATP enzyme activity within the first 48h of the reaction for enhancing the transportation and transformation of TBBPA.

Laccase induction by synthetic dyes in Pycnoporus sanguineus and their possible use for sugar cane bagasse delignification.

The use of synthetic dyes for laccase induction in vivo has been scarcely explored. We characterized the effect of adding different synthetic dyes to liquid cultures of Pycnoporus sanguineus on laccase production. We found that carminic acid (CA) can induce 722 % and alizarin yellow 317 % more laccase than control does, and they promoted better fungal biomass development in liquid cultures. Aniline blue and crystal violet did not show such positive effect. CA and alizarin yellow were degraded up to 95 % during P. sanguineus culturing (12 days). With this basis, CA was selected as the best inducer and used to evaluate the induction of laccase on solid-state fermentation (SSF), using sugarcane bagasse (SCB) as substrate, in an attempt to reach selective delignification. We found that laccase induction occurred in SSF, and a slight inhibition of cellulase production was observed when CA was added to the substrate; also, a transformation of SCB under SSF was followed by the 13C cross polarization magic angle spinning (CPMAS) solid-state nuclear magnetic resonance (NMR). Results showed that P. sanguineus can selectively delignify SCB, decreasing aromatic C compounds by 32.67 % in 16 days; O-alkyl C region (polysaccharides) was degraded less than 2 %; delignification values were not correlated with laccase activities. Cellulose-crystallinity index was increased by 27.24 % in absence of CA and 15.94 % when 0.01 mM of CA was added to SCB; this dye also inhibits the production of fungal biomass in SSF (measured as alkyl C gain). We conclude that CA is a good inducer of laccase in liquid media, and that P. sanguineus is a fungus with high potential for biomass delignification.

Eichhornia crassipes: Agro-waster for a novel thermostable laccase production by Pycnoporus sanguineus SYBC-L1.

The aim of this study was to explore the utilization of an intractable waster of Eichhornia crassipes in laccase production by Pycnoporus sanguineus SYBC-L1. E. crassipes as the sole carbon and nitrogen source was confirmed to produce laccase (7.26 U/g dry substrate). The fermentation medium for the maximum enzyme production was optimized and the laccase was then purified and characterized. The optimized culture medium was 25.1% E. crassipes, 13.9% sawdust, 1.5 mM CuSO4, 40 µM gallic acid, 65% moisture content and initial pH 6.0. A maximum laccase activity of 32.02 U/g dry substrate was detected at 9th day, which was 4.5-fold compared to the initial medium. The molecular mass of the purified Lac-S was 58.4 kDa. The optimum activity of Lac-S for DMP was at pH 3.0 and 70°C. Lac-S showed not only high catalytic activities at low temperature, but also good stabilities toward pH and temperature. The residual catalytic activities of Lac-S were 30%, 40% and 50% at 0°C, 10°C and 20°C, and the half-lives at 50°C, 60°C and 70°C were 21.7, 9.7 and 1.5 h, respectively. The results provide a significant basis for E. crassipes further utilization and Lac-S specific application in harsh industry.

Improving the bioremoval of sulfamethoxazole and alleviating cytotoxicity of its biotransformation by laccase producing system under coculture of Pycnoporus sanguineus and Alcaligenes faecalis.

The occurrence of sulfamethoxazole (SMX) in aquatic environment is a health concern. The presence of SMX significantly inhibited the laccase activity of Pycnoporus sanguineus with a lower removal efficiency of SMX. Although a laccase system with 2,20-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) eliminated 100% SMX within 6h, ABTS might cause an environmental issue. An alternative to SMX elimination is the coculture of Alcaligenes faecalis and P. sanguineus. The SMX removal efficiency at 48h under the coculture with vitamins was higher than that under their pure culture alone, indicating that a coculture was more efficient in eliminating SMX than a pure culture. Only 1% SMX was detected in mycelia, indicating that SMX elimination is achieved primarily through biotransformation rather than adsorption. Laccase production by the coculture effectively inhibited the accumulations of N4-acetyl-SMX and N-hydroxy-SMX and alleviated the cytotoxicity of SMX transformation products. The mixture of SMX and sulfadiazine inhibited their removal efficiency.

Bioreduction of Precious Metals by Microorganism: Efficient Gold@N-Doped Carbon Electrocatalysts for the Hydrogen Evolution Reaction.

The uptake of precious metals from electronic waste is of environmental significance and potential commercial value. A facile bioreductive synthesis is described for Au nanoparticles (ca. 20 nm) supported on N-doped carbon (Au@NC), which was derived from Au/Pycnoporus sanguineus cells. The interface and charge transport between Au and N-doped carbon were confirmed by HRTEM and XPS. Au@NC was employed as an electrocatalyst for the hydrogen evolution reaction (HER), exhibiting a small onset potential of -54.1 mV (vs. RHE), a Tafel slope of 76.8 mV dec(-1) , as well as robust stability in acidic medium. Au@NC is a multifunctional electrocatalyst, which demonstrates high catalytic activity in the oxygen reduction reaction (ORR), as evidenced by an onset potential of +0.97 V, excellent tolerance toward methanol, and long-term stability. This work exemplifies dual recovery of precious Au and fabrication of multifunctional electrocatalysts in an environmentally benign and application-oriented manner.

Evaluation of productivity and antioxidant profile of solid-state cultivated macrofungi Pleurotus albidus and Pycnoporus sanguineus.

The aim of this study was to investigate the production profile of Pleurotus albidus and Pycnoporus sanguineus on different waste substrates containing natural phenolics, and also to investigate whether phenolic-rich substrates can improve the phenolic content of these macrofungi. The medium formulated with Pinus sp. sawdust (PSW) made possible the highest yields (2.62±0.73%) of P. sanguineus. However, the supplementation of PSW with apple waste (AW) resulted in better P. albidus yields (23.94±2.92%). The results indicated that the substrate composition affected macrofungi production, also the chemical composition and the presence of phenolic compounds in the production media influence phenolic content and antioxidant activity in macrofungi.

Adsorption of cinnabarinic acid from culture fluid with magnetic microbeads.

In this study, antimicrobial pigment cinnabarinic acid (CA) was produced from Pycnoporus cinnabarinus in laboratory-scale batch cultures. Magnetic poly(ethylene glycol dimethacrylate-N-methacryloyl-l-tryptophan methyl ester) [m-poly(EGDMA-MATrp)] beads (average diameter = 53-103 µm) were synthesized by copolymerizing of N-methacryloyl-l-tryptophan methyl ester (MATrp) with ethylene glycol dimethacrylate (EGDMA) in the presence of magnetite (Fe3O4) and used for the adsorption of CA. The m-poly(EGDMA-MATrp) beads were characterized by N2 adsorption/desorption isotherms (Brunauer Emmet Teller), X-ray photoelecron spectroscopy, scanning electron microscopy, infrared spectroscopy, thermal gravimetric analysis, electron spin resonance and swelling studies. The efficiency of m-poly(EGDMA-MATrp) beads for separation of CA from culture fluid was evaluated. The effects of pH, initial concentration, contact time and temperature on adsorption were analyzed. The maximum CA adsorption capacity of the m-poly(EGDMA-MATrp) beads was 272.9 mg g(-1) at pH 7.0, 25 °C. All the isotherm data can be fitted with the Langmuir, Freundlich and Dubinin-Radushkevich isotherm models. The adsorption process obeyed pseudo-second-order kinetic model. Thermodynamic parameters ΔH = 5.056 kJ mol(-1), ΔS = 52.44 J K(-1) mol(-1) and ΔG = -9.424 kJ mol-(1) to -11.27 kJ mol-(1) with the rise in temperature from 4 to 40 °C indicated that the adsorption process was endothermic and spontaneous.

Strain-dependent response to Cu(2+) in the expression of laccase in Pycnoporus coccineus.

The effects of Cu(2+) on the activity and expression of laccase were investigated in seven different strains of Pycnoporus coccineus collected from different regions in Korea. Cu(2+) was toxic to mycelial growth at concentrations greater than 0.5 mM CuSO4 and showed complete growth inhibition at 1 mM in the liquid culture. However, Cu(2+) significantly upregulated the extracellular laccase activity at 0.2 mM in five strains of P. coccineus, IUM4209, IUM0032, IUM0450, IUM0470, and IUM4093, whereas two strains, IUM0253 and IUM0049, did not respond to Cu(2+), despite being closely related to the other five strains. Subsequent RT-PCR analysis also showed that the laccase mRNA was highly expressed only in the former five strains in the presence of Cu(2+). Taken together, these results indicate that Cu(2+) regulates expression of the laccase gene in a strain-dependent manner. The five strains commonly produced a single predominant laccase protein with a molecular weight of 68 kDa. Peptide sequencing revealed that the laccase was a homolog of Lcc1 of P. coccineus, which was isolated in China. The Cu(2+)-induced culture supernatants exhibited high degradation of polycyclic aromatic hydrocarbons, indicating that the 68-kDa laccase is the primary extracellular degradative enzyme in P. coccineus.

Magnetic Pycnoporus sanguineus-loaded alginate composite beads for removing dye from aqueous solutions.

Dye pollution in wastewater is a severe environmental problem because treating water containing dyes using conventional physical, chemical, and biological treatments is difficult. A conventional process is used to adsorb dyes and filter wastewater. Magnetic filtration is an emerging technology. In this study, magnetic Pycnoporus sanguineus-loaded alginate composite beads were employed to remove a dye solution. A white rot fungus, P. sanguineus, immobilized in alginate beads were used as a biosorbent to remove the dye solution. An alginate polymer could protect P. sanguineus in acidic environments. Superparamagnetic nanomaterials, iron oxide nanoparticles, were combined with alginate gels to form magnetic alginate composites. The magnetic guidability of alginate composites and biocompatibility of iron oxide nanoparticles facilitated the magnetic filtration and separation processes. The fungus cells were immobilized in loaded alginate composites to study the influence of the initial dye concentration and pH on the biosorption capacity. The composite beads could be removed easily post-adsorption by using a magnetic filtration process. When the amount of composite beads was varied, the results of kinetic studies of malachite green adsorption by immobilized cells of P. sanguineus fitted well with the pseudo-second-order model. The results indicated that the magnetic composite beads effectively adsorbed the dye solution from wastewater and were environmentally friendly.

Growth inhibition and antioxidative response of wood decay fungi exposed to plant extracts of Casearia species.

UNLABELLED: Ligninolytic fungi take part in critical processes in ecosystems such as nutrient recycling; however, some fungal species can be pathogenic to forest and urban trees and deteriorate wood products. The tropical flora is an important source of antimicrobial compounds environmentally safer than traditional wood preservatives. Therefore, this study aimed to evaluate the inhibitory activity of ethanol plant extracts of Casearia sylvestris and Casearia decandra on the white-rot wood decay basidiomycetes Trametes villosa and Pycnoporus sanguineus. In addition, the effect of the extracts on the fungal antioxidative metabolism was studied. Among the different substances present in the extracts, the phytochemical analyses identified a clerodane diterpenoid (C. sylvestris) and cinnamic acid, hydroquinone and ß-sitosterol (C. decandra). The extracts inhibited the fungi up to 70% and caused hyphal morphology changes. The extracts triggered oxidative stress process as indicated by the increased levels of the antioxidant enzymes catalase and glutathione reductase. Therefore, the Casearia extracts are a potential source of natural biocides to control wood decay fungi, and one of the mechanisms of action is the oxidative stress. SIGNIFICANCE AND IMPACT OF THE STUDY: The Casearia plant extracts exhibited important antifungal activity on wood decay fungi and triggered oxidative stress process, an inhibitory mechanism rarely studied in filamentous fungi exposed to plant extracts. Therefore, a starting point was provided for the development of natural compounds-based products as an alternative to chemical fungicides. In addition, subsidies were given to further studies in order to elucidate in more detail how compounds present in extracts of native tropical plants affect the physiology of fungi.

A first insight into Pycnoporus sanguineus BAFC 2126 transcriptome.

Fungi of the genus Pycnoporus are white-rot basidiomycetes widely studied because of their ability to synthesize high added-value compounds and enzymes of industrial interest. Here we report the sequencing, assembly and analysis of the transcriptome of Pycnoporus sanguineus BAFC 2126 grown at stationary phase, in media supplemented with copper sulfate. Using the 454 pyrosequencing platform we obtained a total of 226,336 reads (88,779,843 bases) that were filtered and de novo assembled to generate a reference transcriptome of 7,303 transcripts. Putative functions were assigned for 4,732 transcripts by searching similarities of six-frame translated sequences against a customized protein database and by the presence of conserved protein domains. Through the analysis of translated sequences we identified transcripts encoding 178 putative carbohydrate active enzymes, including representatives of 15 families with roles in lignocellulose degradation. Furthermore, we found many transcripts encoding enzymes related to lignin hydrolysis and modification, including laccases and peroxidases, as well as GMC oxidoreductases, copper radical oxidases and other enzymes involved in the generation of extracellular hydrogen peroxide and iron homeostasis. Finally, we identified the transcripts encoding all of the enzymes involved in terpenoid backbone biosynthesis pathway, various terpene synthases related to the biosynthesis of sesquiterpenoids and triterpenoids precursors, and also cytochrome P450 monooxygenases, glutathione S-transferases and epoxide hydrolases with potential functions in the biodegradation of xenobiotics and the enantioselective biosynthesis of biologically active drugs. To our knowledge this is the first report of a transcriptome of genus Pycnoporus and a resource for future molecular studies in P. sanguineus.