Search

Heterogous expression of lignin peroxidase (LiP) from Phanerochaete chrysosporium was performed in by E. coli prokaryotic expression system, and pure LiP was prepared by washing, refolding, and purification. The enzyme activity was measured by the resveratrol oxidation method. The effects of different chemicals on LiP activity were explored by adding different kinds of metal ions, acids/phenols, and surfactants. The optimal pH and temperature are 4.2 and 40 °C. The single-factor screening experiment showed that adding 1 mM Mn 2+ , 0.1 mM DL-lactic acid, and 2% PEG-4000 had the best promotion effect on the enzyme activity of recombinant LiP, which was 160.61%, 188.46%, and 247.83%, respectively. Further, the synergistic addition of Mn 2+ and PEG-4000 achieved the best enzyme activity promotion effect of 277.51%. In addition, the addition of DL-lactic acid alone could promote LiP activity. However, the co-addition of lactic acid with Mn 2+ and PEG-4000 contributed only 247.87%, which indicated that the addition of DL-lactic acid had an inhibitory effect when applied synergistically. For the first time, it was found that PEG-4000 increased LiP enzyme activity obviously and had a synergistic effect with Mn 2+ , serving as a reference for LiP in studies and applications pertaining to lignin breakdown.

The distribution, characteristics of extracellular polymeric substances (EPS) of Phanerochaete chrysosporium under Pb 2+ stress and the influence on Pb removal were investigated. Polysaccharides was found to be the main composition in both soluble EPS (SEPS) and bounded EPS (BEPS). More polysaccharides and protein in BEPS were detected with the increased Pb 2+ concentration. The ratio of Pb amount distributed in BEPS to the total Pb removed by the fungal biomass gradually decreased from 91.66 to 61.27% in group with 50 mg/L of initial Pb 2+ , but kept at about 35% or 25% in groups with higher Pb 2+ . It implies that BEPS played a certain role in the lead removal process, and the role of BEPS was relatively more important in the removal of lower concentration of Pb 2+ and in the initial period of Pb removal. With FTIR analysis and Pb 2+ adsorption experiment, more effective functional groups and better Pb 2+ adsorption capacity was demonstrated in BEPS than in SEPS. SEM-EDS analysis demonstrated that part of Pb immobilized in BEPS was in the form of Pb precipitation. The increased molecular weight in SEPS and more polysaccharides in BEPS were probably beneficial for the adhesion of Pb precipitation.

Nanodiamonds (NDs) are produced with large scale and applied in many areas, thus the environmental impacts and hazards of NDs should be systematically investigated. In this study, we evaluated the interaction between detonation NDs and white rot fungus Phanerochaete chrysosporium and the impact on the fungus decompositions activities. NDs did not influence the biomass gain of P. chrysosporium and the culture medium pH values. The mycelia of P. chrysosporium were destroyed upon the direct contact with NDs, while the rest retained the fibrous structure. Ultrastructural observations suggested that small aggregates of NDs seldom entered the fungus cells, but the break of cell wall and the loss of cytoplasm were induced by NDs. Under both optical and electron microscopes, the aggregation of colloidal ND particles was observed, which was the possible reason of low toxicity. High concentrations of NDs inhibited the laccase activity and manganese peroxidase activity of P. chrysosporium, which led to the decrease of decomposition activity for pollutants. Colloidal ND particles were not well dispersed in sawdust degradation evaluations, so no inhibitive effect was observed for wood degradation. The toxicological mechanism of NDs was assigned to oxidative stress. The results collectively suggested that NDs had low toxicity to white rot fungi and could be applied safely. The colloid dispersion/aggregation of nanoparticles in biological systems should be carefully considered during the design of safe nanomaterials., Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, influences of cations (Na + , K + , Ca 2+ , and Mg 2+ ), anions (NO 3 - , Cl - , and SO 4 2- ), and humic acid (HA) on the antimicrobial efficacy of silver nanoparticles (AgNPs)/Ag + against Phanerochaete chrysosporium were investigated by observing cell viability and total Ag uptake. K + enhanced the antimicrobial toxicity of AgNPs on P. chrysosporium, while divalent cations decreased the toxicity considerably, with preference of Ca 2+ over Mg 2+ . Impact caused by a combination of monovalent and divalent electrolytes was mainly controlled by divalent cations. Compared to AgNPs, however, Ag + with the same total Ag content exhibited stronger antimicrobial efficacy towards P. chrysosporium, regardless of the type of electrolytes. Furthermore, HA addition induced greater microbial activity under AgNP stress, possibly originating from stronger affinity of AgNPs over Ag + to organic matters. The obtained results suggested that antimicrobial efficacy of AgNPs was closely related to water chemistry: addition of divalent electrolytes and HA reduced the opportunities directly for AgNP contact and interaction with cells through formation of aggregates, complexes, and surface coatings, leading to significant toxicity reduction; however, in monovalent electrolytes, the dominating mode of action of AgNPs could be toxic effects of the released Ag + on microorganisms due to nanoparticle dissolution., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The root-knot nematode Meloidogyne incognita has a wide host range and it is one of the most economically important crop parasites worldwide. Biological control has been a good approach for reducing M. incognita infection, for which many nematophagous fungi are reportedly applicable. However, the controlling effects of Phanerochaete chrysosporium strain B-22 are still unclear. In the present study we characterized the parasitism of this strain on M. incognita eggs, second-stage juveniles (J2), and adult females. The highest corrected mortality was 71.9% at 3 × 108 colony forming units (CFU) mL-1 and the estimated median lethal concentration of the fungus was 0.96 × 108 CFU mL-1. Two days after treatment with Phanerochaete chrysosporium strain B-22 eggshells were dissolved. A strong lethal effect was noted against J2, as the fungal spores developed in their body walls, germinated, and the resulting hyphae crossed the juvenile cuticle to dissolve it, thereby causing shrinkage and deformation of the juvenile body wall. The spores and hyphae also attacked adult females, causing the shrinkage and dissolution of their bodies and leakage of contents after five days. Greenhouse experiments revealed that different concentrations of the fungal spores effectively controlled M. incognita. In the roots, the highest inhibition rate for adult females, juveniles, egg mass, and gall index was 84.61%, 78.91%, 84.25%, and 79.48%, respectively. The highest juvenile inhibition rate was 89.18% in the soil. Phanerochaete chrysosporium strain B-22 also improved tomato plant growth, therefore being safe for tomato plants while effectively parasitizing M. incognita. This strain is thus a promising biocontrol agent against M. incognita., Competing Interests: The authors declare no conflict of interests.

The present study focused on the bioaccumulation and cytotoxicities of Cd 2+ , CdSe quantum dots (QDs) and CdSe/ZnS QDs in Escherichia coli (E. coli, represents prokaryotic system) and Phanerochaete chrysosporium (P. chrysosporium, represents eukaryotic system), respectively. Two types of QDs were characterized by transmission electron microscopy (TEM) and dynamic light scattering. The inductively coupled plasma optical emission spectrometer results showed that the bioaccumulation amounts of CdSe QDs by E. coli and P. chrysosporium were larger than those of CdSe/ZnS QDs due to the smaller particle size and less negative surface charges of CdSe QDs. Confocal microscopy and TEM results showed that there was an interaction between QDs and cells, and QDs have entered into the cells eventually, leading to the change of cell morphology. Plasma membrane fluidities and membrane H + -ATPase activities of E. coli and P. chrysosporium decreased gradually with the increasing concentrations of Cd 2+ , CdSe and CdSe/ZnS QDs. Results of the cell viabilities and intracellular reactive oxygen species levels indicated that the induced cytotoxicities were decreased as follows: CdSe QDs > CdSe/ZnS QDs > Cd 2+ . These findings suggested that the cytotoxicity of QDs was not only attributed to their heavy metal components, but also related to their nanosize effects which could induce particle-specific toxicity. The above results offer valuable information for exploring the cytotoxicity mechanism of QDs in prokaryote and eukaryote., (Copyright © 2019 Elsevier Inc. All rights reserved.)

As a persistent organic pollutant listed in the Stockholm Convention, perfluorooctane sulfonate (PFOS) is extremely refractory to degradation under ambient conditions. Its potential ecotoxicity has aroused great concerns and research interests. However, little is known about the toxicity of PFOS on fungus. In this study, the white rot fungus Phanerochaete chrysosporium (P. chrysosporium) was adopted to assess the toxicity of PFOS in liquid culture. The addition of 100 mg/L PFOS potassium salt significantly decreased the fungal biomass by up to 76.4% comparing with un-amended control during the incubation period. The hyphostroma of P. chrysosporium was wizened and its cell membrane was thickened, while its vesicle structure was increased, based on the observation with scanning electron microscope (SEM) and transmission electron microscope (TEM). Nevertheless, the PFOS dosage of below 100 mg/L did not show a considerable damage to the growth of P. chrysosporium. The degradation of malachite green (MG) and 2,4-dichlorophenol (2,4-DCP) by P. chrysosporium was negatively affected by PFOS. At the initial dosage of 100 mg/L PFOS, the decolorization efficiency of MG and the degradation efficiency of 2,4-DCP decreased by 37% and 20%, respectively. This might be attributed to the inhibition of PFOS on MnP and LiP activities. The activities of MnP and LiP decreased by 20.6% and 43.4%, respectively. At a high dosage PFOS (100 mg/L), P. chrysosporium could show a high adsorption of MG but lose its pollutant degradation ability. Transcriptome analysis indicated that PFOS contamination could lead to the change of gene expression in the studied white rot fungus, and the genes regulating membrane structure, cell redox process, and cell transport, synthesis and metabolism were impacted. Membrane damage and oxidative damage were the two main mechanisms of PFOS' toxicity to P. chrysosporium., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Hydrogen sulfide (H 2 S), an important cellular signaling molecule, plays vital roles in mediating responses to biotic/abiotic stresses. Influences of H 2 S on metal removal, cell viability, and antioxidant response of Phanerochaete chrysosporium upon exposure to heavy metals and silver nanoparticles (AgNPs) in the present study were investigated. An enhancement in Pb(ΙΙ) removal with an increase in concentration of the H 2 S donor sodium hydrosulfide (NaHS) was observed, and the maximum removal efficiencies increased by 31% and 17% under 100 and 200 mg/L Pb(ΙΙ) exposure, respectively, in the presence of 500 μM NaHS. Application of 500 μM NaHS increased the cell viability by 15%-39% under Pb(II) stress (10-200 mg/L) with relative to the untreated control. Increase in total Ag uptake and cell survival was also elicited by NaHS in a concentration-dependent manner under AgNP stress. Meanwhile, activities of superoxide dismutase and catalase were significantly enhanced with the introduction of NaHS under stresses of Pb(II), Cd(II), Cu(II), Zn(II), Ni(II), and AgNPs. The inhibition in lipid peroxidation and oxidative stress was observed in P. chrysosporium cells exposed to these toxicants following NaHS pretreatment, which could be attributed to the upregulation in antioxidant enzymes. The results obtained suggest that H 2 S can alleviate heavy metals and AgNP-induced toxicity to P. chrysosporium and improve the removal efficiency of these toxicants from wastewater., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

In order to evaluate the potential ecological risk and the toxic effect of landfill leachate (LL), Phanerochaete chrysosporium was exposed to LL and their biochemical response was observed by using antioxidant parameters. Phanerochaete chrysosporium, ME 446, was kept at 4 °C after being sub-cultured at 28 °C on Sabouraud Dextrose Agar (SDA). Superoxide dismutase (SOD), catalase (CAT) activities, and malaondialdehyde (MDA) and glutathione (GSH) levels of P. chrysosporium exposed to different dilution rates of leachate (1/10 and 1/20) for 24 and 96 h were analyzed by using the ELISA method. The physiochemical parameters such as pH, conductivity, total dissolved solids (TDS), dissolved oxygen (DO), chemical oxygen demand (COD) of leachate, and reference water were analyzed by using the YSI Professional Plus handheld multiparameter meter. In this study, SOD activities were decreased in the application groups compared with the Control Group at the 24th and 96th hours. CAT activities and GSH levels increased in the application groups compared with the Control Group at the 24th hour but decreased at the 96th hours. MDA levels increased in all of the application groups when compared with the Control Group for both 24 and 96 h. Different concentration of LL induces oxidative stress in P. chrysosporium, increased CAT activity and MDA levels, and decreased SOD activity and GSH levels.

Algal blooms and toxins have become serious ecological problems. White-rot fungi have been demonstrated to be a feasible means of control, but the genotoxicity mechanisms involved have not been reported. In this study, Cryptomonas obovata FACHB-1301, Oscillatoria sp. FACHB-1083, and Scenedesmus quadricauda FACHB-507 were co-cultured with Phanerochaete chrysosporium under optimal conditions of 250 mg-l at 25 °C with DO 7.0 mg-l for 1, 3, 5 and 7 d. Compared to the control groups, the values for tadpoles exposed to algae treated with Phanerochaete chrysosporium were only increased from 1.95 ± 0.09, 2.78 ± 0.08 and 2.37 ± 0.13 to 2.45 ± 0.07, 3.56 ± 0.08 and 2.54 ± 0.10, and the frequency of nuclear anomalies reached 6.45 ± 0.06, 11.14 ± 0.05 and 7.85 ± 0.10 to 7.68 ± 0.08, 13.12 ± 0.06 and 8.57 ± 0.12 in the experimental groups after 7 d. What's more, the tail lengths were only increased to 36.77 ± 0.54, 41.58 ± 0.78 and 35.38 ± 0.66, and the comet length reached 55.67 ± 0.68, 68.56 ± 0.85 and 51.43 ± 0.82. The results demonstrated that Phanerochaete chrysosporium effectively decreased genotoxicity effects in Fejervarya multistriat tadpoles. These results could provide new ideas for inhibiting water blooms, and lay a theoretical foundation for promoting the deepening of water eutrophication., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Carbon nanotubes (CNTs) are widely used in diverse areas with increasing annual production, thus the environmental impact of CNTs needs thorough investigation. In this study, we evaluated the effect of pristine multi-walled CNTs (p-MWCNTs) and oxidized multi-walled CNTs (o-MWCNTs) on white rot fungus Phanerochaete chrysosporium, which is the decomposer in carbon cycle and also has many applications in environmental remediation. Both p-MWCNTs and o-MWCNTs had no influence on the dry weight increase of P. chrysosporium and the pH value of culture system. The fibrous structure of P. chrysosporium was disturbed by p-MWCNTs seriously, while o-MWCNTs had litter influence. The ultrastructural changes were more evident for P. chrysosporium exposed to p-MWCNTs and only p-MWCNTs could penetrate into the cell plasma. The chemical composition of P. chrysosporium was nearly unchanged according to the infrared spectra. The laccase activity was suppressed by p-MWCNTs, while o-MWCNTs showed stimulating effect. The decoloration of reactive brilliant red X-3B was not affected by both CNT samples. However, serious inhibition of wood degradation was observed in the p-MWCNTs exposed groups, suggesting the potential threat of CNTs to the decomposition of carbon cycle. The implication to the environmental risks and safe applications of carbon nanomaterials is discussed., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Extensive evidence showed that the efficiency of fungal wood degradation is closely dependent on their ability to cope with the myriad of putative toxic compounds called extractives released during this process. By analysing global gene expression of Phanerochaete chrysosporium after short oak extractive treatment (1, 3 and 6 h), we show that the early molecular response of the fungus concerns first mitochondrial stress rescue followed by the oxidation and finally conjugation of the compounds. During these early responses, the lignolytic degradative system is not induced, rather some small secreted proteins could play an important role in cell protection or signaling. By focusing on the functional characterization of an hitherto uncharacterized glutathione transferase, we show that this enzyme interacts with wood molecules suggesting that it could be involved in the detoxification of some of them, or act as a scavenger to prevent their cytosolic toxicity and favour their transport., (© 2018 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Due to the widespread application of white-rot fungi for the treatment of pollutants, it's crucial to exploit the special effects of pollutants on the microbes. Here, we studied the effects of cadmium on calcium homeostasis in the most studied white-rot fungus Phanerochaete chrysosporium. The response of P. chrysosporium to cadmium stress is concentration-dependent. A high concentration of cadmium caused the release of calcium from P. chrysosporium, while a hormesis effect was observed at a lower cadmium concentration (10 μM), which resulted in a significant increase in calcium uptake and reversed the decrease in cell viability. Calcium (50 μM) promoted cell viability (127.2% of control), which reflects that calcium can protect P. chrysosporium from environmental stress. Real-time changes in the Ca 2+ and Cd 2+ fluxes of P. chrysosporium were quantified using the noninvasive microtest technique. Ca 2+ influx decreased significantly under cadmium exposure, and the Ca 2+ channel was involved in Ca 2+ and Cd 2+ influx. The cadmium and/or calcium uptake results coupled with the real-time Ca 2+ and Cd 2+ influxes microscale signatures can enhance our knowledge of the homeostasis of P. chrysosporium with respect to cadmium stress, which may provide useful information for improving the bioremediation process., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Potential transformations of silver nanoparticles (AgNPs) upon interaction with naturally ubiquitous organic ligands in aquatic environments influence their transport, persistence, bioavailability, and subsequent toxicity to organisms. In this study, differential behaviors of AgNPs and silver ions (Ag + ) towards cysteine (Cys), an amino acid representative of thiol ligands that easily coordinate to Ag + and graft to nanoparticle surfaces, were investigated in the aspects of bioremediation and their toxicity to Phanerochaete chrysosporium. Total Ag removal, 2,4-dichlorophenol (2,4-DCP) degradation, extracellular protein secretion, and cellular viability were enhanced to some extent after supplement of various concentrations of cysteine under stress of AgNPs and Ag + . However, an obvious decrease in total Ag uptake was observed after 5-50 μM cysteine addition in the groups treated with 10 μM AgNPs and 1 μM Ag + , especially at a Cys:Ag molar ratio of 5. More stabilization in uptake pattern at this ratio was detected under Ag + exposure than that under AgNP exposure. Furthermore, in the absence of cysteine, all Ag + treatments stimulated the generation of reactive oxygen species (ROS) more significantly than high-dose AgNPs did. However, cysteine supply under AgNP/Ag + stress aggravated ROS levels, albeit alleviated at 100 μM Ag + , indicating that the toxicity profiles of AgNPs and Ag + to P. chrysosporium could be exacerbated or marginally mitigated by cysteine. The results obtained were possibly associated with the lability and bioavailability of AgNP/Ag + -cysteine complexes., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Cd-induced stress response of Phanerochaete chrysosporium during the biodegradation of BDE-47 was investigated in this study, with the goal of elucidating the tolerance behavior and the detoxification mechanisms of P. chrysosporium to resist the Cd stress in the course of BDE-47 biodegradation, which has implications for expanding the application of P. chrysosporium in the bioremediation of Cd and BDE-47 combined pollution. The results suggested that single BDE-47 exposure did not induce obvious oxidative stress in P. chrysosporium, but coexistent Cd significantly triggered ROS generation, both intracellular ROS level and H 2 O 2 content showed positive correlation with Cd concentration. The activities of SOD and CAT were enhanced by low level of Cd (≤ 1 mg/L), but Cd of higher doses (＞1 mg/L) depressed the expression of these two antioxidant enzymes at the later exposure period (3-5 days). The intracellular content of GSH along with GSH/GSSG ratio also exhibited a bell-shaped response with a maximum value at Cd of 1 mg/L. Furthermore, Cd-induced ROS generation resulted in the lipid peroxidation, as indicated by a noticeable increment of MDA content found after 3 days. Moreover, the study also indicated that Cd less than 1 mg/L promoted the production of extracellular protein and quickened the decrease of pH value in the medium, while excessive Cd (＞1 mg/L) would lead to inhibition. These findings obtained demonstrated that P. chrysosporium had a certain degree of tolerance to Cd within a specific concentration range via regulating the antioxidant levels, inducing the synthesis of extracellular protein as well as stimulating the production of organic acids, and 1 mg/L is suggested to be the tolerance threshold of this strains under Cd stress during BDE-47 biodegradation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In this study, the effect of chloramine T (Chl-T) on the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and glutathione S-transferase (GST); the levels of reduced (GSH) and oxidised glutathione (GSSG) and their ratios; and also membrane lipid peroxidation (LPO) levels in Phanerochaete chrysosporium were investigated in a dose- (0.25-1 mmol/L) and time-dependent (1.5-9 h) manner. The highest SOD activity was observed in 0.5 mmol/L Chl-T at 6th hour as 1.48-fold of its control. The observed highest level in CAT activities was 4.6-fold of control in 0.5 and 0.75 mmol/L at the 6th hour. The GSH levels that were over the control showed decreasing tendency from the beginning of incubation, except 0.25 mmol/L. In contrast with GSH level variations, GSSG levels reached 10.0-fold of its control by showing increasing tendency with the increases in concentration and time. While the GSH/GSSG ratios were over the control at 0.25 mmol/L during all incubation, they fell under the control values at the earlier hours of incubation with the increasing concentrations of Chl-T. Glutathione-related enzymes GSH-Px, GR and GST were also induced with Chl-T treatment, and the highest activities were 3.29-, 7.5- and 6.56-fold of their controls, respectively. On the other hand, the increases in LPO levels with increasing concentration and time up to 5.27-fold of its control showed that the inductions observed in antioxidant system could not prevent the Chl-T-based oxidative stress.

This study investigated the influence of silver ions (Ag + ) on the cytotoxicity of silver nanoparticles (AgNPs) in Phanerochaete chrysosporium and noted the degree of extracellular secretions in response to the toxicant's stress. Oxalate production was elicited with moderate concentrations of 2,4-dichlorophenol (2,4-DCP) and AgNPs reaching a plateau at 10 mg/L and 10 μM, respectively. Increased oxalate accumulation was accompanied by higher activities of manganese peroxidase (MnP) and lignin peroxidase (LiP). However, the secretion of oxalate, MnP and LiP was significantly inhibited owing to Ag + incorporation into AgNP solution. Production of extracellular polymeric substances (EPS) significantly elevated with an increase in 2,4-DCP concentrations; however, after 24 h of exposure to 100 mg/L 2,4-DCP, an obvious decrease in EPS occurred, indicating that part of EPS could be consumed as carbon and energy sources to ameliorate biological tolerance to toxic stress. Furthermore, AgNP-induced "particle-specific" cytotoxicity was substantially enhanced with additional Ag + as evidenced by its significant negative impact on cellular growth, plasma membrane integrity, and morphological preservation compared with AgNPs at equal Ag concentration., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Metals released into the environment continue to be of concern for human health. Using white-rot fungi as biosorbents for heavy metals removal is an attractive alternative owing to its good performance and low cost. However, the molecular mechanism underlying heavy metal tolerance in white-rot fungi has not yet been fully elucidated. This study identified and analyzed the lead (Pb)-induced transcriptional changes in Phanerochaete chrysosporium, a well-known heavy metal hyperaccumulating white-rot fungus. The results confirmed its outstanding ability in Pb tolerance and effective defense system. By comparative analysis of gene expression profiles obtained from cDNA-amplified fragment length polymorphism (cDNA-AFLP), we isolated 43 transcript-derived fragments (TDFs) differentially regulated by Pb exposure in P. chrysosporium, and 23 TDFs presented significant similarities to genes encoding known or putative proteins which belong to different functional categories involving ion binding, energy and carbohydrate metabolism, and signal transduction. The detailed characterization of these Pb-responsive genes was presented and the expression patterns of some interesting genes were validated by quantitative RT-PCR. This work provides the first evidence of Pb-responsive genes along with their putatively functional annotations in P. chrysosporium, which may help to understand the mechanism underlying heavy metal accumulation and tolerance in P. chrysosporium., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Since the determination of the fermentation kinetics is one of the main challenges in solid state fermentation, the quantitative measurement of biomass growth during microbial pretreatment by FTIR spectroscopy in Attenuated Total Reflectance mode was evaluated. Peaks at wave numbers of 1651 cm -1 and 1593 cm -1 showed to be affected during pretreatment of poplar wood particles by Phanerochaete chrysosporium MUCL 19343. Samples with different microbial biomass fractions were obtained from two different experiments, i.e., shake flask and fixed-bed reactor experiments. The glucosamine concentration was compared to the normalized absorbance ratio of the 1651 cm -1 to 1593 cm -1 peak, measured by FTIR-ATR, and resulted in a linear relationship. The application of a normalized absorbance ratio in function of time provided a graph that was similar to the microbial growth curve. Application of FTIR in ATR mode to follow-up kinetics during solid state fermentation seems to be a fast and easy alternative to laborious measurement techniques, such as glucosamine determination., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The present work investigated the effect of lead (Pb) on the growth, metal accumulation, oxidative stress, and antioxidant response in Phanerochaete chrysosporium, which is a well-known hyperaccumulating species for heavy metal with appreciable bioaccumulation capacity. Results revealed that P. chrysosporium exhibited a good ability in Pb accumulation and tolerance over a concentration range of 50-100 mg L -1 Pb. The removal rate of Pb decreased with the increasing levels of Pb and reached a maximum of 91.3% at 50 mg L -1 . Both extracellular adsorption and intracellular bioaccumulation contributed to the removal of Pb, with the maximum of 123.8 mg g -1 and 162.5 mg g -1 dry weight, respectively. Pb may exert its toxicity to P. chrysosporium by impairing oxidative metabolism, as evidenced by the enhanced accumulation of hydrogen peroxide (H 2 O 2 ) and lipid peroxidation product malonaldehyde (MDA). P. chrysosporium evolved an antioxidant system by elevating the activity of superoxide dismutase (SOD) and the level of reduced glutathione (GSH) in response to Pb stress, whereas decreasing the activities of catalase (CAT) and peroxidase (POD). Moreover, Pearson correlation analysis demonstrated a good correlation between oxidative stress biomarkers and enzymatic antioxidants. The preset work suggested that P. chrysosporium exhibited an outstanding accumulation of Pb and tolerance of Pb-induced oxidative stress by the effective antioxidant defense mechanism., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Lignolytic fungi initiate lignocellulose decay by producing extracellular oxidative enzymes. For better understanding the enzymatic degradation of lignocellulose by white-rot fungi, we investigated the effect of manganese on the organic matter loss, manganese peroxidase (MnP) activity, and manganese peroxidase gene (mnp) transcription levels during solid-state fermentation of rice straw with Phanerochaete chrysosporium. The results showed that the addition of manganese improved MnP activity and made it reach the peak earlier, promoted fungal growth at the early period (0-9 days), and enhanced the degradation of lignocellulosic waste. The total organic matter loss had a good correlation with fungal biomass during 30 days of cultivation, and manganese amendment promoted the ability of P. chrysosporium to degrade lignocellulose. Quantitative real-time RT-PCR revealed the differential expression of mnp1, mnp2, and mnp3: manganese amendment increased the transcription of mnp1 and mnp2 but not mnp3. The results indicated that manganese stimulated mnp transcription levels and played a post-transcriptional role in MnP production. These findings provide opportunity of development in enzymatic degradation of lignocellulosic waste by P. chrysosporium amended with manganese.

The glucuronoyl esterases (GEs) that have been identified so far belong to family 15 of the carbohydrate esterases in the CAZy classification system and are presumed to target ester bonds between lignin alcohols and (4-O-methyl-)D-glucuronic acid residues of xylan. Few GEs have been cloned, expressed and characterised to date. Characterisation has been done on a variety of synthetic substrates; however, the number of commercially available substrates is very limited. We identified novel putative GEs from a wide taxonomic range of fungi and expressed the enzymes originating from Acremonium alcalophilum and Wolfiporia cocos as well as the previously described PcGE1 from Phanerochaete chrysosporium. All three fungal GEs were active on the commercially available compounds benzyl glucuronic acid (BnGlcA), allyl glucuronic acid (allylGlcA) and to a lower degree on methyl glucuronic acid (MeGlcA). The enzymes showed pH stability over a wide pH range and tolerated 6-h incubations of up to 50 °C. Kinetic parameters were determined for BnGlcA. This study shows the suitability of the commercially available model compounds BnGlcA, MeGlcA and allylGlcA in GE activity screening and characterisation experiments. We enriched the spectrum of characterised GEs with two new members of a relatively young enzyme family. Due to its biotechnological significance, this family deserves to be more extensively studied. The presented enzymes are promising candidates as auxiliary enzymes to improve saccharification of plant biomass.

Mechanisms of silver nanoparticles-mediated toxicity to Phanerochaete chrysosporium and the influence of silver nanoparticles (AgNPs) on the biodegradation of 2,4-dichlorophenol (2,4-DCP) have been systematically investigated. AgNPs at low doses (0-60μM) have greatly enhanced the degradation ability of P. chrysosporium to 2,4-DCP with the maximum degradation rates of more than 94%, exhibiting excellent synergies between AgNPs and P. chrysosporium in the degradation of 2,4-DCP. Meanwhile, removal of total Ag was also at high levels and highly pH dependent. However, significant inhibition was highlighted on 2,4-DCP biodegradation and Ag removal upon treatment with AgNPs at high doses and AgNO 3 at low-level exposure. Results also suggested that AgNPs-induced cytotoxicity could arise from the "Trojan-horse" mechanism executing particle effects, ion effects, or both, ruling out extracellularly released Ag + . Moreover, under relatively low concentrations of AgNPs exposure, 2,4-DCP was broken into linear chain organics, and eventually turned into CO 2 and H 2 O through reductive dechlorination and reaction with hydroxyl radicals. FTIR analysis showed that amino, carboxyl, carbonyl, and sulfur-containing functional groups played crucial roles in Ag transportation and the reduction of Ag + to Ag 0 ., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Clothianidin (CLO) is a member of the neonicotinoid pesticides, which have been widely used worldwide over the last two decades. However, its toxicity for bees and neurological toxicity for humans are urgent problems. Here, the degradation of CLO by the white-rot fungus Phanerochaete sordida was examined in nitrogen-limited liquid medium. After incubation for 20days at 30°C, 37% of CLO was degraded in the cultures. High-resolution ESI-MS and NMR analyses of the culture supernatant identified N-(2-chlorothiazol-5-yl-methyl)-N'-methylurea (TZMU) as a metabolite of CLO degradation. The addition of cytochrome P450 inhibitors to the culture medium markedly reduced the degradation of CLO by P. sordida. And manganese peroxidase, a major ligninolytic enzyme secreted by this fungus, were not carried out CLO degradation. The effects of CLO and TZMU on the viability of the neuronal cell line Neuro2a demonstrated that P. sordida effectively degrades CLO into a metabolite that lacks neurotoxicity., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The removal of Cd(2+) from aqueous solutions by agricultural residues rice straw combined with white rot fungus Phanerochaete chrysosporium (P. chrysosporium) was investigated. The results showed that over 99% of the total Cd(2+) (initial concentration of 150mgL(-1)) was removed at the optimal operating conditions (pH 5.0 at 35°C). We also found that P. chrysosporium could survive under Cd(2+) stress even with an initial Cd(2+) concentration of 250mgL(-1). But when Cd(2+) concentration increased to 250mgL(-1), fungus growth and reproduction were remarkably restrained, and as a result, Cd(2+) removal dropped to 59.2%. It was observed that the fungus biomass and activities of ligninolytic enzymes decreased at some degree under high concentration of Cd(2+) (above 100mgL(-1)). Also, we found that a moderate Cd(2+) stress (below 150mgL(-1)) could stimulate P. chrysosporium's production of the heavy metals chelator - oxalate. This study will provide useful information for the application of biological removal of heavy metal irons from wastewater., (Copyright © 2016 Elsevier Inc. All rights reserved.)

With the wide production and applications of graphene and its derivatives, their toxicity to the environment has received much attention nowadays. In this study, we investigated the toxicity of graphene oxide (GO) to white rot fungus (Phanerochaete chrysosporium). GO was prepared by modified Hummers method and well characterized before use. P. chrysosporium was exposed to GO at the concentrations of 0-4 mg/mL for 7 d. The fresh and dry weights, pH values of culture media, structures, ultrastructures, IR spectra and activities of the decomposition of pollutants were measured to reveal the hazards of GO to P. chrysosporium. Our results indicated that low concentrations of GO stimulated the growth of P. chrysosporium. The exposure to GO induced more acidic pH values of the culture media after 7 d. GO induced the disruption of the fiber structure of P. chrysosporium, while at 4 mg/mL some very long and thick fibers were formed. Such changes were reflected in the scanning electron microscopy investigations, where the disruption of fibers was observed. In the ultrastructural investigations, the shape of P. chrysosporium cells changed and more vesicles were found upon the exposure to GO. The infrared spectroscopy analyses suggested that the chemical compositions of mycelia were not changed qualitatively. Beyond the toxicity, GO did not alter the activities of P. chrysosporium at low concentrations, but led to the complete loss of activity at high concentrations. The implication to the ecological safety of graphene is discussed., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Due to the particular activation and inhibition behavior of silver nanoparticles (AgNPs) on microbes at various concentrations, it's crucial to exploit the special concentration effect in environment. Here, we studied the viability variation of Phanerochaete chrysosporium (P. chrysosporium) under exposure to citrate-coated AgNPs (Citrate-AgNPs) in the presence of different sulfide sources (an inorganic sulfide, NaHS and an organic sulfide, thioacetamide (TAA)). The results indicated that both NaHS and TAA can promote activation of P. chrysosporium by Citrate-AgNPs at a higher concentration, which was initial at toxic level. Treatment with various concentrations of Citrate-AgNPs (0-9 mg/L) demonstrated a maximum activation concentration (MAC) at 3 mg/L. With the increase in sulfide concentration, MAC transferred to higher concentration significantly, indicating the obvious "toxicity to activation" transformation at a higher concentration. Ag(+) testing exhibited that variations in sulfide-induced Ag(+) concentration (3-7 μg/L Ag(+)) accounted for the "toxicity to activation" transformation. In addition, the similar results were observed on antibacterial application using Escherichia coli as the model species. Based on the research results, the application of this transformation in improving antibacterial activity was proposed. Therefore, the antibacterial activity of AgNPs can be controlled, even at concentration, via adjusting for the sulfide concentration.

This study examines the growth, metabolism of Phanerochaete chrysosporium (P. chrysosporium) and route of lignin degradation in response to cadmium (Cd) stress in solid-state fermentation of rice straw. Less living fungi biomass was found under Cd exposure, suggesting that Cd had strong toxicity to P. chrysosporium. The maximum values of lignin peroxidase and manganese peroxidase were 0.34 and 5.21 U g(-1) at the Cd concentration of 32 mg kg(-1), respectively, lower than that in control, which indicated Cd stress would inhibit ligninolytic enzymes. The production of reactive oxygen species (ROS) including hydroxyl radicals (OH), superoxide anion radical (O2(-)) and hydrogen peroxide (H2O2) increased after Cd exposure. Higher concentration of oxalate was detected at high Cd concentrations. Cd stress also had influence on the rates of lignocelluloses degradation and the route of lignin degradation. Partial Cd could be removed by P. chrysosporium., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

When exposed to higher Pb(2+) concentration, Phanerochaete chrysosporium secreted higher content of oxalate and thiol compounds. An earlier and faster increase in oxalate was observed after short-term exposure, comparing with a gentle increase in the thiol compounds. In the extracellular polymeric substances (EPS) extract, more oxalate and T-SH were detected when the initial Pb(2+) was higher, and the variations were different from the situation in the culture medium. In EPS solution, the oxalate amount was more closely related with Pb than that of thiol compounds. pH condition in the whole Pb removal process by P. chrysosporium ranged from 4 to 6.5 and was more beneficial for the binding of Pb(2+) to carboxylic groups in the oxalic acid. More Pb(2+) induced more EPS amount, and the increase of EPS amount influenced the immobilized oxalate more seriously. The present study can supply more comprehensive information about the metal passivation mechanism in white-rot fungi and provide meaningful references for an enhanced removal of heavy metals.

Immobilized Phanerochaete chrysosporium loaded with TiO2 nanoparticles (PTNs) are novel high-value bioremediation materials for adsorbing cadmium and for degrading 2,4-dichlorophenol (2,4-DCP). The real-time changes in H(+) and O2 fluxes were measured using the noninvasive microtest technique (NMT). The H(+) influx increased after the addition of 2,4-DCP, and shifted to efflux following the addition of Cd(2+). The O2 flux decreased after the addition of both 2,4-DCP and Cd(2+). A larger Cd(2+) flux was immediately observed after exposure to 0.5mM Cd(2+) (-351.25 pmol cm(-2) s(-1)) than to 0.1 mM Cd(2+) (-107.47 pmol cm(-2) s(-1)). The removal of Cd(2+) by the PTNs increased more after treatment with the 0.5 mM exposure solution (27.6 mg g(-1)) than with the 0.1 mM exposure solution (3.49 mg g(-1)). The enzyme activities were analyzed to review the antioxidative defense system of PTNs in a solution containing various concentrations of Cd(2+). The activities of the coenzyme nicotinamide adenine dinucleotide (NADH) oxidase as well as the enzyme catalase (CAT) plateaued at 6.5 U g(-1) FW and 9.7 U g(-1) FW, respectively, after exposure to 0.25 mM Cd(2+). The activity of superoxide dismutase (SOD) increased gradually in solutions containing 0.1-0.6 mM Cd(2+), and eventually reached a maximum (68.86 U g(-1) FW). These results illustrate how the antioxidative defense system and the physiological fluxes of PTNs respond to the stress caused by toxic pollutants., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

A series of organosilicon Bola-form bi-quaternary ammonium amphiphiles, [OH5C3(C2H5)2N+-(CH2)3-Si(CH2)3-O-(Si(CH3)2O)n-Si(CH2)3⋯(CH2)3-N+(C2H5)2C3H5O]Cl2- (SinN2Cl2, n=0, 4, 6, 8), with the same headgroups and different length of hydrophobic linkage has been synthesized. The critical micelle concentration (CMC) of each amphiphiles was determined by equilibrium surface tension. With the increased length of hydrophobic siloxane spacer, the CMC values follow the order of Si8N2Cl2

Despite the knowledge about increasing discharge of silver nanoparticles (AgNPs) into wastewater and its potential toxicity to microorganisms, the interaction of AgNPs with heavy metals in the biological removal process remains poorly understood. This study focused on the effect of AgNPs (hydrodynamic diameter about 24.3±0.37 nm) on the removal of cadmium (Cd(II)) by using a model white rot fungus species, Phanerochaete chrysosporium. Results showed that the biological removal capacity of Cd(II) increased with the concentration of AgNPs increasing from 0.1 mg/L to 1 mg/L. The maximum removal capacity (4.67 mg/g) was located at 1 mg/L AgNPs, and then decreased with further increasing AgNPs concentration, suggesting that an appropriate concentration of AgNPs has a stimulating effect on the removal of Cd(II) by P. chrysosporium instead of an inhibitory effect. Results of Ag(+) and total Ag concentrations in the solutions together with those of SEM and XRD demonstrated that added AgNPs had undergone oxidative dissolution and transported from the solution to the surface of fungal mycelia (up to 94%). FTIR spectra confirmed that amino, carboxyl, hydroxyl, and other reducing functional groups were involved in Cd(II) removal, AgNPs transportation, and the reduction of Ag(+) to AgNPs., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The ability of Phanerochaete chrysosporium to reduce the oxidized forms of selenium, selenate and selenite, and their effects on the growth, substrate consumption rate, and pellet morphology of the fungus were assessed. The effect of different operational parameters (pH, glucose, and selenium concentration) on the response of P. chrysosporium to selenium oxyanions was explored as well. This fungal species showed a high sensitivity to selenium, particularly selenite, which inhibited the fungal growth and substrate consumption when supplied at 10 mg L(-1) in the growth medium, whereas selenate did not have such a strong influence on the fungus. Biological removal of selenite was achieved under semi-acidic conditions (pH 4.5) with about 40 % removal efficiency, whereas less than 10 % selenium removal was achieved for incubations with selenate. P. chrysosporium was found to be a selenium-reducing organism, capable of synthesizing elemental selenium from selenite but not from selenate. Analysis with transmission electron microscopy, electron energy loss spectroscopy, and a 3D reconstruction showed that elemental selenium was produced intracellularly as nanoparticles in the range of 30-400 nm. Furthermore, selenite influenced the pellet morphology of P. chrysosporium by reducing the size of the fungal pellets and inducing their compaction and smoothness.

Prevalence of heavy metals in the living environment causes chemical stress and reactive oxygen species (ROS) formation in Phanerochaete chrysosporium (P. chrysosporium). However, the mechanisms involved in ROS defense are still under investigation. In the present study, we evaluated the effect of lead- and cadmium-induced oxidative stress on the activities of catalase (CAT), peroxidase (POD), lignin peroxidase (LiP), and manganese peroxidase (MnP). A time-dependent change in all enzyme activities was observed following exposure to 50 μM cadmium and 25 μM lead. The lowest values were recorded at 4 h after exposure. Both cadmium and lead inhibited CAT and POD. The cytochrome P450 (CYP450) levels increased under 50-100 μM cadmium or lead exposure and decreased when heavy metal concentration was under 50 μM; this suggested that ROS is not the only factor that alters the CYP450 levels. The cadmium removal rate in the sample containing 900 μM taxifolin (inhibitor of CYP450) and 100 μM cadmium was reduced to 12.34 %, 9.73 % lower than that of 100 μM cadmium-induced sample, indicating CYP450 may play an indirect but key role in the process of clearance of heavy metals. The pH of the substrate solution decreased steadily during the incubation process.

This study examines the role of oxalic acid in the uptake of Cd and participation in detoxification process in Phanerochaete chrysosporium. Cd-induced oxalic acid secretion was observed with growth inhibition and enzyme inactivation (LiP and MnP) of P. chrysosporium. The peak value of oxalic acid concentration was 16.6 mM at initial Cd concentration of 100 mg L(-1). During the short-term uptake experiments, the uptake of Cd was enhanced and accelerated in the presence of oxalic acid and resulted in alleviated growth and enzyme inhibition ratios. The formation of a metal-oxalate complex therefore may provide a detoxification mechanism via effect on metal bioavailability, whereby many fungi can survive and grow in environments containing high concentrations of toxic metals. The present findings will advance the understanding of fungal resistance to metal stress, which could show promise for a more useful application of microbial technology in the treatment of metal-polluted waste.

The first steps of wood degradation by fungi lead to the release of toxic compounds known as extractives. To better understand how lignolytic fungi cope with the toxicity of these molecules, a transcriptomic analysis of Phanerochaete chrysosporium genes was performed in the presence of oak acetonic extracts. It reveals that in complement to the extracellular machinery of degradation, intracellular antioxidant and detoxification systems contribute to the lignolytic capabilities of fungi, presumably by preventing cellular damages and maintaining fungal health. Focusing on these systems, a glutathione transferase (P. chrysosporium GTT2.1 [PcGTT2.1]) has been selected for functional characterization. This enzyme, not characterized so far in basidiomycetes, has been classified first as a GTT2 compared to the Saccharomyces cerevisiae isoform. However, a deeper analysis shows that the GTT2.1 isoform has evolved functionally to reduce lipid peroxidation by recognizing high-molecular-weight peroxides as substrates. Moreover, the GTT2.1 gene has been lost in some non-wood-decay fungi. This example suggests that the intracellular detoxification system evolved concomitantly with the extracellular ligninolytic machinery in relation to the capacity of fungi to degrade wood., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

In this study, the H2S donor, sodium hydrosulfide (NaHS) was used to pretreat Phanerochaete chrysosporium in order to improve its ability to degrade 2,4-dichlorophenol (2,4-DCP). When pretreated with 100μM NaHS, P. chrysosporium was able to degrade 2,4-DCP completely in 24h, whereas the degradation efficiency of the untreated control was only 57%. The 2,4-DCP-induced oxidative stress was alleviated by NaHS, and the percentage of surviving cells increased by 32%. H2S or HS(-), rather than other compounds derived from NaHS, were responsible for promoting 2,4-DCP degradation by P. chrysosporium. The results of this study suggest that H2S treatment is a potential strategy to alleviate environmental stress and improve the efficiency of the biological removal of pollutants from wastewater., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Phanerochaete chrysosporium are known to be vital hyperaccumulation species for heavy metal removal with admirable intracellular bioaccumulation capacity. This study analyzes the heavy metal-induced glutathione (GSH) accumulation and the regulation at the intracellular heavy metal level in P. chrysosporium. P. chrysosporium accumulated high levels of GSH, accompanied with high intracellular concentrations of Pb and Cd. Pb bioaccumulation lead to a narrow range of fluctuation in GSH accumulation (0.72-0.84 μmol), while GSH plummeted under Cd exposure at the maximum value of 0.37 μmol. Good correlations between time-course GSH depletion and Cd bioaccumulation were determined (R (2) > 0.87), while no significant correlations have been found between GSH variation and Pb bioaccumulation (R (2) < 0.38). Significantly, concentration-dependent molar ratios of Pb/GSH ranging from 0.10 to 0.18 were observed, while molar ratios of Cd/GSH were at the scope of 1.53-3.32, confirming the dominant role of GSH in Cd chelation. The study also demonstrated that P. chrysosporium showed considerable hypertolerance to Pb ions, accompanied with demand-driven stimulation in GSH synthesis and unconspicuous generation of reactive oxygen stress. GSH plummeted dramatically response to Cd exposure, due to the strong affinity of GSH to Cd and the involvement of GSH in Cd detoxification mechanism mainly as Cd chelators. Investigations into GSH metabolism and its role in ameliorating metal toxicity can offer important information on the application of the microorganism for wastewater treatment.

Objective: By analyzing the function and mechanism of nitric oxide in initiating producing lignin peroxidases by phanerochaete chrysosporium, we studied the regulation mechanism triggering the secondary metabolism of white-rot fungi., Methods: Mutant (pcR5305) and wild-type (pc530) strains of phanerochaete chrysosporium were respectively cultured under both the conditions of nitrogen limitation and nitrogen sufficiency. To compare their lignin peroxidases (LiP)-production and nitric oxide(NO)-production kinetics and their different influences on producing LiP after the NO donor Sodium Nitroprusside (SNP) and scavenger cPTIO were respectively added to the nitrogen limitation or sufficiency culture medium to show the function and mechanism of nitric oxide in initiating production of lignin peroxidases by white-rot fungi., Results: Both strains produced nitric oxide (NO) under the two opposite nutritional conditions, but the levels of NO produced were related with the type of strain and the nutritional conditions. Strain pc530 produced NO requiring nutrition depletion and producing of NO was strongly delayed and reduced when it was cultured under nitrogen sufficiency condition. On the contrary, pcR5305 did not require nitrogen depletion to trigger and the levels of NO were higher than that of pc530. The results indicate that LiP content had positive correlation with NO value except the occurrence time of LiP peak value was later than that of NO. The ability of producing LiP was promoted after the NO donor SNP added, but SNP affected more on pc530 than pcR5305 in promoting producing LiP. 15mM cPTIO would greatly repress producing LiP, but could not completely restrain the synthesis of LiP for both strains., Conclusion: By producing NO, Phanerochaete chrysosporium triggers LiP synthesis. However, the evidences do not indicate that NO participates or effect directly in LiP synthesis. It is more likely that NO is reacting as an upstream signal molecule. Besides NO, there are other signal molecules that have a positive effect on NO levels also involving in the regulation producing LiP. The mechanism of the resistance to nutritional repression of pcR5305 in synthesizing lignin degrading peroxidases may be the answer to the different NO production mechanism of pcR5305 from pc530.

In this study the kinetics and equilibrium of phenol biosorption were studied from aqueous solution using batch technique at an initial pH of 5.5. The biosorption was studied on Ca-alginate beads, on non-living mycelial pellets of Phanerochaete chrysosporium immobilized on Ca-alginate, and on free fungal biomass. Ph. chrysosporium was grown in a liquid medium containing mineral and vitamin materials with complex composition. The biosorption process followed pseudo second-order kinetics on all bioadsorbents. The bioadsorption-equilibrium on blank Ca-alginate, free and immobilized fungal biomass can be described by Langmuir, anti-Langmuir and Freundlich isotherm models using nonlinear least-squares estimation., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Interest in environmental-pollutant-induced oxidative stress and knowledge of the interactions between reactive oxygen species and cellular systems have increased in toxicology and microbial ecology considerably in recent decades. These reactive oxidants are produced by a variety of environmental sources: ionizing radiations, ultraviolet light, redox cycling drugs, hyperoxia, ischemia and redox-active xenobiotics or during metabolism of environmental pollutants, such as heavy metals in mining industries, dyes in wastewater of textile industries, pesticides and polycyclic hydrocarbons, i.e. foreign materials. In this study, the effect of dye on the antioxidative defence system of Phanerochaete chrysosporium was investigated, and we showed the ability of Phanerochaete chrysosporium to antioxidative response and defence system exposed to Astrazone Red FBL. Catalase, glutathione reductase, glutathione s-transferase activities and level of glutathione decreased, depending on the period of growth in each exposure to low and high concentration group (20 and 50 ppm) compared with the control group., (Copyright © 2012 John Wiley & Sons, Ltd.)

The expression of three ribosome binding proteins namely; polyadenylate-binding protein, splicing factor RNPS1 and ATP-dependent RNA helicase of Phanerochaete chrysosporium exposed to lead were analyzed by real-time PCR. The mRNA level of splicing factor RNPS1 showed 2.7 (p < 0.05), 2.6 (p < 0.05) and 4.9-fold (p < 0.001) increase when the cells were exposed to 25, 50 and 100 μM lead, respectively. 50 and 100 μM lead exposure resulted in almost 2-fold (p < 0.01and p < 0.05, respectively) increase in the expression of ATP-dependent RNA helicase. Polyadenylate-binding protein mRNA levels did not reveal any significant increase when cells exposed to the concentrations of lead employed. However, the mRNA level of polyadenylate-binding protein and splicing factor RNPS1 within a period of 1 and 2 h temporal exposure to 100 μM lead showed 2.5 (p < 0.001) and 3.4-fold (p < 0.001) increase, respectively. Expression level of ATP-dependent RNA helicase was not affected from the period of exposure to this metal.

The white-rot fungus Phanerochaete chrysosporium has been widely used for the treatment of waste streams containing heavy metals and toxic organic pollutants. The development of fungal-based treatment technologies requires detailed knowledge of the relationship between bulk water quality and the physiological responses of fungi. A noninvasive microtest technique was used to quantify real-time changes in proton, oxygen, and cadmium ion fluxes following the exposure of P. chrysosporium to environmental toxic (2,4-dichlorophenol and cadmium). Significant changes in H(+) and O(2) flux occurred after exposure to 10 mg/L 2,4-dichlorophenol and 0.1 mM cadmium. Cd(2+) flux decreased with time. Reactive oxygen species formation and antioxidant levels increased after cadmium treatment. Superoxide dismutase activity correlated well with malondialdehyde levels (r(2) = 0.964) at low cadmium concentrations. However, this correlation diminished and malondialdehyde levels significantly increased at the highest cadmium concentration tested. Real-time microscale signatures of H(+), O(2), and Cd(2+) fluxes coupled with oxidative stress analysis can improve our understanding of the physiological responses of P. chrysosporium to toxic pollutants and provide useful information for the development of fungal-based technologies to improve the treatment of wastes cocontaminated with heavy metals and organic pollutants.

Introduction: The toxic effect of the oxidation hair dyes on Phanerochaete chrysosporium was investigated by exposure of this fungus in a nitrogen-limited culture medium to various concentrations of the oxidation hair dyes., Results: The results showed that both the size and the dry weight of the mycelial pellets of P. chrysosporium could be reduced when the concentration of the oxidation hair dyes was higher than 300 mg/L. By using the AFLP analysis and the UPGMA dendrogram, the DNA damage of P. chrysosporium by the oxidation hair dyes was also detected. Comparing with that in the control, the percent polymorphism under different concentrations of the oxidation hair dyes increased. In the meantime, the DNA similarity was decreased, which meant that the DNA damage was aggravated with an increase in the concentrations of the oxidation hair dyes., Conclusion: Thus, as an environmental pollutant, the oxidation hair dyes have a toxic effect on P. chrysosporium at both cellular and molecular levels.

Effects and mechanisms of heavy metals Cu, Cd and Pb on the growth of white-rot fungus P. chrysosporium and its ability of BDE-209 degradation were studied. The results showed that low concentrations of heavy metals (< or = 1 mg x L(-1)) stimulated the growth of P. chrysosporium with the order of Cd > Pb > Cu when 1 mg x L(-1) of each heavy metal was concerned, while high concentrations (> 1 mg x L(-1)) depressed it. P. chrysosporium degraded BDE-209 efficiently with degradation efficiency reaching 69.7% after 7 days. The presence of heavy metals significantly influenced the capability of P. chrysosporium to decompose BDE-209 (P < 0.05). Low concentrations of Cu (< or = 1 mg x L(-1)) and Cd (< or = 0.5 mg x L(-1)) accelerated the degradation of BDE-209 and the degradation efficiency was enhanced from 69.7% to 84.4% when 1 mg x L(-1) Cu was present, while Pb had negative effect. On the other hand, all three metals under high concentrations (> 1 mg x L(-1)) showed depressed effects on the degradation in the order of Cd > Pb > Cu. The growth of P. chrysosporium did not completely positively correlate with the degradation capability of BDE-209. The degradation of BDE-209 by P. chrysosporium conformed to the first-order kinetic model. The reaction rate constant k raised firstly and then declined with increasing concentrations of Cu and Cd, and the highest k of 0.321 2 achieved in the presence of 1 mg x L(-1) Cu. By contrast, the constant k declined all the way when Pb existed. A further investigation into the effects of heavy metals on degradation of BDE-209 by extracellular enzymes derived from P. chrysosporium was conducted, and the distance correlation analysis of the degradation by extracellular enzymes and the whole cell was carried out. The results demonstrated that the degradation by extracellular enzymes and the whole cell was 63.7%, 69.7% separately, showing no significant difference of degradation capability between them, which proved that the extracellular enzymes played dominating role in the degradation of BDE-209. Furthermore, the distance correlation coefficient R were all greater than 0.9 when three heavy metals existed, certificating that heavy metals affected the degradation of BDE-209 through the interaction between extracellular enzymes and heavy metals.

In this study, the impact of bacterial and fungal processes on (14)C-hexadecane mineralisation was investigated in weathered hydrocarbon contaminated soil. The extent of (14)C-hexadecane mineralisation varied depending on the bioremediation strategy employed. Under enhanced natural attenuation conditions, (14)C-hexadecane mineralisation after 98 days was 8.5 ± 3.7% compared to <1.2% without nitrogen and phosphorus additions. (14)C-hexadecane mineralisation was further enhanced through Tween 80 amendments (28.9 ± 2.4%) which also promoted the growth of a Phanerochaete chyrsosporium fungal mat. Although fungal growth in weathered hydrocarbon contaminated soil could be promoted through supplementing additional carbon sources (Tween 80, sawdust, compost, pea straw), fungal (14)C-hexadecane mineralisation was negligible when sodium azide was added to soil microcosms to inhibit bacterial activity. In contrast, when fungal activity was inhibited through nystatin additions, (14)C-hexadecane mineralisation ranged from 6.5 ± 0.2 to 35.8 ± 3.8% after 98 days depending on the supplied amendment. Bacteria inhibition with sodium azide resulted in a reduction in bacterial diversity (33-37%) compared to microcosms supplemented with nystatin or microcosms without inhibitory supplements. However, alkB bacterial groups were undetected in sodium azide supplemented microcosms, highlighting the important role of this bacterial group in (14)C-hexadecane mineralisation., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The occurrence of cytochrome P450 and P450-mediated pentachlorophenol oxidation in a white rot fungus Phanerochaete chrysosporium was demonstrated in this study. The carbon monoxide difference spectra indicated induction of P450 (103±13 pmol P450 per mg protein in the microsomal fraction) by pentachlorophenol. The pentachlorophenol oxidation by the microsomal P450 was NADPH-dependent at a rate of 19.0±1.2 pmol min(-1) (mg protein)(-1), which led to formation of tetrachlorohydroquinone and was significantly inhibited by piperonyl butoxide (a P450 inhibitor). Tetrachlorohydroquinone was also found in the cultures, while the extracellular ligninases which were reported to be involved in tetrachlorohydroquinone formation were undetectable. The formation of tetrachlorohydroquinone was not detectable in the cultures added with either piperonyl butoxide or cycloheximide (an inhibitor of de novo protein synthesis). These results revealed the pentachlorophenol oxidation by induced P450 in the fungus, and it should be the first time that P450-mediated pentachlorophenol oxidation was demonstrated in a microorganism. Furthermore, the addition of the P450 inhibitor to the cultures led to obvious increase of pentachlorophenol, suggesting that the relationship between P450 and pentachlorophenol methylation is worthy of further research.

The in vivo biodegradation of the diazo dye Reactive Black 5 (RB5) by Phanerochaete chrysosporium immobilised on cubes of nylon sponge and on sunflower-seed shells (SS) in laboratory-scale bioreactors was investigated. The SS cultivation led to the best results with a decolouration percentage of 90.3% in 72 h for an initial RB5 concentration of 100 mg/L. It was found that the addition of 0.4 mM veratryl alcohol (VA) into the medium considerably increased the decolouration rate in SS cultivation. However, the addition of VA had no effect in the nylon cultivation. Thin layer chromatography (TLC) revealed that RB5 was transformed into one metabolite after 24 h. UV-vis spectroscopy and Fourier Transform Infrared (FT-IR) also confirmed the biodegradation of RB5. Toxicity of RB5 solutions before and after fungal treatment was assayed using Sinorhizobium meliloti as a sensitive soil microorganism. P. chrysosporium transformed the toxic dye RB5 into a non-toxic product., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Lignocellulosic biomass contains cellulose and xylan as major structural components, and starch as a storage polysaccharide. In the present study, we have used comparative secretomic analysis to examine the effects of xylan and starch on the expression level of proteins secreted by the basidiomycete Phanerochaete chrysosporium grown on cellulose,. Forty-seven spots of extracellular proteins expressed by P. chrysosporium separated by two-dimensional electrophoresis were identified by liquid chromatography-tandem mass spectrometry analysis. Addition of starch to the cellulolytic culture did not affect fungal growth significantly, but did decrease the production of total extracellular enzymes, including cellulases and xylanases. In contrast, addition of xylan increased mycelial volume and the production of extracellular proteins. Xylan increased synthesis of several glycoside hydrolase (GH) family 10 putative endoxylanases and a putative glucuronoyl esterase belonging to carbohydrate esterase family 15, for which plant cell wall xylan may be a substrate. Moreover, cellobiose dehydrogenase and GH family 61 proteins, which are known to promote cellulose degradation, were also increased in the presence of xylan. These enzymes may contribute to degradation by the fungus of not only cellulose but also complex carbohydrate components of the plant cell wall., (© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

Aims: To overproduce manganese peroxidase (MnP) using Phanerochaete chrysosporium. Effects of carbon sources, agricultural by-products and small molecules were tested to enhance the MnP productivity., Methods and Results: Various carbon sources and agricultural by-products were compared as the basal medium. A MnP activity of 4.7 ± 0.31 U ml⁻¹ was obtained using mannose as a carbon source. The enzyme productivity further reached 7.36 ± 0.05 and 8.77 ± 0.23 U ml⁻¹ when the mannose medium was supplemented with cyclic adenosine monophosphate (cAMP) and S-adenosylmethionine, respectively., Conclusions: This study revealed the highest MnP productivity obtained by optimizing the carbon sources and supplementing small molecules. It can provide insight for: (i) making high quantities of enzymes by optimizing media resources and (ii) engineering the global regulatory genes in P. chrysosporium for the onsite production of MnP., Significance and Impact of the Study:  As MnP is an important enzyme to hydrolyse lignin polymers which protects the plant cell wall from exposing of cellulose fibres, the production of the enzyme is a current demand for biofuel biotechnology. The knowledge generated from this study can help to advance the technology for stable production of MnP., (© 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.)