Use of Phanerochaete Chrysosporium in a Semibatch Reactor for Removal of Indigo Carmine from Synthetic Textile Wastewater

Abstract The textile industry demonstrates a polluting potential from the planting of cotton to the release of wastewater. The presence of dyes in water bodies decreases the passage of sun rays and directly affects the photosynthetic organisms and the ecosystem. Fungi have potential in the treatment of wastewater containing dyes with complex organic structures due to enzymes that they produce. This study evaluated the use of Phanerochaete chrisosporium in the treatment of synthetic effluent from textile industry containing indigo carmine (20 mg/L). The fungus was immobilized in a semibatch reactor. Glucose was the cosubstrate employed in the experiment and it was used in the system at 1g/L at the beginning of the process and 0.5 g /L after 24 hours of reaction. Average dye removal was 84±10% and chemical oxygen demand removal was 79±14%. For nitrogen compounds, the removal efficiencies were 87±11%, 81±11% and 91±9% for ammonia, nitrite and nitrate, respectively. The pH of the medium remained in the acidic range (2.57 to 5.00) throughout the process, with the lowest values recorded in the effluent of each cycle, justified by the release of organic acids from fungi metabolism. There was contamination of the medium by bacteria (710,000 CFU/mL), but the colonies count showed a predominance of fungi (1,365,000 CFU/mL). With the use of the semibatch system after reading of glucose it was observed that the efficiency of dye removal evolved from 72±17% to 84±10%, producing a final effluent with 3.35±1.99 mg/L of indigo, which proves that treatment configuration analyzed is satisfactory for dye removal.

Diversity of Wood-Inhabiting Polyporoid and Corticioid Fungi in Odaesan National Park, Korea

Polyporoid and corticioid fungi are among the most important wood-decay fungi. Not only do they contribute to nutrient cycling by decomposing wood debris, but they are also valuable sources for natural products. Polyporoid and corticioid wood-inhabiting fungi were investigated in Odaesan National Park. Fruit bodies were collected and identified based on morphological and molecular analyses using 28S and internal transcribed spacer regions of DNA sequences. As a result, a total of 149 species, 69 genera, 22 families, and 11 orders were recognized. Half (74 species) of the species were polypores, and the other half (75 species) were corticioid fungi. Most of the species belonged to Polyporales (92 species) followed by Hymenochaetales (33 species) and Russulales (11 species). At the genus level, a high number of species was observed from Steccherinum, Hyphodontia, Phanerochaete, Postia, and Trametes. Concerning distribution, almost all the species could be found below 1,000 m, and only 20% of the species were observed from above 1,000 m. Stereum subtomentosum, Trametes versicolor, T. hirsuta, T. pubescens, Bjerkandera adusta, and Ganoderma applanatum had wide distribution areas. Deciduous wood was the preferred substrate for the collected species. Sixty-three species were new to this region, and 21 species were new to Korea, of which 17 species were described and illustrated.

Effect of pretreatment by solid-state fermentation of sawdust on the pelletization and pellet's properties / 生物工程学报

We pretreated sawdust (Castanopsis fissa Rehd.et Wils) by solid state fermentation (SSF) with Phanerochaete chrysosporium, and then compressed it into pellets with the moisture content of 15% and the pressure of 98 MPa, to solve the problem of low density, low Meyer hardness, high water uptake, and short storage period of pellet in the woody pellet industry. We studied the effects of fermentation time on pelletization and pellets's characteristics (including energy consumption, density, Meyer hardness, and hydrophobicity). SSF affected the heating values of pellet. Compared with fresh sawdust, SSF consumed more energy at the maximal value by 6.98% but saved extrusion energy by 32.19% at the maximum. Meanwhile, SSF could improve the density, Meyer hardness and hydrophobicity of pellet. Pellet made of sawdust pretreated by SSF for 48 d had best quality, beneficial for long-term transportation and storage of pellets.

Bioremediation of dyes by fungi isolated from contaminated dye effluent sites for bio-usability

Biodegradation and detoxification of dyes, Malachite green, Nigrosin and Basic fuchsin have been carried out using two fungal isolates Aspergillus niger, and Phanerochaete chrysosporium, isolated from dye effluent soil. Three methods were selected for biodegradation, viz. agar overlay and liquid media methods; stationary and shaking conditions at 25 °C. Aspergillus niger recorded maximum decolorization of the dye Basic fuchsin (81.85%) followed by Nigrosin (77.47%), Malachite green (72.77%) and dye mixture (33.08%) under shaking condition. Whereas, P. chrysosporium recorded decolorization to the maximum with the Nigrosin (90.15%) followed by Basic fuchsin (89.8%), Malachite green (83.25%) and mixture (78.4%). The selected fungal strains performed better under shaking conditions compared to stationary method; moreover the inoculation of fungus also brought the pH of the dye solutions to neutral from acidic. Seed germination bioassay study exhibited that when inoculated dye solutions were used, seed showed germination while uninoculated dyes inhibited germination even after four days of observation. Similarly, microbial growth was also inhibited by uninoculated dyes. The excellent performance of A. niger and P. chrysporium in the biodegradation of textile dyes of different chemical structures suggests and reinforces the potential of these fungi for environmental decontamination.

Heterologous Expression of Phanerochaete chrysoporium Glyoxal Oxidase and its Application for the Coupled Reaction with Manganese Peroxidase to Decolorize Malachite Green

cDNA of the glx1 gene encoding glyoxal oxidase (GLX) from Phanerochaete chrysosporium was isolated and expressed in Pichia pastoris. The recombinant GLX (rGLX) produces H2O2 over 7.0 nmol/min/mL using methyl glyoxal as a substrate. Use of rGLX as a generator of H2O2 improved the coupled reaction with recombinant manganese peroxidase resulting in decolorization of malachite green up to 150 microM within 90 min.

Heterologous Expression of Endo-1,4-beta-xylanaseA from Phanerochaete chrysosporium in Pichia pastoris

The cDNA of endo-1,4-beta-xylanaseA, isolated from Phaenerocheate chrysosporium was expressed in Pichia pastoris. Using either the intrinsic leader peptide of XynA or the alpha-factor signal peptide of Saccharomyces cerevisiae, xylanaseA is efficiently secreted into the medium at maximum concentrations of 1,946 U/L and 2,496 U/L, respectively.

The PcACE1 transcription factor from Phanerochaete chrysosporium contains a Cys- and Ser-rich transactivation domain

Transcription factor Ace1 from Saccharomyces cerevisiae regulates the expression of target genes when the copper concentration reaches 200 ìÌ levels. We are studying the ortholog of Ace1 from fungus Phanerochaete chrysosporium PcACE1, isolated by complementation in yeast. In this report we show the localization of the transactivation region of PcACE1. Different PcACE1 fragments were ligated in frame to the GAL4 DNA-binding domain by site-directed mutagenesis in a suitable yeast expression vector. Transformation of an appropriate Saccharomyces cerevisiae strain was used as host. This strain contains the fusion GAL1:lacZ in its genome under the control of promoter sequences recognized by GAL4. Finally, we measured â-galactosidase activity in each yeast clone. The activation of the reporter gene is proportional to the transactivation capacity of the transcription factor PcACE1. The results obtained indicate that PcACE1 transactivation domain is located in the carboxy terminal half and contains an array of cysteines in the form of Cys-X-Cys and Cys-X2-Cys and a 60% of Ser. Therefore, these results show that this type of Cys motif can function as transcription activating domain not only in transcription factors that respond to minimal copper concentrations but also in those that respond to high copper concentrations. This is the first transactivation domain reported in a basidiomycete fungus.

Environmental Factors and Bioremediation of Xenobiotics Using White Rot Fungi

This review provides background information on the importance of bioremediation approaches. It describes the roles of fungi, specifically white rot fungi, and their extracellular enzymes, laccases, ligninases, and peroxidises, in the degradation of xenobiotic compounds such as single and mixtures of pesticides. We discuss the importance of abiotic factors such as water potential, temperature, and pH stress when considering an environmental screening approach, and examples are provided of the differential effect of white rot fungi on the degradation of single and mixtures of pesticides using fungi such as Trametes versicolor and Phanerochaete chrysosporium. We also explore the formulation and delivery of fungal bioremedial inoculants to terrestrial ecosystems as well as the use of spent mushroom compost as an approach. Future areas for research and potential exploitation of new techniques are also considered.

Biodegradation of reactive orange 16 by Phanerochaete chrysosporium fungus; application in a fluidized bed bioreactor

Application of a fluidized bed bioreactor working for treatment of colored wastewaters has been studied using Phanerochaete chrysosporium fungus immobilized in calcium alginate biogel beads. The working volume of the bioreactor was 1 L; experiments were performed at room temperature and pH of culture medium was initially adjusted to 4. Manganese Peroxidase activity, glucose and ammonium concentrations have been assayed daily along the 7 operating days. Azo dye Reactive Orange 16 was added to the bioreactor after 7 days of incubation and decolorization was assayed by spectrophotometer for 1 h intervals. Maximum Manganese peroxidase activity of 96 +/- 8 U/L was obtained on day 7, and 70 +/- 3% decolorization was achieved after 6 h of dye addition. The results were compared to free cell cultures from previous studies and the role of agitation and immobilization of cells in increasing of the efficiency of decolorization was discussed. The mechanism and morphology of the immobilization of cells in ca-alginate beads were studied and the relationship between glucose and ammonium consumption and ligninolytic activity of fungi were discussed

Relationship between cultivation mode of white rot fungi and their efficiency for olive oil mill wastewaters treatment

Four white rot fungi (WRF) strains, Phanerochaete chrysosporium, Trametes versicolor, Coriolopsis polyzona and Pycnoporus coccineus, were tested for efficiency of treatment of Olive Oil mill wastewaters (OOMW) in relation with their cultivation mode, i.e. under the form of free mycelium, mycelium immobilized in alginate beads and solid state cultivation on Petri dishes. Study of biodegradation of phenolic compounds, chemical oxygen demand (COD) decrease and decolourisation of OOMW have shown that Coriolopsis polyzona and Pycnoporus coccineus degradation performances were apparently only slightly affected by the cell cultivation procedures experienced here. In contrast, Phanerochaete chrysosporium and Trametes versicolor showed respectively marked preferences for solid state and alginate immobilisation procedures. Both mono and polyphenolics were reduced to different extent during incubation depending on the strain, as shown by gel filtration analysis. Final pH obtained after fungal treatment of the OOMW based medium (initial pH of 5.0) was measured in order to evaluate the possibility of releasing friendly the treated wastewater in the environment. Laboratory studies as reported here may be useful for orienting the choice of a strain for treating pollution by OOMW in a particular real situation.

Effect of carbon and nitrogen source amendment on synthetic dyes decolourizing efficiency of white-rot fungus, Phanerochaete chrysosporium.

Decolourization activity of Phanerochaete chrysosporium for three synthetic dyes viz., congo red, malachite green and crystal violet and impact of additional carbon and nitrogen supply on decolourization capacity of fungus were investigated. Maximum decolourizing capacity was observed up to 15 ppm. Addition of urea as nitrogen source and glucose as carbon source significantly enhanced decolourizing capacity (up to 87%) of fungus. In all the cases, both colour and COD were reduced more in non-sterilized treatments as compared to sterilized ones. Significant reductions in COD content of dye solutions (79-84%) were recorded by fungus supplied with additional carbon and nitrogen. A highly significant correlation (r = 0.78, p < 0.001) between colour and COD of dye solutions was recorded. Thus, a readily available carbon and nitrogen source is imperative to enhance the bioremediation activity of this fungus which has been the most suitable for synthetic dyes and textile industry wastewater treatment.

Biodegradation of gaseous chlorobenzene by white-rot fungus Phanerochaete chrysosporium / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate the effect of white rot fungus Phanerochaete chrysosporium on removal of gaseous chlorobenzene.</p><p><b>METHODS</b>Fungal mycelium mixed with a liquid medium was placed into airtight bottles. A certain amount of chlorobenzene was injected into the headspace of the bottles under different conditions. At a certain interval, the concentrations in the headspace were analyzed to evaluate the degradation of chlorobenzene by P. chrysosporium.</p><p><b>RESULTS</b>The degradation effects of P. chrysosporium on chlorobenzene under different conditions were investigated. The difference in the optimum temperature for the growth of the fungi and chlorobenzene degradation was observed. The data indicated that a lower temperature (28 degrees C) would promote the degradation of chlorobenzene than the optimum temperature for the growth of the fungi (37 degrees C). A low nitrogen source concentration (30 mg N/L) had a better effect on degrading chlorobenzene than a high nitrogen source concentration (higher than 100 mg N/L). A high initial concentration (over 1100 mg/m3) of chlorobenzene showed an inhibiting effect on degradation by P. chrysosporium. A maximum removal efficiency of 95% was achieved at the initial concentration of 550 mg/m3.</p><p><b>CONCLUSION</b>P. chrysosporium has a rather good ability to remove gaseous chlorobenzene. A low nitrogen source concentration and a low temperature promote the removal of chlorobenzene by P. chrysosporium. However, a high initial chlorobenzene concentration can inhibit chlorobenzene degradation.</p>

Decolorization of remazol yellow RR gran by white rot fungus Phanerochaete chrysosporium.

In this study, the removal of color, chemical oxygen demand (COD) and aromatic group from one of the azo dyes, Remazol Yellow RR Gran, had been carried out by using one of the white rot fungi, Phanerochaete chrysosporium. Experimental studies were performed in growth media containing different amounts of dye and glucose. Color measurements were done at 436nm wavelength using spectrophotometer while aromatic group measurements were done at 280 nm wavelength using UV/Visible spectrophotometer. As a result of this study the values of the removable color concentration were determined as 10 mgl(-1) and lower. The optimum medium glucose concentration was determined to be 2 gl(-1) during color removal processes, aromatic group measurements were done in samples in the UV region at 280 nm wavelength. As a result of the measurements, it was shown that certain amount of aromatic group remained in the model wastewater at the end of the process.

Microbial decolorization and bioremediation of melanoidin containing molasses spent wash.

Molasses spent wash from cane-molasses based distilleries contains a brown coloured recalcitrantpolymer melanoidin, which if disposed untreated poses a great threat to environment. Microbial decolorization and chemical oxygen demand (COD) reduction was found to be dependent on specific carbon and nitrogen source. Under optimal condition of pH, carbon and nitrogen concentration for each treatment, it was found that Bacillus sp isolated from soil was capable of removing COD (85. 35%) and colour (81.10%) from distillery waste to the maximum extent after 9 days atpH 7 in the medium containing 0.5% peptone, 2% glucose and 10% (v/v), followed by Phanerochaete chrysosporium and lowest reduction was obtained by using native microbial consortium.

Enzymatic transformation of crystalline structure of copolymer poly (ethylene-co-vinyl alcohol) (EVOH)

Changes in the crystalline structure of the copolymer poly(ethylene-co-vinyl alcohol) (EVOH) that occurs during the enzymatic fermentation system with the fungus Phanerochaete chrysosoporium were studied. This fungus has a powerful group of oxidative enzymes that had been used for several processes of environmental pollutants degradation. In order to use the enzymatic system of the fungus to improve degradation, films of the copolymer are inoculated and maintained in a solid state fermentation (SSF) system using as substrate a mixture of agroindustrial corn wastes with an initial humidity of 90 por ciento, during forty days. Analyses of the films were made at different times of fermentation. The effect on the chemical structure of the polymer was studied by using FTIR-spectroscopy with attenuated total reflection (ATR) and changes in the crystalline structure were determined by differential scan calorimetry DSC and X-ray diffraction technique. The lignin peroxidase (LiP) activity was followed by UV-spectrophotometry. We concluded that after 20 days of fermentation the copolymer has lost 50 percent of the initial crystalline structure.

Evaluation of White-rot Fungi for Biopulping of Wood

Ergosterol involves in fungal cell growth as a major component in fungal cell membranes. It can be an indicator that shows the fungal activity, and its content depends on the fungal strains, culture, growth conditions and so on. In this study, fungal activities and growth patterns of three white-rot fungi strains isolated in Korea were evaluated by determination of ergosterol contents during the incubation. Wood decay test and chemical analyses of wood were also performed to verify the relationship between fungal activity and wood degrading capacity of white-rot fungi for 60 days. In the results of experiments, it is considered that the test strains selectively degrade large amount of lignin in wood at the early stage of decay. Especially, Phanerochaete chrysosporium showed the best capability on selective degradation of lignin among the test fungi. It is suggested that the determination of ergosterol content in the fungal culture during the incubation is the simple and effective screening method of white-rot fungi for the application to biopulping of wood.

Vinasse biodegradation by Phanerochaete chrysosporium.

Vinasse is a colored recalcitrant wastewater of the distillery industry. The aim of this work was to study the use of Phanerochaete chrysosporium for the vinasse degradation under two different growth conditions. Vinasse was treated by P. chrysosporium in a liquid inoculum form, during 32 days at room temperature (approximately 25 degrees C) and at 39 degres C. Chemical oxygen demand (COD), total phenol concentration and color removal were measured and there8 was a decrease in COD, phenolic concentration and color of 47.48%, 54.72% and 45.10% respectively, at room temperature and a decrease in 54.21%, 59.41% and 56.8 1% respectively at 39 degrees C.

Taxonomic study on Korean Aphyllophorales (5) - on some unrecorded genera and species -

A total of 149 species and 209 strains of Korean Aphyllophorales in Seoul National University Fungus Collection (SFC) were analyzed by taxonomic and phylogenetic methods. Among those examined fungal specimens, 9 genera Abundisporus, Antrodiella, Cyphellopsis, Dendrothele, Dichomitus, Laxitextum, Piloderma, Skeletocutis and Tubulicrinis, and 23 species, Abundisporus fuscopurpureus, Antrodiella semisupina, Auriporia pileata, Cantharellus subalbidus, Clavulina cinerea, Cyphellopsis confusa, Dendrothele acerina, Dichomitus campestris, Haplotrichum aureum, Heterobasidion annosum, Hyphoderma argillaceum, Hyphodontia tropica, Inonotus dryophilus, Ischnoderma benzoinum, Laxitextum bicolor, Phanerochaete radicata, Phellinus lonicericola, Piloderma byssinum, Skeletocutis nivea, Tomentella terrestris, Trametes elegans, Trametes tenuis, and Tubulicrinis accedens were confirmed as new to Korea and registered here with descriptions.

Cloning and functional characterization of the gene encoding the transcription factor Acel in the basidiomycete Phanerochaete chrysosporium

In this report we describe the isolation and characterization of a gene encoding the transcription factor Acel (Activation protein of cup 1 Expression) in the white rot fungus Phanerochaete chrysosporium. Pc-acel encodes a predicted protein of 633 amino acids containing the copper-fist DNA binding domain typically found in fungal transcription factors such as Acel, Macl and Haal from Saccharomyces cerevisiae. The Pc-acel gene is localized in Scaffold 5, between coordinates 220841 and 222983. A S. cerevisiae acel null mutant strain unable to grow in high-copper medium was fully complemented by transformation with the cDNA of Pc-acel. Moreover, Northern blot hybridization studies indicated that Pc-acel cDNA restores copper inducibility of the yeast cup 1 gene, which encodes the metal-binding protein metallothionein implicated in copper resistance. To our knowledge, this is first report describing an Acel transcription factor in basidiomycetes.

Bioremediation of pulp and paper mill effluent with Phanerochaete chrysosporium.

The present study reports on the treatment of pulp and paper mill effluent by Phanerochaete chlysosporium and the same has been compared at two different pH, 5.5 and 8.5. At both the pH, colour, COD, lignin content and total phenols of the effluent significantly declined after bioremediation. However, greater decolourisation and reduction in COD, lignin content and total phenols were observed at pH 5.5. Such bioremediated effluent of pulp and paper mill could gainfully be utilized for crop irrigation.