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.

Azo Dye Mineralization by Phanerochaete Chysosporium in a Sequencing Bath Reactor

ABSTRACT The mineralization of the azo dye congo red by the fungi Phanerochaete chrysosporium was studied in two sequential batch bioreactors (R1 and R2), operated in cycles of 48 h (step I) and 24 h (step II). In step I, glucose concentration was 1 g.L-1 in both reactors and in step II, 1 g.L-1 of glucose was maintained in R1, but R2 received no addition of glucose. In step I, the average dye removal efficiencies were 76 ± 29 % (R1) and 53 ± 15% (R2), while in step II the averages recorded for dye removal for R1 and R2 were 84 ± 15 and 70 ± 28%, respectively. The rates of dye removal were 0.04 h-1 in R1 and 0.03 h-1 in R2 in step I. Higher rates were obtained in step II, 0,07 h-1 and 0,02 h-1 for R1 and R2, respectively. The highest dye removal occurred in R1 and, in R2, the residual dye was further removed. Laccase was the oxidised at higher amount, in step I was 54 μmol.min-1 for R1 and 38 μmolmin-1 for R2. The proposed treatment system was very effective in removing the azo dye, however the mineralization may not be complete and some by-products may have been formed, according to spectrofotometric analysis, were the peak corresponding to benzene, 220 nm, persisted.

Assessment of biodegradability of PVC containing cellulose by white rot fungus

Aims: Polyvinyl chloride (PVC) is the most widely used and environmentally damaging plastic. Processing, production and disposal of PVC cause release of toxic chlorine based compounds into environment. The objective of the present study was to assess the biodegradability of cellulose blended PVC by white rot fungi i.e. Phanerochaete chrysosporium. Methodology and results: Biodegradability of the strain for the polymer was tested on plate assay, sturm test, soil burial and shake flask experiments. The biodegradability of the polymer was determined by visual changes, plate assay and carbon dioxide production. Morphological changes in the polymer such as pits, extensive spotting, clear surface erosion, fungal attachment, roughening and deterioration of some parts were observed using scanning electron microscopy. Chemical changes like appearance and shortening of peaks using fourier transform infrared spectroscopy also confirmed the biodegradability of the polymer. Conclusion, significance and impact of study: The present study confirmed that mixing of small amount of cellulose increases the hydrophilicity of the polymer and lead to its microbial degradation and Phanerochaete chrysosporium has great potential for the treatment of solid waste containing plastics.

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.

Evaluation of Potential Fungal Species for the in situ Simultaneous Saccharification and Fermentation (SSF) of Cellulosic Material

Three fungal species were evaluated for their abilities to saccharify pure cellulose. The three species chosen represented three major wood-rot molds; brown rot (Gloeophyllum trabeum), white rot (Phanerochaete chrysosporium) and soft rot (Trichoderma reesei). After solid state fermentation of the fungi on the filter paper for four days, the saccharified cellulose was then fermented to ethanol by using Saccharomyces cerevisiae. The efficiency of the fungal species in saccharifying the filter paper was compared against a low dose (25 FPU/g cellulose) of a commercial cellulase. Total sugar, cellobiose and glucose were monitored during the fermentation period, along with ethanol, acetic acid and lactic acid. Results indicated that the most efficient fungal species in saccharifying the filter paper was T. reesei with 5.13 g/100 g filter paper of ethanol being produced at days 5, followed by P. chrysosporium at 1.79 g/100 g filter paper. No ethanol was detected for the filter paper treated with G. trabeum throughout the five day fermentation stage. Acetic acid was only produced in the sample treated with T. reesei and the commercial enzyme, with concentration 0.95 and 2.57 g/100 g filter paper, respectively at day 5. Lactic acid production was not detected for all the fungal treated filter paper after day 5. Our study indicated that there is potential in utilizing in situ enzymatic saccharification of biomass by using T. reesei and P. chrysosporium that may lead to an economical simultaneous saccharification and fermentation process for the production of fuel ethanol.

Overview of parameters influencing biomass and bioreactor performance used for extracellular ligninase production from Phanerochaete chrysosporium

The production of extracellular enzymes is gaining momentum as commercial interests seek alternative ways to improve the productivity in the biotechnology and pharmaceutical industries. Early research studies looked at improving batch bioreactor operational challenges; however, the use of continuous cultures was indicated to be favourable. This led to a new approach developed to produce extracellular enzymes continuously using fixed-film bioreactors from biofilms immobilised on polymeric and inorganic membranes. In this review, the performance of P. chrysosporium biomass, evaluated in terms of ligninase production using different bioreactor operation conditions, is highlighted. Furthermore, the limitations related to the implementation of optimised batch culture conditions to continuous fixed-film bioreactors are discussed. DO transportation, trace element toxicity and lipid peroxidation effects on P. chrysosporium biomass in fixed-film bioreactors operated for elongated periods, are also discussed.

Inmovilización de hongos ligninolíticos para la remoción del colorante negro reactivo 5 / Immobilising lignilolytic fungus for removing reactive black 5 dye

La presencia de colorantes azoicos en aguas residuales de la industria textil es un problema ambiental y sanitario, porque muchos de estos compuestos son cancerígenos. Los tratamientos biológicos son una alternativa para la remoción de ese tipo de colorantes. En el presente trabajo se evaluó el efecto de tres hongos de podredumbre blanca, Trametes versicolor, Pleurotus ostreatus y Phanerochaete chrysosporium sobre la decoloración de un agua que contiene colorante negro reactivo 5 (NR5), ampliamente usado en la industria textil. Se estudió la inmovilización de estos hongos en dos soportes, espuma de poliuretano y estropajo (L. cylíndrica) para seleccionar el mejor soporte y el hongo con mayor capacidad para la decoloración. Ambos soportes fueron igualmente efectivos, pero se seleccionó estropajo por ser un producto natural. El hongo que generó los mayores porcentajes de decoloración en 4 días fue Trametes versicolor, con 96%, 98% y 98% para agua con concentración de NR5 300 ppm, 150 ppm y 75 ppm, respectivamente. La actividad lacasa para cada concentración de NR5 fue 8 U L-1, 7 U L-1 y 5 U L-1.

Waste water from the textile industry represents a major environmental and health problem because it contains azo dyes whose carcirogenic effect has been tested in research. Biological treatment represents a valuable alternative for removing these dyes. The effect of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium rot fungi on decoloration of water containing reactive black five (NR5) textile dye was evaluated in this work. Immobilising the fungi on polyurethane foam and luffa sponge (Luffa cylindrica) supports was studied in order to select the best support and the fungi having the best decolorisation. Both supports were equally effective; however, the luffa sponge was selected as being a natural product. Trametes versicolor produced the highest decolorisation percentages in four days (96%, 98% and 98% for 300 ppm, 150 ppm and 75 ppm NR5 concentrations, respectively) while lacase enzyme activity was 8 UL-1, 7 UL-1 and 5 UL-1 for each of them.

Enhanced production of manganese peroxidase by Phanerochaete chrysosporium

Production of manganese-dependent peroxidase (MnP) by the white-rot fungus Phanerochaete chrysosporium BKM-F-1767 (ATCC 24725) was monitored during growth in different media and growth conditions. The effect of some activators of MnP production, Mn2+, Tween 80, phenylmethylsulphonylfloride (PMSF), oxygen, temperature, pH, glycerol and nitrogen was studied. Supplementing the cultures with Tween 80 (0.05 percent, v/v) and Mn2+ (174 µM) resulted a maximum MnP activity of 356 U/L which was approximately two times higher than that obtained in the control culture (without Tween 80). Decolourisation of Direct Blue 15 and Direct Green 6 (50 mg/L) was also achieved with MnP.

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.