Phanerochaete Chrysosporium Strain Research Articles

The effect of microorganisms on the biomodification of montan resin from lignite

Montan resin (MR) is an industrial by-product or solid waste generated during the production of refined montan wax and is not typically reused. In this paper, a bio-modification method using three strains of microorganisms, Acinetobacter venetianus (AV), Pseudomonas aeruginosa (PA), and Phanerochaete chrysosporium (PC), was studied to promote the performance and bio-function of MR so that MR could be recycled. MR can be degraded by these three microorganisms, and their weight loss rates were similar over the treatment period of 15 days. Compared with the original MR, the hydrophilicity of modified MRs was improved, which was related to the increase in apparent oil-water partition coefficients (Kows) and oxygen-containing and hydrophilic groups in modified MRs based on IR and GC-MS analysis. The bio-function of modified MRs by the three strains in terms of promoting maize seed germination and seedling growth was greater compared with untreated MR. Overall, these findings indicate that biomodified MRs might have useful agriculture applications.

Decolorization and detoxification of different azo dyes by Phanerochaete chrysosporium ME-446 under submerged fermentation.

Azo dyes are widely used in the textile industry due to their resistance to light, moisture, and oxidants. They are also an important class of environmental contaminant because of the amount of dye that reaches natural water resources and because they can be toxic, mutagenic, and carcinogenic. Different technologies are used for the decolorization of wastewater containing dyes; among them, the biological processes are the most promising environmentally. The aim of this study was to evaluate the potential of Phanerochaete chrysosporium strain ME-446 to safely decolorize three azo dyes: Direct Yellow 27 (DY27), Reactive Black 5 (RB5), and Reactive Red 120 (RR120). Decolorization efficiency was determined by ultraviolet-visible spectrophotometry and the phytotoxicity of the solutions before and after the fungal treatment was analyzed using Lactuca sativa seeds. P. chrysosporium ME-446 was highly efficient in decolorizing DY27, RB5, and RR120 at 50 mg L-1, decreasing their colors by 82%, 89%, and 94% within 10 days. Removal of dyes was achieved through adsorption on the fungal mycelium as well as biodegradation, inferred by the changes in the dyes' spectral peaks. The intensive decolorization of DY27 and RB5 corresponded to a decrease in phytotoxicity. However, phytotoxicity increased during the removal of color for the dye RR120. The ecotoxicity tests showed that the absence of color does not necessarily translate to an absence of toxicity.

Phanerochaete chrysosporiumstrain B-22, a nematophagous fungus parasitizing Meloidogyne incognita.

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.

Effect of Different Carbon Sources on Decolourisation of an Industrial Textile Dye Under Alkaline–Saline Conditions

White-rot fungal strains of Trametes versicolor and Phanerochaete chrysosporium were selected to study the decolourisation of the textile dye, Reactive Black 5, under alkaline-saline conditions. Free and immobilised T. versicolor cells showed 100% decolourisation in the growth medium supplemented with 15gl(-1) NaCl, pH 9.5 at 30°C in liquid batch culture. Continuous culture experiments were performed in a fixed-bed reactor using free and immobilised T. versicolor cells and allowed 85-100% dye decolourisation. The immobilisation conditions for the biomass and the additional supply of carbon sources improved the decolourisation performance during a long-term trial of 40days. Lignin peroxidase, laccase and glyoxal oxidase activities were detected during the experiments. The laccase activity varied depending on carbon source utilized and glycerol-enhanced laccase activity compared to sucrose during extended growth.

Biodegradation of atrazine by Phanerochaete chrysosporium in optimized conditions

Atrazine is an organochloride herbicide widely used for controlling grassy weeds in agriculture. In this study, biodegradation of atrazine with Phanerochaete chrysosporium strain and the optimum conditions of biodegradation were investigated. The amount of atrazine in the medium was determined through HPLC analysis. The biodegradation experiments were carried out indicated that the optimum incubation time was 5 days and the optimum medium pH was 3.0. In addition, the optimum incubation temperature was determined as 30◦C and the optimum agitation rate was determined as 150 rpm in the relevant experiments. The results of these experiments showed that the lignin peroxidase and manganese peroxidase enzymes do not have any effects on atrazin biodegradation.

An oligopeptide transporter gene family in Phanerochaete chrysosporium

Oligopeptide transporters are believed to translocate their substrates across the membrane from the extracellular environment into cell. In the present study, sixteen oligopeptide transporter (OPT) orthologs (PcOPT1 to PcOPT16) were identified in Phanerochaete chrysosporium strain BKMF1767. They were predicted to encode integral membrane proteins with 13 to 16 transmembrane domains. The cDNA and genomic sequences of these genes were confirmed by clone sequencing and Transcriptome Sequencing. Based on sequence similarities, the PcOPT genes can be grouped into three different subfamilies. Comprehensive analysis revealed correlations between gene groupings and gene structures. The PcOPT genes showed differential expression patterns at different metabolic stages or in various stress conditions, suggesting that they may play different functional roles. PcOPT14 was able to mediate the uptake of tetrapeptide KLGL when heterologously expressed in Saccharomyces cerevisiae. This study provides some basic genomic information for OPT genes in P. chrysosporium and will be useful for elucidating their roles in growth, stress response and survival of this organism. Moreover, the results can help us to better understand the complexity of metabolism of P. chrysosporium.

Validation and application of HPLC–ESI-MS/MS method for the quantification of RBBR decolorization, a model for highly toxic molecules, using several fungi strains

A novel analytical method using HPLC–MS/MS operating in selected reaction monitoring (SRM) for evaluation of fungi efficacy to decolorize Remazol Brilliant Blue R (RBBR) dye solution was developed, validated and applied. The method shows high sensibility allowing the detection of 4.6pM of RBBR. Four fungal strains were tested in liquid medium, three strains of Aspergillus (Aspergillus aculeatus, Aspergillus flavus and Aspergillus fumigatus) and Phanerochaete chrysosporium. All fungi were able to degrade the dye, with efficiencies ranging from 40% for P. chrysosporium up to 99% for A. flavus during a 30-day incubation period. During the experiment, increased accumulation of degradation products was observed in A. flavus cultures containing RBBR. Through the use of full scan HPLC–MS technique it was possible to propose the biogenesis of the microbial metabolic degradation pathway. Screening using microorganisms and RBBR may be hereafter used to investigate microbial biodegradation of high toxicity molecules such as dioxins.

Cloning, expression and characterization of an aryl-alcohol dehydrogenase from the white-rot fungus Phanerochaete chrysosporium strain BKM-F-1767

BackgroundThe white-rot fungus Phanerochaete chrysosporium is among the small group of fungi that can degrade lignin to carbon dioxide while leaving the crystalline cellulose untouched. The efficient lignin oxidation system of this fungus requires cyclic redox reactions involving the reduction of aryl-aldehydes to the corresponding alcohols by aryl-alcohol dehydrogenase. However, the biochemical properties of this enzyme have not been extensively studied. These are of most interest for the design of metabolic engineering/synthetic biology strategies in the field of biotechnological applications of this enzyme.ResultsWe report here the cloning of an aryl-alcohol dehydrogenase cDNA from the white-rot fungus Phanerochaete chrysosporium, its expression in Escherichia coli and the biochemical characterization of the encoded GST and His6 tagged protein. The purified recombinant enzyme showed optimal activity at 37°C and at pH 6.4 for the reduction of aryl- and linear aldehydes with NADPH as coenzyme. NADH could also be the electron donor, while having a higher Km (220 μM) compared to that of NADPH (39 μM). The purified recombinant enzyme was found to be active in the reduction of more than 20 different aryl- and linear aldehydes showing highest specificity for mono- and dimethoxylated Benzaldehyde at positions 3, 4, 3,4 and 3,5. The enzyme was also capable of oxidizing aryl-alcohols with NADP + at 30°C and an optimum pH of 10.3 but with 15 to 100-fold lower catalytic efficiency than for the reduction reaction.ConclusionsIn this work, we have characterized the biochemical properties of an aryl-alcohol dehydrogenase from the white-rot fungus Phanerochaete chrysosporium. We show that this enzyme functions in the reductive sense under physiological conditions and that it displays relatively large substrate specificity with highest activity towards the natural compound Veratraldehyde.

Biochemical and Molecular Characterization of Secreted α-Xylosidase from Aspergillus niger

α-Linked xylose is a major component of xyloglucans in the cell walls of higher plants. An α-xylosidase (AxlA) was purified from a commercial enzyme preparation from Aspergillus niger, and the encoding gene was identified. The protein is a member of glycosyl hydrolase family 31. It was active on p-nitrophenyl-α-d-xyloside, isoprimeverose, xyloglucan heptasaccharide (XXXG), and tamarind xyloglucan. When expressed in Pichia pastoris, AxlA had activity comparable to the native enzyme on pNPαX and IP despite apparent hyperglycosylation. The pH optimum of AxlA was between 3.0 and 4.0. AxlA together with β-glucosidase depolymerized xyloglucan heptasaccharide. A combination of AxlA, β-glucosidase, xyloglucanase, and β-galactosidase in the optimal proportions of 51:5:19:25 or 59:5:11:25 could completely depolymerize tamarind XG to free Glc or Xyl, respectively. To the best of our knowledge, this is the first characterization of a secreted microbial α-xylosidase. Secreted α-xylosidases appear to be rare in nature, being absent from other tested commercial enzyme mixtures and from the genomes of most filamentous fungi.

A quantitative method for fungal ligninolytic enzyme screening studies

AbstractScreening is conducted to select a fungus with desired characteristics intended for various applications, e.g. bioremediation and enzyme production. A qualitative method was used in this study for screening of fungal ligninolytic enzyme activities. The fungal ligninolytic activity was correlated with its growth from the screenings to identify a suitable fungus for solid substrate fermentation. Four strains of fungi, namely, Phanerochaete chrysosporium, Pycnoporus sanguineus, Phlebia radiata and Pleurotus sajor‐caju, were screened for their ligninolytic enzyme activities using guaiacol and remazol brilliant blue‐R (RBB‐R) as screening reagents. The screenings were conducted at both, the optimal growth temperature of each fungus and 35 °C. All the fungi, except P. radiata, showed positive guaiacol oxidation and RBB‐R decolourisation activities. On the basis of the results, P. sajor‐caju may be a suitable fungus for use in bioconversion of lignocellulose because it maintained guaiacol oxidation and RBB‐R decolourisation activities at 35 °C. Its halo‐to‐colony area ratios of guaiacol oxidation and RBB‐R decolourisation after 6 days were 7.0 and 2.7, respectively. In the subsequent growth study, the measured colony area was closely correlated with the dry fungal cell weight with a regression value (R2) of 0.971. Thus, the colony size could be used as an indicator of fungal growth if the screening media do not significantly slow the growth of fungus. Copyright © 2010 Curtin University of Technology and John Wiley & Sons, Ltd.

Ligninolytic Enzymes Produced by Phanerohaete Chrysosporium 1038 and Biotransformation of Lignin

ABSTRACTThe influence of nitrogen and Mn2+ ions concentrations in growth medium on the activity of extracellular lignin degradation enzymes: lignin peroxidase (LiP—EC 1.11.1.14), manganese peroxidase (MnP—EC 1.11.1.13) and laccase (Lac—EC 1.10.3.2), produced by Phanerochaete chrysosporium strain 1038, was studied. Evidence for de lignification processes was provided by FTIR spectroscopy after 24 h incubation of lignin samples with concentrated extracellular fluids of activated strain 1038. FTIR spectra showed decrease of carboxylic groups and increase of hydroxylated and methoxylated aliphatic groups, which indicates an oxidative cleavage. The resulting modifications of the lignin structure are due to the low specificity of the ligninolytic enzyme system, consisting of extracellular LiP, MnP and Lac. The addition of 3.5mg/L Mn2+, 63 mg/L Cu2+ and 2.5mM veratryl alcohol is necessary for the activation of lignolytic complex and obtaining of optimal conditions for delignification process after incubation of lignin samples with fluids from the strain. The effect of growth conditions on the enzyme activities of Phanerochaete chrysosporium 1038 and efficiency of delignification procedure are discussed.

Application of improved chemical–mechanical pulp from poplar wood in the packing paper composition

Populus deltoides variety Onda wood was used for obtaining of improved chemical–mechanical pulp (CMP) under laboratory conditions by preliminary treatment of wood chips with three types of enzymes, such as lignin peroxidase, manganese peroxidase and laccase, produced by Phanerochaete chrysosporium strain 1038.

Lignocellulolytic enzyme production from submerged fermentation of paddy straw

Five strains of cellulolytic bacteria and four strains of Phanerochaete chrysosporium were evaluated for the lignocellulolytic enzyme production during submerged fermentation (SmF) of paddy straw. Extra-cellular enzyme assay for CMCase, FPase, Cellobiase, Xylanase, Lignin peroxidase and Laccase enzymes was performed after 7 and 15 days of submerged fermentation. Cellulomonas cellulans MTCC 23, Cytophaga hutchinsonii NCIM 2338 and Phanerochaete chrysosporium MTCC 787 were found to produce higher lignocellulolytic enzyme activities than rest of the cultures after 15 days of fermentation.

Decolorization of semisolid olive residues of “alperujo” during the solid state fermentation by Phanerochaete chrysosporium, Trametes versicolor, Pycnoporus cinnabarinus and Aspergillus niger

Studies were carried out on decolorization of semisolid olive mill residues called “alperujo” (AL), by four strains of Phanerochaete chrysosporium, Trametes versicolor, Pycnoporus cinnabarinus and Aspergillus niger in solid state frementation (SSF). Fungal strains were selected by their ability to grow on olive mill waste water. The treatment of AL with P. chrysosporium led to higher removal of organic matter and decolorization than P. cinnabarinus and T. versicolor. The strain A. niger leads to a relative improvement of the biodegradability of OMW. Using P. chrysosporium, the treatment of AL substrate in SSF showed an efficient decolorization and an appreciable COD and phenolic content removal only in the presence of a support. Sugarcane bagasse used as support improved the oxygen transfer in the culture. The extracellular fluid of P. chrysosporium (composed mainly of LiP) exhibited high ability to decolorize AL showing the efficiency of the enzyme produced. A percentage of sugarcane bagasse equal to 30% was an optimal condition to improve the growth and the decolorization of AL. The results showed good prospects of using the three basidiomycetes, in particular P. chrysosporium, for the Decolorization of AL.

Decolorization of high concentrations of synthetic dyes by the white rot fungus Bjerkandera adusta strain CCBAS 232

Among thirty different basidiomycetes screened, Bjerkandera adusta strain CCBAS 232, Phanerochaete chrysosporium strain CCBAS 571 and Pleurotus ostreatus strain CCBAS 473 showed the best decolorization properties. B. adusta strain CCBAS 232 was able to decolorize a number of chemically different synthetic dyes (Orange G, Amaranth, Remazol Brilliant Blue R, Cu-phthalocyanine and Poly R-478) at relatively high concentrations of 2–4 g L −1 both on solid and in liquid medium. This strain also grew easily and produced biomass under these conditions. Unlike P. chrysosporium and P. ostreatus, B. adusta was able to efficiently decolorize all tested dyes. These properties and also a good ligninolytic enzyme production predetermine B. adusta strain CCBAS 232 for biotechnological applications.

Synthetic dye decolorization by white rot fungus, Ganoderma sp. WR-1

Decolorization of recalcitrant dyes by an indigenous strain of white rot fungus isolated from bark of dead tree, WR-1 identified as Ganoderma sp. was investigated. The fermentation medium was optimized using a combination of one factor at a time and orthogonal array method. Maximum decolorization (96%) of 100 ppm amaranth was achieved in 8 h with optimized medium containing 2% starch and 0.125% yeast extract. Rate of dye decolorization by the indigenous isolate Ganoderma sp. WR-1 was very high compared to the most widely used strains of Trametes versicolor and Phanerochaete chrysosporium. The broad-spectrum decolorization efficiency of the isolate was assessed using chemically different dyes. The isolate was further evaluated for the decolorization of industrial effluent. Complete decolorization was achieved in 12 days.

Production of multiple laccase isoforms by Phanerochaete chrysosporium grown under nutrient sufficiency

Phanerochaete chrysosporium strain ME446 produced laccase when grown in a medium with high nitrogen (24 mM), glucose, acetate buffer and high copper (0.4 mM CuSO4). Copper appeared to serve as an inducer. The highest amount of laccase activity was found in 2–3 day old cultures. Both intracellular and extracellular laccases eluted as four peaks (L1–L4) on a MonoQ (anion exchange) column. All four peaks from intracellular fluid appear to comprise multiple polypeptides that gave rise to a ∼400 kDa peak on a gel filtration column. Extracellular laccases had a non-denaturing Mr of 100 kDa.

Assessment of wood-inhabiting Basidiomycetes for biokraft pulping of softwood chips

Wood-inhabiting Basidiomycetes have been screened for various applications in the pulp and paper industry and it is evident that different fungi need to be used to suit the specific requirements of each application. This study assessed the suitability of 278 strains of South African wood-decay fungi for the pre-treatment of softwood chips for kraft pulping. The influence of these fungi on kappa number, yield and strength properties of pulp was evaluated. A number of these strains were more efficient in reducing kappa number than the frequently used strains of Phanerochaete chrysosporium and Ceriporiopsis subvermispora. Six strains of Stereum hirsutum and a strain of an unidentified species were able to reduce the kappa number significantly without a significant influence on the pulp yield. Treatment of wood with two strains of S. hirsutum, one strain of Peniophora sp. and a strain of an unidentified species resulted in paper with improved strength properties.

Genome sequence of the lignocellulose degrading fungus Phanerochaete chrysosporium strain RP78.

White rot fungi efficiently degrade lignin, a complex aromatic polymer in wood that is among the most abundant natural materials on earth. These fungi use extracellular oxidative enzymes that are also able to transform related aromatic compounds found in explosive contaminants, pesticides and toxic waste. We have sequenced the 30-million base-pair genome of Phanerochaete chrysosporium strain RP78 using a whole genome shotgun approach. The P. chrysosporium genome reveals an impressive array of genes encoding secreted oxidases, peroxidases and hydrolytic enzymes that cooperate in wood decay. Analysis of the genome data will enhance our understanding of lignocellulose degradation, a pivotal process in the global carbon cycle, and provide a framework for further development of bioprocesses for biomass utilization, organopollutant degradation and fiber bleaching. This genome provides a high quality draft sequence of a basidiomycete, a major fungal phylum that includes important plant and animal pathogens.

Roles of two white-rot basidiomycete fungi in decolorisation and detoxification of olive mill waste water.

A Phanerochaete chrysosporium strain was isolated from Moroccan olive mill waste water (OMW) and its ability to degrade OMW in different culture conditions was investigated and compared to that of Pleurotus ostreatus. The results indicated that Ph. chrysosporium isolate is more efficient than Pl. ostreatus in decolorising and detoxifying OMW in the presence of added nutrients. Ph. chrysosporium is able to remove more than 50% of the colour and phenols from OMW within 6 days of incubation, whereas Pl. ostreatus needs more than 12 days to reach similar results in the same conditions. Many factors affecting the treatment of diluted OMW (20%) by Ph. chrysosporium were studied, including the effects of added nutrients, initial pH, temperature and inoculated biomass. Once the optimisation of 20% OMW biodegradation process had been set up, higher OMW concentrations (50%) were tested. The results show that the fungus is capable of reducing all parameters analysed (colour A395, phenol content and chemical oxygen demand) by at least 60%, after only 9 days of growth.