Search

Coriolopsis trogii is a basidiomycete fungus which utilizes a large array of lignin-modifying enzymes to colonize and decompose dead wood. Its extracellular enzymatic arsenal includes laccases, i.e., polyphenol oxidases of relevant interest for different industrial applications thanks to their ability to oxidize a diverse range of natural and synthetic compounds. In this work, the production of laccases in C. trogii MUT3379 was explored and improved. From an initial production of ca. 10,000 U L−1, the fermentation process was gradually optimized, reaching a final yield of ca. 200,000 U L−1. An SDS-PAGE analysis of the secretome highlighted the presence of a main protein of ca. 60 kDa showing laccase activity, which was designated as Lac3379-1 once its primary sequence was established by tandem mass spectrometry. The characterization of Lac3379-1 revealed a remarkable enzymatic stability in the presence of surfactants and solvents and a diversified activity on a broad range of substrates, positioning it as an interesting tool for diverse biotechnological applications. The high-yield and robust production process indicates C. trogii MUT3379 as a promising cell factory for laccases, offering new perspectives for industrial applications of lignin-modifying enzymes. [ABSTRACT FROM AUTHOR]

The genome of Trametes versicolor encodes multiple laccase isozymes, the expression of which is responsive to various conditions. Here, we set out to investigate the potential of orange peel extract as an inducer of laccase production in this white-rot fungus, in comparison to the previously identified inducing chemical compound, veratryl alcohol. For four geographically distinct T. versicolor strains, a positive correlation has been observed between their oxidative activity and incubation time in liquid cultures. The addition of 20% orange peel extract or 5 mM veratryl alcohol caused a rapid increase in the oxidative potential of T. versicolor M99 after 24 h, with a more pronounced effect observed for the orange peel extract. To elucidate the underlying molecular mechanisms of the induced laccase activity, a transcriptional gene expression analysis was performed for the seven individual laccase genes in T. versicolor, revealing the upregulation of several laccase genes in response to the addition of each inducer. Notably, the gene encoding TvLac5 demonstrated a substantial upregulation in response to the addition of 20% orange peel extract, likely contributing to the observed increase in its oxidative potential. In conclusion, our results demonstrate that orange peels are a promising agro-industrial side stream for implementation as inducing agents in large-scale laccase production with T. versicolor. [ABSTRACT FROM AUTHOR]

Multi-walled carbon nanotubes (MWCNTs) have significant environmental concerns for soil, water and ecosystems. Since their harmful effects on human health, including respiratory disease, they are considered dangerous substances. Thus, it is required to discover a constructive solution to decrease of MWCNT toxicity. Due to their high degradability, white rot and soft rot fungi have been used in the biodegradation of various pollutants. In this study, we evaluated the ability of native Iranian white and soft rot fungi to degrade MWCNTs mainly by means of dynamic light scattering (DLS) and surface charge measurement. In this regard, growth kinetics, CO2 and protein production, size, surface charge and pH change were measured. Continuous Raman spectroscopy, scanning electron microscope (SEM) and transmission electron microscope (TEM) imaging were carried out to detect the process of degradation. The results showed that the morphology, arrangement, size and diameter of MWCNTs were modified due to fungal degradation. The cytotoxicity of treated MWCNTs was determined by the MTT test. The results indicated that native Iranian white and soft rot fungi have a significant ability for degradation of MWCNTs. Furthermore, it could be concluded that fungal treatment could reduce the toxicity of MWCNTs and the best result was achieved in 500 ppm MWCNTs for Trichoderma sp. The activities of oxidative enzymes such as laccase, manganese peroxidase (MnP) and lignin peroxidase (LiP) were measured to determine the mechanism of degradation. Enzyme assays have suggested that these oxidising enzymes especially laccase might play a key role in the degradation of MWCNTs in white rot and soft rot fungi. The overall results confirmed the reproductive ability of Trichoderma sp. WF29 as a soft rot and two white rot fungi Irpex lacteus WF36 and Trametes versicolor to degrade MWCNT. [ABSTRACT FROM AUTHOR]

Laccases are multicopper oxidases able to oxidize several phenolic compounds and find application in numerous industrial applications. Among laccase producers, white-rot fungi represent a valuable source of multiple isoforms and isoenzymes of these multicopper oxidases. Here we describe the identification, biochemical characterization, and application of laccase 2 from Trametes polyzona (TP-Lac2), a basidiomycete fungus emerged among others that have been screened by plate assay. This enzyme has an optimal temperature of 50 °C and in acidic conditions it is able to oxidize both phenolic and non-phenolic compounds. The ability of TP-Lac2 to decolorize textile dyes was tested in the presence of natural and synthetic mediators at 30 °C and 50 °C. Our results indicate that TP-Lac2 most efficiently decolorizes (decolorization rate > 75%) malachite green oxalate, orange G, amido black10B and bromocresol purple in the presence of acetosyringone and 2,2′-azinobis (3-ethylbenzthiazoline-6-sulfonate)—ABTS. Overall, the laccase mediator system consisting of TP-Lac2 and the natural mediator acetosyringone has potential as an environmentally friendly alternative for wastewater treatment in the textile industry. Key points: A novel laccase from T. polyzona was induced by the addition of veratryl alcohol TP-Lac2 was heterologous expressed in S. cerevisiae and biochemically characterized TP-Lac2 efficiently decolorizes textile dyes using acetosyringone as mediator [ABSTRACT FROM AUTHOR]

To fully harness the potential of laccase in the efficient decolorization and detoxification of single and mixed dyes with diverse chemical structures, we carried out a systematic study on the decolorization and detoxification of single and mixed dyes using a crude laccase preparation obtained from a white-rot fungus strain, Pleurotus eryngii. The crude laccase preparation showed efficient decolorization of azo, anthraquinone, triphenylmethane, and indigo dyes, and the reaction rate constants followed the order Remazol Brilliant Blue R > Bromophenol blue > Indigo carmine > New Coccine > Reactive Blue 4 > Reactive Black 5 > Acid Orange 7 > Methyl green. This laccase preparation exhibited notable tolerance to SO42− salts such as MnSO4, MgSO4, ZnSO4, Na2SO4, K2SO4, and CdSO4 during the decolorization of various types of dyes, but was significantly inhibited by Cl− salts. Additionally, this laccase preparation demonstrated strong tolerance to some organic solvents such as glycerol, ethylene glycol, propanediol, and butanediol. The crude laccase preparation demonstrated the efficient decolorization of dye mixtures, including azo + azo, azo + anthraquinone, azo + triphenylmethane, anthraquinone + indigo, anthraquinone + triphenylmethane, and indigo + triphenylmethane dyes. The decolorization kinetics of mixed dyes provided preliminary insight into the interactions between dyes in the decolorization process of mixed dyes, and the underlying reasons and mechanisms were discussed. Importantly, the crude laccase from Pleurotus eryngii showed efficient repeated-batch decolorization of single-, two-, and four-dye mixtures. This crude laccase demonstrated high stability and reusability in repeated-batch decolorization. Furthermore, this crude laccase was efficient in the detoxification of different types of single dyes and mixed dyes containing different types of dyes, and the phytotoxicity of decolorized dyes (single and mixed dyes) was significantly reduced. The crude laccase efficiently eliminated phytotoxicity associated with single and mixed dyes. Consequently, the crude laccase from Pleurotus eryngii offers significant potential for practical applications in the efficient decolorization and management of single and mixed dye pollutants with different chemical structures. [ABSTRACT FROM AUTHOR]

Being an abundant renewable source of aromatic compounds, lignin is an important component of future bio-based economy. Currently, biotechnological processing of lignin through low molecular weight compounds is one of the conceptually promising ways for its valorization. To obtain lignin fragments suitable for further inclusion into microbial metabolism, it is proposed to use a ligninolytic system of white-rot fungi, which mainly comprises laccases and peroxidases. However, laccase and peroxidase genes are almost always represented by many non-allelic copies that form multigene families within the genome of white-rot fungi, and the contributions of exact family members to the overall process of lignin degradation has not yet been determined. In this article, the response of the Trametes hirsuta LE-BIN 072 ligninolytic system to the presence of various monolignol-related phenolic compounds (veratryl alcohol, p-coumaric acid, vanillic acid, and syringic acid) in culture media was monitored at the level of gene transcription and protein secretion. By showing which isozymes contribute to the overall functioning of the ligninolytic system of the T. hirsuta LE-BIN 072, the data obtained in this study will greatly contribute to the possible application of this fungus and its ligninolytic enzymes in lignin depolymerization processes. [ABSTRACT FROM AUTHOR]

Lentinula edodes (shiitake mushroom) is one of the most widely cultivated edible mushrooms and is primarily cultivated using sawdust medium. While there have been improvements in the cultivation technology, the mechanism of mycelial block cultivation, such as mycelial growth and enzymatic sawdust degradation, has not been clarified. In this study, the mycelium was elongated longitudinally in the bottle sawdust culture for 27 days, and the cultivated sawdust medium was divided into three sections (top, middle, and bottom parts). To determine spatial heterogeneity in the enzyme secretion, the enzymatic activities of each part were analyzed. Lignocellulose degradation enzymes, such as endoglucanase, xylanase, and manganese peroxidase were highly secreted in the top part of the medium. On the other hand, amylase, pectinase, fungal cell wall degradation enzyme (β-1,3-glucanase, β-1,6-glucanase, and chitinase), and laccase activities were higher in the bottom part. The results indicate that the principal sawdust degradation occurs after mycelial colonization. Proteins with the laccase activity were purified from the bottom part of the medium, and three laccases, Lcc5, Lcc6 and Lcc13, were identified. In particular, the expression of Lcc13 gene was higher in the bottom part compared with the level in the top part, suggesting Lcc13 is mainly produced from the tip region and have important roles for mycelial spread and nutrient uptake during early stage of cultivation. [ABSTRACT FROM AUTHOR]

Phlebia radiata is a widespread white-rot basidiomycete fungus with significance in diverse biotechnological applications due to its ability to degrade aromatic compounds, xenobiotics, and lignin using an assortment of oxidative enzymes including laccase. In this work, a chemical screen with 480 conditions was conducted to identify chemical inducers of laccase expression in P. radiata. Among the chemicals tested, phenothiazines were observed to induce laccase activity in P. radiata, with promethazine being the strongest laccase inducer of the phenothiazine-derived compounds examined. Secretomes produced by promethazine-treated P. radiata exhibited increased laccase protein abundance, increased enzymatic activity, and an enhanced ability to degrade phenolic model lignin compounds. Transcriptomics analyses revealed that promethazine rapidly induced the expression of genes encoding lignin-degrading enzymes, including laccase and various oxidoreductases, showing that the increased laccase activity was due to increased laccase gene expression. Finally, the generality of promethazine as an inducer of laccases in fungi was demonstrated by showing that promethazine treatment also increased laccase activity in other relevant fungal species with known lignin conversion capabilities including Trametes versicolor and Pleurotus ostreatus. [ABSTRACT FROM AUTHOR]

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The effect of farnesol, a sesquiterpene alcohol, on the production of laccases by Trametes versicolor and Pycnoporus sanguineus in pineapple waste solid-state fermentation was evaluated. Extracellular laccase production reached a maximum of 77.88 ± 5.62 U/g (236% above control) in farnesol-induced cultures of T. versicolor on the 17th day, whereas in a similar P. sanguineus culture, a maximal laccase activity of 130.95 ± 2.20 U/g (159% increase) was obtained on the 17th day. A single 45 KDa laccase was produced by both fungi under the influence of farnesol. These and other data allow us to conclude that farnesol acted as an inducer of the same form of laccase in both fungi. Farnesol disfavored fungal growth by increasing the lag phase, but it also clearly improved the oxidative state of the cultures. Contrary to the results obtained previously in submerged cultures, farnesol did not promote hyperbranching in the fungal mycelia. This is the first demonstration that farnesol is an excellent inducer of laccases in T. versicolor and P. sanguineus in solid-state cultivation. In quantitative terms, the results can be regarded as an excellent starting point for developing industrial or at least pre-industrial procedures to produce laccases using T. versicolor and P sanguineus under the stimulus of farnesol. [ABSTRACT FROM AUTHOR]

Background: Trametes gibbosa, which is a white-rot fungus of the Polyporaceae family found in the cold temperate zone, causes spongy white rot on wood. Laccase can oxidize benzene homologs and is one of the important oxidases for white rot fungi to degrade wood. However, the pathway of laccase synthesis in white rot fungi is unknown. Results: The peak value of laccase activity reached 135.75 U/min/L on the 9th day. For laccase activity and RNA-seq data, gene expression was segmented into 24 modules. Turquoise and blue modules had greater associations with laccase activity (positively 0.94 and negatively -0.86, respectively). For biology function, these genes were concentrated on the cell cycle, citrate cycle, nicotinate, and nicotinamide metabolism, succinate dehydrogenase activity, flavin adenine dinucleotide binding, and oxidoreductase activity which are highly related to the laccase synthetic pathway. Among them, gene_8826 (MW199767), gene_7458 (MW199766), gene_61 (MW199765), gene_1741 (MH257605), and gene_11087 (MK805159) were identified as central genes. Conclusion: Laccase activity steadily increased in wood degradation. Laccase oxidation consumes oxygen to produce hydrogen ions and water during the degradation of wood. Some of the hydrogen ions produced can be combined by Flavin adenine dinucleotide (FAD) to form reduced Flavin dinucleotide (FADH2), which can be transmitted. Also, the fungus was starved of oxygen throughout fermentation, and the NADH and FADH2 are unable to transfer hydrogen under hypoxia, resulting in the inability of NAD and FAD to regenerate and inhibit the tricarboxylic acid cycle of cells. These key hub genes related to laccase activity play important roles in the molecular mechanisms of laccase synthesis for exploring industrial excellent strains. [ABSTRACT FROM AUTHOR]

Laccase activity secreted by Pleurotus eryngii Han 1787 and Lentinus edodes Han 1788 grown on six types of agroindustrial wastes was investigated. Maximum laccase activity of P. eryngii Han 1787 on Ulmus pumila, Juniperus formosana, Pinus tabuliformis, cottonseed shell, corncob, and leaf of corncob was nearly 5.77-fold, 2.37-fold, 2.78-fold, 2.81-fold, 11.53-fold, and 6.73-fold higher than that of L. edodes Han1788 on corresponding agroindustrial wastes. In general, the capacity of secreting laccase of P. eryngii Han 1787 was superior to that of L. edodes Han 1788. Furthermore, laccase activity of P. eryngii Han 1787 on the leaf of corncob, the corncob, Ulmus pumila, and Juniperus formosana was relatively stable during the whole fermentation process. Different fungi showed different preferences in different agroindustrial wastes to secret laccase on whole fermentation stage. The presence of leaf of corncob was useful for improving laccase activity of P. eryngii Han 1787, while L. edodes Han 1788 was more preferred to produce laccase along with the presence of Juniperus formosana. These results were preliminary conducive in laying the foundation for increasing industrial laccaseproducing strains and producing low-cost laccase. [ABSTRACT FROM AUTHOR]

Lignocellulosic biomass is widely used in the field of laccase production because it has the advantages of low price and easy availability. Thus, a comparative analysis was performed of the laccase secretion ability of five white-rot fungi in submerged fermentation using single or mixtures of lignocellulosic biomass. Maximum laccase activity of Trametes gibbosa An 360, Vanderbylia fraxinea An 369, Perenniporia pyricola Han 202, Coriolopsis trogii Han 474, and Trametes versicolor Han 1504 was 55.83 ± 0.28 U/L on the mixture of corncob and cottonseed hull, 77.96 ± 1.60 U/L on corncob, 443.33 ± 15.49 U/L on corncob, 686.57 ± 16.49 U/L on corncob, and 162.04 ± 11.33 U/L on cottonseed hull. The mixed lignocellulosic material effectively improved the laccase activity of T. gibbosa An 360 compared with other fungal strains. However, the presence of corncob contributed to the secretion of laccase activity for V. fraxinea An 369, P. pyricola Han 202, and C. trogii Han 474. Meanwhile, cottonseed hull was conducive to the secretion of laccase of T. versicolor Han 1504. Laccase activity of P. pyricola Han 202 was very stable throughout most of the fermentation time. The results from this study provide new methods and increase fungal strains to improve laccase activity. [ABSTRACT FROM AUTHOR]

Basidiomycetes are attracting a lot of interest due to their remarkable ability to degrade phthalic acid esters (PAEs). In this article, the degradation process of benzyl butyl phthalate (BBP) and diisobutyl phthalate (DiBP) by the fungus Trametes hirsuta was studied. It was shown that T. hirsuta destroyed >95% of the initial BBP during 24 h, with complete destruction (>99%) occurring by the 3rd day of cultivation. DiBP was destroyed more slowly: 95% of the initial DiBP was destroyed by 10 days. The oxidase activity in the presence of PAEs increased by about 6.0 times, and the esterase activity by 2.0–2.5 times compared to the control. Enzymes involved in the process of PAEs degradation were identified on day 10: manganese peroxidases (MnP2, MnP5, MnP7), versatile peroxidase (VP2), lignin peroxidase (LiP9) and laccase (LacA). For the DiBP, the proportion of MnP5 and LiP9 increased, while the proportion of MnP2 and MnP7 decreased markedly. For the BBP, the proportion of MnP7 increased, while the proportion of VP2 significantly decreased. The GC–MS analysis of PAEs decomposition products were performed. For both PAEs, unique metabolites were found. A toxicity assessment revealed a pronounced toxic effect of metabolites formed during the degradation of BBP. [ABSTRACT FROM AUTHOR]

The textile industry generated a series of synthetic dyestuffs that threatened environmental protection. Azo dyes, widely utilized in textile, paper, fruit, leathers, cosmeceuticals and pharmaceutical fields, account for most of the dyestuffs made. Since they have colour fastness properties, stability, and susceptibility to oxidation, existing effluent treatment methods cannot entirely strip different dyes from effluents. Under certain environmental factors, bacteria decolourize and degrade dyes. The treatment process is cheap, environmentally safe, and can be used on various dyes. However, textile plant wastewater can produce many polluting chemicals and dyes. Environmental legislation is increasingly being enacted to regulate mainly azo-based dyes in the environment. The potential of the microbes for the decolourization of dyes and metabolizing them is long-known knowledge. The toxic components of dyes challenge a potential threat to all the living forms of life. Though both natural and synthetic dyes are used for the colourization of textiles, only synthetic ones are challenging to decolourize. Microbial-based bioremediation of dyes has been studied and reviewed primarily to accelerate dye degradation. The various piece of the literature revealed that the majority of these dye removal microbes belong to mainly white-rot fungi, a consortium of anaerobic bacteria. In addition to this, there are several (genetically engineered microorganisms) GEMs that remediate dyes efficiently. Here in the current review, the authors have tried to bridge the existing gap in the bioremediation of dyestuff. Moreover, the authors have also tried to provide the latest trend in this field. This study will surely benefit the industries and researchers related to dyestuffs by maintaining eco-friendly approaches. [ABSTRACT FROM AUTHOR]

White-rot basidiomycete fungi are a unique group of organisms that evolved an unprecedented arsenal of extracellular enzymes for an efficient degradation of all components of wood such as cellulose, hemicelluloses and lignin. The exoproteomes of white-rot fungi represent a natural enzymatic toolbox for white biotechnology. Currently, only exoproteomes of a narrow taxonomic group of white-rot fungi—fungi belonging to the Polyporales order—are extensively studied. In this article, two white-rot fungi, Peniophora lycii LE-BIN 2142 from the Russulales order and Trametes hirsuta LE-BIN 072 from the Polyporales order, were compared and contrasted in terms of their enzymatic machinery used for degradation of different types of wood substrates—alder, birch and pine sawdust. Our findings suggested that the studied fungi use extremely different enzymatic systems for the degradation of carbohydrates and lignin. While T. hirsuta LE-BIN 072 behaved as a typical white-rot fungus, P. lycii LE-BIN 2142 demonstrated substantial peculiarities. Instead of using cellulolytic and hemicellulolytic hydrolytic enzymes, P. lycii LE-BIN 2142 primarily relies on oxidative polysaccharide-degrading enzymes such as LPMO and GMC oxidoreductase. Moreover, exoproteomes of P. lycii LE-BIN 2142 completely lacked ligninolytic peroxidases, a well-known marker of white-rot fungi, but instead contained several laccase isozymes and previously uncharacterized FAD-binding domain-containing proteins. [ABSTRACT FROM AUTHOR]

Most of the sequenced wood-rotting edible mushroom produce fruiting body at relatively low temperatures. Little information has been known about the high-temperature wood-rotting mushroom. Here, we performed de novo sequencing and assembly of the genome of a high-temperature edible mushroom Pleurotus giganteus from a monokaryotic strain zhudugu2 using the Illumina and Pac-Bio CLR sequencing technologies. P. giganteus, also known as Zhudugu in China, is a well-known culinary edible mushroom that has been widely distributed and cultivated in China, Southeast Asia, and South Asia. The genome consists of 40.00 Mb in 27 contigs with a contig N50 of 4.384 Mb. Phylogenetic analysis reveals that P. giganteus and other strains in Pleurotus clustered in one clade. Phylogenetic analysis and average nucleotide identity analysis indicated that the P. giganteus genome showed a closer relationship with other Pleurotus species. Chromosome collinearity analysis revealed a high level of collinearity between P. ostreatus and P. giganteus. There are 12,628 protein-coding genes annotated in this monoploid genome. A total of 481 enzymes accounting for 514 carbohydrateactive enzymes (CAZymes) terms were identified in the P. giganteus genome, including 15 laccases and 10 class II peroxidases predicted in the genome, which revealed the robustness of lignocellulose degradation capacity of P. giganteus. The mating-A type locus of P. giganteus consisted of a pair of homeodomain mating-type genes HD1 and HD2. The mating-B type locus of P. giganteus consisted of at least four pheromone receptor genes and three pheromone genes. The genome is not only beneficial for the genomeassisted breeding of this mushroom but also helps us to understand the high-temperature tolerance of the edible mushroom. [ABSTRACT FROM AUTHOR]

Coloured wastewater from the textile industry is a very serious global problem. Among 16 different white-rot fungal isolates, Trametes hirsuta PW17-41 revealed high potential for decolourisation of mixed textile dyes (Navy EC-R, Ruby S3B and Super Black G) from real industrial wastewater samples. The efficiency of dye decolourisation was evaluated using the American Dye Manufacturers' Institute (ADMI) standard methodology. The suitable support for fungal mycelium immobilisation was nylon sponges. The optimal dye decolourisation (95.39%) was achieved by using palm sugar and ammonium nitrate as carbon and nitrogen sources, respectively. The initial pH was 5 and the agitation speed was 100 rpm at 30 °C. The ADMI values of textile dyes decreased from 2475 to 114 within two days, reducing the treatment time from seven days before optimisation. The major mechanism of dye decolourisation was biodegradation, which was confirmed by UV–visible and FTIR spectra. Manganese peroxidase (MnP) (4942 U L−1) was found to be the main enzyme during the decolourisation process at an initial dye concentration of 21,200 ADMI. The results indicated the strong potential of immobilised fungal cells to remove high concentrations of textile dyes from industrial wastewater and their potential ability to produce high MnP and laccase activities that can be used in further application. [ABSTRACT FROM AUTHOR]

Ciprofloxacin (CIP), a widely used antibiotic, is frequently detected in the environment due to insufficient wastewater and water treatment. Hence, novel, green and cost-effective technologies are required to enhance the removal of these pollutants. The potency of crude enzymes, especially laccases, produced by white-rot fungi was tested to assess their effectiveness to degrade CIP from water. Crude laccase alone could not oxidize CIP. The addition of syringaldehyde, a redox mediator, resulted in a decrease in antibiotic concentration up to 68.09±0.12% in 24 h, which was the highest removal efficiency achieved with 0.15 mg/mL syringaldehyde and 2 mg/mL of crude laccase (0.1 U/ml). Crude laccase oxidation of CIP was inhibited after 6 h of treatment. To compare, a pure enzyme with the same activity as the crude one removed 86% of CIP in 24 h. No inhibitory effect during the treatment was observed. The estimation of antimicrobial efficiency revealed that after 6 h of treatment, the toxicity towards Escherichia coli decreased by 30%. The wastewater treatment by the crude laccase-mediated system was estimated to significantly reduce the cost of enzymatic treatment. [ABSTRACT FROM AUTHOR]

A laccase from Oudemansiella canarii was immobilized using the crosslinked enzyme aggregate (CLEA) methodology and applied in the degradation of the anthraquinonic dye Remazol Brilliant Blue R (RBBR). The immobilized laccase was superior to the free laccase in both thermal and storage stabilities. Both immobilized and free laccase decolourized 100 mg/L RBBR within 24 h at 30 °C and pH 5.0, but the former was still efficient in degrading the dye after at least 6 cycles. The relationship between the decolorization rate and the RBBR concentration obeyed Michaelis–Menten kinetics, with KM of 0.126 ± 0.044 mmol/L and Vmax of 1.412 ± 0.295 µmol/min for free laccase and KM of 0.159 ± 0.050 mmol/L and Vmax of 1.214 ± 0.242 µmol/min for immobilized laccase. Fourier transform infra-red spectroscopy (FTIR) and mass spectrometry allowed to conclude that the O. canarii laccase acts not only on the chromophore group of the dye, but that it also cleaves other covalent bonds, causing an effective fragmentation of the molecule. The Microtox assay detected a significant diminution in toxicity, a finding corroborated by the phytotoxicity test performed with lettuce seeds. Our results indicate that the immobilized laccase from O. canarii could be useful in biological strategies aiming at degrading unwanted dyes in the environment. [ABSTRACT FROM AUTHOR]

Ganoderma lucidum is a representative white-rot fungus that has great potential to degrade lignocellulose biomass. Laccase is recognized as a class of the most important lignin-degrading enzymes in G. lucidum. However, the comprehensive regulatory mechanisms of laccase are still lacking. Based on the genome sequence of G. lucidum, 15 laccase genes were identified and their encoding proteins were analyzed in this study. All of the laccase proteins are predicted to be multicopper oxidases with conserved copper-binding domains. Most laccase proteins were secreted enzymes in addition to Lac14 in which the signal peptide could not be predicted. The activity of all laccases showed the highest level at pH 3.0 or pH 7.0, with total laccase activity of approximately 200 U/mg protein. Silencing PacC resulted in a 5.2 fold increase in laccase activity compared with WT. Five laccase genes (lac1, lac6, lac9, lac10 and lac14) showed an increased transcription levels (approximately 1.5–5.6 fold) in the PacC-silenced strains versus that in WT, while other laccase genes were downregulated or unchanged. The extracellular pH value was about 3.1, which was more acidic in the PacC-silenced strains than in the WT (pH 3.5). Moreover, maintaining the fermentation pH resulted in a downregulation of laccase activity which is induced by silencing PacC. Our findings indicate that in addition to its function in acidification of environmental pH, PacC plays an important role in regulating laccase activity in fungi. [ABSTRACT FROM AUTHOR]

Laccases (E.C. 1.10.3.2) produced by white-rot fungi (WRF) can be widely used, but the high cost prevents their use in large-scale industrial processes. Finding a solution to the problem could involve laccase production by solid-state fermentation (SSF) simulating the natural growth conditions for WRF. SSF offers several advantages over conventional submerged fermentation (SmF), such as higher efficiency and productivity of the process and pollution reduction. The aim of this review is therefore to provide an overview of the current state of knowledge about the laccase production by WRF under SSF conditions. The focus is on variations in the up-stream process, fermentation and down-stream process and their impact on laccase activity. The variations of up-stream processing involve inoculum preparation, inoculation of the medium and formulation of the propagation and production media. According to the studies, the production process can be shortened to 5–7 days by the selection of a suitable combination of lignocellulosic material and laccase producer without the need for any additional components of the culture medium. Efficient laccase production was achieved by valorisation of wastes as agro-food, municipal wastes or waste generated from wood processing industries. This leads to a reduction of costs and an increase in competitiveness compared to other commonly used methods and/or procedures. There will be significant challenges and opportunities in the future, where SSF could become more efficient and bring the enzyme production to a higher level, especially in new biorefineries, bioreactors and biomolecular/genetic engineering. [ABSTRACT FROM AUTHOR]

The main purpose of this work was to use pineapple crowns as substrate for optimizing laccase production by Trametes versicolor in lab-scale experiments. One-factor-at-the-time analysis and response surface methodology were used to optimize production. A single laccase with molecular weight of 45 kDa was the main protein produced. A maximal laccase activity of 60.73 ± 1.01 U/g was obtained in 7-day cultures, representing a 6.7-fold increase compared to non-optimized conditions. The optimized conditions were temperature: 28 °C; initial moisture: 90%; glucose: 8.38%; yeast extract: 2.86%. Combining activity and stability, the best conditions for using this laccase during the long periods required by large-scale processes are pH 4.0–5.0 and temperature of 40–50 °C. Under these conditions, the crude laccase was efficient in detoxifying the dye malachite green with a KM of 14.33 ± 1.94 µM and a Vmax of 0.482 ± 0.029 µM/min with 0.1 units/mL. It can be concluded that pineapple crown leaves can be effectively used as substrate by T. versicolor for producing laccase under solid-state culture conditions. Laccase is an industrially relevant enzyme and its production with concomitant valorization of pineapple crowns as substrate offers highly interesting perspectives. [ABSTRACT FROM AUTHOR]

The capacity of novel isolated white-rot fungi secreting laccase was evaluated for various kinds of lignocellulosic biomass in submerged fermentation. The laccase secreted by Neofomitella fumosipora Han 386 and Pleurotus pulmonarius Han 527 was significantly faster than that by Coriolopsis trogii Han 751 and Coriolopsis sanguinaria An 282. Maximum laccase from N. fumosipora Han 386 on the four kinds of lignocellulosic biomass tested appeared on the first day. This phenomenon indicated that N. fumosipora Han 386 secreted laccase rapidly compared with other tested strains in this study and showed the superiority in the rate of secreting laccase. Based on the maximum laccase activity, the ability of secreting laccase of C. sanguinaria An 282 was superior to other tested novel isolated strains. On the whole, N. fumosipora Han 386 and P. pulmonarius Han 527 preferred Toona sinensis to produce laccase, C. trogii Han 751 preferred to produce laccase on Populus beijingensis, and C. sanguinaria An 282 grown on Sorghum straw was more suitable for secreting laccase. The results will be helpful for developing bioprocesses using various kinds of lignocellulosic biomass for lignocellulolytic enzyme production and enlarging the number of laccase producing strains for industrial application. [ABSTRACT FROM AUTHOR]

The utilization of rice straw for biofuel production is limited by its composition. The pretreatment process is required to improve the enzymatic accessibility of polysaccharides in the biomass prior to enzymatic saccharification. In this study, simultaneous biological pretreatment and saccharification (SPS) of rice straw starting from laccase production by Panus neostrigosus I9 was operated in a 2-L fermenter. It was found that fungal physiology was strongly influenced by the agitation, and that the highest laccase production was obtained at an agitation speed of 750 rpm (209.96 ± 0.34 U/L). The dilution rate of 0.05 h-1 was set in continuous fermentation which resulted in laccase activity of 678.49 ± 20.39 U/L, approximately three times higher than that in batch culture. Response surface methodology (RSM) was applied to achieve the condition for maximum percentage of delignification. The maximum percentage of delignification of 45.55% was accomplished after pretreatment of rice straw with laccase enzyme 39.40 U/g rice straw at 43.70 °C for 11.19 h. Reducing sugar of 3.85 ± 0.15 g/L was obtained from the digested rice straw in a SPS reactor, while nonpretreated rice straw gave only 1.13 ± 0.10 g/L within 12 h of incubation. The results indicated that simultaneous biological pretreatment and saccharification (SPS) of rice straw by laccase helped to improve the accessibility of cellulose by cellulolytic enzymes. [ABSTRACT FROM AUTHOR]

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The catalytic depolymerization of biological enzymes during biological liquefaction of coal is very important. In this study, the effects of medium types, lignite addition, and mixed culture on lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (Lac) secreted by white-rot fungi are investigated. Fifteen strains of white-rot fungi are used for microbial liquefaction of the Zhaotong lignite in Yunnan. The results show that the activities of LiP, MnP, and Lac are highest in the Czapek-Dox medium (DOX), followed by the Sabouraud Maltose Broth (SMB) medium, and are almost zero in the Potato Dextrose Agar (PDA) medium. In the early stage of fungal growth, the medium contains sufficient nutrients, and lignite addition induces an inhibitory effect on extracellular enzyme secretion, whereas in the late growth stage characterized by nutrient deficiency, it exhibits an enhancing effect. For fungi pair combinations, if only single enzyme activity was considered, the positive synergistic effect of Lac in the mixed culture of s.commune and SL203 would be the highest, with the synergistic coefficient of 233.33. While three enzymes were considered, the integrated effects of ZTW07 and SL203 are the highest, with Lac, MnP, and LiP producing synergistic coefficients of 158.795, 1.217, and 1.006, respectively. It can be seen that the mixed culture can greatly improve the enzyme activity of white rot fungi and is one of the effective methods to improve the liquefaction efficiency. [ABSTRACT FROM AUTHOR]

Xenobiotics are hazardous compounds that are foreign to natural ecosystems. The production and use of xenobiotic compounds continue to increase worldwide, which in turn causes environmental pollution and has adverse effects on humans. Degradation of such compounds, therefore, needs urgent awareness and attention. The physicochemical approaches to treat the contaminants are expensive. Bioremediation is a concept that exploits organisms to manage the environment with less manpower and time. White-rot fungi-mediated bioremediation offers inexpensive, environmentally sustainable and potential degradation mechanisms for different recalcitrant chemicals. White-rot fungi secrete lignolytic enzymes that have extensive substrate specificities and are involved in the transformation and solubilization of lignin-like structural contaminants. The main lignolytic enzymes occurring in white-rot fungi are laccases, lignin peroxidase, manganese peroxidase, and other peroxidases. Such lignolytic enzymes permit white-rot fungi to endure high toxic levels. This review describes the opportunities to use white-rot fungi and their enzyme systems in the biodegradation of multiple xenobiotic contaminants. [ABSTRACT FROM AUTHOR]

A new wild-type Ganoderma resinaceum isolate was cultivated on glucose-enriched liquid cultures with olive mill wastewaters (OMWs) in initial phenolic compounds concentrations 0.0 (control), 0.5, 0.8, and 1.5 g/l. The effect of the fungus on the reduction of phenolics and color was assessed, whereas biomass production, glucose consumption, intra-cellular (IPS) and extra-cellular (EPS) polysaccharides biosynthesis, antioxidant activity of the biomass, and laccase synthesis were monitored. Results showed that significant phenolic reduction (94.5%) and decolorization (76.5%) occurred, 14.6 g/l of biomass was produced, glucose was almost totally consumed, EPS were produced in sufficient amounts (0.79 g/l), whereas the presence of OMWs enhanced the synthesis of IPS (maximum absolute values 4.0–5.2 g/l corresponding to 35–42% w/w). Kinetic analysis demonstrated that EPS and IPS values fluctuated with time, regardless of the available amount of glucose in the media, showing a maximum in the 17th day of culture. Laccase was highly synthesized in the middle of the fermentation, reaching the maximum value of 14 U/ml. Little growth was however observed at 1.5 g/l phenolics. Strong correlation between total phenolic content and free radical scavenging activity has been noticed in the methanolic extracts of the mycelium. Results strongly suggest the potentiality of G. resinaceum utilization in the OMW waste treatment. [ABSTRACT FROM AUTHOR]

Water pollution by dyes represents from dyestuff industry becomes an environmental concern. Finding new isolates capable of decolorizing these dyes is important. The study aimed to assess the new isolates of white-rot fungi (WRF) as decolorizing agent of anthraquinone and azo dyes. Decolorization assay were conducted in agar plates and liquid medium. During the decolorization, laccase activities produced by the fungal strains were analyzed. Identification of the fungal strains were investigated using molecular DNA analysis. The results showed that isolates M3, H18, and GP1 were able to decolorize anthraquinone and azo dyes in agar and liquid medium. Based on DNA analysis, isolates M3, H18, and GP1 have the similarity to Trametes sanguinea, Trametes polyzona, and Neofomitella guangxiensis, respectively. Among the fungi, T. polyzona H18 exhibited high decolorization ability (70-90%) to the dyes (100 mg/L) after 96 h incubation. Laccase activity was fluctuated during the reactions with tendency to increase at the beginning until its peak, then decreased at the end of incubation. This study demonstrated the potential of the new isolates from Indonesia to decolorize anthraquinone and azo dyes. The results of the study can provide an alternative for bioremediation agents of contaminated water by synthetic dyes. [ABSTRACT FROM AUTHOR]

Laccases (EC. 1.10.3.2) have a broad range of industrial applications. In order to overcome problems associated with loss of activity, storage and reusability of the free enzyme, a laccase from Ganoderma lucidum was immobilised successfully on monoaminoethyl-N-aminoethyl (MANAE)-agarose with high immobilisation yield and high activity recovery. The immobilisation improved significantly the thermal stability, with half-life 2.1- and 5.0-fold higher at 45 and 55 °C, respectively, in comparison with the free laccase. The storage stability at 4–8 °C of the immobilised laccase was also significantly higher than that the free form. Kinetic studies revealed that both free and immobilised G. lucidum laccases obeyed the Michaelis-Menten equation. Immobilisation increased Km and Vmax of the enzyme, reflecting a higher catalytic efficiency at high substrate concentrations. However, the Vmax/Km ratios were decreased by immobilisation, indicating a diminished catalytic efficiency at low substrate concentrations. The immobilised laccase was slightly more efficient than free laccase in the biodegradation of BPA. After 1 h, the immobilised laccase degraded 96.1 ± 3.0% of BPA at the initial concentration of 100 mg.L−1, whereas the free laccase removed 80.9 ± 3.5%. The immobilised G. lucidum laccase also efficiently degraded bisphenol A for at least 15 cycles of reuse. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the potential of white-rot fungi, namely Pycnoporus cinnabarinus, Pleurotus ostreatus and Trametes hirsuta, for the mono azo dye Allura Red AC (AR) removal from aqueous solutions. AR belongs to the hardly degradable xenobiotic associated with a neurotoxic effect on humans and animals. Our results suggested that degradation processes driven by the activity of laccases were not involved in the process of AR removal and the predominant mechanism of dye elimination was biosorption. The surface of fungal biomass was analyzed by Fourier transform infrared spectroscopy (FTIR) and Langmuir and Freundlich models of absorption isotherms were applied to describe the biosorption isotherms. Langmuir model fitted the equilibrium data better than Freundlich isotherm according to the corrected Akaike Information Criterion (AICc). From Langmuir model, dead biomass of P. ostreatus modified by heat was the most suitable biosorbent with the maximum sorption capacity of 118.3 ± 9.9 mg/g dried biomass. Obtained results suggest that biomass of white-rot fungi can be used as a suitable and low-cost biosorbent for the removal of azo dyes from contaminated waters. [ABSTRACT FROM AUTHOR]

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The aim of the present study was to assess the ability of different white-rot fungi to tolerate polychlorinated biphenyls (PCBs) using predictive mycology, by relating fungal growth inhibition to ligninolityc enzyme secretion. Fungal strains were grown in the presence of PCBs in solid media and their radial growth values were modelled through the Dantigny-logistic like function in order to estimate the time required by the fungal colonies to attain half their maximum diameter. The principal component analysis (PCA) revealed an inverse correlation between strain tolerance to PCBs and the laccase secretion over time, being laccase production closely associated with fungal growth capacity. Finally, a PCA was run to regroup and split between resistant and sensitive fungi. Simultaneously, a function associated with a model predicting the tolerance to PCBs was developed. Some of the assayed isolates showed a promising capacity to be applied in PCB bioremediation. Abbreviations: Polychlorinated biphenyls (PCBs), white-rot fungi (WRF) [ABSTRACT FROM AUTHOR]

A novel laccase from a newly isolated white-rot fungus Trametes sp. MA-X01 was characterized. The sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the molecular weight of the laccase produced by Trametes sp. MA-X01 in liquid medium was approximately 62 kDa. The optimal temperature was 60 °C. The enzyme was quite stable at high temperature and maintained about 85% of its highest activity after incubation at 60 °C for 2 h. The laccase could work efficiently over a wide pH range (3.0-5.0) with no significant difference in enzyme activity. Among the metals tested, Mg2+, Mn2+, Zn2+ and Cu2+ could promote the laccase activity, whereas the laccase activity was strongly inhibited by β-mercaptoethanol, L-cysteine and dithiothreitol. Furthermore, the laccase from Trametes sp. MA-X01 exhibited strong ability to decolorize different structural dyes, including azo, heterocyclic and triphenylmethane dyes, in the absence of a redox mediator. [ABSTRACT FROM AUTHOR]

BACKGROUND: Organosolv pretreatment produces a hemicellulose‐rich stream with limited applications due to its toxicity. In this study, beech wood organosolv liquor was valorized through the production of ligninolytic enzymes, the extraction of ganoderic acid and the detoxification of this process stream. RESULTS: Laccase and manganese peroxidase (MnP) were produced in 30 L bioreactors by the white‐rot fungi Ganoderma lucidum and Irpex lacteus, respectively, using the hemicellulose‐rich liquor as carbon source. Both fungi caused a significant reduction in the hemicellulose stream toxicity (between 6.1 and 8.3 times lower). The extraction of ganoderic acid (24.3 mg g−1 of G. lucidum biomass) as a fungal metabolite with high pharmacological activity greatly increased the profitability of the process. Finally, the economic viability of the proposed valorization strategy was analyzed on the basis of the operational costs estimated for the process scale‐up. The specific costs of MnP and laccase production were €2.27 kU−1and €1.08 kU−1, respectively, which are 1109 and 78 times lower than the average price of commercially available enzymes. CONCLUSION: It was found that the integration of the proposed valorization strategy into an organosolv‐based biorefinery would help to increase the economic feasibility of this pretreatment technology. © 2018 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Soil organic matter (SOM) turnover by ligninolytic fungi is a large‐scale process that controls organic carbon geochemistry in terrestrial ecosystems. However, the role of certain oxidative enzymes (e.g., laccase) in humus degradation remains unclear, as well as the molecular structure and recalcitrance of SOM components. In order to address these questions, the degradation of forest soil humic acid (HA) in the presence of laccase and a versatile peroxidase (VP) has been studied in the liquid culture of Lentinus tigrinus. Contrary to the evolving views on humus structure, we have found that alkali‐extractable and acid‐insoluble constituents of SOM (HA) contain true macromolecular components, stable in the presence of 0.1% sodium dodecyl sulfate but degradable/resynthesizable by oxidative enzymes acting on covalent linkages. The HA degradation in the presence of laccase (high N medium) occurs at slower initial rate than in the presence of VP (low N medium). However, each of the enzymes caused about 60% color loss and almost complete degradation of HA into smaller molecules (gel‐filtration data) within 2 weeks of cultivation. Depolymerization of HA in the culture liquid in the presence of laccase was accompanied by polymerization of degradation products on mycelium. Our results show that (1) humus macromolecules are not stable to oxidative enzymes once desorbed from the mineral phase, (2) laccase of Lentinus tigrinus is comparable by its degradation potential to VP, and (3) interfacial secondary synthesis reactions occur during humus decay in the presence of laccase. Our results highlight the important role of laccases in SOM sequestration in soils. Key Points: Soil humic acid is decomposed in the presence of laccase as efficiently as in the presence of VP in the culture liquid of Lentinus tigrinusCell‐associated secondary synthesis reactions occur during humic acid decay in the presence of laccaseAlkali‐extractable humus components are macromolecular but not stable to oxidative enzymes once desorbed from the mineral phase [ABSTRACT FROM AUTHOR]

The aim of this research is to isolate and identify fungi with high lignin-degrading abilities that are autochthonous to southern Serbian region. Two novel fungal isolates identified as Trametes hirsuta F13 and Stereum gausapatum F28 were selected to assess their ligninolytic enzyme activities and the efficiency of lignin removal from beech wood sawdust. Obtained results show that both isolates are good sources of industrially valuable enzymes with a potential for application in various biotechnological and industrial processes. Both isolates showed laccase, manganese-dependent peroxidase, and versatile peroxidase activities, while only S. gausapatum F28 had lignin peroxidase activity. This is the first record of the ability of S. gausapatum species to produce lignin peroxidase. T. hirsuta F13 showed higher laccase activity than S. gausapatum F28, while S. gausapatum F28 had higher manganese peroxidase activity. Also, T. hirsuta F13 exhibited much higher laccase activity under submerged cultivation conditions than solid-state cultivation conditions, which is rare for fungi. This is important for industrial processes since the submerged fermentation is a dominant technique in industry. The test of the efficiency of lignin removal showed that both isolates are efficient lignin decomposers. After five weeks of incubation on beech wood sawdust, the total lignin losses were 33.84 % with T. hirsuta F13 and 28.8 % with S. gausapatum F28. [ABSTRACT FROM AUTHOR]

Phanerochaete chrysosporium was selected as the production strain of laccase, and the effects of stirring speed, ventilation volume, culture temperature, inoculation amount and initial pH of medium on laccase production by liquid fermentation in cylinder were studied. On the basis of single factor test, an orthogonal test was carried out to find optimal conditions for laccase production P. ckrysosporium through liquid fermentation. These results showed that the stirring speed of fermentation cylinder had the highest effect on laccase production, and the optimal conditions were shown as follows: the temperature at 28 t, the rotating speed at 300 r/min, the ventilation volume of 5 L/min (ventilation ratio of 1.0wm), the initial pH of medium of 5, and the inoculation amount of 15%, which gave the highest laccase level of 14.86 U/ml. [ABSTRACT FROM AUTHOR]

White-rot fungi (WRF) and their ligninolytic enzymes (laccases and peroxidases) are considered promising biotechnological tools to remove lignin related Persistent Organic Pollutants from industrial wastewaters and contaminated ecosystems. A high diversity of the genus Ganoderma has been reported in Cuba; in spite of this, the diversity of ligninolytic enzymes and their genes remained unexplored. In this study, 13 native WRF strains were isolated from decayed wood in urban ecosystems in Havana (Cuba). All strains were identified as Ganoderma sp. using a multiplex polymerase chain reaction (PCR)-method based on ITS sequences. All Ganoderma sp. strains produced laccase enzymes at higher levels than non-specific peroxidases. Native-PAGE of extracellular enzymatic extracts revealed a high diversity of laccase isozymes patterns between the strains, suggesting the presence of different amino acid sequences in the laccase enzymes produced by these Ganoderma strains. We determined the diversity of genes encoding laccases and peroxidases using a PCR and cloning approach with basidiomycete-specific primers. Between two and five laccase genes were detected in each strain. In contrast, only one gene encoding manganese peroxidase or versatile peroxidase was detected in each strain. The translated laccases and peroxidases amino acid sequences have not been described before. Extracellular crude enzymatic extracts produced by the Ganoderma UH strains, were able to degrade model chromophoric compounds such as anthraquinone and azo dyes. These findings hold promises for the development of a practical application for the treatment of textile industry wastewaters and also for bioremediation of polluted ecosystems by well-adapted native WRF strains. [ABSTRACT FROM AUTHOR]

Regulation of alkaline-resistant laccase from Perenniporia tephropora KU-Alk4 was proved to be controlled by several factors. One important factor was the initial pH, which drove the fungus to produce different kinds of ligninolytic enzymes. P. tephropora KU-Alk4 could grow at pH 4.5, 7.0, and 8.0. The fungus produced laccase and MnP at pH 7.0, but only laccase at pH 8.0. The specific activity of laccase in the pH 8.0 culture was higher than that in the pH 7.0 culture. At pH 8.0, glucose was the best carbon source for laccase production but growth was better with lactose. Low concentrations of glucose at 0.1% to 1.0% enhanced laccase production, while concentrations over 1% gave contradictory results. Veratryl alcohol induced the production of laccase. A trace concentration of copper ions was required for laccase production. Biomass increased with an increasing rate of aeration of shaking flasks from 100 to 140 rpm; however, shaking at over 120 rpm decreased laccase quantity. Highest amount of laccase produced by KU-Alk4, 360 U/mL, was at pH 8.0 with 1% glucose and 0.2 mM copper sulfate, unshaken for the first 3 days, followed by addition of 0.85 mM veratryl alcohol and shaking at 120 rpm. The crude enzyme was significantly stable in alkaline pH 8.0~10.0 for 24 hr. After treating the pulp mill effluent with the KU-Alk4 system for 3 days, pH decreased from 9.6 to 6.8, with reduction of color and chemical oxygen demand at 83.2% and 81%, respectively. Laccase was detectable during the biotreatment process. [ABSTRACT FROM AUTHOR]

Bioremediation refers to cost-effective and environment-friendly method for converting the toxic, recalcitrant pollutants into environmentally benign products through the action of various biological treatments. Fungi play a major role in bioremediation owing to their robust morphology and diverse metabolic capacity. The review focuses on different fungal groups from a variety of habitats with their role in bioremediation of different toxic and recalcitrant compounds; persistent organic pollutants, textile dyes, effluents from textile, bleached kraft pulp, leather tanning industries, petroleum, polyaromatic hydrocarbons, pharmaceuticals and personal care products, and pesticides. Bioremediation of toxic organics by fungi is the most sustainable and green route for cleanup of contaminated sites and we discuss the multiple modes employed by fungi for detoxification of different toxic and recalcitrant compounds including prominent fungal enzymes viz., catalases, laccases, peroxidases and cyrochrome P450 monooxygeneses. We have also discussed the recent advances in enzyme engineering and genomics and research being carried out to trace the less understood bioremediation pathways. [ABSTRACT FROM AUTHOR]

White-rot fungi have potential in organic pollutant degradation. Immobilization of microorganisms has been successfully used for bioremediation. In this work, 25 isolates of Argentinean white-rot fungi were tested for their tolerance toward up to 10 mM phenol in agar plates. Seven isolates were further evaluated for their ability to grow on plates with 2,6-dimethoxyphenol, gallic acid, 2,4-dichlorophenol, or guaiacol (7.5 mM), or with phenol as sole carbon source. Best results were obtained with Trametes versicolor, Irpex lacteus, Lentinus tigrinus, and Pleurotus lindquistii. The ability of immobilized cultures of T. versicolor BAFC 2234 to remove phenol, was studied. Silica-alginate-fungus biocomposites resulted in phenol removal (10 mM) of up to 48% (mainly attributable to biosorption) within 14 days. Immobilized on Luffa aegyptica, it removed between 62 and 74% of phenol (15 mM) in three repeated cycles over a period of 23 days. Laccase was the main oxidative enzyme detected, and the purified enzyme oxidized 84% of phenol (0.5 mM) in vitro within 4 h, while 43% was converted by a purified Mn-peroxidase. The phenol phytotoxicity decreased noticeably. The concentrations of phenol removed are among the highest reported so far, thus this strain of T. versicolor may have good prospects for application in industrial wastewater treatment. [ABSTRACT FROM AUTHOR]

Coculturing of two white-rot fungi, Dichomitus squalens and Ceriporiopsis subvermispora, was explored for the optimization of cultivation media for simultaneous augmentation of laccase and peroxidase activities by response surface methodology ( RSM). Nutrient parameters chosen from our previous studies with the monocultures of D. squalens and C. subvermispora were used to design the experiments for the cocultivation study. Glucose, arabinose, sodium nitrate, casein, copper sulfate (CuSO4), and manganese sulfate (MnSO4) were combined according to central composite design and used as the incubation medium for the cocultivation. The interaction of glucose and sodium nitrate resulted in laccase and peroxidase activities of approximately 800 U/g protein. The addition of either glucose or sodium nitrate to the medium also modifies the impact of other nutrients on the ligninolytic activity. Both enzyme activities were cross-regulated by arabinose, casein, CuSO4, and MnSO4 as a function of concentrations. Based on RSM, the optimum nutrient levels are 1% glucose, 0.1% arabinose, 20 mM sodium nitrate, 0.27% casein, 0.31 mM CuSO4, and 0.07 mM MnSO4. Cocultivation resulted in the production of laccase of 1,378 U/g protein and peroxidase of 1,372 U/g protein. Lignin (16.9%) in wheat straw was degraded by the optimized enzyme mixture. [ABSTRACT FROM AUTHOR]

Free laccase and fungal biomass from white-rot fungi were compared in the thermokinetics study of the laccase-catalyzed decolorization of an azo dye, i.e., Trypan Blue. The decolorization in both systems followed a first-order kinetics. The apparent first-order rate constant, k′, value increases with temperature. Apparent activation energy of decolorization was similar for both systems at ∼22 kJ mol, while energy for laccase inactivation was 18 kJ mol. Although both systems were endothermic, fungal biomass showed higher enthalpy, entropy, and Gibbs free energy changes for the decolorization compared to free laccase. On the other hand, free laccase showed reaction spontaneity over a wider range of temperature (Δ T = 40 K) as opposed to fungal biomass (Δ T = 15 K). Comparison of entropy change (Δ S) values indicated metabolism of the dye by the biomass. [ABSTRACT FROM AUTHOR]

The laccase gene of Polyporus brumalis was genetically transformed to overexpress its laccase. The transformants exhibited increased laccase activity and effective decolorization of the dye Remazol Brilliant Blue R than the wild type. When the transformants were pretreated with wood chips from a red pine (softwood) and a tulip tree (hardwood) for 15 and 45 days, they showed higher lignin-degradation activity as well as higher wood-chip weight loss than the wild type. When the wood chips treated with the transformant were enzymatically saccharified, the highest sugar yields were found to be 32.5 % for the red pine wood and 29.5 % for the tulip tree wood, on the basis of the dried wood weights, which were 1.6-folds higher than those for the wild type. These results suggested that overexpression of the laccase gene from P. brumalis significantly contributed to the pretreatment of lignocellulose for increasing sugar yields. [ABSTRACT FROM AUTHOR]

A white-rot basidiomycete, isolated from decayed acacia wood (from Northwest of Tunisia) and identified as Trametes sp, was selected in a broad plate screening because of its ability to decolorize and dephenolize olive oil mill wastewater (OMW) efficiently. The major laccase was purified and characterized as a monomeric protein with apparent molecular mass of 61 kDa (SDS-PAGE). It exhibits high enzyme activity over broad pH and temperature ranges with optimum activity at pH 4.0 and a temperature of 60 °C. The purified laccase is stable at alkaline pH values. The enzyme retained 50 % of its activity after 90 min of incubation at 55 °C. Using ABTS, this laccase presented K and V values of 0.05 mM and 212.73 μmoL min mg, respectively. It has shown a degrading activity towards a variety of phenolic compounds. The purified laccase was partially inhibited by Fe, Zn, Cd and Mn, while Cu acted as inducer. EDTA (10 mM) and NaN (10 mM) were found to completely inhibit its activity. 73 % OMW was dephenolized after 315 min incubation at 30 °C with 2 U mL of laccase and 2 mM HBT. [ABSTRACT FROM AUTHOR]

The lignin-degrading, biopulping white-rot fungus Physisporinus rivulosus secretes several laccases of distinct features such as thermostability, extremely low pH optima and thermal activation for oxidation of phenolic substrates. Here we describe the cloning, heterologous expression and structural and enzymatic characterisation of two previously undescribed P. rivulosus laccases. The laccase cDNAs were expressed in the methylotrophic yeast Pichia pastoris either with the native or with Saccharomyces cerevisiae α-factor signal peptide. The specific activity of rLac1 and rLac2 was 5 and 0.3 μkat/μg, respectively. However, mutation of the last amino acid in the rLac2 increased the specific laccase activity by over 50-fold. The recombinant rLac1 and rLac2 enzymes demonstrated low pH optima with both 2,6-dimethoxyphenol (2,6-DMP) and 2,2′-azino-bis(3-ethylbenzathiazoline-6-sulfonate). Both recombinant laccases showed moderate thermotolerance and thermal activation at +60 °C was detected with rLac1. By homology modelling, it was deduced that Lac1 and Lac2 enzymes demonstrate structural similarity with the Trametes versicolor and Trametes trogii laccase crystal structures. Comparison of the protein architecture at the reducing substrate-binding pocket near the T1-Cu site indicated the presence of five amino acid substitutions in the structural models of Lac1 and Lac2. These data add up to our previous reports on laccase production by P. rivulosus during biopulping and growth on Norway spruce. Heterologous expression of the novel Lac1 and Lac2 isoenzymes in P. pastoris enables the detailed study of their properties and the evaluation of their potential as oxidative biocatalysts for conversion of wood lignin, lignin-like compounds and soil-polluting xenobiotics. [ABSTRACT FROM AUTHOR]

A wood-rot fungus was isolated and investigated for its laccase production in submerged cultures. Based on sequence comparison and phylogenetic analysis of the 18S rDNA genes with reference taxa, the wild-type strain HLS-2 with high laccase production was identified as Psathyrella candolleana with GenBank Accession no. HQ215597. The production of laccase by P. candolleana HLS-2 can be enhanced by adding several agricultural wastes to the medium. Among them, wheat bran had the most significant effect and the highest laccase activity of nearly 12000 U/L. Extracellular laccase formations by this fungus can also be induced by veratryl alcohol, guaiacol, vanillic acid, coumaric acid, 2-2'-azinobis(3-ethyl- benzthiazoline-6-sulfonate) (ABTS), and copper sulfate. The maximal laccase activity, approximately 23000 U/L, was obtained from the shake-flask culture containing 1 mM Cu2+ without phenolic or aromatic inducers. Laccase activity of the extracellular liquid was stable at pH 8.0 and retained over 90% activity at 4 °C after 40 days. [ABSTRACT FROM AUTHOR]

Yellow passion fruit waste (YPFW) is an abundant food waste in Brazil, rich in carbohydrates. The aim of the present work was to obtain useful oxidative and hydrolytic enzymes. YPFW solid-state cultures were done using the food-grade white-rot fungi Pleurotus ostreatus, Pleurotus pulmonarius, Macrocybe titans, Ganoderma lucidum, and Grifola frondosa. Under the conditions used in this work, the main enzymes produced by the fungi were laccases, pectinases, and aryl-β- d-glycosidases (β-glucosidases, β-xylosidases, and β-galactosidases). Laccases were produced by all fungi, and in this respect, the YPFW was as good as substrate as wheat bran, the most commonly substrate used for white-rot fungi cultivation. M. titans was the best producer of pectinase in YPFW cultures, while P. ostreatus and P. pulmonarius were the best producers of aryl-β-glycosidases in both YPFW and wheat bran cultures. [ABSTRACT FROM AUTHOR]