Laccase is predominantly found in lignin degrading filamentous white rot fungi, where it is involved in the oxidative degradation of this recalcitrant heteropolymer. In brown rot fungi it is much less prevalent: laccases from only a few brown rots have been detected and only two have been characterized. This study tries to understand the role of this ligninolytic enzyme in brown rots by investigating the catalytic properties of laccases secreted by Fomitopsis pinicola FP58527 SS1. When grown on either poplar or spruce wood blocks, several laccases were detected in the secretome. Two of them (FpLcc1 and FpLcc2) were heterologously produced using Trichoderma reesei QM9414 Δxyr1 as expression host and purified to homogeneity by consecutive steps of hydrophobic interaction, anion exchange and size exclusion chromatography. With the substrates 2,2-azino-bis(3-ethylthiazoline-6-sulfonate) (ABTS), 2,6-dimethoxyphenol (2,6-DMP) and guaiacol both laccases showed similar, low pH-optima below 3 for ABTS and 2,6-DMP and at pH 3.5 for guaiacol which is at the acidic end of laccases isolated from white rot fungi. The determined KM values were low while kcat values measured at acidic conditions were comparable to those reported for other laccases from white rot fungi. While both enzymes showed a moderate decrease in activity in the presence of oxalic and citric acid FpLcc2 was activated by acetic acid up to 3.7 times. This activation effect is much more pronounced at pH 5.0 compared to pH 3.0 and could already be observed at a concentration of 1 mM acetic acid.
- MeSH
- Coriolaceae * genetika MeSH
- Hypocreales MeSH
- lakasa * genetika MeSH
- lignin MeSH
- Publikační typ
- časopisecké články MeSH
Laccases have been widely explored for their ligninolytic capability in bioethanol production and bioremediation of industrial effluents. However, low reaction rates have posed a major challenge to commercialization of such processes. This study reports the first evidence of laccase inhibition by two types of lignin degradation intermediates - fungal-solubilized lignin and alkali-treated lignin - thus offering a highly plausible explanation for low reaction rates due to buildup of inhibitors during the actual process. Reversed-phase high-performance liquid chromatography revealed the presence of similar polar compounds in both lignin samples. A detailed kinetic study on laccase, using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as the substrate, was used to calculate the Michaelis constant (Km) and maximum reaction rate (Vmax). With an increase in the concentration of lignin degradation intermediates, Vmax remained nearly constant, while Km increased from 1.3 to 4.0 times that of pure laccase, revealing that the inhibition was competitive in nature. The kinetic studies reported here and the insight gained into the nature of inhibition can help design process strategies to mitigate this effect and improve overall process efficiency. This work is applicable to processes that employ laccase for delignification of biomass, such as second-generation biofuels processes, as well as for industrial effluent treatment in paper and pulp industries.
- MeSH
- biodegradace MeSH
- biokatalýza MeSH
- fungální proteiny chemie MeSH
- kinetika MeSH
- lakasa chemie MeSH
- lignin chemie MeSH
- Trametes chemie enzymologie genetika MeSH
- Publikační typ
- časopisecké články MeSH
Most known brown rot-producing species of Polyporales belong to the so-called "Antrodia clade" that largely consists of poroid species. In this study, we use three genetic markers to revise Antrodia s. str., the core group of this clade. We show that a corticioid species with a smooth hymenophore, Phlebia griseoflavescens, belongs to Antrodia s. str. Accordingly, we revise the generic concept of Antrodia s. str. to accommodate this species and two recently described poroid taxa, A. tenerifensis and A. multiformis. In addition, we describe two new poroid species within Antrodia s. str., A. latebrosa from Africa and A. peregrina from East Asia, and provide new documentation for the Southeast Asian species A. parvula based on recent collections from the type location.
- MeSH
- Antrodia klasifikace cytologie genetika izolace a purifikace MeSH
- DNA fungální chemie genetika MeSH
- fylogeneze MeSH
- mezerníky ribozomální DNA chemie genetika MeSH
- mikrobiologie životního prostředí MeSH
- mikroskopie MeSH
- ribozomální DNA chemie genetika MeSH
- sekvenční analýza DNA MeSH
- shluková analýza MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Laccases are enzymes with a broad range of biotechnological applications and have, for example, the ability to oxidize many xenobiotics including synthetic dyes. In order to obtain an efficient laccase for the decolorization of dyes which spoil wastewater from the textile industry, genes encoding three various laccase enzymes were expressed in Saccharomyces cerevisiae. The expression of laccases from ascomycete Myceliophthora thermophila (MtL), and two basidiomycetes Trametes versicolor (TvL) and Trametes trogii (TtL) was optimized via selection of plasmids, promoters, media composition, and cultivation conditions. For the first time, the activity of the three secreted laccases was directly compared with the use of various substrates, including different dyes and a wastewater sample. A strong constitutive ADH1 promoter, minimal growth medium, optimized combination of copper and organic nitrogen source, and low cultivation temperature were shown to significantly increase the yields and relative activities of secreted laccases. Heterologous expression of three fungal laccases was successfully achieved in S. cerevisiae being the highest for MtL and the lowest for TvL. MtL, and particularly TtL, showed the decolorization capacity. This is the first report which compared decolorization of synthetic dyes and wastewater by several recombinant laccases and suggested MtL and TtL to be applicable in the ecofriendly enzymatic treatment of colored industry effluent. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:69-80, 2018.
- MeSH
- barvicí látky chemie MeSH
- dusík chemie MeSH
- kultivační média chemie MeSH
- lakasa chemie genetika MeSH
- odpadní voda chemie MeSH
- oxidace-redukce MeSH
- Saccharomyces cerevisiae enzymologie genetika MeSH
- textilní průmysl MeSH
- Trametes chemie enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The key to obtaining an optimum performance of an enzyme is often a question of devising a suitable enzyme and optimisation of conditions for its immobilization. In this study, laccases from the native isolates of white rot fungi Fomes fomentarius and/or Trametes versicolor, obtained from Czech forests, were used. From these, cross-linked enzyme aggregates (CLEA) were prepared and characterised when the experimental conditions were optimized. Based on the optimization steps, saturated ammonium sulphate solution (75 wt.%) was used as the precipitating agent, and different concentrations of glutaraldehyde as a cross-linking agent were investigated. CLEA aggregates formed under the optimal conditions showed higher catalytic efficiency and stabilities (thermal, pH, and storage, against denaturation) as well as high reusability compared to free laccase for both fungal strains. The best concentration of glutaraldehyde seemed to be 50 mM and higher efficiency of cross-linking was observed at a low temperature 4 °C. An insignificant increase in optimum pH for CLEA laccases with respect to free laccases for both fungi was observed. The results show that the optimum temperature for both free laccase and CLEA laccase was 35 °C for T. versicolor and 30 °C for F. fomentarius. The CLEAs retained 80% of their initial activity for Trametes and 74% for Fomes after 70 days of cultivation. Prepared cross-linked enzyme aggregates were also investigated for their decolourisation activity on malachite green, bromothymol blue, and methyl red dyes. Immobilised CLEA laccase from Trametes versicolor showed 95% decolourisation potential and CLEA from Fomes fomentarius demonstrated 90% decolourisation efficiency within 10 h for all dyes used. These results suggest that these CLEAs have promising potential in dye decolourisation.
- MeSH
- azosloučeniny chemie MeSH
- barva MeSH
- barvicí látky chemie MeSH
- bromthymolová modř chemie MeSH
- enzymy imobilizované chemie MeSH
- glutaraldehyd chemie MeSH
- katalýza MeSH
- lakasa chemie MeSH
- Polyporales enzymologie MeSH
- reagencia zkříženě vázaná chemie MeSH
- rosanilinová barviva chemie MeSH
- síran amonný chemie MeSH
- teplota MeSH
- Trametes enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Taxonomy and phylogeny of the Antrodia serialis group are revised with morphological, ecological, and geographic data, partial translation elongation factor 1-α (tef1) gene sequences, and nuc rDNA ITS1-5.8S-ITS2-28S sequences. The group contains 13 species found in boreal and temperate zones of the Northern Hemisphere. The species are limited to certain geographic areas within Eurasia and North America. The traditional morphology-based concept of A. serialis covers at least four closely related species: A. serialis s. str. in Eurasia, A. angusta, sp. nov., in East Asia, A. serrata, sp. nov., in the American Northeast, and A. calcitrosa, sp. nov., in the American Northwest. They all are associated mostly with Picea spp. and show small, but stable morphological differences from each other. In addition, A. morganii, comb. nov., inhabiting wood of Populus spp., occurs in North America, and Antrodia alaskana, comb. nov., a large-pored species, macroscopically similar to A. variiformis, is distributed along the Pacific coast of North America. The pine-dwelling A. flavimontis, sp. nov., similar to A. primaeva from Eurasia, is so far known only from the eastern part of the Rocky Mountains (Utah and Wyoming).
- MeSH
- Antrodia klasifikace genetika MeSH
- DNA fungální genetika MeSH
- elongační faktor 1 genetika MeSH
- fylogeneze * MeSH
- mezerníky ribozomální DNA genetika MeSH
- mykologické určovací techniky MeSH
- ribozomální DNA genetika MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
An investigation was carried out on the effect of inoculation methods on the compost of an organic fraction of municipal solid waste. Three types of white-rot fungi (Phanerochaete chrysosporium, Trametes versicolor and Fomes fomentarius), and a consortium of these fungi, were used. The study assessed their influence on microbial enzymatic activities and the quality of the finished compost. It was found that the addition of white-rot fungi to municipal solid waste (after 37days of composting) could be a useful strategy for enhancing the properties of the final compost product. In comparison with the control sample (compost without inoculation), it accelerates degradation of solid waste as indicated by changes in C/N, electrical conductivity and pH. However, the effectiveness of waste degradation and compost maturation depends on the type of microorganism used for inoculation. The presence of inoculants, such as Trametes versicolor and Fomes fomentarius, led to a higher degrading ratio and a better degree of maturity. This resulted in an increase of enzymatic activities (especially dehydrogenase and protease) and a germination index in comparison with inoculation using Phanerochaete chrysosporium or a consortium of fungi.
- MeSH
- biodegradace MeSH
- Coriolaceae metabolismus MeSH
- dusík metabolismus MeSH
- enzymy metabolismus MeSH
- houby metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- odpadky - odstraňování metody MeSH
- Phanerochaete metabolismus MeSH
- půda * MeSH
- půdní mikrobiologie * MeSH
- Trametes metabolismus MeSH
- tuhý odpad * MeSH
- uhlík metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Ligninolytic enzymes, such as laccase, lignin peroxidase and manganese peroxidase, are biotechnologically-important enzymes. The ability of five white-rot fungal strains Daedaleopsis confragosa, Fomes fomentarius, Trametes gibbosa, Trametes suaveolens and Trametes versicolor to produce these enzymes has been studied. Three different copper(II) complexes have been prepared ((Him)[Cu(im)₄(H₂O)₂](btc)·3H₂O, where im = imidazole, H₃btc = 1,3,5-benzenetricarboxylic acid, [Cu₃(pmdien)₃(btc)](ClO₄)₃·6H₂O) and [Cu₃(mdpta)₃(btc)](ClO₄)₃·4H₂O, where pmdien = N,N,N',N'',N''-pentamethyl-diethylenetriamine and mdpta = N,N-bis-(3-aminopropyl)methyl- amine), and their potential application for laccase and peroxidases induction have been tested. The enzyme-inducing activities of the complexes were compared with that of copper sulfate, and it has been found that all of the complexes are suitable for the induction of laccase and peroxidase activities in white-rot fungi; however, the newly-synthesized complex M1 showed the greatest potential for the induction. With respect to the different copper inducers, this parameter seems to be important for enzyme activity, which depends also on the fungal strains.
- MeSH
- enzymová indukce MeSH
- fungální proteiny biosyntéza genetika MeSH
- komplexní sloučeniny chemie farmakologie MeSH
- lakasa biosyntéza genetika MeSH
- měď chemie MeSH
- peroxidasy biosyntéza genetika MeSH
- regulace genové exprese enzymů MeSH
- Trametes enzymologie MeSH
- vodíková vazba MeSH
- Publikační typ
- časopisecké články MeSH
The tinder polypore, Fomes fomentarius, is a wood-decaying macrofungus well known for its potential use in a wide range of biotechnological applications. The existence of 3 distinct internal transcribed spacer lineages/sublineages among its strains has been clearly established. Sublineage A1 consists of strains isolated from North America, whereas sublineage A2 consists of strains only from Europe. Lineage B comprises strains from Europe and Asia. A better understanding of the biological features of F. fomentarius lineages/sublineages could lead to improved characterization, leading to better biotechnological applications. The medicinal value of F. fomentarius is discussed.
In recent years, there has been an increase in efforts to improve wastewater treatment as the concentration of dangerous pollutants, such as endocrine disrupting chemicals, in wastewater increases. These compounds, which mimic the effect of hormones, have a negative impact on human health and are not easily removed from water. One way to effectively eliminate these pollutants is to use enzymatically activated materials. In this study, we report on the use of laccase from the white rot fungus Trametes versicolor immobilized onto polyamide 6/chitosan (PA6/CHIT) nanofibers modified using two different spacers (bovine serum albumin and hexamethylenediamine). We then tested the ability of the PA6/CHIT-laccase biocatalysts to eliminate a mixture containing 50μM of two endocrine disrupting chemicals: bisphenol A and 17α-ethinylestradiol. The PA6/CHIT nanofiber matrix used in this study not only proved to be a suitable carrier for immobilized and modified laccase but was also efficient in the removal of a mixture of endocrine disrupting chemicals in three treatment cycles.
- MeSH
- biodegradace MeSH
- chitosan chemie MeSH
- endokrinní disruptory metabolismus MeSH
- enzymy imobilizované metabolismus MeSH
- fungální proteiny metabolismus MeSH
- kaprolaktam analogy a deriváty chemie MeSH
- lakasa metabolismus MeSH
- lidé MeSH
- nanovlákna chemie ultrastruktura MeSH
- polymery chemie MeSH
- Trametes enzymologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
