Degradation of bisphenol A by different fungal laccases and identification of its degradation products

29 p.-5 fig.Different fungal laccases were used as biocatalysts for the biotransformation of bisphenol A (BPA). The quantitative analysis by gas chromatography-mass spectrometry (GCeMS) showed that BPA is more rapidly oxidized by Coriolopsis gallica laccase among the different fungal laccases tested. Carboxylic acid derivatives such as tartaric acid was found as BPA degradation products resulting from reactions catalyzed by 1 U ml 1 of laccase from C. gallica in the presence of 1 mM of the laccase-mediator 1-hydroxybenzotriazole (HBT), while b-hydroxybutyric acid resulted from oxidative reactions without HBT. BPA was completely removed within 3 h and pyroglutamic acid was found as a supplementary oxidative degradation product from HBT when identified by GCeMS. Laccase played a critical role in BPA biodegradation and catalyzed a cross-coupling reactionThis work was supported by the Ministry of Higher Education, Scientific Research (Tunisia) and the Comunidad de Madrid project S2013/MAE-2907Peer reviewe

Phylogenetic and metabolic diversity of Tunisian forest wood-degrading fungi: a wealth of novelties and opportunities for biotechnology

16 p.-2 fig.-5 tab.In this study, 51 fungal strains were isolated from decaying wood samples collected from forests located in the Northwest of Tunisia in the vicinity of Bousalem, Ain Draham and Kef. Phylogenetic analysis based on the sequences of the internal transcribed spacers of the ribosomal DNA showed a high diversity among the 51 fungal isolates collection. Representatives of 25 genera and 29 species were identified, most of which were members of one of the following phyla (Ascomycota, Basidiomycota and Zygomycota). In addition to the phylogenetic diversity, a high diversity of secreted enzyme profiles was also detected among the fungal isolates. All fungal strains produced at least one of the following enzymes: laccase, cellulase, protease and/or lipase.This project received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under Grant Agreement No. 245268 (to L.B.). This study was also supported by the SwissBOL project, financed by the Swiss Federal Office for the Environment (Grant holder L.B.), the Swiss State Secretariat for Education and Research (L.B. Grant Reference: SER No. C09.0139), and the European Union for the COST action FP0801 ‘‘Established and Emerging Phytophthora: Increasing Threats to Woodland and Forest Ecosystems in Europe’’.Peer reviewe

Autochthonous fungal strains with high ligninolytic activities from Tunisian biotopes

6 p.-2 tab.-3 fig.This work represents the first report on the ability of autochthonous fungi of Tunisia to produce ligninolytic enzymes. Three hundred fifteen fungal strains were isolated from different Tunisian biotopes. These fungal strains were first screened for lignin-modifying enzymes on solid media containing Poly R-478 or ABTS. Of the 315 tested strains, 49 exhibited significant ABTS-oxidation activity, expressed within the first week of incubation and only 18 strains decolourised the Poly R-478. These positive strains were further screened in liquid culture and laccase, and lignin and Mn2+-oxidizing peroxidases activities were assayed. Of the 67 strains grown on liquid medium, 28 produced at least one of these 3 enzymes. The 8 highest producers of ligninolytic activities were identified by molecular techniques and 3 among them produced Lac, MnP and LiP simultaneously. New isolates reported in this work as fungi with significant ligninolytic activities includes Oxyporus, Stereum and Trichoderma. The isolated Trametes trogii CTM 10156 was the best Lac producer. Culture conditions and medium composition were optimised for this strain and resulted in high Lac production of 110 U ml-1 within 15 days of incubation (367 times higher than control medium).This research was supported by EU contract N° ICA3-CT-1999-00010 and “Contrats Programmes SERST, Tunisia.Peer reviewe

Autochthonous fungal strains with high ligninolytic activities from Tunisian biotopes

This work represents the first report on the ability of autochthonous fungi of Tunisia to produce ligninolytic enzymes. Three hundred fifteen fungal strains were isolated from different Tunisian biotopes. These fungal strains were first screened for lignin-modifying enzymes on solid media containing Poly R-478 or ABTS. Of the 315 tested strains, 49 exhibited significant ABTS-oxidation activity, expressed within the first week of incubation and only 18 strains decolourised the Poly R-478. These positive strains were further screened in liquid culture and laccase, and lignin and Mn2+-oxidizing peroxidases activities were assayed. Of the 67 strains grown on liquid medium, 28 produced at least one of these 3 enzymes. The 8 highest producers of ligninolytic activities were identified by molecular techniques and 3 among them produced Lac, MnP and LiP simultaneously. New isolates reported in this work as fungi with significant ligninolytic activities includes Oxyporus, Stereum and Trichoderma. The isolated Trametes trogii CTM 10156 was the best Lac producer. Culture conditions and medium composition were optimised for this strain and resulted in high Lac production of 110 U ml-1 within 15 days of incubation (367 times higher than control medium).African Journal of Biotechnology Vol. 4 (5), pp. 431-436, 200

Decolorization of the azo dye Acid Orange 51 by laccase produced in solid culture of a newly isolated Trametes trogii strain

11 p.-5 fig.-6 tab.This study concerns the decolorization and detoxification of the azo dye Acid Orange 51 (AO51) by crude laccase from Trametes trogii produced in solid culture using sawdust as support media. A three-level Box–Behnken factorial design with four factors (enzyme concentration, 1-hydroxybenzotriazole (HBT) concentration, dye concentration and reaction time) combined with response surface methodology was applied to optimize AO51 decolorization. A mathematical model was developed showing the effect of each factor and their interactions on color removal. The model predicted that Acid Orange 51 decolorization above 87.87 ± 1.27 % could be obtained when enzyme concentration, HBT concentration, dye concentration and reaction time were set at 1 U/mL, 0.75 mM, 60 mg/L and 2 days, respectively. The experimental values were in good agreement with the predicted ones and the models were highly significant, the correlation coefficient (R 2) being 0.9. Then the desirability function was employed to determine the optimal decolorization condition for each dye and minimize the process cost simultaneously. In addition, germination index assay showed that laccase-treated dye was detoxified; however in the presence of HBT, the phytotoxicity of the treated dye was increased. By using cheap agro-industrial wastes, such as sawdust, a potential laccase was obtained. The low cost of laccase production may further broaden its application in textile wastewater treatment.Peer reviewe

Laccase purification and characterization from Trametes trogii isolated in Tunisia: decolorization of textile dyes by the purified enzyme

8 páginas, 3 figuras, 5 tablas -- PAGS nros. 141-148A white-rot basidiomycete, isolated from decayed acacia wood (from Northwest of Tunisia) and identified as Trametes trogii, was selected in a broad plate screening because of its ability to degrade commercial dyes. In liquid cultures using a glucose–peptone medium, the sole ligninolytic activity detected was laccase. The highest laccase levels were obtained in presence of CuSO4 as inducer (around 20000 U/l). Two isoenzymes, were purified using anion-exchange and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with Mw around 62 kDa and isoelectric points of 4.3 and 4.5, showing similar stability at pH and temperature, optimum pH and substrate specificity. The highest oxidation rate was obtained at pH 2 and 2.5 for ABTS and DMP, respectively. They were stable up to 50 °C for 24 h and the stability was higher at alkaline pH. Activity increased by the addition of 10 mM Ni, Mo or Mn but it was not affected by Cd, Al, Li and Ca. Identical N-terminal sequences were determined in both laccases. The crude enzyme, as well as the purified laccase, was able to decolorize dyes from the textile industryThis research has been funded by a Cooperation project between Spain and Tunisia (31p/02 – Spanish “Ministerio de Asuntos Exteriores” and “MRSTDC”), and in part by a grant from “Contrats Programmes MRSTCD” (Tunisia)Peer reviewe

Laccase purification and characterization from Trametes trogii isolated in Tunisia: decolorization of textile dyes by the purified enzyme

8 páginas, 3 figuras, 5 tablas -- PAGS nros. 141-148A white-rot basidiomycete, isolated from decayed acacia wood (from Northwest of Tunisia) and identified as Trametes trogii, was selected in a broad plate screening because of its ability to degrade commercial dyes. In liquid cultures using a glucose–peptone medium, the sole ligninolytic activity detected was laccase. The highest laccase levels were obtained in presence of CuSO4 as inducer (around 20000 U/l). Two isoenzymes, were purified using anion-exchange and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with Mw around 62 kDa and isoelectric points of 4.3 and 4.5, showing similar stability at pH and temperature, optimum pH and substrate specificity. The highest oxidation rate was obtained at pH 2 and 2.5 for ABTS and DMP, respectively. They were stable up to 50 °C for 24 h and the stability was higher at alkaline pH. Activity increased by the addition of 10 mM Ni, Mo or Mn but it was not affected by Cd, Al, Li and Ca. Identical N-terminal sequences were determined in both laccases. The crude enzyme, as well as the purified laccase, was able to decolorize dyes from the textile industryThis research has been funded by a Cooperation project between Spain and Tunisia (31p/02 – Spanish “Ministerio de Asuntos Exteriores” and “MRSTDC”), and in part by a grant from “Contrats Programmes MRSTCD” (Tunisia)Peer reviewe

Optimization of the Decolorization of the Reactive Black 5 by a Laccase-like Active Cell-Free Supernatant from Coriolopsis gallica

The textile industry generates huge volumes of colored wastewater that require multiple treatments to remove persistent toxic and carcinogenic dyes. Here we studied the decolorization of a recalcitrant azo dye, Reactive Black 5, using laccase-like active cell-free supernatant from Coriolopsis gallica. Decolorization was optimized in a 1 mL reaction mixture using the response surface methodology (RSM) to test the influence of five variables, i.e., laccase-like activity, dye concentration, redox mediator (HBT) concentration, pH, and temperature, on dye decolorization. Statistical tests were used to determine regression coefficients and the quality of the models used, as well as significant factors and/or factor interactions. Maximum decolorization was achieved at 120 min (82 ± 0.6%) with the optimized protocol, i.e., laccase-like activity at 0.5 U mL−1, dye at 25 mg L−1, HBT at 4.5 mM, pH at 4.2 and temperature at 55 °C. The model proved significant (ANOVA test with p < 0.001): coefficient of determination (R²) was 89.78%, adjusted coefficient of determination (R²A) was 87.85%, and root mean square error (RMSE) was 10.48%. The reaction conditions yielding maximum decolorization were tested in a larger volume of 500 mL reaction mixture. Under these conditions, the decolorization rate reached 77.6 ± 0.4%, which was in good agreement with the value found on the 1 mL scale. RB5 decolorization was further evaluated using the UV-visible spectra of the treated and untreated dyes

Biotransformation of the fluoroquinolone, levofloxacin, by the white-rot fungus Coriolopsis gallicaCoriolopsis\ gallica

International audienceThe wastewater from hospitals, pharmaceutical industries and more generally human and animal dejections leads to environmental releases of antibiotics that cause severe problems for all living organisms. The aim of this study was to investigate the capacity of three fungal strains to biotransform the fluoroquinolone levofloxacin. The degradation processes were analyzed in solid and liquid media. Among the three fungal strains tested, $Coriolopsis\ gallica$ strain CLBE55 (BRFM 3473) showed the highest removal efficiency, with a 15% decrease in antibiogram zone of inhibition for Escherichia coli cultured in solid medium and 25% degradation of the antibiotic in liquid medium based on high-performance liquid chromatography (HPLC). Proteomic analysis suggested that laccases and dye-decolorizing peroxidases such as extracellular enzymes could be involved in levofloxacin degradation, with a putative major role for laccases. Degradation products were proposed based on mass spectrometry analysis, and annotation suggested that the main product of biotransformation of levofloxacin by $Coriolopsis\ gallica$ is an N-oxidized derivative