Optimized decolorization of two poly azo dyes Sirius Red and Sirius Blue using laccase-mediator system.

Factors, namely pH, laccase-like activity, dyes concentration as well as 1-Hydroxybenzotriazole (HBT) concentration was examined. The results indicated that the maximum decolorization yield and rate reached 98.30 ± 0.10% and 5.84 ± 0.01%/min, respectively for Sirius Blue, and 99.34 ± 0.47% and 5.85 ± 0.12%/min, respectively for Sirius Red after 4 h. The presence of the redox mediator 1-hydroxybenzotriazole (HBT) greatly improved the decolorization levels. The optimum concentrations of HBT, dyes, and laccase were 0.62 mM, 50 mg/L, and 0.89 U/mL respectively at pH 4.58 for both dyes. Phytotoxicity tests using treated and untreated dyes proved that the applied treatment slightly decreased the toxicity of the by-products. However, the germination index (GI) increased from 14.6 to 36.08% and from 31.6 to 36.96% for Sirius Red and Sirius Blue, respectively. The present study focused on the treatment of two recalcitrant azo dyes, namely: Sirius Blue (Direct Blue 71) and Sirius Red (Direct Red 80). The decolorization was performed using cell-free supernatant from Coriolopsis gallica culture with high laccase activity. Response surface methodology (RSM) and Box-Behnken design were applied to optimize the decolorization of the two tested dyes. The effect of four.

Genome sequencing of Porostereum spadiceum to study the degradation of levofloxacin.

Despite various plans to rationalize antibiotic use, antibiotic resistance in environmental bacteria is increasing due to the accumulation of antibiotic residues in the environment. This study aimed to test the ability of basidiomycete fungal strains to biotransform the antibiotic levofloxacin, a widely-used third-generation broad-spectrum fluoroquinolone, and to propose enzyme targets potentially involved in this biotransformation. The biotransformation process was performed using fungal strains. Levofloxacin biotransformation reached 100% after 9 days of culture with Porostereum spadiceum BS34. Using genomics and proteomics analyses coupled with activity tests, we showed that P. spadiceum produces several heme-peroxidases together with H2O2-producing enzymes that could be involved in the antibiotic biotransformation process. Using UV and high-resolution mass spectrometry, we were able to detect five levofloxacin degradation products. Their putative identity based on their MS2 fragmentation patterns led to the conclusion that the piperazine moiety was the main target of oxidative modification of levofloxacin by P. spadiceum, leading to a decrease in antibiotic activity.

Efficient Decolorization of the Poly-Azo Dye Sirius Grey by Coriolopsis gallica Laccase-Mediator System: Process Optimization and Toxicity Assessment.

The textile industry produces high volumes of colored effluents that require multiple treatments to remove non-adsorbed dyes, which could be recalcitrant due to their complex chemical structure. Most of the studies have dealt with the biodegradation of mono or diazo dyes but rarely with poly-azo dyes. Therefore, the aim of this paper was to study the biodegradation of a four azo-bond dye (Sirius grey) and to optimize its decolorization conditions. Laccase-containing cell-free supernatant from the culture of a newly isolated fungal strain, Coriolopsis gallica strain BS9 was used in the presence of 1-hydroxybenzotriazol (HBT) to optimize the dye decolorization conditions. A Box-Benken design with four factors, namely pH, enzyme concentration, HBT concentration, and dye concentration, was performed to determine optimal conditions for the decolorization of Sirius grey. The optimal conditions were pH 5, 1 U/mL of laccase, 1 mM of HBT, and 50 mg/L of initial dye concentration, ensuring a decolorization yield and rate of 87.56% and 2.95%/min, respectively. The decolorized dye solution showed a decrease in its phytotoxicity (Germination index GI = 80%) compared to the non-treated solution (GI = 29%). This study suggests that the laccase-mediator system could be a promising alternative for dye removal from textile wastewater.

Isolation and characterization of a plant growth­promoting rhizobacterium strain MD36 that promotes barley seedlings and growth under heavy metals stress.

Plant growth, promoting, bacteria, (PGPB) can improve plant germination and growth in heavy metal-contaminated land and enhance heavy metal removal efficiency. In this study, we isolated PGPR bacterial strains which can withstand heavy metal pollution and tested their ability to improve barley germination under heavy metal stress. Out of 16 multi-resistant heavy metal isolates, strain MD36 was identified by 16S rRNA sequencing and shared close relation to different species of the genus Glutamicibacter. The new isolated strain showed other important PGPR activities, mainly IAA production and salt tolerance. The effect of adding the strain MD36 to barley grains under heavy metal stress enhanced their germination up to 100%, while the percentage of germination ranged between 0 and 20% for non-inoculated grains. In addition, in these conditions, MD36 can significantly enhance barley growth by reducing the heavy metal effect. This study strongly recommends the use of MD36 as seed coatings trials in the field to enhance growth and yield in soils contaminated with heavy metals, as well as in bioremediation of HM-contaminated salt-containing soils and water.

Biotransformation of the Fluoroquinolone, Levofloxacin, by the White-Rot Fungus Coriolopsis gallica.

The 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.

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.

Sawdust waste as a low-cost support-substrate for laccases production and adsorbent for azo dyes decolorization.

BACKGROUND: Laccases are multicopper oxidases with high potential for environmental and industrial applications. Low-cost laccase production could be achieved by solid state fermentation on agro-industrial by-products. METHODS: A number of agro-industrial solid wastes were tested as support-substrate for laccase production by Coriolopsis gallica under solid-state fermentation (SSF) conditions. Response surface methodology (RSM) was used to optimize the medium composition for laccase production. Initial screening by Plackett-Burman design was performed to select the major variables out of 20 tow medium components fellowing this Central composite design (CCD) was employed to optimize the level of the selected variables. RESULTS: Sawdust waste was shown to be the best support-substrate for laccase production by the C. gallica. Peptone as source of organic nitrogen, Cd(2+) as laccase inducer and liquid/solid (L/S) ratio were found to have significant effects on laccase production. Operating at optimum concentrations of the most significant variables (peptone, 4.5 g L(-1), L/S ratio, 5.0 and Cd(+2) 1.0 mM) extracellular laccase activity was enhanced from 1480 U L(-1) (60.5 U g(-1)), to 4880 U L(-1) (200 U g(-1)) which meant a 3.2-fold increase in laccase activity. On the other hand, sawdust waste was studied as a low cost adsorbent to remove the azo dyes Reactive Black 5 (RB5) and Acid Orange 51 (AO51). Decolorization percentages around 67 and 75 % were obtained in 24 h for RB5 and AO51, respectively. CONCLUSION: When used as a support substrate, sawdust yielded the highest laccase production which was increased 3.2 times using RMS optimization.

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

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.

Biosynthesis of single-cell biomass from olive mill wastewater by newly isolated yeasts.

The aim of this study was to assess the potential of newly isolated yeast strains Schwanniomyces etchellsii M2 and Candida pararugosa BM24 to produce yeast biomass on olive mill wastewater (OMW). Maximum biomass yield was obtained at 75% (v/v) OMW, after 96 h of incubation at 30 °C and 5% (v/v) inoculum size. The optimal carbon/nitrogen (C/N) ratio was in the range of 8:1 to 10:1, and ammonium chloride was selected as the most suitable nitrogen source. Under these conditions, a maximum biomass production of 15.11 and 21.68 g L(-1) was achieved for Schwanniomyces etchellsii M2 and Candida pararugosa BM24, respectively. Proteins were the major constituents of yeast cells (35.9-39.4% dry weight), lipids were 2.8-5% dry weight, and ash ranged from 4.8 to 9.5 % dry weight. Besides biomass production, yeast strains were also able to reduce toxicity and polluting parameter levels of the spent OMW-based medium. The practical results presented show that pH rose from initial value of 5.5 to 7.24-7.45 after fermentation. Approximately 23.1-41.4% of the chemical oxygen demand (COD) and 15.4-19.2% of the phenolic compounds were removed. The removal of phenolic compounds was associated with their biodegradation and their partial adsorption on yeast cells.

Olive oil mill wastewaters: phenolic content characterization during degradation by Coriolopsis gallica.

Olive mill wastewaters (OMW) pose a serious environmental concern owing to high polyphenol content. Decolorization and degradation of phenolic compounds (PC) by Coriolopsis gallica was demonstrated in our laboratory as a potential biotreatment of OMW in solid and liquid media. High performance liquid chromatography coupled to electrospray time-of-flight mass spectrometry was used to analyze the evolution of the main phenolic compounds during the C. gallica biodegradation process. Amongst total the compounds characterized in methanolic extracts of OMW, 12 were unknown, 15 were from different polyphenolic families, and 27 were other non-phenolic compounds. The evolution of PC content during the degradation process indicated that, despite the complexity of the OMW phenolic fraction, C. gallica was able to grow on OMW-based media using PC as sources of carbon and energy, particularly acids, alcohols, lignans and flavones. Complete dephenolization of OMW was obtained.

Purification and biochemical characterization of a new alkali-stable laccase from Trametes sp. isolated in Tunisia: role of the enzyme in olive mill waste water treatment.

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 m and V max values of 0.05 mM and 212.73 µmoL min(-1) mg(-1), respectively. It has shown a degrading activity towards a variety of phenolic compounds. The purified laccase was partially inhibited by Fe(2+), Zn(2+), Cd(2+) and Mn(2+), while Cu(2+) acted as inducer. EDTA (10 mM) and NaN3 (10 mM) were found to completely inhibit its activity. 73 % OMW was dephenolized after 315 min incubation at 30 °C with 2 U mL(-1) of laccase and 2 mM HBT.

Decolorization and detoxification of two textile industry effluents by the laccase/1-hydroxybenzotriazole system.

The aim of this work was to determine the optimal conditions for the decolorization and the detoxification of two effluents from a textile industry-effluent A (the reactive dye bath Bezactive) and effluent B (the direct dye bath Tubantin)-using a laccase mediator system. Response surface methodology (RSM) was applied to optimize textile effluents decolorization. A Box-Behnken design using RSM with the four variables pH, effluent concentration, 1-hydroxybenzotriazole (HBT) concentration, and enzyme (laccase) concentration was used to determine correlations between the effects of these variables on the decolorization of the two effluents. The optimum conditions for pH and concentrations of HBT, effluent and laccase were 5, 1 mM, 50 % and 0.6 U/ml, respectively, for maximum decolorization of effluent A (68 %). For effluent B, optima were 4, 1 mM, 75 %, and 0.6 U/ml, respectively, for maximum decolorization of approximately 88 %. Both effluents were treated at 30 °C for 20 h. A quadratic model was obtained for each decolorization through this design. The experimental and predicted values were in good agreement and both models were highly significant. In addition, the toxicity of the two effluents was determined before and after laccase treatment using Saccharomyces cerevisiae, Bacillus cereus, and germination of tomato seeds.

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

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 (R2) 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.

Application of response surface methodology to optimize decolourization of dyes by the laccase-mediator system.

Response surface methodology (RSM) was applied to optimize the decolourization of 3 dyes belonging to 3 dye families such as reactive black 5 (diazoic), indigo carmine (indigoid) and aniline blue (anthraquinonic). Crude laccase from Trametes trogii and the laccase-mediator 1-hydroxybenzotriazole (HBT) were used in this study. Box-Behnken design using RSM with six variables namely pH, temperature, enzyme concentration, HBT concentration, dye concentration and incubation time was used in this study to optimize significant correlation between the effects of these variables on the decolourization of reactive black 5 (RB5), indigo carmine (IC) and aniline blue (AB). The optimum of pH, temperature, laccase, HBT, RB5 and reaction time were 4.5, 0.5 U ml(-1), 0.5 mM, 100 mg ml(-1) and 150 min respectively, for a maximum decolourization of RB5 (about 92.92% ± 7.21). Whereas, the optimum decolourization conditions of both IC (99.76% ± 7.75) and AB (98.44% ± 10) were: pH 4.5, temperature of 45 °C, enzyme concentration of 0.1 U ml(-1) and 0.5 U ml(-1), HBT concentration of 0.9 mM and 0.5 mM, dye concentration of 60 mg l(-1) and reaction time of 150 and 90 min, respectively. The experimental values were in good agreement with the predicted ones and the models were highly significant, the correlation coefficient (R(2)) being 0.864, 0.663 and 0.776 for RB5, IC and AB, respectively. In addition, when the kinetic parameters for the three dyes decolourization were calculated according to Hannes-Wolf plot, the following values were obtained: Km of 268.4, 47.94 and 44.64 mg l(-1) then V(max) of 35.58, 10.43 and 9.23 mg l(-1) min for the RB5, IC and AB decolourizations by laccase, respectively.