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Continued widespread use of antibiotics, especially fluoroquinolones, raises environmental concerns, as its driving bacterial resistance and disrupts microbial ecosystems. Here we investigate the biodegradation of ten fluoroquinolone antibiotics (six for medical use and four for veterinary use) by ligninolytic fungi, including Trametes versicolor, Bjerkandera adusta, Porosterum spadiceum, Irpex lacteus, Pleuroteus ostreatus, Phanerochaete chrysosporium, Pycnoporus cinnabarinus, Ganoderma lucidum, and Gloeophyllum trabeum. The results show significant variations between strains in the efficiency of antibiotic transformation. B. adusta and P. spadiceum were the fungi that most efficiently reduced antibiotic concentrations and were able to totally degrade eight and six antibiotics, respectively, within a 15-day period. T. versicolor and P. ostreatus also showed the ability to effectively degrade antibiotics. Specifically, T. versicolor degraded six out of the ten fluoroquinolone antibiotics by more than 70 %, while P. ostreatus degraded the tested antibiotics between 43 % and 100 %. The remaining antibiotic activity did not always correlate with a reduction in antibiotic concentrations, which points to the presence of post-transformation antimicrobial metabolites. This study also explores the potential mechanisms used by these fungi to remove selected models of fluroquinolones via enzymatic routes, such as oxidation by laccases, heme-peroxidases, and cytochrome P450, or via adsorption on fungal biomass.
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Due to widespread overuse, pharmaceutical compounds, such as antibiotics, are becoming increasingly prevalent in greater concentrations in aquatic ecosystems. In this study, we investigated the capacity of the white-rot fungus, Coriolopsis gallica (a high-laccase-producing fungus), to biodegrade ampicillin under different cultivation conditions. The biodegradation of the antibiotic was confirmed using high-performance liquid chromatography, and its antibacterial activity was evaluated using the bacterial growth inhibition agar well diffusion method, with Escherichia coli as an ampicillin-sensitive test strain. C. gallica successfully eliminated ampicillin (50 mg L-1) after 6 days of incubation in a liquid medium. The best results were achieved with a 9-day-old fungal culture, which treated a high concentration (500 mg L-1) of ampicillin within 3 days. This higher antibiotic removal rate was concomitant with the maximum laccase production in the culture supernatant. Meanwhile, four consecutive doses of 500 mg L-1 of ampicillin were removed by the same fungal culture within 24 days. After that, the fungus failed to remove the antibiotic. The measurement of the ligninolytic enzyme activity showed that C. gallica laccase might participate in the bioremediation of ampicillin.
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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.
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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.
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Compostos Azo , Lacase , Polyporaceae , Compostos Azo/toxicidade , Biodegradação Ambiental , Corantes/toxicidade , Poli ARESUMO
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.
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Peróxido de Hidrogênio , Levofloxacino , Polyporales , Antibacterianos/química , Fluoroquinolonas/química , Fungos/metabolismoRESUMO
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.
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Water is a dwindling natural resource, and potable water is wrongly considered an unlimited resource. Dialysis, particularly hemodialysis, is a water-hungry treatment that impacts the environment. The global annual water use of hemodialysis is approximately 265 million m3/yr. In this reference estimate, two-thirds of this water is represented by reverse osmosis reject water discharged into the drain. In this review, we would like to draw attention to the complexity and importance of water saving in hemodialysis. We propose that circular water management may comply with the "3R" concept: reduce (reduce dialysis need, reduce dialysate flow, and optimize reverse osmosis performance), reuse (reuse wastewater as potable water), and recycle (dialysis effluents for agriculture and aquaponic use). Awareness and sustainability should be integrated to create positive behaviors. Effective communication is crucial for water savings because local perspectives may lead to global opportunities. Besides the positive environmental impacts, planet-friendly alternatives may have significant financial returns. Innovative policies based on the transition from linear to circular water management may lead to a paradigm shift and establish a sustainable water management model. This review seeks to support policymakers in making informed decisions about water use, avoiding wasting, and finding solutions that may be planet friendly and patient friendly in dialysis, especially in hemodialysis treatments.
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Água Potável , Purificação da Água , Humanos , Diálise Renal/efeitos adversos , Ingestão de Líquidos , PlanetasRESUMO
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.
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In this paper, we assessed the ability of two strains of Saccharomyces cerevisiae, in viable and dead forms, to remove ochratoxin A (OTA) from an artificially contaminated synthetic grape juice medium (SGM) (10 µg OTA/L) and a naturally contaminated grape juice (6.64 µg OTA/L). The first strain, named Levulin FB, is a commercial yeast used in making wine. The second, named SC5, is an autochthonous strain isolated from table grapes. OTA concentrations in juices before and after their contact with yeast cells were assessed. A significant decrease in OTA level (p < 0.05) in the SGM medium and in the natural grape juice was observed after 1 h of adding yeast cells (20 g/L) in viable and heat-treated forms. It was inferred that the dead forms of the two strains were more able to eliminate OTA than their viable forms in both media. This study demonstrates the potential application of an autochthonous yeast for the natural decontamination of grape juice from fungal toxins.
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Ocratoxinas , Vitis , Vinho , Meios de Cultura , Ocratoxinas/análise , Saccharomyces cerevisiae , Vitis/microbiologia , Vinho/análiseRESUMO
Plant growth promoting rhizobacteria (PGPR) arouse an increasing interest as an eco-friendly solution for improving crop tolerance to environmental stresses. In this study, we report the characterization of a novel halotolerant PGPR strain (named C2) identified in a screen of rhizospheric bacterial isolates from southeast of Tunisia. Phylogenetic analysis showed that strain C2 is most likely affiliated to the genus Siccibacter with Siccibacter turicensis as the closest species (98.19%). This strain was able to perform phosphate solubilization and production of indole acetic acid (IAA), siderophores, hydrogen cyanide (HCN), as well as different hydrolytic enzymes (proteases, amylases, cellulases, and lipases). The potential of strain C2 in enhancing salt stress tolerance of Hordeum vulgare was also investigated. Our greenhouse inoculation assays showed that strain C2 promotes barley growth in the presence of 400 mM NaCl by increasing biomass, root length, and chlorophyll contents. It has a positive effect on the photosynthetic efficiency, concomitantly with lower intercellular CO2 contents, compared to non-inoculated plants. Moreover, barley inoculation with strain C2 under salt stress, resulted in higher accumulation of proline and soluble sugars and alleviate the oxidative stress by decreasing hydrogen peroxide and malondialdehyde contents. Remarkably, this positive effect corroborates with a significant activation in the expression of a subset of barley stress responsive genes, including HVA1, HvDREB1, HvWRKY38 and HvP5CS. In summary, Siccibacter sp. strain C2 is able to enhance barley salt stress tolerance and should be leveraged in developing sustainable practices for cereal crop production.
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Hordeum , Filogenia , Desenvolvimento Vegetal , Tolerância ao Sal/fisiologia , Estresse FisiológicoRESUMO
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.
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In the current investigation, the capacity of different yeast strains to decolorize reactive black 5 (RB-5) was assessed. A comparative study between the different strains demonstrated that Saccharomyces cerevisiae X19G2 exhibited the highest decolorization rate (69.20 ± 1.16%) after 48 h of incubation. This strain was selected to optimize the medium components' concentrations for maximum RB-5 decolorization. Response-surface methodology (RSM) was tested for the most significant parameters (glucose, yeast extract and RB-5 dye concentrations) that were previously determined by Plackett-Burman design. A dye decolorization rate of 99.59 ± 0.24% was achieved within 48 h using a maximum RB-5 concentration (0.15 g/L) with glucose and yeast extract concentrations equalling to 10.5 g/L and 1 g/L, respectively. Experimental data results proved to fit well with the pseudo-second order kinetics model. The phytotoxicity assessment was carried out using Raphanus sativus seeds to determine the toxicity of RB-5 before and after treatment by S. cerevisiae. Results suggested that germination rate and the length of seeds radical irrigated with 0.15 g/L of RB-5 decreased by 30 and 53%, compared to those irrigated with treated solution. Therefore, metabolites derived from decolorization of RB-5 by S. cerevisiae X19G2 were significantly less toxic than the original dye.
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The effect of different pretreatment approaches based on alkali (NaOH)/hydrogen peroxide (H2O2) on willow sawdust (WS) biomass, in terms of delignification efficiency, structural changes of lignocellulose and subsequent fermentation toward ethanol, was investigated. Bioethanol production was carried out using the conventional yeast Saccharomyces cerevisiae, as well as three non-conventional yeasts strains, i.e., Pichia stipitis, Pachysolen tannophilus, Wickerhamomyces anomalus X19, separately and in co-cultures. The experimental results showed that a two-stage pretreatment approach (NaOH (0.5% w/v) for 24 h and H2O2 (0.5% v/v) for 24 h) led to higher delignification (38.3 ± 0.1%) and saccharification efficiency (31.7 ± 0.3%) and higher ethanol concentration and yield. Monocultures of S. cerevisiae or W. anomalus X19 and co-cultures with P. stipitis exhibited ethanol yields in the range of 11.67 ± 0.21 to 13.81 ± 0.20 g/100 g total solids (TS). When WS was subjected to H2O2 (0.5% v/v) alone for 24 h, the lowest ethanol yields were observed for all yeast strains, due to the minor impact of this treatment on the main chemical and structural WS characteristics. In order to decide which is the best pretreatment approach, a detailed techno-economical assessment is needed, which will take into account the ethanol yields and the minimum processing cost.
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Biocombustíveis , Etanol/metabolismo , Fermentação , Madeira , Leveduras/metabolismo , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/metabolismo , Análise Espectral , Madeira/química , Madeira/ultraestruturaRESUMO
This study is to test the capacity of the white rot fungus Coriolopsis gallica for the biodegradation of Diesel Fuel hydrocarbons (DHs). Using the experimental face centered central composite design (FCCCD), culture conditions of the Diesel-mended medium were optimized to reach 110.43% of DHs removal rate, and l5267.35 U L-1 of laccase production by C. gallica, simultaneously. The optimal combination of the cultural parameters was: Diesel concentration range of 2.95-3.14%, inoculum size of 3%, incubation time of 15 days, Tween 80 concentration of 0.05%, and the ratio glucose/peptone (G/P) range of 10.15-10.27. Further, the degradation ability of C. gallica for Diesel Fuel was evaluated through mycelial pellets uptake and oxidative action of fungal enzymes in the optimized degrading-medium using gas chromatography-mass spectrometry (GC-MS). Cyclosiloxanes and C20 PAHs detected as the major compound in Diesel Fuel (46%) was completely bio-transformed to simple metabolites including, essentially benzoic acid ester (71%), alcohols (1.52%) epoxy alkane (1.07%), carboxylic acids (1.24%) and quinones (0.33%). Germination rate and root elongation, as a rapid phytotoxicity test demonstrated that toxicity of Diesel's PAHs is minimized by fungal treatment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02769-w.
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The functional diversity of the New Caledonian mangrove sediments was examined, observing the distribution of fungal dye-decolorizing peroxidases (DyPs), together with the complete biochemical characterization of the main DyP. Using a functional metabarcoding approach, the diversity of expressed genes encoding fungal DyPs was investigated in surface and deeper sediments, collected beneath either Avicennia marina or Rhizophora stylosa trees, during either the wet or the dry seasons. The highest DyP diversity was observed in surface sediments beneath the R. stylosa area during the wet season, and one particular operational functional unit (OFU1) was detected as the most abundant DyP isoform. This OFU was found in all sediment samples, representing 51-100% of the total DyP-encoding sequences in 70% of the samples. The complete cDNA sequence corresponding to this abundant DyP (OFU 1) was retrieved by gene capture, cloned, and heterologously expressed in Pichia pastoris. The recombinant enzyme, called DyP1, was purified and characterized, leading to the description of its physical-chemical properties, its ability to oxidize diverse phenolic substrates, and its potential to decolorize textile dyes; DyP1 was more active at low pH, though moderately stable over a wide pH range. The enzyme was very stable at temperatures up to 50 °C, retaining 60% activity after 180 min incubation. Its ability to decolorize industrial dyes was also tested on Reactive Blue 19, Acid Black, Disperse Blue 79, and Reactive Black 5. The effect of hydrogen peroxide and sea salt on DyP1 activity was studied and compared to what is reported for previously characterized enzymes from terrestrial and marine-derived fungi.
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Only a few studies have examined how marine-derived fungi and their enzymes adapt to salinity and plant biomass degradation. This work concerns the production and characterisation of an oxidative enzyme identified from the transcriptome of marine-derived fungus Stemphylium lucomagnoense. The laccase-encoding gene SlLac2 from S. lucomagnoense was cloned for heterologous expression in Aspergillus niger D15#26 for protein production in the extracellular medium of around 30 mg L-1. The extracellular recombinant enzyme SlLac2 was successfully produced and purified in three steps protocol: ultrafiltration, anion-exchange chromatography, and size exclusion chromatography, with a final recovery yield of 24%. SlLac2 was characterised by physicochemical properties, kinetic parameters, and ability to oxidise diverse phenolic substrates. We also studied its activity in the presence and absence of sea salt. The molecular mass of SlLac2 was about 75 kDa, consistent with that of most ascomycete fungal laccases. With syringaldazine as substrate, SlLac2 showed an optimal activity at pH 6 and retained nearly 100% of its activity when incubated at 50°C for 180 min. SlLac2 exhibited more than 50% of its activity with 5% wt/vol of sea salt.