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Laccase production by Pleurotus ostreatus using cassava waste and its application in remediation of phenolic and polycyclic aromatic hydrocarbon-contaminated lignocellulosic biorefinery wastewater
2022
Kumar, Vaidyanathan Vinoth | Venkataraman, Swethaa | Kumar, P Senthil | George, Jenet | Rajendran, Devi Sri | Shaji, Anna | Lawrence, Nicole | Saikia, Kongkona | Rathankumar, Abiram Karanam
The treatment of contaminants from lignocellulosic biorefinery effluent has recently been identified as a unique challenge. This study focuses on removing phenolic contaminants and polycyclic aromatic hydrocarbons (PAHs) from lignocellulosic biorefinery wastewater (BRW) applying a laccase-assisted approach. Cassava waste was used as a substrate to produce the maximum yield of laccase enzyme (3.9 U/g) from Pleurotus ostreatus. Among the different inducers supplemented, CuSO₄ (0.5 mM) showed an eight-fold increase in enzyme production (30.8 U/g) after 240 h of incubation. The catalytic efficiency of laccase was observed as 128.7 ± 8.47 S⁻¹mM⁻¹ for syringaldazine oxidation at optimum pH 4.0 and 40 °C. Laccase activity was completely inhibited by lead (II) ion, mercury (II) ion, sodium dodecyl sulphate, sodium azide and 1,4 dithiothretiol and induced significantly by manganese (II) ion and rhamnolipid. After treating BRW with laccase, the concentrations of PAHs and phenolic contaminants of 1144 μg/L and 46160 μg/L were reduced to 96 μg/L and 16100 μg/L, respectively. The ability of laccase to effectively degrade PAHs in the presence of different phenolic compounds implies that phenolic contaminants may play a role in PAHs degradation. After 240 h, organic contaminants were removed from BRW in the following order: phenol >2,4-dinitrophenol > 2-methyl-4,6-dinitrophenol > 2,3,4,6-tetrachlorophenol > acenaphthene > fluorine > phenanthrene > fluoranthene > pyrene > anthracene > chrysene > naphthalene > benzo(a)anthracene > benzo(a)pyrene > benzo(b)fluoranthene > pentachlorophenol > indeno(1,2,3-cd)pyrene > benzo(j) fluoranthene > benzo[k]fluoranthène. The multiple contaminant remediation from the BRW by enzymatic method, clearly suggests that the laccase can be used as a bioremediation tool for the treatment of wastewater from various industries.
Show more [+] Less [-]Fungal biodegradation of the N-nitrosodimethylamine precursors venlafaxine and O-desmethylvenlafaxine in water
2019
Llorca, Marta | Castellet-Rovira, Francesc | Farré, María-José | Jaén-Gil, Adrián | Martínez-Alonso, Maira | Rodríguez-Mozaz, Sara | Sarrà, Montserrat | Barceló, Damià
Antidepressant drugs such as Venlafaxine (VFX) and O-desmethylvenlafaxine (ODMVFX) are emerging contaminants that are commonly detected in aquatic environments, since conventional wastewater treatment plants are unable to completely remove them. They can be precursors of hazardous by-products, such as the carcinogenic N-nitrosodimethylamine (NDMA), generated upon water chlorination, as they contain the dimethylamino moiety, necessary for the formation of NDMA. In this study, the capability of three white rot fungi (Trametes versicolor, Ganoderma lucidum and Pleurotus ostreatus) to remove both antidepressants from water and to decrease NDMA formation potential was investigated. Furthermore, transformation by-products (TPs) generated along the treatment process were elucidated and also correlated with their NDMA formation potential.Very promising results were obtained for T. versicolor and G. lucidum, both being able to remove up to 100% of ODMVFX. In the case of VFX, which is very recalcitrant to conventional wastewater treatment, a 70% of removal was achieved by T. versicolor, along with a reduction in NDMA formation potential, thus decreasing the associated problems for human health and the environment. However, the NDMA formation potential remained practically constant during treatment with G. lucidum despite of the equally high VFX removal (70%). This difference was attributed to the generation of different TPs during both fungal treatments. For example, G. lucidum generated more ODMVFX, which actually has a higher NDMA formation potential than the parent compound itself.
Show more [+] Less [-]Transformation of lamotrigine by white-rot fungus Pleurotus ostreatus
2019
Chefetz, Benny | Marom, Rotem | Salton, Orit | Oliferovsky, Mariana | Mordehay, Vered | Ben-Ari, Julius | Hadar, Y. (Yitzhak)
One of the most persistent pharmaceutical compounds commonly found in treated wastewater is lamotrigine (LTG). It has also been detected in soils and crops irrigated with treated wastewater. Here we focused on the ability of the white-rot edible mushroom Pleurotus ostreatus to remove and transform LTG in liquid cultures. At concentrations of environmental relevance (1 and 10 μg L−1) LTG was almost completely removed from the culture medium within 20 days. To elucidate the mechanism of LTG removal and transformation, we applied a physiological-based approach using inhibitors and a competing agent. These experiments were conducted at a higher concentration for metabolites detection. Based on identification of sulfur-containing metabolites and LTG N2-oxide and the effect of specific inhibitors, cytochrome P450 oxidation is suggested as one of the reaction mechanisms leading to LTG transformation. The variety and number of transformation products (i.e., conjugates) found in the current study were larger than reported in mammals. Moreover, known conjugates with glucuronide, glutathione, or cysteine/glycine, were not found in our system. Since the majority of the identified transformation products were conjugates of LTG, this study highlights the persistence of LTG as an organic pollutant in ecosystems exposed to wastewater.
Show more [+] Less [-]Laccase-mediated transformation of triclosan in aqueous solution with metal cations and humic acid
2017
Sun, Kai | Kang, Fuxing | Waigi, Michael Gatheru | Gao, Yanzheng | Huang, Qingguo
Triclosan (TCS) is a broad-spectrum antimicrobial agent that is found extensively in natural aquatic environments. Enzyme-catalyzed oxidative coupling reactions (ECOCRs) can be used to remove TCS in aqueous solution, but there is limited information available to indicate how metal cations (MCs) and natural organic matter (NOM) influence the environmental fate of TCS during laccase-mediated ECOCRs. In this study, we demonstrated that the naturally occurring laccase from Pleurotus ostreatus was effective in removing TCS during ECOCRs, and the oligomerization of TCS was identified as the dominant reaction pathway by high-resolution mass spectrometry (HRMS). The growth inhibition studies of green algae (Chlamydomonas reinhardtii and Scenedesmus obliquus) proved that laccase-mediated ECOCRs could effectively reduce the toxicity of TCS. The presence of dissolved MCs (Mn²⁺, Al³⁺, Ca²⁺, Cu²⁺, and Fe²⁺ ions) influenced the removal and transformation of TCS via different mechanisms. Additionally, the transformation of TCS in systems with NOM derived from humic acid (HA) was hindered, and the apparent pseudo first-order kinetics rate constants (k) for TCS decreased as the HA concentration increased, which likely corresponded to the combined effect of both noncovalent (sorption) and covalent binding between TCS and humic molecules. Our results provide a novel insight into the fate and transformation of TCS by laccase-mediated ECOCRs in natural aquatic environments in the presence of MCs and NOM.
Show more [+] Less [-]Integration of transcriptomic and proteomic reveals the toxicological molecular mechanisms of decabromodiphenyl ethane (DBDPE) on Pleurotus ostreatus
2022
Li, Wanlun | Wang, Shutao | Chen, Yangyang | Liu, Lu | Hou, Shuying | You, Hong
Decabromodiphenyl ethane (DBDPE), as one of the most widely used new brominated flame retardants (NBFRs), can pose a potential threat to human health and the environment. An integrated transcriptome and proteome was performed for investigating the toxicological molecular mechanisms of Pleurotus ostreatus (P. ostreatus) during the biodegradation of DBDPE at the concentrations of 5 and 20 mg/L. A total of 1193/1018 and 92/126 differentially expressed genes/proteins (DEGs/DEPs) were found, respectively, with DBDPE exposure at 5 and 20 mg/L. These DEGs and DEPs were mainly involved in the cellular process as well as metabolic process. DEPs for oxidation-reduction process and hydrolase activity were up-regulated, and those for membrane, lipid metabolic process and transmembrane transport were down-regulated. The DEGs and DEPs related to some key enzymes were down-regulated, such as NADH dehydrogenase/oxidoreductase, succinate dehydrogenase, cytochrome C1 protein, cytochrome-c oxidase/reductase and ATP synthase, which indicated that DBDPE affected the oxidative phosphorylation as well as tricarboxylic acid (TCA) cycle. Cytochrome P450 enzymes (CYPs) might be involved in DBDPE degradation through hydroxylation and oxidation. Some stress proteins were induced to resist DBDPE toxicity, including major facilitator superfamily (MFS) transporter, superoxide dismutase (SOD), molecular chaperones, heat shock proteins (HSP20, HSP26, HSP42), 60S ribosomal protein and histone H4. The findings help revealing the toxicological molecular mechanisms of DBDPE on P. ostreatus, aiming to improve the removal of DBDPE.
Show more [+] Less [-]Effectiveness of Dyes Removal by Mixed Fungal Cultures and Toxicity of Their Metabolites
2013
Przystaś, Wioletta | Zabłocka-Godlewska, Ewa | Grabińska-Sota, Elżbieta
Decolorization of brilliant green (0.06 g/L), Evans blue (0.15 g/L), and their mixture (total concentration 0.08 g/L, proportion 1:1 w/w) by fungi was studied. Fungal strains [Pleurotus ostreatus (BWPH), Gloeophyllum odoratum (DCa), and Fusarium oxysporum (G1)] were used separately and as a mixture of them. Zootoxicity (Daphnia magna) and phytotoxicity (Lemna minor) changes were estimated after the end of experiment. Mixtures of fungal strains were less effective in decolorization process than the same strains used separately (as a single strains). After 96 h of experiment, living biomass of strain BWPH removed up to 95.5 %; DCa, up to 84.6 %; G1, up to 79.2 % where mixtures BWPH + DCa removed up to 74.3 %; and BWPH + G1, only up to 32.2 % of used dyes. High effectiveness of dyes removal not always corresponded with decrease of toxicity. The highest decrease of zootoxicity and phytotoxicity (from V to III toxicity class or to even nontoxic) was noticed for single strains, while no changes or slight toxicity decrease was noticed in samples with strains mixtures.
Show more [+] Less [-]Biological Removal of Azo and Triphenylmethane Dyes and Toxicity of Process By-Products
2012
Przystaś, Wioletta | Zabłocka-Godlewska, Ewa | Grabińska-Sota, Elżbieta
Increasing environmental pollution is connected with broad applications of dyes and imperfection of dyeing technology. Decolourization of triphenylmethane brilliant green and disazo Evans blue by bacterial and fungal strains and toxicity (phyto- and zootoxicity) of degradation by-products were investigated. Influence of incubation method on dyes removal was evaluated (static, semi-static, shaken). Dead biomass was used for sorption estimation. Toxicity of treated dyes was measured to estimate possible influence on aquatic ecosystems. The zootoxicity test was done with Daphnia magna and phytotoxicity with Lemna minor. Samples were classified according to ACE 89/BE 2/D3 Final Report Commission EC. The best results of removal for all tested strains were reached in shaken samples. In opposite to fungi, bacterial strains decolourized brilliant green more effectively than Evans blue. The most effective bacterial strain was Erwinia spp. (s12) and fungal strains were Polyporus picipes (RWP17) and Pleurotus ostreatus (BWPH and MB). Decolourization of brilliant green was connected with decrease of zootoxicity (D. magna) and phytotoxicity (L. minor). Removal of Evans blue was connected with no changes in zootoxicity and decrease of phytotoxicity in most of samples.
Show more [+] Less [-]Conversion of brewers’ spent grain into proteinaceous animal feed using solid state fermentation
2022
Eliopoulos, Christos | Arapoglou, Dimitrios | Chorianopoulos, Nikos | Markou, Giorgos | Haroutounian, Serkos A.
Brewers’ spent grain (BSG) represents the 85% of the total residue produced during the beer brewing process, with a global annual production volume exceeding 30 Mtons. The current study concerns the application of solid state fermentation (SSF) as a bioprocess where the nutritional value of BSG is improved for further use as animal feed with increased value. The investigated SSF procedure was initiated by the edible fungi Pleurotus ostreatus, which constitutes a natural source of proteins, β-glucans, and various metabolites (vitamins, nutrients, etc.). Herein, the SSF of BSG resulted in a significant increase of protein content by 49.49%, a 10-fold increase of 1,3-1,6 β-glucans, and a respective reduction of cellulose by 11.42%. The application of this method is expected to provide some useful information on the utilization of BSG as substrate for fungi-initiated SSF, a bioprocess allowing the significant reduction of the environmental impact caused by the beer brewing industry and simultaneously producing animal feed with higher protein content and improved nutritional characteristics. Such studies contribute to confront the unavailability of proteinaceous animal feed observed in the last decade.
Show more [+] Less [-]Laccase-Catalyzed Oxidative Coupling Reaction of Triclosan in Aqueous Solution
2016
Sun, Kai | Huang, Qingguo | Gao, Yanzheng
Triclosan (TCS) is an antimicrobial agent that is extensively used in personal care products (PCPs), and its residue is frequently reported in aquatic environments. In this study, we investigated the reaction behavior of TCS during enzyme-catalyzed oxidative coupling reactions (ECOCRs) by laccase from Pleurotus ostreatus and determined how the presence of natural organic matter (NOM) influenced the formation of the products. Results indicated that the optimum pH for TCS transformation was 6.0 in laccase-mediated ECOCRs. At pH values below 5.0 and above 7.0, the pseudo first-order kinetic rate constants (k) of TCS transformation declined significantly. Moreover, the k values of TCS transformation increased as the laccase activity increased (0.1179–0.5757 h⁻¹). A total of four product peaks were generated, and they were more hydrophobic than TCS. High-resolution mass spectrometry (HRMS) analysis indicated that these products could be the oligomers resulting from TCS self-coupling reactions. The relative peak areas of these oligomers displayed strong linear correlations with the different initial TCS concentrations, and the saturation point of laccase (3.0 U mL⁻¹), when the binding with TCS was 40 μmol L⁻¹. In the presence of NOM (i.e., humates and fulvates), humates in particular strongly inhibited TCS transformation and lowered the extent of its self-coupling, which likely resulted from the cross-coupling between TCS and NOM. Our study improves a better understanding of the reaction behavior of TCS in the natural aquatic environment during laccase-mediated ECOCRs.
Show more [+] Less [-]Two birds with one stone: oyster mushroom mediated bimetallic Au-Pt nanoparticles for agro-waste management and anticancer activity
2021
Chaturvedi, Vivek K | Yadav, Navneet | Rai, Neeraj K. | Bohara, Raghvendra A. | Rai, Sachchida Nand | Aleya, Lotfi | Singh, Mohan P.
Agriculture has the most significant contribution in fulfilling the basic human need, sustaining life, and strengthening the economy of any country. To feed the exploding population of the world, there has been a quantum jump in the production of agricultural commodities, which has led to the production of a substantial considerable quantity of agricultural and agro-industrial wastes. The bulks of these wastes are lignocellulosic in nature and consist of three main polymeric constituents, i.e., cellulose, hemicellulose, and lignin, which are recalcitrant. The primary significant portions of these remain unutilized and are burnt in the field, leading to severe environmental aggression and wastage of resource. Farmers across the globe, including India, burn these agricultural wastes in their thousands of acre land, which contribute to spoiling the air quality index (AQI). This is very harmful, especially to children, pregnant women, old adults, and for patients suffering from respiratory diseases. The current manuscript sets up an agro-waste management platform by using paddy straw as a substrate for the production of nutritionally and medically rich oyster mushroom, Pleurotus florida (Pf) and which is further used in the green synthesis of bimetallic (gold-platinum) Au-Pt nanoparticle. Yield performance and biological efficiency of Pf were calculated from the degraded paddy straw. The green synthesized Au-Pt NPs were structurally characterized by ultraviolet-visible (UV-Vis), X-ray diffraction (XRD), transmission electron microscopy (TEM), energy dispersive spectroscopy (EDS), and zeta potential analysis. The prepared NPs showed a face-centered cubic crystal structure, icosahedral shape with a mean particle size of 16 nm. Furthermore, we examined the cytotoxic activity of Au-Pt NPs using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, intracellular reactive oxygen species (ROS) generation, and apoptosis by propidium iodide assay. We found that Au-Pt NPs exerted apoptotic activity on the human colon cancer cell line (HCT 116) in a dose-dependent manner from 12.5 to 200 μg/mL. Overall, our findings create a prototype and open a new door to synthesizing functional nanoparticle by using oyster mushroom as the substrate for paddy straw agro-waste management and the applicability of Pf in the synthesis of eco-friendly Au-Pt NPs. This is the first kind of approach that kills two birds with one stone.
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