Effect of Micro-Nanobubbles on Arsenic Removal by Trichoderma atroviride for Bioscorodite Generation.

The global environmental issue of arsenic (As) contamination in drinking water is a significant problem that requires attention. Therefore, the aim of this research was to address the application of a sustainable methodology for arsenic removal through mycoremediation aerated with micro-nanobubbles (MNBs), leading to bioscorodite (FeAsO4·2H2O) generation. To achieve this, the fungus Trichoderma atroviride was cultivated in a medium amended with 1 g/L of As(III) and 8.5 g/L of Fe(II) salts at 28 °C for 5 days in a tubular reactor equipped with an air MNBs diffuser (TR-MNBs). A control was performed using shaking flasks (SF) at 120 rpm. A reaction was conducted at 92 °C for 32 h for bioscorodite synthesis, followed by further characterization of crystals through Fourier-Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and X-ray diffraction (XRD) analyses. At the end of the fungal growth in the TR-MNBs, the pH decreased to 2.7-3.0, and the oxidation-reduction potential (ORP) reached a value of 306 mV at 5 days. Arsenic decreased by 70%, attributed to possible adsorption through rapid complexation of oxidized As(V) with the exchangeable ferrihydrite ((Fe(III))4-5(OH,O)12), sites, and the fungal biomass. This mineral might be produced under oxidizing and acidic conditions, with a high iron concentration (As:Fe molar ratio = 0.14). The crystals produced in the reaction using the TR-MNBs culture broth and characterized by SEM, XRD, and FTIR revealed the morphology, pattern, and As-O-Fe vibration bands typical of bioscorodite and römerite (Fe(II)(Fe(III))2(SO4)4·14H2O). Arsenic reduction in SF was 30%, with slight characteristics of bioscorodite. Consequently, further research should include integrating the TR-MNBs system into a pilot plant for arsenic removal from contaminated water.

Nanotechnology as an efficient and effective alternative for wastewater treatment: an overview.

The increase in the surface and groundwater contamination due to global population growth, industrialization, proliferation of pathogens, emerging pollutants, heavy metals, and scarcity of drinking water represents a critical problem. Because of this problem, particular emphasis will be placed on wastewater recycling. Conventional wastewater treatment methods may be limited due to high investment costs or, in some cases, poor treatment efficiency. To address these issues, it is necessary to continuously evaluate novel technologies that complement and improve these traditional wastewater treatment processes. In this regard, technologies based on nanomaterials are also being studied. These technologies improve wastewater management and constitute one of the main focuses of nanotechnology. The following review describes wastewater's primary biological, organic, and inorganic contaminants. Subsequently, it focuses on the potential of different nanomaterials (metal oxides, carbon-based nanomaterials, cellulose-based nanomaterials), membrane, and nanobioremediation processes for wastewater treatment. The above is evident from the review of various publications. However, nanomaterials' cost, toxicity, and biodegradability need to be addressed before their commercial distribution and scale-up. The development of nanomaterials and nanoproducts must be sustainable and safe throughout the nanoproduct life cycle to meet the requirements of the circular economy.

Removal of orange G dye by Aspergillus niger and its effect on organic acid production.

Azo dyes have been found in wastewater from textile industries. These compounds continuously persist in the environment for long periods of time and may be toxic for living beings. An alternative treatment for dye removal that has proven to be effective is aerobic treatment with fungi. In this study, Aspergillus niger was investigated as a mechanism to remove orange G (OG). Removal of 200 mg/L of OG by A. niger biomass was carried out in solid and liquid medium, which showed a positive correlation between A. niger growth and dye removal. In liquid media what was proved is that the efficiency of OG removal by A. niger depends on its concentration; at 200 mg/L of OG remove by degradation and at 400 mg/L by processes as sorption and degradation. During OG removal, the generation of organic acids by A. niger was modified compared to constitutive generation, one of the modifications was the increase of gluconic acid production and the decrease of acids involved in the Krebs cycle, as well as the null detection of oxalic acid. The monitoring of organic acids by high-performance liquid chromatography (HPLC) was important because some of them have been linked to dye removal.

Bioremediation of an agricultural saline soil contaminated with endosulfan and Escherichia coli by an active surface agent induced in a Penicillium crustosum culture.

This study evaluates the production of a biological active surface agent (BASA) through its surface tension (ST) and emulsifying activity (E24) for endosulfan degradation (ED) and Escherichia coli growth inhibition (EcGI) in an agricultural saline soil. The fungus, identified as Penicillium crustosum was isolated from the Citrus sinensis peel (CsP), then the surface properties were evaluated in 9 culture media through a Taguchi L9 experimental design. The culture conditions included: stirring speed, pH, carbon (C) and nitrogen (N) sources; being glucose, NH4N03, 120 rpm and pH of 5, the most significant parameters in the BASA production. The BASA identified as a lipopeptide type, showed a ST = 38 mN m-1 and E24=71%. Both properties were stable at 80 °C, while ST presented stability in the pH range of 2 - 12, and a saline concentration of 200 g L-1; E24 was also stable at a pH between 8-12. Further application of BASA and fungal inoculum to a contaminated agricultural saline soil presented an EcGI of 99.8% on the 8th day, and ED of 92.9 ± 4.7% in 30 days, respectively; being the first report that uses this fungus for pesticide and bacteria elimination from an agricultural saline soil.

Effect of fungal co-cultures on ligninolytic enzyme activities, H2O2 production, and orange G discoloration.

In this study, the effects of Aspergillus niger in coculture with the basidiomycetes, Trametes versicolor, T. maxima, and Ganoderma spp., were studied to assess H2O2 production and laccase (Lac), Lignin Peroxidase (LiP), and manganese peroxidase (MnP) activities. The results indicated that maximum discoloration was of 97%, in the T. maxima and A. niger coculture, where the concentration of H2O2 was 5 mg/L and 6.3 mg/L in cultures without and with dye, respectively. These concentrations of H2O2 were 1.6- and 1.8-fold higher than monocultures of T. maxima (3.37 mg/L) and A. niger (3.87 mg/L), respectively. In the same coculture, the LiP and MnP enzyme activities also increased 12-fold, (from 0.08 U/mg to 0.99 U/mg), and 67-fold, (from 0.11 U/mg to 7.4 U/mg), respectively. The Lac activity increased 1.7-fold (from 13.46 U/mg to 24 U/mg). Further, a Box-Behnken experimental design indicated a 1.8-fold increase of MnP activity (from 7.4 U/mg to 13.3 U/mg). In addition, dye discoloration regression model obtained from the Box-Behnken experimental design showed a positively correlation with H2O2, (R2 = 0.58) and a negatively correlation with Lac activity (R2 = -0.7).

Removal of phenanthrene in aqueous solution containing photon competitors by TiO2-C-Ag film supported on fiberglass.

Surface interactions with pollutants and photons are key factors that affect the applications of TiO2 in environmental remediation. In this study, the solubilizing agents dimethylsulfoxide and polyoxyethylene sorbitan monooleate, which act as photon competitors, had no effect on the photocatalytic activity of TiO2-C-Ag film in phenanthrene (PHE) removal. Fiberglass with TiO2-C-Ag coating removed 91.1 ± 5.2 and 99.7 ± 0.4% of PHE in treatments using UVA (365-465 nm) and UVC (254 nm) irradiation, respectively. The use of fiberglass as a support increased the superficial area, thus allowing PHE sorption. C and Ag, which are electrically active impurities in TiO2, enhanced its photocatalytic activity and thus the attraction of the pollutant to its surface. The use of high-frequency UV light (UVC) decreased the amount of carbon species deposited on the TiO2CAg film surface. X-ray photoelectron spectroscopy of the TiO2-C-Ag film revealed extensive oxidation of the carbon deposited on the film under UVC light and loss of electrons from Ag clusters by conversion of Ag0 to Ag3+.

Production of a halotolerant biofilm from green coffee beans immobilized on loofah fiber (Luffa cylindrica) and its effect on phenanthrene degradation in seawater.

A biofilm developed from low quality green coffee beans was tested for its capacity to degrade the polynuclear aromatic hydrocarbon (PAH), phenanthrene (Phe), in seawater. Microorganisms were immobilized on two types of Luffa cylindrica (with three and four placental cavities), and the effects of moisture content (20, 30 and 40% of water holding capacity) and particle size (<0.42 mm, 0.42-0.86 mm and 0.86-2.0 mm) of green coffee beans on microbial activity were considered. Biofilm growth determined by respirometry showed a highest microbial activity at a moisture content of 40% and particle size of 0.42-0.86 mm. The loofah fiber with three placental cavities showed the highest adherence of microorganisms. The kinetics of microbial growth in both seawater and distilled water and the scanning electron microscopies indicated that the microorganisms associated with green coffee beans are halotolerant. In fact, I-GCB-SW-G biofilm degraded 67.56% of Phe (50 mg L-1) in seawater, at a significantly higher rate than in distilled water (I-GCB-DW-W).

Partial removal and detoxification of Malachite Green and Crystal Violet from laboratory artificially contaminated water by Pleurotus ostreatus / Eliminación parcial y desintoxicación de Verde Malaquita y Cristal Violeta del laboratorio con agua contaminada artificialmente por Pleurotus ostreatus / Extração e desintoxicação de Verde Malaquita e Cristal Violeta de água contaminada artificialmente em laboratório por Pleurotus ostreatus

Abstract The triphenylmethane Malachite Green (MG) and Crystal Violet (CV) dyes are cationic dyes and mix with domestic wastewater when dumped; increasing, among others, the chemical and biological oxygen demand and can cause acute toxicity at different trophic levels. Promoting the removal (decolorization) of MG and CV, and laccase activity (54.8 ± 8.9 and 30.6 ± 2.9 UL-1 respectively) by using P. ostreatus viable biomass needed parameters such as pH (4.5 and 6.0), temperature (25 to 30 °C), stirring speed (120 rpm), percentage of inoculum (2% v/v), and dye concentration (20 and 10 mg L-1). In adsorption studies, it was showed that an acidic pH favors the adsorption of both dyes and the model of pseudo-second order describes best the phenomenon of adsorption. Finally, the germination index (GI), using Lactuca sativa seeds for the initial dyes solutions, was < 50%; demonstrating its high phytotoxic effect. When dye solutions were treated with viable biomass, the GI increased, leaving open the possibility to perform future research to determine if the aqueous solutions, post-treated with P. ostreatus, could be used in treatments that generate less toxic water which could be used in processes that do not require potable water.

Resumen Los colorantes trifenilmetánicos Verde Malaquita (MG) y Crystal Violeta (CV) son catiónicos y al ser vertidos se mezclan con aguas residuales domésticas, incrementando, entre otros, la demanda química y biológica de oxígeno; pudiendo causar toxicidad aguda en diferentes niveles tróficos. En este estudio se encontró que los parámetros pH (4,5 y 6,0), temperatura (25 y 30 °C), velocidad de agitación (120 r.p.m.), porcentaje de inóculo (2 % v/v) y concentración de colorante (20 y 10 mgL-1), presentaron un efecto significativo (p < 0.05) para favorecer la remoción (decoloración) de MG y CV, así como la actividad lacasa (54,76 ± 8,91 y 30,59 ± 2,89 UL-1 respectivamente) al utilizar biomasa viable de P. ostreatus. En los estudios de adsorción se evidenció que pH ácidos favorecen la adsorción de ambos colorantes y que el modelo de Pseudo-segundo orden describe mejor el fenómeno de quimisorción. Finalmente los índices de germinación (IG) empleando semillas de Lactuca sativa, para los colorantes iniciales fueron < 50 %; demostrando su efecto fitotóxico elevado. Cuando las soluciones de colorantes fueron tratadas con biomasa viable, el IG aumentó, dejando abierta la puerta para la realización de investigaciones futuras con la intensión de determinar si las soluciones acuosas, postratadas con P ostreatus, pueden ser utilizadas en tratamientos que generen aguas menos tóxicas y que estas puedan ser empleadas en otros procesos que no requieran agua potable.

Resumen Os corantes de tipo trifenilmetano Verde Malaquita (VM) e Cristal Violeta (CV) são corantes catiônicos e se misturam com águas residuais domésticas quando descartadas; aumentando, entre outros, as demandas químicas e biológicas de oxigênio, podendo causar toxicidade aguda em diferentes níveis tróficos. Promoveu-se a remoção (descoloração) de VM e CV, e atividade da lacase (54.8 ± 8.9 e 30.6 ± 2.9 UL-1 respectivamente) utilizando como parâmetros necessários para a biomassa viável de P. ostreatus como pH (4,5 e 6,0), temperatura (25 a 30 °C), velocidade de agitação (120 RPM), porcentagem de inócuo (2 % v/v), e concentração de corante (20 e 10 mg L-1). Em estudos de absorção, se demonstrou que um pH mais ácido favorece a absorção de ambos corantes e o modelo de pseudo-segunda ordem descreve melhor o fenômeno da absorção. Finalmente, o índice de germinação (IG), utilizando sementes de Lactuca sativa para as soluções iniciais dos corantes, foi < 50 %; demonstrando assim seu alto efeito fitotóxico. Quando as soluções de corante foram tratadas com a biomassa viável, o IG aumentou, deixando em aberto a possibilidade de realizar futuras investigações para determinar se as soluções aquosas, tratadas com P. ostreatus, poderiam ser utilizadas em tratamentos que gerem águas menos tóxicas, que poderia ser utilizada em processos que não requerem água potável.

Atrazine degradation by fungal co-culture enzyme extracts under different soil conditions.

This investigation was undertaken to determine the atrazine degradation by fungal enzyme extracts (FEEs) in a clay-loam soil microcosm contaminated at field application rate (5 µg g(-1)) and to study the influence of different soil microcosm conditions, including the effect of soil sterilization, water holding capacity, soil pH and type of FEEs used in atrazine degradation through a 2(4) factorial experimental design. The Trametes maxima-Paecilomyces carneus co-culture extract contained more laccase activity and hydrogen peroxide (H2O2) content (laccase = 18956.0 U mg protein(-1), H2O2 = 6.2 mg L(-1)) than the T. maxima monoculture extract (laccase = 12866.7 U mg protein(-1), H2O2 = 4.0 mg L(-1)). Both extracts were able to degrade atrazine at 100%; however, the T. maxima monoculture extract (0.32 h) achieved a lower half-degradation time than its co-culture with P. carneus (1.2 h). The FEE type (p = 0.03) and soil pH (p = 0.01) significantly affected atrazine degradation. The best degradation rate was achieved by the T. maxima monoculture extract in an acid soil (pH = 4.86). This study demonstrated that both the monoculture extracts of the native strain T. maxima and its co-culture with P. carneus can efficiently and quickly degrade atrazine in clay-loam soils.

Effect of Cd⁺² on phosphate solubilizing abilities and hydrogen peroxide production of soil-borne micromycetes isolated from Phragmites australis-rhizosphere.

The aims of this work were to evaluate the phosphate-solubilization and hydrogen peroxide (H2O2) production by the soil-borne micromycetes, Aspergillus japonicus, Penicillium italicum and Penicillium dipodomyicola, isolated from Phragmites australis rhizosphere and to study the effect of several concentrations of Cadmium (Cd(2+)) on both variables. Our results showed that P. italicum achieved a higher P-solubilization and H2O2 production than A. japonicus and P. dipodomyicola, as only P. italicum showed a positive correlation (R(2) = 0.71) between P-solubilization and H2O2 production. In dose-response assays, P. italicum was also more tolerant to Cd(2+) (0.31 mM) in comparison to A. japonicus (0.26 mM). Analysis of the 2(4) factorial experimental design showed that P-solubilization by P. italicum was negatively affected by increases in Cd(2+) (p = 0.04) and yeast extract (p = 0.02) in the culture medium. The production of H2O2 was positively affected only by glucose (p = 0.002). Fungal biomass production was reduced significantly (p = 0.0009) by Cd(2+) and increased (p = 0.0003) by high glucose concentration in the culture medium. The tolerance and correlation between P-solubilization and H2O2 production in the presence of Cd(2+) was strain and species dependent. The effects of Cd(2+), glucose, ammonium sulfate and yeast extract on those variables were evaluated through a two-level factorial design. P. italicum is promising for P-solubilization in soils contaminated with Cd(2+) and may be an alternative for manufacture of biofertilizers to replace chemical fertilizers.

The Fatty Acid Profile Analysis of Cyperus laxus Used for Phytoremediation of Soils from Aged Oil Spill-Impacted Sites Revealed That This Is a C18:3 Plant Species.

The effect of recalcitrant hydrocarbons on the fatty acid profile from leaf, basal corm, and roots of Cyperus laxus plants cultivated in greenhouse phytoremediation systems of soils from aged oil spill-impacted sites containing from 16 to 340 g/Kg total hydrocarbons (THC) was assessed to investigate if this is a C18:3 species and if the hydrocarbon removal during the phytoremediation process has a relationship with the fatty acid profile of this plant. The fatty acid profile was specific to each vegetative organ and was strongly affected by the hydrocarbons level in the impacted sites. Leaf extracts of plants from uncontaminated soil produced palmitic acid (C16), octadecanoic acid (C18:0), unsaturated oleic acids (C18:1-C18:3), and unsaturated eichosanoic (C20:2-C20:3) acids with a noticeable absence of the unsaturated hexadecatrienoic acid (C16:3); this finding demonstrates, for the first time, that C. laxus is a C18:3 plant. In plants from the phytoremediation systems, the total fatty acid contents in the leaf and the corm were negatively affected by the hydrocarbons presence; however, the effect was positive in root. Interestingly, under contaminated conditions, unusual fatty acids such as odd numbered carbons (C15, C17, C21, and C23) and uncommon unsaturated chains (C20:3n6 and C20:4) were produced together with a remarkable quantity of C22:2 and C24:0 chains in the corm and the leaf. These results demonstrate that weathered hydrocarbons may drastically affect the lipidic composition of C. laxus at the fatty acid level, suggesting that this species adjusts the cover lipid composition in its vegetative organs, mainly in roots, in response to the weathered hydrocarbon presence and uptake during the phytoremediation process.

Evaluation of hydrocarbons and organochlorine pesticides and their tolerant microorganisms from an agricultural soil to define its bioremediation feasibility.

The concentrations of hydrocarbons and organochlorine pesticides (OCPs), nutrients and tolerant microorganisms in an agricultural soil from a locality in Tepeaca, Puebla, Mexico, were determined to define its feasibility for bioremediation. The OCPs detected were heptachlor, aldrin, trans-chlordane, endosulfán I, endosulfán II, 1,1,1-bis-(4-chlorophenyl)-2,2-trichloroethane (4,4'-DDT), 1,1-bis-(4-chlorophenyl)-2,2-dichloroethene (4,4'-DDE) and endrin aldehyde, with values of 0.69-30.81 ng g(-1). The concentration of hydrocarbons in the soil of Middle Hydrocarbons Fraction (MHF), C10 to C28, was 4608-27,748 mg kg(-1) and 1117-19,610 mg kg(-1) for Heavy Hydrocarbons Fraction (HHF), C28 to C35, due to an oil spill from the rupture of a pipeline. The soil was deficient in nitrogen (0.03-0.07%) and phosphorus (0 ppm), and therefore it was advisable to fertilize to bio-stimulate the native microorganisms of soil. In the soil samples, hydrocarbonoclast fungi 3.72 × 10(2) to 44.6 × 10(2) CFU g(-1) d.s. and hydrocarbonoclast bacteria (0.17 × 10(5) to 8.60 × 10(5) CFU g(-1) d.s.) were detected, with a tolerance of 30,000 mg kg(-1) of diesel. Moreover, pesticideclast fungi (5.13 × 10(2) to 42.2 × 10(2) CFU g(-1) d.s.) and pesticideclast bacteria (0.15 × 10(5) to 9.68 × 10(5) CFU g(-1) d.s.) were determined with tolerance to 20 mg kg(-1) of OCPs. Fungi and bacteria tolerant to both pollutants were also quantified. Therefore, native microorganisms had potential to be stimulated to degrade hydrocarbons and pesticides or both pollutants. The concentration of pollutants and the microbial activity analyzed indicated that bioremediation of the soil contaminated with hydrocarbons and pesticides using bio-stimulation of native microorganisms was feasible.

Computational analysis and low-scale constitutive expression of laccases synthetic genes GlLCC1 from Ganoderma lucidum and POXA 1B from Pleurotus ostreatus in Pichia pastoris.

Lacasses are multicopper oxidases that can catalyze aromatic and non-aromatic compounds concomitantly with reduction of molecular oxygen to water. Fungal laccases have generated a growing interest due to their biotechnological potential applications, such as lignocellulosic material delignification, biopulping and biobleaching, wastewater treatment, and transformation of toxic organic pollutants. In this work we selected fungal genes encoding for laccase enzymes GlLCC1 in Ganoderma lucidum and POXA 1B in Pleurotus ostreatus. These genes were optimized for codon use, GC content, and regions generating secondary structures. Laccase proposed computational models, and their interaction with ABTS [2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] substrate was evaluated by molecular docking. Synthetic genes were cloned under the control of Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase (GAP) constitutive promoter. P. pastoris X-33 was transformed with pGAPZαA-LaccGluc-Stop and pGAPZαA-LaccPost-Stop constructs. Optimization reduced GC content by 47 and 49% for LaccGluc-Stop and LaccPost-Stop genes, respectively. A codon adaptation index of 0.84 was obtained for both genes. 3D structure analysis using SuperPose revealed LaccGluc-Stop is similar to the laccase crystallographic structure 1GYC of Trametes versicolor. Interaction analysis of the 3D models validated through ABTS, demonstrated higher substrate affinity for LaccPost-Stop, in agreement with our experimental results with enzymatic activities of 451.08 ± 6.46 UL-1 compared to activities of 0.13 ± 0.028 UL-1 for LaccGluc-Stop. This study demonstrated that G. lucidum GlLCC1 and P. ostreatus POXA 1B gene optimization resulted in constitutive gene expression under GAP promoter and α-factor leader in P. pastoris. These are important findings in light of recombinant enzyme expression system utility for environmentally friendly designed expression systems, because of the wide range of substrates that laccases can transform. This contributes to a great gamut of products in diverse settings: industry, clinical and chemical use, and environmental applications.

Response of ligninolytic macrofungi to the herbicide atrazine: dose-response bioassays / Respuesta de macrohongos ligninolíticos al herbicida atrazina: bioensayos dosis-respuesta

The effect of atrazine concentrations on mycelial growth and ligninolytic enzyme activities of eight native ligninolytic macrofungi isolated in Veracruz, México, were evaluated in a semi-solid culture medium. Inhibition of mycelial growth and growth rates were significantly affected (p = 0.05) by atrazine concentrations (468, 937, 1875, and 3750 mg/l). In accordance with the median effective concentration (EC50), Pleurotus sp. strain 1 proved to be the most tolerant isolate to atrazine (EC50 = 2281.0 mg/l), although its enzyme activity was not the highest. Pycnoporus sanguineus strain 2, Daedalea elegans and Trametes maxima showed high laccase activity (62.7, 31.9, 29.3 U mg/protein, respectively) without atrazine (control); however, this activity significantly increased (p < 0.05) (to 191.1, 83.5 and 120.6 U mg/protein, respectively) owing to the effect of atrazine (937 mg/l) in the culture medium. Pleurotus sp. strain 2 and Cymatoderma elegans significantly increased (p < 0.05) their manganese peroxidase (MnP) activities under atrazine stress at 468 mg/l. The isolates with high EC50 (Pleurotus sp. strain 1) and high enzymatic activity (P. sanguineus strain 2 and T. maxima) could be considered for future studies on atrazine mycodegradation. Furthermore, this study confirms that atrazine can increase laccase and MnP activities in ligninolytic macrofungi.

Se evaluó el efecto de diferentes concentraciones de atrazina sobre el crecimiento micelial y la actividad enzimática de ocho macrohongos ligninolíticos aislados en Veracruz, México. La inhibición del crecimiento micelial y la tasa de crecimiento diaria fueron significativamente (p < 0,05) afectadas por todas las dosis de atrazina (468, 937, 1875 y 3750 mg/l) adicionadas al medio de cultivo. De acuerdo con la concentración efectiva media (CE50), Pleurotus sp. cepa 1 fue el aislamiento más tolerante a la atrazina (CE50 = 2281 mg/l), aunque sus actividades enzimáticas no fueron altas. Pycnoporus sanguineus cepa 2, Daedalea elegans y Trametes maxima mostraron actividades altas de lacasa (62,7, 31,9 y 29,3 U mg/proteína, respectivamente) en ausencia de atrazina (control); estas actividades se incrementaron (p < 0,05) significativamente (191,1, 83,5 y 120,6 U mg/proteína, respectivamente) en presencia de atrazina (937 mg/l) en el medio de cultivo. Pleurotus sp. cepa 2 y Cymatoderma elegans incrementaron significativamente (p < 0,05) sus actividades de manganeso peroxidasa (MnP) bajo la concentración de 468 mg/l de atrazina. Los aislamientos con alta CE50 (Pleurotus sp. cepa 1) y alta actividad enzimática (P. sanguineus cepa 2 y T. maxima) podrían ser considerados para futuros estudios en la micodegradación de atrazina. Además, el presente estudio confirma que la atrazina puede incrementar las actividades lacasa y MnP en macrohongos ligninolíticos.

Response of ligninolytic macrofungi to the herbicide atrazine: dose-response bioassays / Respuesta de macrohongos ligninolíticos al herbicida atrazina: bioensayos dosis-respuesta

The effect of atrazine concentrations on mycelial growth and ligninolytic enzyme activities of eight native ligninolytic macrofungi isolated in Veracruz, México, were evaluated in a semi-solid culture medium. Inhibition of mycelial growth and growth rates were significantly affected (p = 0.05) by atrazine concentrations (468, 937, 1875, and 3750 mg/l). In accordance with the median effective concentration (EC50), Pleurotus sp. strain 1 proved to be the most tolerant isolate to atrazine (EC50 = 2281.0 mg/l), although its enzyme activity was not the highest. Pycnoporus sanguineus strain 2, Daedalea elegans and Trametes maxima showed high laccase activity (62.7, 31.9, 29.3 U mg/protein, respectively) without atrazine (control); however, this activity significantly increased (p < 0.05) (to 191.1, 83.5 and 120.6 U mg/protein, respectively) owing to the effect of atrazine (937 mg/l) in the culture medium. Pleurotus sp. strain 2 and Cymatoderma elegans significantly increased (p < 0.05) their manganese peroxidase (MnP) activities under atrazine stress at 468 mg/l. The isolates with high EC50 (Pleurotus sp. strain 1) and high enzymatic activity (P. sanguineus strain 2 and T. maxima) could be considered for future studies on atrazine mycodegradation. Furthermore, this study confirms that atrazine can increase laccase and MnP activities in ligninolytic macrofungi.(AU)

Se evaluó el efecto de diferentes concentraciones de atrazina sobre el crecimiento micelial y la actividad enzimática de ocho macrohongos ligninolíticos aislados en Veracruz, México. La inhibición del crecimiento micelial y la tasa de crecimiento diaria fueron significativamente (p < 0,05) afectadas por todas las dosis de atrazina (468, 937, 1875 y 3750 mg/l) adicionadas al medio de cultivo. De acuerdo con la concentración efectiva media (CE50), Pleurotus sp. cepa 1 fue el aislamiento más tolerante a la atrazina (CE50 = 2281 mg/l), aunque sus actividades enzimáticas no fueron altas. Pycnoporus sanguineus cepa 2, Daedalea elegans y Trametes maxima mostraron actividades altas de lacasa (62,7, 31,9 y 29,3 U mg/proteína, respectivamente) en ausencia de atrazina (control); estas actividades se incrementaron (p < 0,05) significativamente (191,1, 83,5 y 120,6 U mg/proteína, respectivamente) en presencia de atrazina (937 mg/l) en el medio de cultivo. Pleurotus sp. cepa 2 y Cymatoderma elegans incrementaron significativamente (p < 0,05) sus actividades de manganeso peroxidasa (MnP) bajo la concentración de 468 mg/l de atrazina. Los aislamientos con alta CE50 (Pleurotus sp. cepa 1) y alta actividad enzimática (P. sanguineus cepa 2 y T. maxima) podrían ser considerados para futuros estudios en la micodegradación de atrazina. Además, el presente estudio confirma que la atrazina puede incrementar las actividades lacasa y MnP en macrohongos ligninolíticos.(AU)

Enhancement of ligninolytic enzyme activities in a Trametes maxima-Paecilomyces carneus co-culture: Key factors revealed after screening using a Plackett-Burman experimental design

Background In the industrial biotechnology, ligninolytic enzymes are produced by single fungal strains. Experimental evidence suggests that co-culture of ligninolytic fungi and filamentous microfungi results in an increase laccase activity. In this topic, only the ascomycete Trichoderma spp. has been studied broadly. However, fungal ligninolytic-filamentous microfungi biodiversity interaction in nature is abundant and poorly studied. The enhancement of laccase and manganese peroxidase (MnP) activities of Trametes maxima as a function of time inoculation of Paecilomyces carneus and under several culture conditions using Plackett-Burman experimental design (PBED) were investigated. Results The highest increases of laccase (12,382.5 U/mg protein) and MnP (564.1 U/mg protein) activities were seen in co-cultures I3 and I5, respectively, both at 10 d after inoculation. This level of activity was significantly different from the enzyme activity in non-inoculated T. maxima (4881.0 U/mg protein and 291.8 U/mg protein for laccase and MnP, respectively). PBED results showed that laccase was increased (P < 0.05) by high levels of glucose, (NH4)2SO4 and MnSO4 and low levels of KH2PO4, FeSO4 and inoculum (P < 0.05). In addition, MnP activity was increased (P < 0.05) by high yeast extract, MgSO4, CaCl2 and MnSO4 concentrations. Conclusions Interaction between indigenous fungi: T. maxima-P. carneus improves laccase and MnP activities. The inoculation time of P. carneus on T. maxima plays an important role in the laccase and MnP enhancement. The nutritional requirements for enzyme improvement in a co-culture system are different from those required for a monoculture system.

Treatment of a textile effluent from dyeing with cochineal extracts using Trametes versicolor fungus.

Trametes versicolor (Tv) fungus can degrade synthetic dyes that contain azo groups, anthraquinone, triphenylmethane polymers, and heterocyclic groups. However, no references have been found related to the degradation of natural dyes, such as the carminic acid that is contained in the cochineal extract. Experiments to determine the decolorization of the effluent used in the cotton dyeing process with cochineal extract by means of Tv fungus were done. Treatments to determine decolorization in the presence or absence of Kirk's medium, glucose, and fungus, with an addition of 50% (v v-1) of nonsterilized effluent were performed. Physicochemical characterization was performed at the start and end of the treatment. Degradation kinetics were determined. A direct relationship was found between the dry weight of fungi, pH, and the decolorization system, with higher decolorization at lower pH levels (pH ~4.3). High decolorization (81% ± 0.09; 88% ± 0.17; and 99% ± 0.04) for three of the eight treatments (Kirk's medium without glucose, Kirk's medium with glucose, and without medium with glucose, respectively) was found. Toxicity tests determined an increase in the initial effluent toxicity (7.33 TU) compared with the final treatment (47.73 TU) in a period of 11 days. For this system, a degradation sequence of the carminic acid structure present in the effluent by the Tv fungus is suggested, in which it is seen that metabolites still containing aromatic structures are generated.

Application of 2III7-3 fractional factorial experimental design to enhance enzymatic activities of Pleurotus ostreatus with high concentrations of polychlorinated biphenyls.

A 2(III)(7-3) fractional factorial experimental design was used to establish 16 culture media, with and without PCBs to enhance the activities of laccase (Lac), manganese peroxidase (MnP), and versatile peroxidase (VP) produced by the white rot fungus Pleurotus ostreatus. The culture was added to 10,000 mg L(-1) of transformer oil, containing 71% of the identified Arochlor 1242. The culture conditions were established with eight variables at two values (levels); pH (4 and 6), agitation (100 and 200 rpm), CuSO(4) (150 and 250 mg L(-1)), MnSO(4) (50 and 200 mg L(-1)), Tween 80 (13 and 3500 mg L(-1)), wheat straw (0 and 2.5 g L(-1)), sugarcane bagasse (0 and 2.5 g L(-1)),and Arochlor 1242 (0 and 7100 mg L(-1)) at 4, 8, 12, 16 and 20 days old culture. Laccase activity was enhanced at a high value of pH and low value of agitation (P<0.001) and correlated positively (R(2)= 0.9; α=0.05) with the removal of polychlorinated biphenyls (PCBs). VP activity was enhanced 27-fold with PCBs, Tween 80 and pH. The MnP activity was increased 1.2-fold with PCBs. The fractional factorial experimental design methodology allowed us to determine the P. ostreatus culture media conditions to enhance Lac and VP activities for efficient removal of Arochlor 1242 (one of the most recalcitrant organochloride pollutants). The factors that shown the greatest effect on Lac activity were: pH, agitation and high concentrations of Arochlor 1242.

Heterologous expression of manganese peroxidase in Aspergillus niger and its effect on phenanthrene removal from soil.

A strain of Aspergillus niger, previously isolated from sugarcane bagasse because of its capacity to degrade phenanthrene in soil by solid culture, was used to express a manganese peroxidase gene (mnp1) from Phanerochaete chrysosporium, aiming at increasing its polycyclic aromatic hydrocarbons degradation capacity. Transformants were selected based on their resistance to hygromycin B and the discoloration induced on Poly R-478 dye by the peroxidase activity. The recombinant A. niger SBC2-T3 strain developed MnP activity and was able to remove 95% of the initial phenanthrene (400 ppm) from a microcosm soil system after 17 days, whereas the wild strain removed 72% under the same conditions. Transformation success was confirmed by PCR amplification using gene-specific primers, and a single fragment (1,348 bp long, as expected) of the recombinant mnp1 was amplified in the DNA from transformants, which was absent from the parental strain.

Desarrollo de un bioadsorbente laminar con Phanerochaete chrysosporium hipertolerante al cadmio, al níquel y al plomo para el tratamiento de aguas / Development of a laminar bioadsorbent with hypertolerant Phanerochaete chrysosporium to Cd, Ni and Pb for wastewater treatment

Antecedentes. El uso de basidiomicetes para la remoción de metales es una alternativa con respecto a los métodos tradicionales, básicamente porque la biomasa actúa como un intercambiador natural al remover metales en solución. Objetivo. Desarrollar un bioadsorbente laminar empleando el hongo basidiomicete con mayor resistencia al cadmio (Cd), al níquel (Ni) y al plomo (Pb). Métodos. Se valoró la tolerancia de Trametes versicolor, Pleurotus ostreatus y Phanerochaete chrysosporium frente a concentraciones ascendentes de sulfato de sulfato de cadmio, acetato de plomo y cloruro de níquel. Se desarrolló un bioadsorbente a base de láminas de polietileno con biomasa fúngica, que se evaluó en columnas de burbujeo empleando agua residual sintética con los 3 metales con una concentración final de 300mg/l. Finalmente, con un experimento complementario en matraces Erlenmeyer, se valoró el efecto de mayor cantidad de biomasa sobre la eficiencia de remoción. Resultados. El hongo P. chrysosporium fue la cepa más tolerante a C4H6O4Pb (10.000mg/l), a Cl2Ni (300mg/l) y CdSO4·8H2O (1.500mg/l). En el reactor y bajo condiciones no ligninolíticas, el hongo removió el 69% de la demanda química de oxígeno, produjo enzimas como lignino peroxidasa (0,01U/l) y manganeso peroxidasa (0,6U/l) y se observó la acumulación de los metales en la pared. Al incrementar la biomasa a 1,6 (p/v) se favoreció la bioadsorción de los metales en la mezcla (el 57% para el Pb, el 74% para el Cd y el 98% para el Ni) y por separado (el 95% para el Pb, el 60% para el Cd y el 56% para el Ni). Se observó una competencia entre el Ni y el Pb por los ligandos de la pared. Conclusión. Se desarrolló un novedoso sistema laminar a base de biomasa viable de P. chrysosporium que posee gran área superficial y tolera elevadas concentraciones de Cd, Ni y Pb, como alternativa para la remoción de metales en aguas(AU)

Background. The use of basidiomycetes for metal removal is an alternative to traditional methods. In this, the biomass acts as a natural ionic exchanger removing metals from solution. Objective. To develop a laminar biosorbent using a basidiomycete fungus resistant to high Cd, Ni and Pb concentrations. Methods. The tolerance of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium was evaluated using increasing concentrations of the heavy metal salts, cadmium sulphate, lead acetate and nickel chloride. A biosorbent system was developed based on polyethylene sheets with a fungal biomass. It was evaluated in bubble columns using synthetic wastewater with the 3 metal salts at a rate of 300mg/l. Finally, in a complementary experiment using shake flasks, the effect of a higher amount of biomass related to the metal removal efficiency was evaluated. Results. P. chrysosporium strain was more tolerant to C4H6O4Pb (10,000mg/l), Cl2Ni (300mg/l) and CdSO4·8H2O (1,500mg/l). In a reactor, under non-ligninolytic conditions, the fungus removed 69% of the chemical oxygen demand and produced enzymes such as LiP (0.01U/l) and MnP (0.6U/l.). An accumulation of metals in the wall was observed. By increasing the biomass to 1.6 (w/v), the metal biosorption was favored in the mixture (57% Pb, 74% Cd, and 98% Ni) and separately (95% Pb, 60% Cd, and 56% Ni). Competition between Ni and Pb by ligands of the wall was observed. Conclusion. A novel laminar system based on P. chrysosporium viable biomass was developed. It has a large surface area and tolerance to high concentrations of Cd, Ni and Pb. It seems to be an alternative for the removal of metals from water(AU)