M. Cea

Degradation of polycyclic aromatic hydrocarbons by free and nanoclay-immobilized manganese peroxidase from Anthracophyllum discolor

Manganese peroxidase (MnP) produced by Anthracophyllum discolor, a Chilean white rot fungus, was immobilized on nanoclay obtained from volcanic soil and its ability to degrade polycyclic aromatic hydrocarbons (PAHs) compared with the free enzyme was evaluated. At the same time, nanoclay characterization was performed. Nanoclay characterization by transmission electronic microscopy showed a particle average size smaller than 100 nm. The isoelectric points (IEP) of nanoclay and MnP from A. discolor were 7.0 and 3.7, respectively, as determined by micro electrophoresis migration and preparative isoelectric focusing. Results indicated that 75% of the enzyme was immobilized on the nanoclay through physical adsorption. As compared to the free enzyme, immobilized MnP from A. discolor achieved an improved stability to temperature and pH. The activation energy (Ea) value for immobilized MnP (51.9 kJ mol(-1)) was higher than that of the free MnP (34.4 kJ mol(-1)). The immobilized enzyme was able to degrade pyrene (>86%), anthracene (>65%), alone or in mixture, and to a less extent fluoranthene (<15.2%) and phenanthrene (<8.6%). Compared to free MnP from A. discolor, the enzyme immobilized on nanoclay enhanced the enzymatic transformation of anthracene in soil. Overall results indicate that nanoclay, a carrier of natural origin, is a suitable support material for MnP immobilization. In addition, immobilized MnP shows an increased stability to high temperature, pH and time storage, as well as an enhanced PAHs degradation efficiency in soil. All these characteristics may suggest the possible use of nanoclay-immobilized MnP from A. discolor as a valuable option for in situ bioremediation purposes.

Bioremediation of soil contaminated with pentachlorophenol by Anthracophyllum discolor and its effect on soil microbial community.

Bioaugmentation is a promising technology to clean up sites contaminated with recalcitrant chemicals. White-rot fungi have proven to be effective in the degradation of pentachlorophenol. Here, we report the bioremediation of soil contaminated with pentachlorophenol (PCP) by Anthracophyllum discolor and its impact on the soil microbial community. In this study three types of microcosms were established: fresh soil (C(0)), fresh soil plus wheat straw (WS(0)) and, fresh soil plus wheat straw inoculated with A. discolor (WSAD(0)). Additionally, similar treatments and a control of sterile soil spiked with PCP (C(250), WS(250) and WSAD(250)) were used to evaluate the remediation and adsorption of PCP. The PCP removal, total microbial activity, and enzymatic activities were evaluated. This study also investigated the structure of soil microbial community by denaturing gradient gel electrophoresis (DGGE), identifying some of the dominant bacterial and fungal species. The results showed that PCP was effectively degraded in soils by A. discolor and by indigenous soil microorganisms. The addition of wheat straw increased the PCP degradation and enzymatic activities. Only laccase activity was negatively affected by PCP contamination. The PCP degradation was associated with changes in microbial communities, mainly stimulation of members of bacterial phylum Proteobacteria (Xanthomonadaceae, Burkholderiaceae and Enterobacteriaceae), and fungal phylum Ascomycota and Basidiomycota. This study shows the ability of A. discolor to degrade PCP from contaminated soil, and demonstrates that agricultural residues, such as wheat straw, can be used as growth substrate by microorganisms in PCP-contaminated soil, demonstrating a great potential of autochthonous microorganisms for soil remediation.

Chlorpyrifos degradation in a biomixture of biobed at different maturity stages

The biomixture is a principal element controlling the degradation efficacy of the biobed. The maturity of the biomixture used in the biobed affects its overall performance of the biobed, but this is not well studied yet. The aim of this research was to evaluate the effect of using a typical composition of Swedish biomixture at different maturity stages on the degradation of chlorpyrifos. Tests were made using biomixture at three maturity stages: 0 d (BC0), 15 d (BC15) and 30 d (BC30); chlorpyrifos was added to the biobeds at final concentration of 200, 320 and 480 mg kg(-1). Chlorpyrifos degradation in the biomixture was monitored over time. Formation of TCP (3,5,6-trichloro-2-pyrinidol) was also quantified, and hydrolytic and phenoloxidase activities measured. The biomixture efficiently degraded chlorpyrifos (degradation efficiency >50%) in all the evaluated maturity stages. However, chlorpyrifos degradation decreased with increasing concentrations of the pesticide. TCP formation occurred in all biomixtures, but a major accumulation was observed in BC30. Significant differences were found in both phenoloxidase and hydrolytic activities in the three maturity stages of biomixture evaluated. Also, these two biological activities were affected by the increase in pesticide concentration. In conclusion, our results demonstrated that chlorpyrifos can be degraded efficiently in all the evaluated maturity stages.

Kinetic and thermodynamic study of chlorophenol sorption in an allophanic soil.

The sorption of 2,4-dichlorophenol, 2,4,6-trichlorophenol, and pentachlorophenol by a allophanic soil was studied in a series of batch experiments. Chlorophenol sorption behavior was evaluated as a function of reaction time (0-96h) and input concentration at a fixed ionic strength (0.1mol L(-1) KCl) at 25, 35, and 45 degrees C. Sorption results for the various reaction temperatures were used in calculating thermodynamic parameters. Chlorophenol sorption increased with temperature, suggesting an endothermic process. The Elovich equation was used to describe the kinetic data. Data from the isotherm experiments were described by the Triple-Layer Model in which monodentate outer- and inner-sphere complexes were formed between deprotonated organic molecules and active sites on the variable-charge soil. The calculated thermodynamic parameters suggest that chlorophenol sorption is a spontaneous (DeltaG<0), endothermic (DeltaH>0) and entropy-driven reaction (DeltaS>0).

BIOSTIMULATION OF AGRICULTURAL BIOBEDS WITH NPK FERTILIZER ON CHLORPYRIFOS DEGRADATION TO AVOID SOIL AND WATER CONTAMINATION

Degradation of the insecticide chlorpyrifos (160 a.i mg kg -1 ) using a biomix of a biobed system biostimulated with inorganic fertilizer (NPK) was investigated. Three concentrations of the fertilizer (0.1%, 0.5% and 1.0% ww -1 ) were evaluated on chlorpyrifos degradation, TCP (3, 5, 6-trichloro-2-pyrinidol) accumulation and biological activity of the biomix. The chlorpyrifos was dissipated efficiently (>75%) after 40 days of incubation and no additional dissipation was obtained with increasing concentration of NPK after 20 days of incubation. TCP accumulation occurred in all evaluated NPK concentrations and its concentration increased with the increment of NPK addition raising the probability of leaching of this compound. Biological activity (FDA and ligninolytic enzyme activity) in the biomix increased by the NPK presence in all evaluated concentrations. The DGGE analyses showed that combined treatments with lower amounts of NPK (0% and 0.1%) and chlorpyrifos showed no significant modifications in the microbial community in the biomix. However, combined overdoses of NPK (0.5 and 1.0%) and chlorpyrifos caused significant modifications in the bacterial communities that could be associated with TCP degradation reduction in the biomix. In conclusion, the obtained results demonstrated that the biomix prepared with Andisol and biostimulated with NPK nutrient can be recommended in biobeds as a viable alternative of chlorpyrifos dissipation avoiding soil and water contamination probability.

Evaluation of biodegradable polymers as encapsulating agents for the development of a urea controlled-release fertilizer using biochar as support material.

Biochar constitutes a promising support material for the formulation of controlled-release fertilizers (CRFs). In this study we evaluated the effect of different polymeric materials as encapsulating agents to control nitrogen (N) leaching from biochar based CRFs. Nitrogen impregnation onto biochar was performed in a batch reactor using urea as N source. The resulting product was encapsulated by using sodium alginate (SA), cellulose acetate (CA) and ethyl cellulose (EC). Leaching potential was studied in planted and unplanted soil columns, monitoring nitrate, nitrite, ammonium and urea concentrations. After 90 days, plants were removed from the soil columns and plant yield was evaluated. It was observed that the ammonium concentration in leachates presented a maximum concentration for all treatments at day 22. The highest concentration of N in the leachates was the nitrate form. The crop yield was negatively affected by all developed CRFs using biochar compared with the traditional fertilization.

In situ biodiesel production from greasy sewage sludge using acid and enzymatic catalysts.

This study proposes to select the most appropriate sewage sludge (greasy, primary and secondary) for in situ transesterification and to compare the technical, economic and energetic performance of an enzymatic catalyst (Novozym®435) with sulfuric acid. Greasy sludge was selected as feedstock for biodiesel production due to its high lipid content (44.4%) and low unsaponifiable matter. Maximum methyl esters yield (61%) was reached when processing the wet sludge using sulfuric acid as catalyst and n-hexane, followed by dried-greasy sludge catalyzed by Novozym®435 (57% methyl esters). Considering the economic point of view, the process using acid catalyst was more favorable compared to Novozym®435 catalyst due to the high cost of lipase. In general, greasy sludge (wet or dried) showed high potential to produce biodiesel. However, further technical adjustments are needed to make biodiesel production by in situ transesterification using acid and enzymatic catalyst feasible.

Performance of an enzymatic extract in Botrycoccus braunii cell wall disruption.

Microalgae can produce and contain lipids, proteins and carbohydrates, which can be extracted and marketed as potential novel added-value bio-products. However, microalgae cell wall disruption is one of the most important challenges involved while processing this type of biomass. In this context, white-rot fungi, responsible for the biodegradation of lignin present in wood due to non-specific extracellular enzymes, could be applied for promoting microalgae cell wall degradation. Therefore, the aim of this study was to evaluate the use of an enzymatic extract produced by the white-rot fungi Anthracophyllum discolor as a biotechnological tool for Botryococcus braunii cell wall disruption. The fungus was inoculated in wheat grains and manganese peroxidase (MnP) activity was monitored while obtaining the enzymatic extract. Then, cell wall disruption trials with different MnP activity were evaluated by the biochemical methane potential (BMP). In relation to cell wall disruption, it was observed that the optimal value was obtained with enzymatic concentration of 1000 U/L with a BMP of 521 mL CH4/g VS. Under these conditions almost 90% of biomass biodegradability was observed, increasing in 62% compared to the microalgae without treatment. Therefore, the results indicate that enzymes secreted by A. discolor promoted the attack of the different cell wall components finally weakening it. Therefore, the application of this treatment could be a promissory biotechnological approach to decrease the energetic input required for the cell wall disruption step.

Influence of Aluminum on the Growth and Organic Acid Exudation in Alfalfa Cultivars Grown in Nutrient Solution

ABSTRACT A study was conducted to evaluate the effects of aluminum (Al) in nutrient solutions on the dry weight (DW) yield, Al and phosphorus (P) contents, and organic acid exudation in alfalfa (Medicago sativa L.). Four alfalfa cultivars (‘Robust’, ‘Sceptre’, ‘Aquarius’, and ‘California-55’) were grown in nutrient solution at pH 4.5 and 6.0, with (50 and 100 μM) and without Al. The results revealed that Al caused a significant reduction in DW, especially in pH 4.5 treatment. Organic acid exudation was affected by pH and Al treatments. Citrate and succinate exudation increased with the high Al treatment at pH 4.5. However, no relationship between pH and carboxylate exudation was observed at pH 6.0. Accumulation of P and Al in roots suggests the existence of an exclusion mechanism for Al in alfalfa. Selection of cultivars with enhanced organic exudation capacity in response to Al might be useful for alfalfa production in moderately acidic soils.

Biosorption of pentachlorophenol by Anthracophyllum discolor in the form of live fungal pellets

Pentachlorophenol (PCP) is an extremely dangerous pollutant for every ecosystem. In this study we have detected how PCP concentration and pH levels can influence PCP adsorption by Anthracophyllum discolor in the form of live fungal pellets. PCP adsorption was evaluated after 24 hours in KCl 0.1 M electrolyte solution with initial PCP concentrations of 5 and 10 mg L (-1) and with pH values between 4 and 9 (at intervals of 0.5). Fourier Transform Infrared Spectroscopy (FTIR) was used to identify functional groups of fungal biomass that can interact with PCP. The amount of PCP that was adsorbed by A. discolor was >80% at pH values between 5 and 5.5, whatever the concentration tested. PCP adsorption significantly decreased in liquid medium of pH > 6.0. FTIR results showed that amides, alkanes, carboxylates, carboxyl and hydroxyl groups may be important to the PCP adsorption for pellets of A. discolor. Live fungal pellets of A. discolor may be used as a natural biosorbent for liquid solutions contaminated by PCP.