Search

In this work, five different magnetic biofilters, containing magnetic nanoparticles (142 nm), immobilized laccase on nanoparticles (190 nm) and permanent magnetic elements, such as neodymium magnets and metallic meshes, were designed, manufactured and tested. The five types of filters were compared by measuring the decolorization of Congo Red dye inside bioreactors, the half-life of the filters and the amount of magnetic nanoparticle and enzyme lost during multiple cycles of operation. Filters containing laccase immobilized on magnetite (Laccase-magnetite), permanent magnets and metallic mesh presented the highest Congo Red decolorization (27%) and the largest half-life among all types of filters (seven cycles). The overall dye decolorization efficiencies were 5%, 13%, 17%, 23%, and 27% for the paper filter, paper filter with magnetite, paper filter with Laccase-magnetite, paper filter with Laccase-magnetite with magnets and paper filter with Laccase-magnetite with magnets and metallic mesh, respectively. Although the highest losses of magnetite occurred when using the filters containing magnets (57 mg), the use of permanent magnetic elements in the filters increased the half-life of the filter three-fold compared to the filters without enzymatic properties and two-fold compared to the filters with Laccase-magnetite. Results indicate that the novel use of permanent magnetic elements improved the nanoparticle retention in the filters and promoted the mass transfer between the dye and the biocatalyst to enhance wastewater treatment.

The polymeric ouzo effect is an energy-efficient and robust method to create nanoparticles with biologically degradable polymers. Usually, a discontinuous or semi-continuous process is employed due to its low technical effort and the fact that the amount of dispersions needed in a laboratory is relatively small. However, the number of particles produced in this method is not enough to make this process economically feasible. Therefore, it is necessary to improve the productivity of the process and create a controllable and robust continuous process with the potential to control parameters, such as the particle size or surface properties. In this study, nanoparticles were formulated from polycaprolactone (PCL) in a continuous process using additively manufactured micromixers. The main goal was to be able to exert control on the particle parameters in terms of size and zeta potential. The results showed that particle size could be adjusted in the range of 130 to 465 nm by using different flow rates of the organic and aqueous phase and varying concentrations of PCL dissolved in the organic phase. Particle surface charge was successfully shifted from a slightly negative potential of -14.1 mV to a negative, positive, or neutral value applying the appropriate surfactant. In summary, a continuous process of nanoprecipitation not only improves the cost of the method, but furthermore increases the control over the particle's parameters.

The presence of micropollutants in wastewater is one of the most significant environmental challenges. Particularly, pollutants such as pharmaceutical residues present high stability and resistance to conventional physicochemical and biological degradation processes. Thus, we aimed at immobilizing a laccase enzyme by two different methods: the first one was based on producing alginate-laccase microcapsules through a droplet-based microfluidic system; the second one was based on covalent binding of the laccase molecules on aluminum oxide (Al 2 O 3 ) pellets. Immobilization efficiencies approached 92.18% and 98.22%, respectively. Laccase immobilized by the two different methods were packed into continuous flow microreactors to evaluate the degradation efficiency of acetaminophen present in artificial wastewater. After cyclic operation, enzyme losses were found to be up to 75 µg/mL and 66 µg/mL per operation cycle, with a maximum acetaminophen removal of 72% and 15% and a retention time of 30 min, for the laccase-alginate microcapsules and laccase-Al 2 O 3 pellets, respectively. The superior catalytic performance of laccase-alginate microcapsules was attributed to their higher porosity, which enhances retention and, consequently, increased the chances for more substrate-enzyme interactions. Finally, phytotoxicity of the treated water was lower than that of the untreated wastewater, especially when using laccase immobilized in alginate microcapsules. Future work will be dedicated to elucidating the routes for scaling-up and optimizing the process to assure profitability.

The development of microfluidics-based systems in the recent years has provided a rapid and controlled method for the generation of monodisperse microencapsulates for multiple applications. Here, we explore the design, manufacture and characterization of a low-cost microsystem for the encapsulation of the fungal laccase from Pycnoporus sanguineus CS43 in alginate microcapsules. Multiphysics simulations were used to overview the fluid behavior within the device and estimate the resulting capsule size. Polymethylmethacrylate (PMMA) sheets were used for final microsystem manufacture. Different flow rates of the continuous (Q c ) and discrete (Q d ) phases in the ranges of 83-293 mL/h and 1-5 mL/h, respectively, were evaluated for microcapsule fabrication. Universal Serial Bus (USB) microscope and image analysis was used to measure the final particle size. Laccase encapsulation was evaluated using spectrophotometry and with the aid of fluorescent dyes and confocal microscopy. Results showed microcapsule size was in the range of 203.13-716.00 μm and Q c was found as the dominant parameter to control capsule size. There was an effective enzyme encapsulation of 65.94% with respect to the initial laccase solution.