His-tagged lactate oxidase production for industrial applications using fed-batch fermentation.

L-lactate oxidase (LOX) is a biotechnologically important enzyme used in biosensors and colorimetric kits to detect lactate, a key biomarker in clinical diagnostics, sports medicine and the food industry. In this work, we produced a recombinant His-tagged Aerococcus viridans LOX (rLOX) in Escherichia coli and carried out its functional characterization for industrial applications. Our rLOX was evaluated in a colorimetric kit for human diagnostics and in an amperometric biosensor to measure the lactic acid in food products. The rLOX was fully functional for both applications, with a performance comparable to commercial untagged LOXs. As the industrial use of LOX enzyme requires a large-scale production, we scaled up the rLOX production in a fed-batch bioreactor culture and obtained a yield approximately ten times higher than that of the Erlenmeyer scale. The His-tag allowed an easy and highly efficient purification process, and a high-purity rLOX was recovered after this one-step affinity purification. In this study, we described a simple, rapid and cost-competitive approach for the production of a recombinant His-tagged LOX enzyme suitable for industrial use.

Design and performance evaluation of a photocatalytic reactor for indoor air disinfection.

Since COVID-19 pandemic, indoor air quality control has become a priority, and the development of air purification devices effective for disinfecting airborne viruses and bacteria is of outmost relevance. In this work, a photocatalytic device for the removal of airborne microorganisms is presented. It is an annular reactor filled with TiO2-coated glass rings and irradiated internally and externally by UV-A lamps. B. subtilis spores and vegetative cells have been employed as model biological pollutants. Three types of assays with aerosolized bacterial suspensions were performed to evaluate distinct purification processes: filtration, photocatalytic inactivation in the air phase, and photocatalytic inactivation over the TiO2-coated rings. The radiation distribution inside the reactor was analysed by performing Monte Carlo simulations of photon absorption in the photocatalytic bed. Complete removal of a high load of microorganisms in the air stream could be achieved in 1 h. Nevertheless, inactivation of retained bacteria in the reactor bed required longer irradiation periods: after 8 h under internal and external irradiation, the initial concentration of retained spores and vegetative cells was reduced by 68% and 99%, respectively. Efficiency parameters were also calculated to evaluate the influence of the irradiation conditions on the photocatalytic inactivation of bacteria attached at the coated rings.

Photocatalytic inactivation of bioaerosols in a fixed-bed reactor with TiO2-coated glass rings.

The photocatalytic inactivation of Bacillus subtilis spores in air was evaluated employing a fixed-bed reactor with TiO2-coated glass rings, under artificial UV-A radiation. Calculations of the radiation effectively absorbed inside the reactor were carried out by Monte Carlo simulations. The photocatalytic inactivation was assessed by analyzing the viability of the microorganisms retained by the coated glass rings inside the reactor at different irradiation periods. The initial concentration of the spores was reduced by almost 55% at the end of the experiment (12 h). Complementary assays were carried out employing Bacillus subtilis vegetative cells, obtaining a reduction of more than 96% under the same conditions. Two efficiency parameters were computed to assess the reactor performance: the photonic efficiency and the quantum efficiency of inactivation. Results of the efficiency parameters allow an objective comparison of the reactor performance under different experimental conditions and configurations.

Photocatalytic degradation of an emerging pollutant by TiO2-coated glass rings: a kinetic study.

This work presents the photocatalytic degradation of the pharmaceutical drug clofibric acid in a fixed-bed reactor filled with TiO2-coated glass rings. Experiments were carried out under UV radiation. A kinetic model that takes into account radiation absorption by means of the local surface rate of photon absorption (LSRPA) has been developed. The LSRPA was obtained from the results of a radiation model. The Monte Carlo method was employed to solve the radiation model, where the interaction between photons and TiO2-coated rings was considered. Data from experiments carried out with rings with different numbers of catalyst coatings and different irradiation levels were used to estimate the parameters of the kinetic model. A satisfactory agreement was obtained between model simulations and experimental results.

Kinetic modeling of the photocatalytic degradation of clofibric acid in a slurry reactor.

A kinetic study of the photocatalytic degradation of the pharmaceutical clofibric acid is presented. Experiments were carried out under UV radiation employing titanium dioxide in water suspension. The main reaction intermediates were identified and quantified. Intrinsic expressions to represent the kinetics of clofibric acid and the main intermediates were derived. The modeling of the radiation field in the reactor was carried out by Monte Carlo simulation. Experimental runs were performed by varying the catalyst concentration and the incident radiation. Kinetic parameters were estimated from the experiments by applying a non-linear regression procedure. Good agreement was obtained between model predictions and experimental data, with an error of 5.9 % in the estimations of the primary pollutant concentration.