Improvement in the Thermostability of a Recombinant ß-Glucosidase Immobilized in Zeolite under Different Conditions.

ß-Glucosidase is part of the cellulases and is responsible for degrading cellobiose into glucose, a compound that can be used to produce biofuels. However, the use of the free enzyme makes the process more expensive. Enzyme immobilization improves catalytic characteristics and supports, such as zeolites, which have physical-chemical characteristics and ion exchange capacity that have a promising application in the biotechnological industry. This research aimed to immobilize by adsorption a recombinant ß-glucosidase from Trichoderma reesei, obtained in Escherichia coli BL21 (DE3), in a commercial zeolite. A Box Behnken statistical design was applied to find the optimal immobilization parameters, the stability against pH and temperature was determined, and the immobilized enzyme was characterized by SEM. The highest enzymatic activity was determined with 100 mg of zeolite at 35 °C and 175 min. Compared to the free enzyme, the immobilized recombinant ß-glucosidase presented greater activity from pH 2 to 4 and greater thermostability. The kinetic parameters were calculated, and a lower KM value was obtained for the immobilized enzyme compared to the free enzyme. The obtained immobilization parameters by a simple adsorption method and the significant operational stability indicate promising applications in different fields.

Study of coliforms and Clostridium bacteria inactivation in wastewaters by a pilot photolysis process and by the maturation lagoons of a low-cost nature-based WWTP.

The inactivation processes of coliform bacteria (total and fecal) and sulphito-reducing Clostridium bacteria (vegetative species and spores) in water maturation lagoon of a low-cost nature-based wastewater treatment plant using constructed wetlands and through processes of photolysis in a pilot photoreactor have been comparatively studied. The different inactivation mechanisms by photolysis of these bacteria have been studied following the criteria of different statistical and kinetic models. Clostridium disinfection treatments fit models in which two types of bacteria populations coexist, one sensitive (vegetative species) and the other (spores) resistant to the treatment, the sensitive one (94%) with an inactivation rate of k = 0.24 ± 0.07 min-1 and the resistant one (6%) with k = 0.11 ± 0.05 min-1. Total coliform photolytic disinfection also shows two populations with different physiological state. The time required to reduce the first logarithmic decimal cycle of the different types of bacteria (physiological states) are δ1 = 4.2 ± 0.9 and δ2 = 8.3 ± 1.1 min, respectively. For fecal coliform photolytic disinfection, only bacteria population, with k = 1.15 ± 0.19 min-1, is found. The results obtained confirm the photolytic disinfection processes and maturation lagoon are effective systems for Clostridia bacteria removal after water treatment by nature-based systems. Total removal of coliform bacteria is not achieved by maturation lagoons, but their reduction is significant using low doses of cumulative radiation.