Imobilização de neutrase® em suportes a base de quitosana e alginato ativados com glutaraldeído.

Abstract

Neutrase® is a protease that hydrolyses the whey proteins, a highly pollutant byproduct, in intend to obtain derivatives with improved functional properties. The aim of this study was to immobilize the Neutrase® enzyme on the Chitosan and Sodium Alginate basis and characterize them in terms of yield of immobilization, recovered activity, stability factor, and kinetic tests. To this end, the basis were prepared Chitosan 2.5% - Alginate 2.5% and Chitosan 2.5% - Alginate 2.5% - Saccharomyces cerevisiae 5%, activated with glutaraldehyde for 1 hour at 25 °C and fixed at 30 rpm for 3 hours at 25°C, and offered a load of 10 U/g of basis. The activity of each basis was measured using a spectrophotometer at a wavelength of 700 nm, and through it, we found the activity recovered and yield of immobilization. Thermal stability tests at a temperature of 60 °C were conducted with the basis Chitosan 2.5% - Alginate 2.5% and Chitosan 2.5% - Alginate 2.5% - Saccharomyces cerevisiae 5% for 2.5 hours and 6 hours, respectively. Finally, kinetic tests with different concentrations of substrate were performed on both basis. The Chitosan 2.5% - Alginate 2.5% - Saccharomyces cerevisiae 5% showed the best results with immobilization yield of 47.57%; related to the thermal stability, it was found to be 87.86 times more stable than the soluble enzyme and about the kinetic test, it was the basis that best fit to the Michaelis-Menten model. With these results, it was concluded that the basis Chitosan 2.5% - Alginate 2.5% - Saccharomyces cerevisiae 5% is characterized as a better basis for the Neutrase® enzyme immobilization when compared to Chitosan 2.5% - Alginate 2 5%.