Highly reusable invertase nanocatalyzer produced by amyloids nanofibers functionalization

MACHÍN, B.; CHAVES S.; CHEHÍN R.; VERA PINGITORE E.

Santos, Sao Paulo

Congreso; XLII Congress of the Brazilian Biophysical Society (SBBf),; 2017

Brazilian Biophysical Society

The external invertase enzyme (β-fructofuranosid fructohydrolase, EC3.2.1.26) obtained from yeast Saccharomyces cerevisiae is of great importance in the food and beverage industry. This enzyme is used to catalyse the conversion of sucrose to an equimolar mixture of glucose and fructose. The aim of this study was to design a solid nanocatalyst throughout photo-induced immobilization of the enzyme invertase on the amyloid nanofibrils for technological applications. Lysozyme was used to produce the support applying the protocol of the patent developed in our working group. Spectroscopic fluorescence and transmission electron microscopy confirmed the success in formation of amyloids nanofibers. In the present work the functionalization of the amyloid nanofibers was developed using chemical properties of the side chains of the amino acids for their application as nano-supports. In order to optimize the cross-linking and immobilization of the invertase enzyme in the solid support, a molecular model will be constructed and the amino acids and their degree of exposure to the solvent will be analyzed, this will make it possible to understand the feasibility of immobilization process. The immobilization of Saccharomyces cerevisiae invertase on lysozyme nanofibers is produced by a photo-induced chemical crosslinking reaction, which was optimized for this system. The kinetic parameters and stability were determined for the free and immobilized invertase. In addition, were studied the effect of substrate concentration on invertase activity and a thermodynamic study was made for the free and immobilized inversion. The immobilized biocatalyst also showed high operational stability.