Bifunctional chimeric protein of endoxylanase/arabinofuranosidase for agro-industrial wastes hydrolysis and xylooligosaccharides production

Abstract

Xylo-oligosaccharides have prebiotic activity, favoring intestinal function, immunological action and other health benefits. The production of these oligosaccharides can be carried out by enzymatic hydrolysis of lignocellulosic material, a selective, controlled and sustainable method. Genetic engineering tools allow the fusion of enzymes produced by different microorganisms, which have the potential to produce XOS, making it possible to obtain a stable bifunctional chimera capable of efficiently hydrolyzing the hemicellulosic fraction of agro-industrial waste, while reducing the production costs of enzymes. The present work aims the cloning and constitutive expression of arabinofuranosidase gene of A. fumigatus in Pichia pastoris using the pGAP vector, and later fusion with C. flavescens endoxylanase to obtain bifunctional chimeric protein. Hydrolysis profiles and conditions, synergy, specificity and structure of the obtained chimera will be investigated. The hydrolysis products (xylo-oligosaccharides) of lignocellulosic material from agro-industrial residues of corn, orange and soy husks, using the chimeric protein obtained will be quantified, purified and evaluated for antioxidant, prebiotic and antiproliferative activity. The presented project is associated to the FAPESP 2019/08542-3 Process (Enzymatic biotechnology for the production of xylooligosaccharides from lignocellulosic agroindustrial residues aiming at the production of symbiotics), in which we are inserted. (AU)