Expression and purification of recombinants xylanolitics enzymes and study of their activities against wheat and sugar cane arabinoxylans

• Grant number: 18/04504-7
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2018
• End date: April 30, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Richard John Ward
• Grantee: Carolina Victal Garbelotti
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The research for methods that use lignocellulosic biomass for production of second generation ethanol has leaded to the study of enzymes able to hydrolyse this substrate and provide fermentable sugars. However, the heterogenic and recalcitrant composition of lignocellulosic biomass require a group of enzymes acting synergistically for its complete hydrolysis. Hence, this works aims the expression, purification and activity study of the enzymes Feruloil Esterase (Fae), Acetil Xylan Esterase (Axe), Arabinofuranosidase (Ara), Xylanase (Xyl) and ±-Glucuronidase (AGlu), which have as substrate different glycosidic bonds on the polysaccharide arabinoxylan, part of that complex feedstock. pET-28 plasmids containing the enzymes insert will be transformed in Escherichia coli strains. The enzymes will be purified by affinity chromatography and used in the activity essays, alone and combined, against wheat and sugar cane arabinoxylan, aiming the possibility of elaborating a mix of enzymes that could work synergistically and efficiently hydrolyse lignocellulosic biomass.