Obtaining xylanase from Escherichia coli A GH11 and evaluating its immobilization on magnetized grapheno oxide support

• Grant number: 25/17207-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2025
• End date: November 30, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Fernando Masarin
• Grantee: Luiza Jolli Nunes
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated research grant: 21/07058-0 - Immobilization of endoglucanases and xylanases on magnetic supports: evaluation of the hydrolysis potential of agroindustrial by-products in oligosaccharides, AP.R

Abstract

Enzymes are of great interest in various sectors due to their wide commercial and industrial applicability, offering alternatives that are compatible with sustainable practices. Among them, xylanases stand out for their high biotechnological potential and can be applied in different segments to obtain a variety of products or carry out specific processes. However, despite their advantages, the use of enzymes in industrial processes still faces limitations related to the high cost of production and instability under certain operating conditions. To make bioprocess on a large scale economically sustainable and viable, the strategy of enzyme immobilization arises through solid support, especially magnetized ones, to mitigate this problem. This project aims to evaluate the production and purification of the enzyme xylanase from the heterologous expression of Escherichia coli GH11, as well as its immobilization on magnetized graphene oxide (GMO) support. Experiments on cultivating E. coli will be conducted. The Luria-Bertani (LB) medium will be used to obtain inoculation, while the Super Broth (SB) medium will be used to produce xylanase, with the addition of Ampicillin as a selective press. The produced xylanase will be pre-purified/concentrated using precipitation techniques such as ammonium sulfate and ultrafiltration, being subsequently characterized. The pre-purified/concentrated xylanase will be immobilized in the magnetized graphene oxide (GMO) support which will be effective starting from the oxidation of graphite and magnetization with iron. The activities of xylanase are determined, as well as the maximum activity parameters of pH and temperature, and protein level, as well as forms of the xylanase enzyme, free and immobilization. It is hoped that the observable results of this project will contribute to the development of materials and extracts that are more sustainable and economically viable, attending to the demands of the sectors that use xylanase in these processes and promoting positive impacts in the production of bioproducts of economic, social and environmental interest. (AU)