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Optimization of culture conditions in a bench bioreactor to maximize the expression of recombinant alpha-L-arabinofuranosidase

Grant number: 20/04276-4
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): August 01, 2020
Effective date (End): July 31, 2021
Field of knowledge:Engineering - Chemical Engineering
Principal Investigator:Eutimio Gustavo Fernández Núñez
Grantee:Giovanna Vernillo Guth
Home Institution: Escola de Artes, Ciências e Humanidades (EACH). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Considering the scientific community's warnings about global warming, the advantages of using renewable energy sources and the need to guarantee food and energy security, second-generation ethanol presents itself as an alternative to be considered in face of the strong dependence on fossil fuel of the current society. This new category of biofuels produced from lignocellulosic biomass has been the subject of comprehensive studies, mainly focused on the stage that comprises the enzymatic hydrolysis that is essential for the release of fermentable sugars by microorganisms. One of the main constituents of biomass is hemicellulose, which has pentoses linked to its chain, known as L-Arabinoses. The ±-L-arabinofuranosidases are responsible for releasing these monosaccharides and consequently enabling the complete degradation of hemicellulose. Given the significance of this enzyme, the current proposal aims to optimize the operating conditions in a bench-scale bioreactor to maximize the expression of the ±-L-arabinofuranosidase from a recombinant E. coli. To achieve the proposed goal, a bioreactor will be used for the bench-scale production of the enzyme ±-L-arabinofuranosidase. The following parameters will be assessed in this study: dissolved oxygen, nitrogen and glucose concentrations, according to a Box-Behnken experimental design. At the end of the experimental work, it will be possible to create equations through mathematical and statistical modeling performed in Microsoft Office EXCEL 365 and MatLab software, which link the measured parameters with kinetic, yield and productivity factors, thus revealing the best combination that was obtained from them. Therefore, this project opens the possibility of scaling up the optimum conditions obtained for the industrial scale, as well as an eventual contribution in the preparation of enzymatic cocktails.