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New recombinant PMOS from Aspergillus fumigatus: biochemical characterization and its effect on saccharification of sugarcane bagasse with different pre-treatments

Grant number: 19/01165-0
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2019
Effective date (End): June 30, 2022
Field of knowledge:Biological Sciences - Biochemistry
Principal Investigator:Fernando Segato
Grantee:Josman Andrey Velasco Mendoza
Home Institution: Escola de Engenharia de Lorena (EEL). Universidade de São Paulo (USP). Lorena , SP, Brazil

Abstract

The recent discovery of the enzymes called LPMOs or PMOs (Lytic Polysaccharide Monooxygenase or Polysaccharide Monooxygenase) opened new perspectives on the enzymatic mechanisms involved in the deconstruction of lignocellulosic biomass. Similarly, these prospects have significantly impacted the increase in the number of patents filed and related scientific publications. This impact has been motivated by the promising results that these oxidative enzymes present to improve the conventional processes of enzymatic hydrolysis, thus contributing to a greater viability of the biorefinery projects. The present project aims to the kinetic and functional study of three recombinant PMOs of the thermophilic fungus Aspergillus fumigatus var niveus through thermostability, oxidative behavior at different substrates, regioselectivity, preference for different reducing agents, kinetic studies with O2 and H2O2 as co-substrates and other biochemical assays that are necessary for the deep understanding of the catalytic mechanism of these enzymes. The monooxygenases that best contribute to the oxidation / degradation process of cellulose will be evaluated for their potential to increase the efficiency in saccharification of sugarcane bagasse, being added in the commercial enzyme cocktail Celluclast 1.5L (Novozymes). In the saccharification process will be also evaluate the effect caused by the type of pre-treatment used, since it will be tested bagasse subjected to different pretreatment methodologies such as dilute acid, alkaline sulfite and LHW (Liquid Hot Water). The results obtained will increase the knowledge of these enzymes in order to take advantage of their potential in the improvement of enzymatic cocktails that are more efficient in the degradation of the biomass, thus favoring the techno-economic viability of the projects focused on bioeconomics. (AU)