Functional and biological analyses of LPMO (Lytic Polysaccharide Monooxigenases) and accessory proteins (non-CAZymes) for degradation of sugarcane straw by filamentous fungi

• Grant number: 16/16306-0
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): November 01, 2016
• Status: Discontinued
• Field of knowledge: Biological Sciences - Microbiology
• Principal Investigator: André Ricardo de Lima Damasio
• Grantee: César Rafael Fanchini Terrasan
• Home Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 12/20549-4 - Secretion of heterologous glycoproteins in Aspergillus: effect of glycosylation pattern in functional parameters of glycosyl hydrolases, AP.BIOEN.JP
• Associated scholarship(s): 19/08263-7 - Spectroscopic and structural investigations on target fungal Lytic Polysaccharide Monooxygenases, BE.EP.PD

Abstract

Due to the heterogeneity and recalcitrance of plant cell wall, its complete and efficient degradation is a challenge to be overcome aiming the economic viability of second-generation ethanol. The development of new and improved enzyme cocktails requires greater knowledge of the active proteins in order to be optimized for the specific degradation of certain biomass. One of the most significant advancement was the recent recognition of Lytic Polysaccharide Monooxygenases (LPMO), which employ chemical oxidation to disrupt the crystalline structure of polysaccharides, notably improving the degradation by classical hydrolytic enzymes. These enzymes are therefore promising for improvement in processes of plant biomass hydrolysis. Such enzymes are currently part of the class AA (Auxiliary Activity) in the CAZy database (Carbohydrate-Active Enzymes), while another group of non-CAZymes proteins also alters the structure of polymers, being denominated accessory or amorphogenesis inducing proteins. The objective of this project is to analyze the secretome of filamentous fungi such as Aspergillus nidulans, Laetiporus sulphureus, Pycnoporus coccineus e Trametes versicolor grown on pure carbohydrates and sugar cane residues (bagasse and straw), focusing both on LPMOs and accessory proteins that can present important role in biomass degradation. Subsequently, some target proteins will be cloned and expressed in Aspergillus nidulans and then purified, characterized, and evaluated in the hydrolysis of sugarcane straw. In addition, the biological implications of these proteins will be assessed through deletion in A. nidulans, and mutant strains will be evaluated for their growth. The results of this project will allow better understanding of both the physiological function of these proteins and their role in the degradation of lignocellulosic biomass. (AU)