Enzymatic oxidation of sugarcane bagasse 2: exploring the interaction between LPMOs and its redox partner CDHs aiming for the development of more efficient enzymes for insertion into an engineered cell factory

• Grant number: 21/06679-1
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Young Investigators Grants - Phase 2
• Start date: April 01, 2022
• End date: March 31, 2027
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Fernando Segato
• Grantee: Fernando Segato
• Host Institution: Escola de Engenharia de Lorena (EEL). Universidade de São Paulo (USP). Lorena , SP, Brazil
• Associated researchers: Adriane Maria Ferreira Milagres ; Gabriela Leila Berto ; Igor Polikarpov ; Leandro Cristante de Oliveira ; Michael A. Marletta ; Sam Hay
• Associated research grant: 14/18714-2 - Enzymatic oxidation of sugarcane bagasse: discovery, characterization and new application of oxidative enzymes active in carbohydrates, applied to the enhancement of a fungal cell factory, AP.BIOEN.JP
• Associated research grant(s): 24/11129-9 - Exploring the potential of Auxiliary Activity enzymes: hemicellulose oxidation and its biotechnological application as plant biostimulant. Acronym: AAs4Plants, AP.R; 22/00539-6 - Computational analysis and synthetic biology approaches to understand the interaction and electron transfer between CDH and LPMO, AP.R SPRINT; 22/04227-9 - Multiuser equipment granted in the proposal 2021/06679-1: Dionex ICS600 Capillary HPIC system, AP.EMU
• Associated scholarship(s): 24/22624-0 - Enzymatic oxidation of sugarcane bagasse II: exploring the interaction between LPMOs and CDHs aiming for the development of more efficient enzymes for insertion into an engineered cell factory, BP.PD; 25/00367-9 - Functional studies of LPMOs from the AA9 and AA16 family of Thermothielavioides terrestris, BP.IC; 24/07449-8 - Enzymatic oxidation of sugarcane bagasse 2: exploring the interaction between LPMOs and its redox partner CDHs aiming for the development of more efficient enzymes for insertion into an engineered cell factory, BP.PD; + associated scholarships 24/10678-9 - Enzymatic oxidation of sugarcane bagasse: exploring molecular biology tools for the development of more efficient engineered cell factory, BP.MS; 23/08824-4 - Enzymatic oxidation of sugarcane bagasse 2: exploring the interaction between LPMOs and its redox partner CDHs aiming for the development of more efficient enzymes for insertion into an engineered cell factory, BP.PD; 22/14376-1 - Functional analysis of disordered regions associated to lytic polysaccharide monooxygenases (LPMO): Biochemical characterization and interaction with cellobiose dehydrogenase (CDH), BP.IC; 22/09945-7 - Evaluation of interaction sites in the recombinant cellobiohydrolase Cel7D from Phanerochaete chrysosporium with low molecular weight phenolic compounds, BP.IC; 22/04713-0 - Exploring the interaction between lytic polysaccharide monooxygenases and its redox partner cellobiose dehydrogenases, BP.DD - associated scholarships

Abstract

Lignocellulose is an abundant source of polysaccharides that can be used as a renewable raw material for the production of biofuels and other compounds of interest. The conversion of lignocellulose into its subunits is a slow and costly process, therefore, more study is needed to make this process feasible. Until recently, the available enzymatic cocktails were composed of hydrolytic enzymes that act on the cellulose components of lignocellulose. These enzymes cocktails are low and inefficient. In recent years, oxidative proteins have been discovered that act directly on crystalline cellulose have been described, and are named LPMOs. Since the discovery, LPMOs have been added to the commercial cocktails improving their performance, as well as, have been the focus of many studies related to their mode of action, interaction with substrate and electron donors including the enzyme CDH. Through the analysis of amino acid, sequence of LPMOs from different filamentous fungi an extra-region was found to the structure described so far. Previous molecular docking analysis of the interaction between CDHs and LPMOs provided evidenced that these extra-regions serve to anchor location of the CDH for electron transfer pathway. Since we have been developing a filamentous fungus for the enzymes production the current proposal aim to 1) understanding the coupling mechanism between CDH and LPMO, as well as describing the electron transfer pathway to the copper active site; 2) use a multiplex RNAi system to improve the production of proteins in filamentous fungi; 3) insert the synthetically modified proteins from previous computational analysis into our expression system in order to improve the biomass saccharification, as well propose the use of this oxidative enzymes in biosensors. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

OLIVA, BIANCA; FERRAZ, ANDRE; SEGATO, FERNANDO. Biochemical and inhibitor analysis of recombinant cellobiohydrolases from Phanerochaete chrysosporium. BIOTECHNOLOGY FOR BIOFUELS AND BIOPRODUCTS, v. 17, n. 1, p. 17-pg., 2024-11-29. (14/18714-2, 22/01756-0, 19/06663-8, 22/04227-9, 21/06679-1, 19/22284-7)

BARRIOS-GUTIERREZ, SOLANGE GRACE; VELEZ-MERCADO, MARTHA INES; ORTEGA, JULIA RODRIGUES; LIMA, AWANA DA SILVA; SARAIVA, ANA LUIZA DA ROCHA FORTES; BERTO, GABRIELA LEILA; SEGATO, FERNANDO. Oxidative Machinery of basidiomycetes as potential enhancers in lignocellulosic biorefineries: A lytic polysaccharide monooxygenases approach. Bioresource Technology, v. 386, p. 11-pg., 2023-10-01. (22/00539-6, 22/04227-9, 14/18714-2, 19/22284-7, 22/04713-0, 21/06679-1)

OLIVA, BIANCA; VELASCO, JOSMAN; BERTO, GABRIELA LEILA; POLIKARPOV, IGOR; DE OLIVEIRA, LEANDRO CRISTANTE; SEGATO, FERNANDO. Recombinant cellobiose dehydrogenase from Thermothelomyces thermophilus : Its functional characterization and applicability in cellobionic acid production. Bioresource Technology, v. 402, p. 11-pg., 2024-05-03. (14/18714-2, 19/06663-8, 19/01165-0, 22/04227-9, 21/06679-1, 19/22284-7)

BERTO, GABRIELA L.; MATTOS, BRUNO D.; VELASCO, JOSMAN; ZHAO, BIN; SEGATO, FERNANDO; ROJAS, ORLANDO J.; ARANTES, VALDEIR. Endoglucanase effects on energy consumption in the mechanical fibrillation of cellulose fibers into nanocelluloses. International Journal of Biological Macromolecules, v. 243, p. 9-pg., 2023-06-13. (15/02862-5, 19/22284-7, 21/07023-2, 21/06679-1)

VELASCO, JOSMAN; SEPULCHRO, ANA GABRIELA V.; HIGASI, PAULA M. R.; PELLEGRINI, VANESSA O. A.; CANNELLA, DAVID; DE OLIVEIRA, LEANDRO CRISTANTE; POLIKARPOV, IGOR; SEGATO, FERNANDO. Light Boosts the Activity of Novel LPMO from Aspergillus fumigatus Leading to Oxidative Cleavage of Cellulose and Hemicellulose. ACS SUSTAINABLE CHEMISTRY & ENGINEERING, v. 10, n. 50, p. 16-pg., 2022-12-08. (19/22284-7, 18/22300-0, 19/01165-0, 21/06679-1, 14/18714-2, 15/13684-0)