Computational analysis and synthetic biology approaches to understand the interaction and electron transfer between CDH and LPMO

• Grant number: 22/00539-6
• Support Opportunities: Regular Research Grants
• Duration: July 01, 2022 - June 30, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Convênio/Acordo: University of Manchester
• Mobility Program: SPRINT - Projetos de pesquisa - Mobilidade
• Principal Investigator: Fernando Segato
• Grantee: Fernando Segato
• Principal researcher abroad: Sam Hay
• Institution abroad: University of Manchester, England
• Host Institution: Escola de Engenharia de Lorena (EEL). Universidade de São Paulo (USP). Lorena , SP, Brazil
• Associated research grant: 21/06679-1 - 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, AP.BIOEN.JP2

Abstract

Recently, enzymes with oxidative mechanisms acting straight on crystalline cellulose have been described as LPMOs. Since that, these enzymes have been added to the commercial cocktails boosting their performance, making them the focus of several studies, such as: 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 in the structure described so far. Previous computational analysis of the interaction between CDHs and LPMOs provided evidenced that these extra-regions may serve as anchor location to the CDH for electron transfer pathway. This project would combine the talents of two researchers, Dr. Sam Hay of the University of Manchester and Dr. Fernando Segato of the University of São Paulo, to study the coupling mechanism between CDH and LPMO, as well as describe the electron transfer pathway to the copper active site at the LPMO using computational and synthetic biology approaches to create and insert the synthetically modified proteins from previous computational analysis which will be performed by Hay's Lab into the expression system previous developed by Segato's Lab in order to improve the biomass saccharification. Each principal investigator (Pi) and post-doc will visit each other's institution twice. In the first year, the main lab experiments will be conducted and in the second year, each Pi and post-doc will train the other group in their area of specialization. (AU)