Functional and kinetics characterization of sugar transporters in Trichoderma reesei involved in cellulase production

• Grant number: 24/11106-9
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: April 01, 2025
• End date: March 31, 2026
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Roberto do Nascimento Silva
• Grantee: Lucas Matheus Soares Pereira
• Supervisor: Bjorn Panyella Pedersen
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: Aarhus University, Denmark
• Associated to the scholarship: 22/14399-1 - Functional characterization and overexpression of potential cellobiose and xylose transporters in Trichoderma reesei during cellulase production, BP.DR

Abstract

The filamentous fungus Trichoderma reesei can produce high amounts of cellulases and hemicellulases that convert the polysaccharides in lignocellulosic biomass into fermentable sugars. However, to make second-generation ethanol economically viable, optimizing the production of these enzymes is necessary. Sugar transporters in T. reesei are essential for the regulation of cellulases and hemicellulases, either through the uptake of inducing sugars or in the signaling of these molecules. Moreover, the kinetic parameters of these membrane proteins are directly related to their function in cellulase production. Despite the importance of these membrane proteins, only a few have been kinetically characterized in T. reesei. Therefore, this project aims to functionally and kinetically characterize the sophorose and cellobiose transporters Tr44175, CRT1, the uncharacterized Tr128050, identified in proteomic analysis by our research group in the early stages of cellulose culture, as well as the transporter Tr79202. To accomplish this, we will use two techniques: solid-supported membrane (SSM)-based electrophysiology with the semi-automated SURFE2R N1 system as well as Xenopus laevis oocyte uptake assay. Our results will contribute to future usage of renewable carbon from biomass, by understanding the affinity of key sugar transporters, the mechanisms involved in the induction of cellulases, and by identifying potential targets for genetic engineering to increase enzyme production.