HARNESSING EXTREMOTOLERANT GENES TO ENHANCE Saccharomyces cerevisiae RESISTANCE TO PHENOLIC INHIBITORS FROM LIGNOCELLULOSIC BIOMASS

• Grant number: 25/00730-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: September 01, 2025
• End date: February 28, 2027
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Elizabeth Bilsland
• Grantee: Pedro Koliski Coelho Neto
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

In response to the growing global demand for sustainable biofuels, addressing the challenges posed by inhibitors in biofuel production processes has become increasingly important. Second-generation (2G) biofuels, derived from lignocellulosic biomass, hold significant promise for reducing reliance on fossil fuels and promoting environmental sustainability. However, the presence of inhibitory compounds in hydrolysates often impairs the fermentation efficiency of Saccharomyces cerevisiae, a widely used yeast in biofuel production. These inhibitors negatively impact the conversion efficiency of biomass, depleting cellular energy resources such as ATP, NADH, and NADPH. This depletion can prolong fermentation time and increase operational costs. This study aims to enhance S. cerevisiae tolerance to phenolic compounds, a class of inhibitors present in lignocellulosic hydrolysates, thereby improving the overall efficiency of 2G biofuel production. To achieve this objective, extremotolerant genes from tardigrades will be utilized. A synthetic tardigrade cDNA yeast library, containing approximately 300,000 clones, will be employed to identify key genes that can enhance yeast tolerance to common phenolic compounds encountered during fermentation. These genes will be optimized for expression in S. cerevisiae and integrated into an industrial xylose-fermenting yeast strain's genome using CRISPR/Cas9 technology. Additionally, bioinformatic analyses will predict gene functions, and protein-protein interaction assays. Alongside transcriptomic and metabolomic profiling, will elucidate the molecular mechanisms underlying the enhanced tolerance. The expected outcome is that tardigrade-derived genes will significantly increase yeast resistance to common phenolic inhibitors, contributing to the advancement of biofuel technologies and promoting the use of renewable resources.