1 Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2 Fungal Cell Factories, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3 Chalmers University of Technology4 Novozymes A/S5 KTH - Royal Institute of Technology6 Department of Systems Biology, Technical University of Denmark
Ethanol production for use as a biofuel is mainly achieved through simultaneous saccharification and fermentation by yeast. Operating at ≥40°C would be beneficial in terms of increasing efficiency of the process and reducing costs, but yeast does not grow efficiently at those temperatures. We used adaptive laboratory evolution to select yeast strains with improved growth and ethanol production at ≥40°C. Sequencing of the whole genome, genome-wide gene expression, and metabolic-flux analyses revealed a change in sterol composition, from ergosterol to fecosterol, caused by mutations in the C-5 sterol desaturase gene, and increased expression of genes involved in sterol biosynthesis. Additionally, large chromosome III rearrangements and mutations in genes associated with DNA damage and respiration were found, but contributed less to the thermotolerant phenotype.