Engineering Yeast Transcription Machinery for Improved Ethanol Tolerance and Production

Abstract

Global transcription machinery engineering (gTME) is an approach for reprogramming gene transcription to elicit cellular phenotypes important for technological applications. Here we show the application of gTME to Saccharomyces cerevisiae for improved glucose/ethanol tolerance, a key trait for many biofuels programs. Mutagenesis of the transcription factor Spt15p and selection led to dominant mutations that conferred increased tolerance and more efficient glucose conversion to ethanol. The desired phenotype results from the combined effect of three separate mutations in the SPT15 gene [serine substituted for phenylalanine (Phe ¹⁷⁷ Ser) and, similarly, Tyr ¹⁹⁵ His, and Lys ²¹⁸ Arg]. Thus, gTME can provide a route to complex phenotypes that are not readily accessible by traditional methods.