title: 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.
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology, Room 56-469, Cambridge, MA 02139, USA.
authors.forename: Hal
authors.initials: H
authors.lastname: Alper
authors.forename: Joel
authors.initials: J
authors.lastname: Moxley
authors.forename: Elke
authors.initials: E
authors.lastname: Nevoigt
authors.forename: Gerald R
authors.initials: GR
authors.lastname: Fink
authors.forename: Gregory
authors.initials: G
authors.lastname: Stephanopoulos
mesh_terms: D019943:Amino Acid Substitution; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism* / Q000494:pharmacology; D005285:Fermentation; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D016296:Mutagenesis; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D035381:TATA-Binding Protein Associated Factors / Q000235:genetics / Q000378:metabolism; D035181:TATA-Box Binding Protein / Q000235:genetics* / Q000378:metabolism; D035362:Transcription Factor TFIID / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D015854:Up-Regulation
chemical_list: D018797:Cell Cycle Proteins; C115017:SPT15 protein, S cerevisiae; C075582:SPT3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C100497:TAF10 protein, S cerevisiae; D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D000431:Ethanol; D005947:Glucose