Comparative functional genomics identifies an iron-limited bottleneck in a Saccharomyces cerevisiae strain with a cytosolic-localized isobutanol pathway

作     者：Francesca V.Gambacorta Ellen R.Wagner Tyler B.Jacobson Mary Tremaine Laura K.Muehlbauer Mick A.McGee Justin J.Baerwald Russell L.Wrobel John F.Wolters Mike Place Joshua J.Dietrich Dan Xie Jose Serate Shabda Gajbhiye Lisa Liu Maikayeng Vang-Smith Joshua J.Coon Yaoping Zhang Audrey P.Gasch Daniel Amador-Noguez Chris Todd Hittinger Trey K.Sato Brian F.Pfleger 

作者机构：DOE Great Lakes Bioenergy Research CenterUniversity of Wisconsin-MadisonMadisonWIUSA Department of Chemical and Biological EngineeringUniversity of Wisconsin-MadisonMadisonWIUSA Laboratory of GeneticsCenter for Genomic Science InnovationUniversity of Wisconsin-MadisonMadisonWIUSA Department of BacteriologyUniversity of Wisconsin-MadisonMadisonWIUSA Department of ChemistryUniversity of Wisconsin-MadisonMadisonWIUSA Wisconsin Energy InstituteJ.F.Crow Institute for the Study of EvolutionUniversity of Wisconsin-MadisonMadisonWIUSA Department of Biomolecular ChemistryUniversity of Wisconsin-MadisonMadisonWIUSA 

出 版 物：《Synthetic and Systems Biotechnology》 (合成和系统生物技术（英文）)

年 卷 期：2022年第7卷第2期

页      面：738-749页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：National Institutes of Health Biotechnology Training Program Office of the Vice Chancellor for Research and Graduate Education Office of the Vice Chancellor for Research and Graduate Education at the University United States Department of Agriculture National Institute of Food and Agriculture, (1020204) National Science Foundation, NSF, (DEB-1442148, DEB-2110403) National Institutes of Health, NIH, (1907278, DGE-1747503, T32 HG002760-16) U.S. Department of Energy, USDOE National Institute of General Medical Sciences, NIGMS, (T32 GM135066) Wisconsin Alumni Research Foundation, WARF Office of Science, SC Biological and Environmental Research, BER, (DE-SC0018409) Graduate School, University of Oregon Great Lakes Bioenergy Research Center, GLBRC 

主　　题：Saccharomyces cerevisiae Isobutanol Functional genomics analysis Pathway localization Fe incorporation 

摘      要：Metabolic engineering strategies have been successfully implemented to improve the production of isobutanol,a next-generation biofuel,in Saccharomyces ***,we explore how two of these strategies,pathway re-localization and redox cofactor-balancing,affect the performance and physiology of isobutanol producing *** equipped yeast with isobutanol cassettes which had either a mitochondrial or cytosolic localized isobutanol pathway and used either a redox-imbalanced(NADPH-dependent)or redox-balanced(NADH-dependent)ketol-acid reductoisomerase *** then conducted transcriptomic,proteomic and metabolomic analyses to elucidate molecular differences between the engineered *** localization had a large effect on isobutanol production with the strain expressing the mitochondrial-localized enzymes producing 3.8-fold more isobutanol than strains expressing the cytosolic ***-balancing did not improve isobutanol titers and instead the strain with the redox-imbalanced pathway produced 1.5-fold more isobutanol than the balanced version,albeit at low overall pathway *** genomic analyses suggested that the poor performances of the cytosolic pathway strains were in part due to a shortage in cytosolic Fe-S clusters,which are required cofactors for the dihydroxyacid dehydratase *** then demonstrated that this cofactor limitation may be partially recovered by disrupting iron homeostasis with a fra2 mutation,thereby increasing cellular iron *** resulting isobutanol titer of the fra2 null strain harboring a cytosolic-localized isobutanol pathway outperformed the strain with the mitochondrial-localized pathway by 1.3-fold,demonstrating that both localizations can support flux to isobutanol.