Friederike Gutmann1,2, Cosimo Jann3,4, Filipa Pereira5, Andreas Johansson6, Lars M Steinmetz6,7,8, Kiran R Patil9,10. 1. European Molecular Biology Laboratory (EMBL), Structural and Cell Biology Unit, 69117, Heidelberg, Germany. 2. Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), 13125, Berlin, Germany. 3. European Molecular Biology Laboratory (EMBL), Genome Biology Unit, 69117, Heidelberg, Germany. jann@embl.de. 4. Department of Biology, Institute of Biochemistry, ETH Zurich, Zurich, Switzerland. jann@embl.de. 5. European Molecular Biology Laboratory (EMBL), Structural and Cell Biology Unit, 69117, Heidelberg, Germany. filipa.pereira@embl.de. 6. European Molecular Biology Laboratory (EMBL), Genome Biology Unit, 69117, Heidelberg, Germany. 7. Department of Genetics, Stanford University School of Medicine, Stanford, CA, 94305, USA. 8. Stanford Genome Technology Center, Palo Alto, CA, 94304, USA. 9. European Molecular Biology Laboratory (EMBL), Structural and Cell Biology Unit, 69117, Heidelberg, Germany. patil@mrc-tox.cam.ac.uk. 10. MRC Toxicology Unit, University of Cambridge, Cambridge, UK. patil@mrc-tox.cam.ac.uk.
Abstract
BACKGROUND: Baker's yeast is a widely used eukaryotic cell factory, producing a diverse range of compounds including biofuels and fine chemicals. The use of lignocellulose as feedstock offers the opportunity to run these processes in an environmentally sustainable way. However, the required hydrolysis pretreatment of lignocellulosic material releases toxic compounds that hamper yeast growth and consequently productivity. RESULTS: Here, we employ CRISPR interference in S. cerevisiae to identify genes modulating fermentative growth in plant hydrolysate and in presence of lignocellulosic toxins. We find that at least one-third of hydrolysate-associated gene functions are explained by effects of known toxic compounds, such as the decreased growth of YAP1 or HAA1, or increased growth of DOT6 knock-down strains in hydrolysate. CONCLUSION: Our study confirms previously known genetic elements and uncovers new targets towards designing more robust yeast strains for the utilization of lignocellulose hydrolysate as sustainable feedstock, and, more broadly, paves the way for applying CRISPRi screens to improve industrial fermentation processes.
BACKGROUND:Baker's yeast is a widely used eukaryotic cell factory, producing a diverse range of compounds including biofuels and fine chemicals. The use of lignocellulose as feedstock offers the opportunity to run these processes in an environmentally sustainable way. However, the required hydrolysis pretreatment of lignocellulosic material releases toxic compounds that hamper yeast growth and consequently productivity. RESULTS: Here, we employ CRISPR interference in S. cerevisiae to identify genes modulating fermentative growth in plant hydrolysate and in presence of lignocellulosic toxins. We find that at least one-third of hydrolysate-associated gene functions are explained by effects of known toxic compounds, such as the decreased growth of YAP1 or HAA1, or increased growth of DOT6 knock-down strains in hydrolysate. CONCLUSION: Our study confirms previously known genetic elements and uncovers new targets towards designing more robust yeast strains for the utilization of lignocellulose hydrolysate as sustainable feedstock, and, more broadly, paves the way for applying CRISPRi screens to improve industrial fermentation processes.
Authors: Luke A Gilbert; Matthew H Larson; Leonardo Morsut; Zairan Liu; Gloria A Brar; Sandra E Torres; Noam Stern-Ginossar; Onn Brandman; Evan H Whitehead; Jennifer A Doudna; Wendell A Lim; Jonathan S Weissman; Lei S Qi Journal: Cell Date: 2013-07-11 Impact factor: 41.582