Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae
Abstract
Bottlenecks in the efficient conversion of xylose into cost-effective biofuels have limited the widespread use of plant lignocellulose as a renewable feedstock. The yeast Saccharomyces cerevisiae ferments glucose into ethanol with such high metabolic flux that it ferments high concentrations of glucose aerobically, a trait called the Crabtree/Warburg Effect. In contrast to glucose, most engineered S. cerevisiae strains do not ferment xylose at economically viable rates and yields, and they require respiration to achieve sufficient xylose metabolic flux and energy return for growth aerobically. Here, we evolved respiration-deficient S. cerevisiae strains that can grow on and ferment xylose to ethanol aerobically, a trait analogous to the Crabtree/Warburg Effect for glucose. Through genome sequence comparisons and directed engineering, we determined that duplications of genes encoding engineered xylose metabolism enzymes, as well as TKL1, a gene encoding a transketolase in the pentose phosphate pathway, were the causative genetic changes for the evolved phenotype. Reengineered duplications of these enzymes, in combination with deletion mutations in HOG1, ISU1, GRE3, and IRA2, increased the rates of aerobic and anaerobic xylose fermentation. Importantly, we found that these genetic modifications function in another genetic background and increase the rate and yield of xylose-to-ethanol conversion in industrially relevantmore »
- Publication Date:
- Research Org.:
- Great Lakes Bioenergy Research Center (GLBRC), Madison, WI (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF)
- OSTI Identifier:
- 1827679
- Grant/Contract Number:
- SC0018409; FC02-07ER64494; DEB-1442148; DEB-2110403
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Metabolic Engineering
- Additional Journal Information:
- Journal Volume: 68; Journal ID: ISSN 1096-7176
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Crabtree/Warburg effect; Metabolic engineering; Biofuels; Xylose fermentation; Adaptive laboratory evolution; Saccharomyces cerevisiae
Citation Formats
Lee, Sae-Byuk, Tremaine, Mary, Place, Michael, Liu, Lisa, Pier, Austin, Krause, David J., Xie, Dan, Zhang, Yaoping, Landick, Robert, Gasch, Audrey P., Hittinger, Chris Todd, and Sato, Trey K. Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae. United States: N. p., 2021.
Web. doi:10.1016/j.ymben.2021.09.008.
Lee, Sae-Byuk, Tremaine, Mary, Place, Michael, Liu, Lisa, Pier, Austin, Krause, David J., Xie, Dan, Zhang, Yaoping, Landick, Robert, Gasch, Audrey P., Hittinger, Chris Todd, & Sato, Trey K. Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae. United States. https://doi.org/10.1016/j.ymben.2021.09.008
Lee, Sae-Byuk, Tremaine, Mary, Place, Michael, Liu, Lisa, Pier, Austin, Krause, David J., Xie, Dan, Zhang, Yaoping, Landick, Robert, Gasch, Audrey P., Hittinger, Chris Todd, and Sato, Trey K. Mon .
"Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae". United States. https://doi.org/10.1016/j.ymben.2021.09.008. https://www.osti.gov/servlets/purl/1827679.
@article{osti_1827679,
title = {Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae},
author = {Lee, Sae-Byuk and Tremaine, Mary and Place, Michael and Liu, Lisa and Pier, Austin and Krause, David J. and Xie, Dan and Zhang, Yaoping and Landick, Robert and Gasch, Audrey P. and Hittinger, Chris Todd and Sato, Trey K.},
abstractNote = {Bottlenecks in the efficient conversion of xylose into cost-effective biofuels have limited the widespread use of plant lignocellulose as a renewable feedstock. The yeast Saccharomyces cerevisiae ferments glucose into ethanol with such high metabolic flux that it ferments high concentrations of glucose aerobically, a trait called the Crabtree/Warburg Effect. In contrast to glucose, most engineered S. cerevisiae strains do not ferment xylose at economically viable rates and yields, and they require respiration to achieve sufficient xylose metabolic flux and energy return for growth aerobically. Here, we evolved respiration-deficient S. cerevisiae strains that can grow on and ferment xylose to ethanol aerobically, a trait analogous to the Crabtree/Warburg Effect for glucose. Through genome sequence comparisons and directed engineering, we determined that duplications of genes encoding engineered xylose metabolism enzymes, as well as TKL1, a gene encoding a transketolase in the pentose phosphate pathway, were the causative genetic changes for the evolved phenotype. Reengineered duplications of these enzymes, in combination with deletion mutations in HOG1, ISU1, GRE3, and IRA2, increased the rates of aerobic and anaerobic xylose fermentation. Importantly, we found that these genetic modifications function in another genetic background and increase the rate and yield of xylose-to-ethanol conversion in industrially relevant switchgrass hydrolysate, indicating that these specific genetic modifications may enable the sustainable production of industrial biofuels from yeast. We propose a model for how key regulatory mutations prime yeast for aerobic xylose fermentation by lowering the threshold for overflow metabolism, allowing mutations to increase xylose flux and to redirect it into fermentation products.},
doi = {10.1016/j.ymben.2021.09.008},
journal = {Metabolic Engineering},
number = ,
volume = 68,
place = {United States},
year = {Mon Sep 27 00:00:00 EDT 2021},
month = {Mon Sep 27 00:00:00 EDT 2021}
}
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