Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae

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.

doi:10.1016/j.ymben.2021.09.008

Title: Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae

Journal Article · Mon Sep 27 00:00:00 EDT 2021 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2021.09.008· OSTI ID:1827679

^[1]; Tremaine, Mary ^[1]; Place, Michael ^[1]; Liu, Lisa ^[1]; Pier, Austin ^[1]; Krause, David J. ^[1]; Xie, Dan ^[1]; Zhang, Yaoping ^[1];

^[1]; Gasch, Audrey P. ^[1];

^[1];

^[1]

Univ. of Wisconsin, Madison, WI (United States)

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.

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Research Organization:: Great Lakes Bioenergy Research Center (GLBRC), Madison, WI (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF)

Grant/Contract Number:: SC0018409; FC02-07ER64494; DEB-1442148; DEB-2110403

OSTI ID:: 1827679

Journal Information:: Metabolic Engineering, Vol. 68; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Crabtree/Warburg effect
Metabolic engineering
Biofuels
Xylose fermentation
Adaptive laboratory evolution
Saccharomyces cerevisiae

Title: Crabtree/Warburg-like aerobic xylose fermentation by engineered Saccharomyces cerevisiae

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