Synthesis of three advanced biofuels from ionic liquid-pretreated switchgrass using engineered Escherichia coli

Gregory Bokinsky, Pamela P. Peralta-Yahya, Anthe George, Bradley M. Holmes, Eric J. Steen, Jeffrey Dietrich, Taek Soon Lee, Danielle Tullman-Ercek, Christopher A. Voigt, Blake A. Simmons, Jay D. Keasling*

*Corresponding author for this work

Research output: Contribution to journalArticle

248 Scopus citations

Abstract

One approach to reducing the costs of advanced biofuel production from cellulosic biomass is to engineer a single microorganism to both digest plant biomass and produce hydrocarbons that have the properties of petrochemical fuels. Such an organism would require pathways for hydrocarbon production and the capacity to secrete sufficient enzymes to efficiently hydrolyze cellulose and hemicellulose. To demonstrate how one might engineer and coordinate all of the necessary components for a biomass-degrading, hydrocarbon-producing microorganism, we engineered a microorganism naïve to both processes, Escherichia coli, to grow using both the cellulose and hemicellulose fractions of several types of plant biomass pretreated with ionic liquids. Our engineered strains express cellulase, xylanase, beta-glucosidase, and xylobiosidase enzymes under control of native E. coli promoters selected to optimize growth on model cellulosic and hemicellulosic substrates. Furthermore, our strains grow using either the cellulose or hemicellulose components of ionic liquid-pretreated biomass or on both components when combined as a coculture. Both cellulolytic and hemicellulolytic strains were further engineered with three biofuel synthesis pathways to demonstrate the production of fuel substitutes or precursors suitable for gasoline, diesel, and jet engines directly from ionic liquid-treated switchgrass without externally supplied hydrolase enzymes. This demonstration represents a major advance toward realizing a consolidated bioprocess. With improvements in both biofuel synthesis pathways and biomass digestion capabilities, our approach could provide an economical route to production of advanced biofuels.

Original languageEnglish (US)
Pages (from-to)19949-19954
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume108
Issue number50
DOIs
StatePublished - Dec 13 2011

Keywords

  • Consolidated bioprocessing
  • Ionic liquid pretreatment

ASJC Scopus subject areas

  • General

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    Bokinsky, G., Peralta-Yahya, P. P., George, A., Holmes, B. M., Steen, E. J., Dietrich, J., Lee, T. S., Tullman-Ercek, D., Voigt, C. A., Simmons, B. A., & Keasling, J. D. (2011). Synthesis of three advanced biofuels from ionic liquid-pretreated switchgrass using engineered Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America, 108(50), 19949-19954. https://doi.org/10.1073/pnas.1106958108