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Microbial production of fatty-acid-derived fuels and chemicals from plant biomass


Increasing energy costs and environmental concerns have emphasized the need to produce sustainable renewable fuels and chemicals1. Major efforts to this end are focused on the microbial production of high-energy fuels by cost-effective ‘consolidated bioprocesses’2. Fatty acids are composed of long alkyl chains and represent nature’s ‘petroleum’, being a primary metabolite used by cells for both chemical and energy storage functions. These energy-rich molecules are today isolated from plant and animal oils for a diverse set of products ranging from fuels to oleochemicals. A more scalable, controllable and economic route to this important class of chemicals would be through the microbial conversion of renewable feedstocks, such as biomass-derived carbohydrates. Here we demonstrate the engineering of Escherichia coli to produce structurally tailored fatty esters (biodiesel), fatty alcohols, and waxes directly from simple sugars. Furthermore, we show engineering of the biodiesel-producing cells to express hemicellulases, a step towards producing these compounds directly from hemicellulose, a major component of plant-derived biomass.

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Figure 1: Engineered pathways for production of fatty acid-derived molecules from hemicelluloses or glucose and depiction of the synthetic operons used in this study.
Figure 2: Total free fatty acid production and respective theoretical yield by engineered E. coli strains.
Figure 3: Engineered production of FAEEs and fatty alcohols with controlled chain length.
Figure 4: Towards a single cell catalyst: biodiesel (FAEE) production by various strains without exogenous ethanol supplementation.
Figure 5: Consolidated bioprocessing: growth and FAEE production by xylan-using strains.

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E.J.S. was supported by the Tien Scholar Environmental Fellowship and the Synthetic Biology Engineering Research Center (SynBERC). Y.K. and G.B. were supported by a grant from LS9, Inc. (South San Francisco, California) through the University of California Discovery Grant program. This research was performed at the Joint BioEnergy Institute. We thank M. Rude with help on the manuscript and J. Cronan and the LS9 Scientific Advisory Board for technical insight and discussion.

Author Contributions E.J.S., Y.K., G.B., Z.H., A.S., A.M., S.B.d.C. and J.D.K. conceived of the experiments. E.J.S. and Y.K. constructed the strains and metabolic pathways for fatty-acid-derived products and performed the production experiments. LS9 engineered and evaluated FAEE and fatty alcohol producing strains for thioesterase evaluations. G.B. conceived, constructed and performed the xylan-metabolizing pathway growth experiments. E.J.S. and Y.K. constructed the xylan-metabolizing, fatty acid production strain and performed the production experiments. E.J.S., Y.K., A.S., S.B.d.C. and J.D.K. drafted the manuscript. All authors approved the final manuscript.

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Correspondence to Stephen B. del Cardayre or Jay D. Keasling.

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J.D.K. has financial interests in Amyris and LS9, both of which are involved in producing advanced biofuels. Z.H., A.S., A.M. and S.B.d.C. have a financial interest in LS9.

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Steen, E., Kang, Y., Bokinsky, G. et al. Microbial production of fatty-acid-derived fuels and chemicals from plant biomass. Nature 463, 559–562 (2010).

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