Engineering Pseudomonas putida KT2440 for chain length tailored free fatty acid and oleochemical production

Luis E. Valencia, Matthew R. Incha, Matthias Schmidt, Allison N. Pearson, Mitchell G. Thompson, Jacob B. Roberts, Marina Mehling, Kevin Yin, Ning Sun, Asun Oka, Patrick M. Shih, Lars M. Blank, John Gladden, Jay D. Keasling*

*Corresponding author for this work

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Abstract

Despite advances in understanding the metabolism of Pseudomonas putida KT2440, a promising bacterial host for producing valuable chemicals from plant-derived feedstocks, a strain capable of producing free fatty acid-derived chemicals has not been developed. Guided by functional genomics, we engineered P. putida to produce medium- and long-chain free fatty acids (FFAs) to titers of up to 670 mg/L. Additionally, by taking advantage of the varying substrate preferences of paralogous native fatty acyl-CoA ligases, we employed a strategy to control FFA chain length that resulted in a P. putida strain specialized in producing medium-chain FFAs. Finally, we demonstrate the production of oleochemicals in these strains by synthesizing medium-chain fatty acid methyl esters, compounds useful as biodiesel blending agents, in various media including sorghum hydrolysate at titers greater than 300 mg/L. This work paves the road to produce high-value oleochemicals and biofuels from cheap feedstocks, such as plant biomass, using this host.
Original languageEnglish
Article number1363
JournalCommunications Biology
Volume5
Issue number1
Number of pages12
ISSN2399-3642
DOIs
Publication statusPublished - 2022

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