Production of itaconic acid from alkali pretreated lignin by dynamic two stage bioconversion

Joshua R. Elmore, Gara N. Dexter, Davinia Salvachúa, Jessica Martinez-Baird, E. Anne Hatmaker, Jay D. Huenemann, Dawn M. Klingeman, George L. Peabody, Darren J. Peterson, Christine Singer, Gregg T. Beckham and Adam M. Guss ()
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Joshua R. Elmore: Biosciences Division, Oak Ridge National Laboratory
Gara N. Dexter: Biosciences Division, Oak Ridge National Laboratory
Davinia Salvachúa: National Bioenergy Center, National Renewable Energy Laboratory
Jessica Martinez-Baird: Biosciences Division, Oak Ridge National Laboratory
E. Anne Hatmaker: Biosciences Division, Oak Ridge National Laboratory
Jay D. Huenemann: Biosciences Division, Oak Ridge National Laboratory
Dawn M. Klingeman: Biosciences Division, Oak Ridge National Laboratory
George L. Peabody: Biosciences Division, Oak Ridge National Laboratory
Darren J. Peterson: National Bioenergy Center, National Renewable Energy Laboratory
Christine Singer: National Bioenergy Center, National Renewable Energy Laboratory
Gregg T. Beckham: National Bioenergy Center, National Renewable Energy Laboratory
Adam M. Guss: Biosciences Division, Oak Ridge National Laboratory

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract Expanding the portfolio of products that can be made from lignin will be critical to enabling a viable bio-based economy. Here, we engineer Pseudomonas putida for high-yield production of the tricarboxylic acid cycle-derived building block chemical, itaconic acid, from model aromatic compounds and aromatics derived from lignin. We develop a nitrogen starvation-detecting biosensor for dynamic two-stage bioproduction in which itaconic acid is produced during a non-growth associated production phase. Through the use of two distinct itaconic acid production pathways, the tuning of TCA cycle gene expression, deletion of competing pathways, and dynamic regulation, we achieve an overall maximum yield of 56% (mol/mol) and titer of 1.3 g/L from p-coumarate, and 1.4 g/L titer from monomeric aromatic compounds produced from alkali-treated lignin. This work illustrates a proof-of-principle that using dynamic metabolic control to reroute carbon after it enters central metabolism enables production of valuable chemicals from lignin at high yields by relieving the burden of constitutively expressing toxic heterologous pathways.

Date: 2021
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DOI: 10.1038/s41467-021-22556-8

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