Muconic acid production from glucose and xylose in Pseudomonas putida via evolution and metabolic engineering

Chen Ling, George L. Peabody, Davinia Salvachúa, Young-Mo Kim, Colin M. Kneucker, Christopher H. Calvey, Michela A. Monninger, Nathalie Munoz Munoz, Brenton C. Poirier, Kelsey J. Ramirez, Peter C. John, Sean P. Woodworth, Jon K. Magnuson, Kristin E. Burnum-Johnson, Adam M. Guss (), Christopher W. Johnson () and Gregg T. Beckham ()
Additional contact information
Chen Ling: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
George L. Peabody: Agile BioFoundry
Davinia Salvachúa: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Young-Mo Kim: Agile BioFoundry
Colin M. Kneucker: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Christopher H. Calvey: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Michela A. Monninger: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Nathalie Munoz Munoz: Agile BioFoundry
Brenton C. Poirier: Agile BioFoundry
Kelsey J. Ramirez: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Peter C. John: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Sean P. Woodworth: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Jon K. Magnuson: Agile BioFoundry
Kristin E. Burnum-Johnson: Agile BioFoundry
Adam M. Guss: Agile BioFoundry
Christopher W. Johnson: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory
Gregg T. Beckham: Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory

Nature Communications, 2022, vol. 13, issue 1, 1-14

Abstract: Abstract Muconic acid is a bioprivileged molecule that can be converted into direct replacement chemicals for incumbent petrochemicals and performance-advantaged bioproducts. In this study, Pseudomonas putida KT2440 is engineered to convert glucose and xylose, the primary carbohydrates in lignocellulosic hydrolysates, to muconic acid using a model-guided strategy to maximize the theoretical yield. Using adaptive laboratory evolution (ALE) and metabolic engineering in a strain engineered to express the D-xylose isomerase pathway, we demonstrate that mutations in the heterologous D-xylose:H+ symporter (XylE), increased expression of a major facilitator superfamily transporter (PP_2569), and overexpression of aroB encoding the native 3-dehydroquinate synthase, enable efficient muconic acid production from glucose and xylose simultaneously. Using the rationally engineered strain, we produce 33.7 g L−1 muconate at 0.18 g L−1 h−1 and a 46% molar yield (92% of the maximum theoretical yield). This engineering strategy is promising for the production of other shikimate pathway-derived compounds from lignocellulosic sugars.

Date: 2022
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DOI: 10.1038/s41467-022-32296-y

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