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

Ling, Chen; Peabody, George L.; Salvachúa, Davinia; Kim, Young-Mo; Kneucker, Colin M.; Calvey, Christopher H.; Monninger, Michela A.; Munoz, Nathalie Munoz; Poirier, Brenton C.; Ramirez, Kelsey J.; St. John, Peter C.; Woodworth, Sean P.; Magnuson, Jon K.; Burnum-Johnson, Kristin E.; Guss, Adam M.; Johnson, Christopher W.; Beckham, Gregg T.

doi:10.1038/s41467-022-32296-y

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

Journal Article · Mon Aug 22 00:00:00 EDT 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-32296-y· OSTI ID:1883309

^[1]; Peabody, George L. ^[2]; Salvachúa, Davinia ^[1];

^[3]; Kneucker, Colin M. ^[1];

^[4]; Monninger, Michela A. ^[1];

^[3]; Poirier, Brenton C. ^[3];

^[1];

^[3];

^[2];

^[1];

^[1]

National Renewable Energy Lab. (NREL), Golden, CO (United States); USDOE Agile BioFoundry, Emeryville, CA (United States)
USDOE Agile BioFoundry, Emeryville, CA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
USDOE Agile BioFoundry, Emeryville, CA (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
National Renewable Energy Lab. (NREL), Golden, CO (United States)

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.

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Research Organization:: Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Renewable Energy Lab. (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Transportation Office. Bioenergy Technologies Office

Grant/Contract Number:: AC05-76RL01830; AC36-08GO28308; AC05-00OR22725

OSTI ID:: 1883309

Alternate ID(s):: OSTI ID: 1883794; OSTI ID: 1887873

Report Number(s):: PNNL-SA-168423; NREL/JA-2A00-81804

Journal Information:: Nature Communications, Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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applied microbiology
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Title: Muconic acid production from glucose and xylose in Pseudomonas putida via evolution and metabolic engineering

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