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. St. John, Sean P. Woodworth, Jon K. Magnuson, Kristin E. Burnum-Johnson, Adam M. Guss, Christopher W. Johnson, Gregg T. Beckham

Research output: Contribution to journalArticlepeer-review

37 Scopus Citations


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.

Original languageAmerican English
Article numberArticle No. 4925
Number of pages14
JournalNature Communications
Issue number1
StatePublished - Dec 2022

Bibliographical note

Publisher Copyright:
© 2022, The Author(s).

NREL Publication Number

  • NREL/JA-2A00-81804


  • muconic acid
  • performance advantaged bioproducts
  • Pseudomonas putida
  • shikimate


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