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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.
Nature communications, 08/2022, Volume: 13, Issue: 1Journal Article
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.
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