(228bh) Metabolic Modeling and Pathway Engineering of an Industrially Relevant Pseudomonas Putida KT2440 Strain to Produce Muconate from Glucose

The biological production of muconic acid has received significant interest because of the metabolite's facile conversion to adipic acid and terephthalic acid, commodity chemicals used in large quantities as plastic precursors. The latter's traditional production route from petrochemicals involves the generation of toxic byproducts, further motivating microbial production strategies. A key limitation in the production of muconate from biomass has been the relatively low yields achieved from industrially relevant strains. In this talk, we describe a core-carbon metabolic model to guide strain engineering efforts to improve flux to muconate in a genome-integrated Pseudomonas putida KT2440 strain. Using elementary flux modes and constrained minimum cut sets, we determine an optimum set of knockout targets to force carbon flow into the engineered muconate pathway. After further pathway de-bottlenecking, we achieve muconate yields nearly equal to those expected from the stoichiometric model. This work highlights the metabolic flexibility of P. putida, as well as the advantages of the Entner-Doudoroff pathway in coupling growth to product formation.