Bioprocess development and scale-up for cis,cis-muconic acid production from glucose and xylose by Pseudomonas putida†

cis,cis-Muconic acid (MA) is a bio-based chemical that can be converted to direct replacement chemicals or performance-advantaged bioproducts. We recently engineered the bacterium Pseudomonas putida KT2440 for the co-utilization of glucose and xylose to produce MA. This study evaluates the effect of additional genetic modifications, media composition, and bioprocess strategy on MA titer, productivity, and yield in bioreactor cultivations. We achieve a MA titer of 47.2 g L⁻¹, a productivity of 0.49 g L⁻¹ h⁻¹, and a yield of 0.50 C-mol C-mol⁻¹ from glucose and xylose supplemented with 5% (v/v) corn steep liquor with a P. putida strain harboring the deletion of gacS. Additionally, we demonstrate efficient MA production from corn stover-derived sugars and scalability to 150 L bioreactors. Techno-economic analysis and life cycle assessment predict that adipic acid, derived from catalytic hydrogenation of MA, can achieve a selling price as low as $2.60 per kg, approaching cost parity and reducing greenhouse gas emissions by up to 80% relative to fossil carbon-based adipic acid.