Coordinated Two-Stage Dynamic Deregulation of Central Metabolism Improves Malonyl-CoA Biosynthesis

Malonyl-CoA (malonyl-CoA) is a platform chemical that serves as a precursor for a wide range of commercial products and pharmaceutical intermediates. In E. coli, malonyl-CoA levels are tightly regulated to remain at low levels. Two Stage Dynamic Metabolic Control (DMC) is a tool previously demonstrated to improve desired metabolite flux for several products. This work leverages DMC to improve malonyl-CoA fluxes. Specifically, we demonstrate coordinated dynamic reductions in the activities of fabI (enoyl-ACP reductase), gltA (citrate synthase), zwf (glucose-6-phosphate dehydrogenase) and glnB (nitrogen regulatory protein PII-1), during stationary phase lead to synergistic improvements in malonyl-CoA flux and the production of malonyl-CoA dependent products, 1,3,6,8-tetrahydroxynaphthalene (THN), Triacetic Lactone (TAL), and Phloroglucinol (PG). We also discuss the unique set of limitations that were observed for both TAL and PG biosynthesis as well as the strategies that were tested to overcome them. Additionally, we provide a historical review of the challenges associated with the production of Phloroglucinol. Finally, we end with a critical review focused on the bioproduction of an Acetyl-CoA and Succinyl-CoA derived product, Adipic Acid, to give perspective to common challenges associated with biobased product development.