Layered dynamic regulation for improving metabolic pathway productivity in Escherichia coli

Significance Optimization of the engineered pathway for production of D-glucaric acid, a precursor to nylons and detergents, requires strategies that manage pathway competition with glycolysis as well as the stability and activity of the rate-controlling enzyme. To address both of these challenges, two orthogonal, autonomous, and tunable dynamic regulation strategies were layered to produce the highest reported glucaric acid titers in Escherichia coli K-12 strains. The implementation of two regulatory circuits harnesses synthetic biology tools to program and optimize cellular behavior and demonstrates the power of multiplexed dynamic control for strain optimization.