Synthetic biosensor accelerates evolution by rewiring carbon metabolism toward a specific metabolite

Proper carbon flux distribution between cell growth and production of a target compound is important for biochemical production because improper flux reallocation inhibits cell growth, thus adversely affecting production yield. Here, using a synthetic biosensor to couple production of a specific metabolite with cell growth, we spontaneously evolve cells under the selective condition toward the acquisition of genotypes that optimally reallocate cellular resources. Using 3-hydroxypropionic acid (3-HP) production from glycerol in Escherichia coli as a model system, we determine that mutations in the conserved regions of proteins involved in global transcriptional regulation alter the expression of several genes associated with central carbon metabolism. These changes rewire central carbon flux toward the 3-HP production pathway, increasing 3-HP yield and reducing acetate accumulation by alleviating overflow metabolism. Our study provides a perspective on adaptive laboratory evolution (ALE) using synthetic biosensors, thereby supporting future efforts in metabolic pathway optimization.

Automated summary

Proper carbon flux distribution between cell growth and production of a target compound is crucial for biochemical production. Using a synthetic biosensor, cells were evolved under selective conditions towards optimal resource allocation. Mutations in conserved regions of transcriptional regulation proteins rewired central carbon flux, enhancing target compound yield and reducing accumulation of byproducts. This study provides insight into adaptive laboratory evolution using synthetic biosensors for metabolic pathway optimization.