Engineering Metabolic Pathways for Cofactor Self-Sufficiency and Serotonin Production in Escherichia coli

Serotonin is a neurotransmitter that plays an essential regulatory role in numerous cognitive and behavioral functions. Recent advances in synthetic biology have enabled engineering of non-natural biosynthetic pathways for serotonin production in E. coli. Here, an optimized heterologous serotonin biosynthetic pathway was engineered in E. coli and coupled with the biosynthetic and regeneration modules of the endogenous vital cofactor tetrahydrobiopterin (BH4) for efficient serotonin production using whole-cell catalysis. Further metabolic engineering efforts were performed to ensure an adequate endogenous BH4 supply, including enhancements of GTP biosynthesis and intracellular reducing power availability. Using the optimized fed-batch fermentation, an overall maximum serotonin yield of 40.3% (mol/mol) and a peak titer of 1.68 g/L (production rate of 0.016 g/L/h) were achieved. The strategies employed in this study show the promise of using E. coli for pterin self-sufficiency and high-level serotonin production, and the engineered strains hold the potential for use in industrial applications.

Automated summary

In this study, an optimized heterologous serotonin biosynthetic pathway was engineered in E. coli, coupled with the biosynthetic and regeneration modules of tetrahydrobiopterin (BH4). By further metabolic engineering efforts, an adequate endogenous BH4 supply was ensured. Using the optimized fed-batch fermentation, a high level of serotonin production was achieved, demonstrating the potential of E. coli for industrial applications.