(641e) Metabolic Engineering for Bacterial Production of Caffeic Acid Derived Phenethyl Esters and Amides in Escherichia coli

Knowledge-based metabolic engineering of natural products in microbes is a sustainable and cost-effective biomanufacturing alternative to direct extraction from plant tissues or chemical synthesis. Especially, genetic amenability makes the well-characterized microbes like Escherichia coli suitable industrial microbial platform for production of noncognate natural products from simple carbon source. However, the limited availability of known pathways or functional enzymes sometimes presents a significant challenge for metabolic engineering. In the present research, we established artificial biosynthetic pathways for therapeutically important compounds including caffeic acid derived phenethyl esters and amides, whose natural biosynthetic routes remain unknown in nature. We first identified functional acyltransferases capable of generating caffeic acid derived esters or amides from caffeoyl-CoA with counterpart aromatic alcohols or amines. Then we took advantage of this CoA-dependent trans-esterification pathway and constructed an E. coli platform for production of a collection of caffeic acid derived phenethyl esters and amides from glucose. Further production enhancement was achieved by up to 3-fold via host engineering and pathway optimization. This work permitted the bacterial production of natural compounds caffeic acid derived phenethyl esters and amides from simple carbon source, which also expanded the capabilities of microbial production of value-added aromatic esters and amides.