Metabolic division engineering of Escherichia coli consortia for de novo biosynthesis of flavonoids and flavonoid glycosides

Heterologous biosynthesis of natural products with long biosynthetic pathways in microorganisms often suffers from diverse problems, such as enzyme promiscuity and metabolic burden. Flavonoids and their glycosides are important phytochemicals in the diet of human beings , with various health benefits and biological activities . Despite previous efforts and achievements, efficient microbial production of plant-derived flavonoid compounds with long pathways remains challenging. Herein, we applied metabolic division engineering of Escherichia coli consortia to overcome these limitations. By establishing new biosynthetic pathways, rationally adjusting metabolic node intermediates, and engineering different auxotrophic and orthogonal carbon sources for hosts, we established stable two- and three-bacteria co-culture systems to efficiently de novo produce 12 flavonoids (61.15–325.31 mg/L) and 36 corresponding flavonoid glycosides (1.31–191.79 mg/L). Furthermore, the co-culture system was rapidly extended in a plug-and-play manner to produce isoflavonoids , dihydrochalcones, and their glycosides . This study successfully alleviates metabolic burden and overcomes enzyme promiscuity, and provides significant insights that could guide the biosynthesis of other complex secondary metabolites . • Mutualistic co-culture systems produced 12 flavonoids and 36 flavonoid glycosides . • Metabolic division engineering reduces metabolic burden and increases production. • Strategic selection of metabolic splitting nodes could regulate metabolic flow. • First report on the de novo production of flavonoid-di-glycosides. • Extended systems synthesized isoflavonoid, the dihydrochalcone and their glycosides.