Programmable Biosynthesis of Plant-Derived 4′-Deoxyflavone Glycosides by an Unconventional Yeast Consortium

Previous data established 4′-deoxyflavone glycosides (4′-DFGs) as important pharmaceutical components in the roots of rare medical plants like Scutellaria baicalensis Georgi. Extracting these compounds from plants involves land occupation and is environmentally unfriendly. Therefore, a modular (“plug-and-play”) yeast-consortium platform is developed to synthesize diverse 4′-DFGs de novo. By codon-optimizing glycosyltransferase genes from different organisms for Pichia pastoris , six site-specific glycosylation chassis are generated to be capable of biosynthesizing 18 different 4′-DFGs. Cellular factories showed increased 4′-DFG production (up to 18.6-fold) due to strengthened synthesis of UDP-sugar precursors and blocked hydrolysis of endogenous glycosides. Co-culturing upstream flavone-synthesis-module cells with downstream glycoside-transformation-module cells alleviated the toxicity of 4′-deoxyflavones and enabled high-level de novo synthesis of 4′-DFGs. Baicalin is produced at the highest level (1290.0 mg L −1 ) in a bioreactor by controlling the consortium through carbon-source shifting. These results provide a valuable reference for biosynthesizing plant-derived 4′-DFGs and other glycosides with potential therapeutic applications.