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

Abstract 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.