Biosynthesis of catharanthine in engineered Pichia pastoris

Catharanthine can be coupled with vindoline to synthesize vinblastine and vincristine, which have been used clinically as potent anticancer drugs. However, the structural complexity and low abundance in nature hamper bulk chemical synthesis and plant extraction, leading to a limited supply and a high cost of this plant natural product. Here, we engineer the methylotrophic yeast Pichia pastoris for complete biosynthesis of catharanthine from simple carbon sources. Through the selection of stable integration sites, screening of biosynthetic pathway enzymes with higher activity and/or specificity, amplification of flux-limiting enzyme encoding genes, rewiring of cellular metabolism and process optimization, we achieve de novo biosynthesis of catharanthine with a titre as high as 2.57 mg l −1 , which also represents the most complicated molecule heterologously synthesized in a non-model microorganism. Our study establishes P. pastoris as a cell factory for producing plant natural products with complex biosynthetic pathways.