De novo Biosynthesis of Vindoline and Catharanthine in Saccharomyces cerevisiae

Abstract Vinblastine has been used clinically as one of the most potent therapeutics for the treatment of several types of cancer. However, the traditional plant extraction method suffers from unreliable supply, low abundance, and extremely high cost. Here we employ synthetic biology approach to engineer Saccharomyces cerevisiae for de novo biosynthesis of vindoline and catharanthine, which can be coupled chemically or biologically to vinblastine. Based on a platform strain with sufficient supply of precursors and co-factors for biosynthesis, we reconstituted, debottlenecked, and optimized the vindoline and catharanthine biosynthetic pathways, representing the most complicated pathways ever reconstituted in microbial cell factories. Using shake flask fermentation, our engineered yeast strains were able to produce catharanthine and vindoline at a titer of 527.1 μg L-1 and 149.3 μg L-1, respectively, without accumulating detectable amount of pathway intermediates. The present study holds great promise for scalable production of vinblastine and other plant natural products.