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...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 use synthetic biology approach to engineer Saccharomyces cerevisiae for de novo biosynthesis of vindoline and catharanthine,which can be coupled chemically or biologically to vinblastine.On the basis of a platform strain with sufficient supply of precursors and cofactors for biosynthesis,we reconstituted,debottlenecked,and optimized the biosynthetic pathways for the production of vindoline and catharanthine.The vindoline biosynthetic pathway represents one of 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 and 305.1μg·liter^(−1),respectively,without accumulating detectable amount of pathway intermediates.This study establishes a representative example for the production of valuable plant natural products in yeast.展开更多
基金the National Key Research and Development Program of China(2018YFA0901800 and 2021YFC2103200)the Natural Science Foundation of Zhejiang Province(LR20B060003)+1 种基金the Natural Science Foundation of China(22278361)the Fundamental Research Funds for the Central Universities(226-2022-00214).
文摘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 use synthetic biology approach to engineer Saccharomyces cerevisiae for de novo biosynthesis of vindoline and catharanthine,which can be coupled chemically or biologically to vinblastine.On the basis of a platform strain with sufficient supply of precursors and cofactors for biosynthesis,we reconstituted,debottlenecked,and optimized the biosynthetic pathways for the production of vindoline and catharanthine.The vindoline biosynthetic pathway represents one of 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 and 305.1μg·liter^(−1),respectively,without accumulating detectable amount of pathway intermediates.This study establishes a representative example for the production of valuable plant natural products in yeast.
