Production of Plant Sesquiterpene Lactone Parthenolide in the Yeast Cell Factory

Parthenolide, a kind of sesquiterpene lactone, is the direct precursor for the promising anti-glioblastoma drug ACT001. Compared with traditional parthenolide source from plant extraction, de novo biosynthesis of parthenolide in microorganisms has the potential to make a sustainable supply. Herein, an integrated strategy was designed with P450 source screening, nicotinamide adenine dinucleotide phosphate (NADPH) supply, and endoplasmic reticulum (ER) size rewiring to manipulate three P450s regarded as the bottleneck for parthenolide production. Germacrene A oxidase from Cichorium intybus, costunolide synthase from Lactuca sativa, and parthenolide synthase from Tanacetum parthenium have the best efficiency, resulting in a parthenolide titer of 2.19 mg/L, which was first achieved in yeast. The parthenolide titer was further increased by 300% with NADPH supplementation and ER expanding stepwise. Finally, the highest titers of 31.0 mg/L parthenolide and 648.5 mg/L costunolide in microbes were achieved in 2.0 L fed-batch fermentation. This study not only provides an alternative microbial platform for producing sesquiterpene lactones in a sustainable way but also highlights a general strategy for manipulating multiple plant-derived P450s in microbes.

Automated summary

This study successfully manipulated the production of parthenolide, a sesquiterpene lactone and precursor for the anti-glioblastoma drug ACT001, in microorganisms through a comprehensive strategy involving P450 source screening, NADPH supply, and endoplasmic reticulum size rewiring. The highest titers of parthenolide and costunolide achieved in microbes were 31.0 mg/L and 648.5 mg/L, respectively, demonstrating the potential for sustainable production of sesquiterpene lactones and highlighting a general strategy for manipulating multiple plant-derived P450s in microbes.