Construction and Optimization of Nonclassical Isoprenoid Biosynthetic Pathways in Yeast Peroxisomes for (+)-Valencene Production

Isoprenoids are a kind of natural product with variousactivities,but their plant extraction suffers low concentration. The rapid developmentof synthetic biology offers a sustainable route for supply of high-value-addednatural products by engineering microorganisms. However, the complexityof cellular metabolism makes engineering endogenous isoprenoid biosyntheticpathways with metabolic interaction difficult. Here, for the firsttime, we constructed and optimized three types of isoprenoid pathways(the Haloarchaea-type, Thermoplasma-type, and isoprenoid alcohol pathway) in yeast peroxisomes for thesynthesis of sesquiterpene (+)-valencene. In yeast, the Haloarchaea-type MVA pathway is more effective than the classical MVA pathway. MVK and IPK were determined to be the rate-limitingsteps of the Haloarchaea-type MVA pathway, and theproduction of 869 mg/L (+)-valencene under fed-batch fermentationin shake flasks was realized. This work expands isoprenoid synthesisin eukaryotes and provides a more efficient pathway for isoprenoidsynthesis.

Automated summary

Engineering microorganisms to synthesize isoprenoids is an efficient approach to increase their concentration. In this study, three types of isoprenoid pathways were constructed and optimized in yeast peroxisomes to produce the sesquiterpene (+)-valencene. The Haloarchaea-type MVA pathway was found to be more effective, and a production of 869 mg/L (+)-valencene was achieved. This work expands isoprenoid synthesis in eukaryotes and provides a more efficient pathway for isoprenoid production.