Harnessing <i>Yarrowia lipolytica</i> Peroxisomes as a Subcellular Factory for α-Humulene Overproduction

Abstract

The sesquiterpene α-humulene has been shown to have anti-inflammatory and anticancer activities, which has led to its vast application potential in medicine. However, α-humulene production methods including phytoextraction and chemical synthesis currently were limited to low yield, high costs, and expensive catalysts, which cannot meet the increasing market demand. In this study, <i>Yarrowia lipolytica</i> was developed as a robust cell factory for α-humulene production. The peroxisome in <i>Y. lipolytica</i> was first engineered to boost the synthesis of the sesquiterpene α-humulene. By compartmentalization of the α-humulene biosynthesis pathway, improving ATP and acetyl-CoA supply, and optimizing the gene copy numbers of rate-limiting enzymes, the engineered strain GQ2012 could produce 3.2 g/L α-humulene in a 5 L bioreactor, the highest α-humulene titer reported so far. Our study provides a valuable reference for highly sustainable production of terpenoids by peroxisome engineering in <i>Y. lipolytica</i>.