Exploring oleaginous yeast Yarrowia lipolytica as a cell factory for isoprenoids

The oleaginous yeast Yarrowia lipolytica has a remarkable ability to accumulate high levels of lipids and to consume hydrophobic substrates. Moreover, this yeast has a generally regarded as safe status and a long history of industrial applications for the production of lipases, lactone flavors, citric acid, and other products. We explored the potential of Y. lipolytica to produce isoprenoids. Isoprenoids biosynthesis requires a supply of cytosolic acetyl-CoA and NADPH, same as lipids, hence we hypothesized that Y. lipolytica may be naturally superior to the conventional yeast Saccharomyces cerevisiae as an isoprenoid host.

We developed a genetic toolbox, which comprises well-characterized integrative vectors for targeting specific intergenic regions of Yarrowia chromosomes and CRISPR/Cas9-based system for efficient marker-free genome editing, including knock-outs and integrations. In contrast to the previously published works, our system does not require outgrowth to achieve high engineering efficiencies.

Using this toolbox, we implemented biosynthetic pathways towards representative isoprenoids from different classes (monoterpenoids, sesquiterpenoids, diterpenoids, triterpenoids, and tetraterpenoids). Moreover, we engineered the mevalonate pathway to increase the flux towards the isoprenoids. Our results show a great potential of Y. lipolytica to become an industrial platform strain for isoprenoids production.