(41g) Engineering Yarrowia Lipolytica for Fuels and Oleochemicals Applications

Concerns about energy security, the global petroleum supply and climate change have increased interest in the production of sustainable and renewable biofuels.  The oleaginous yeast Yarrowia lipolytica naturally possesses moderate lipid (biodiesel precursor) production and grows on different kinds of biomass (and organic waste). However, production from native, un-engineered strains is not sufficient for an industrial process.  Here, we report on a rational and combinatorial metabolic engineering approach to establish Y. lipolytica as a premier platform for industrial-level, high lipid production. Specifically, several rational gene targets were combined to uncover potential synergistic genetic influencing lipogenesis. This study resulted in the largest collection of genetically modified strains of Y. lipolytica. The lipid content in the best engineered strain exceeded 80% of its dry weight. In parallel with these efforts, an inverse, combinatorial metabolic engineering approach was used to isolate improved lipid production strains. Whole genome re-sequencing of isolated strains revealed a novel lipid enhancer element. Further improvements of lipid production were achieved by combining these two approaches. Through this effort, we enabled extremely high lipid production in Y. lipolytica. This project helps elucidate the mechanism of lipogenesis and establish Y. lipolytica as an oleochemicals platform strain.