(28g) Enabling Genetic Tools for Engineering Oleochemical Production in Yarrowia Lipolytica

Oleaginous yeast have been successfully engineered to produce a wide variety of lipids, and oleochemicals. Their success is tied to the native high flux pathways producing malonyl-CoA and NADPH that serve as better starting points for metabolic engineering than S. cerevisiae. A rapidly improving understanding of Y. lipolytica genetics and metabolism, coupled with increasing tools to engineer Y. lipolytica have opened up engineering opportunities approaching those in S. cerevisiae; however, engineering of highly productive oleochemical pathways are still stymied by an incomplete understanding of the complexity of its cell physiology. I will discuss our work to increase the reliability and complexity of available tools for engineering Y. lipolytica, including the development of responsive promoters, dynamic promoters, gene editing systems, and genome-scale screens that provide insights into the physiology and metabolism. These tools are applied to engineering several pathways that we will discuss, including lipid production, and oleochemical production including fatty alcohols and polyhydroxyalkonoates (PHAs), both utilizing the peroxisome’s unique environment. This suggests a complementary approach to traditional pathway engineering can be used for strain development, leading to a higher ceiling before the easy bottlenecks are exhausted.