Engineering Yarrowia lipolytica for producing plant sterols(β-sitosterol)

Monoterpenes are widely used in food, pharmaceuticals and cosmetics. The monoterpene linalool for example is mainly used in flavors and fragrances, green solvents, platform chemicals and active agents and its market size is projected to surpass USD 1.9 billion by 2024. However, traditional methods of monoterpene production - plant extraction and chemical synthesis - are not sustainable and inefficient, due to cumbersome processes, high energy consumption and low yields. Large-scale and low-cost microbial production of monoterpenes is therefore crucial to establish a sustainable and economically viable supply of monoterpenes. However, despite several research efforts, it has become clear that rational engineering of the heterologous monoterpene pathways alone will not lead to economically viable yields. This is mainly due to (i) fundamental shortcomings of current production strains, which have low pathway fluxes, (ii) monoterpene cytotoxicity and (iii) suboptimal natural monoterpene synthases. This project uses the complementary expertise of both partners to comprehensively tackle these problems, using high-throughput screening methods, cutting-edge synthetic biology tools and enzyme engineering. Ultimately, we will create a portfolio of chassis strains that can serve as ‘plug-and-play' strains for production of a plethora of monoterpenes. As proof-of-concept, we will develop production strains for three high-value monoterpenes: linalool, limonene and cineole.