Metabolic engineering of the yeast Yarrowia lipolytica for monoterpene geraniol production

• Grant number: 25/04277-4
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: August 01, 2025
• End date: March 31, 2029
• Field of knowledge: Agronomical Sciences - Food Science and Technology - Food Engineering
• Principal Investigator: Thiago Olitta Basso
• Grantee: Solange Grace Barrios Gutierrez
• Host Institution: Escola Politécnica (EP). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 22/03052-0 - Metabolic and evolutionary engineering of non-conventional yeasts for bioactive terpenoid production, AP.PNGP.PI

Abstract

Geraniol is a monoterpene widely used as a fragrance in cosmetic and cleaning industry products. Despite the increasing demand, its production is still limited to essential oil extraction through fractional distillation or chemical synthesis, which exhibits low efficiency and high costs. In this context, metabolic engineering emerges as a promising alternative for the sustainable production of this compound using microorganisms. The yeast Yarrowia lipolytica, due to its unique characteristics such as high tolerance to organic compounds and elevated levels of cytosolic acetyl-CoA, a key precursor in monoterpene biosynthesis stands out as an ideal model organism for this purpose. This project aims to apply CRISPR-Cas9 technology for the expression of the Geraniol Synthase (GES) enzyme in Y. lipolytica, under the control of the quasi-constitutive hp4d promoter, which is activated during the stationary phase of microbial growth. Additionally, the fusion of the GES gene with geranyl pyrophosphate (GPP) synthase will be implemented to enhance metabolic channeling and production efficiency. As part of the project stages, adaptive evolution experiments will be conducted to select strains with increased geraniol tolerance, and bioreactor optimizations will be performed using an experimental design that maximizes compound production. This work plan seeks not only to enable the biotechnological production of geraniol but also to contribute to the development of more sustainable and economically competitive technologies for the chemical, food, and fragrance industries. (AU)