Metabolic and evolutionary engineering of non-conventional yeasts for the production of bioactive terpenes

• Grant number: 25/01995-3
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: May 31, 2025
• End date: November 29, 2025
• Field of knowledge: Agronomical Sciences - Food Science and Technology - Food Engineering
• Principal Investigator: Thiago Olitta Basso
• Grantee: Mayra Paula de Souza e Sousa
• Supervisor: Yong-Su Jin
• Host Institution: Escola Politécnica (EP). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of Illinois at Urbana-Champaign, United States
• Associated to the scholarship: 23/18192-5 - Bioprocess engineering for the production of geraniol by Yarrowia lipolytica, BP.MS

Abstract

Geraniol is a monoterpene, commonly extracted from essential oils at high cost, which has properties of interest in the medicinal field. The biotechnology industry offers an efficient alternative to produce this bioactive terpene through the metabolic engineering of microorganisms such as yeasts. Yarrowia lipolytica is a yeast characterized by its rapid growth and ability to accumulate lipids and acetyl-CoA, characteristics that demonstrate advantages on the production of terpenes. An initial strain, BYa3105, was modified through CRISPR-Cas9, where the GES gene from Catharantus roseus (crGES) was integrated into the IntE_1 locus with the hp4d promoter, resulting in the production of geraniol, which is still open to optimization. This project aims to advance the development of two new strains of Y. lipolytica with expression of GES fused to GPPS, applying the integration of a new modified gene, ERG20F88W - N119W together with crGES from the previous strain. The modification will be tested with two different promoters: hp4d (semi-constitutive) and TEFin(constitutive), with integration for the new IntE_3 locus. The new approach aims to explore alternatives with a focus on optimizing new strains for the production of geraniol by Y. lipolytica.