Study of lipid particle formation after insertion of the Q3 and Q4 helices of Coq3p from Saccharomyces cerevisiae into the orthologous protein of Yarrowia Lipolytica

• Grant number: 24/10926-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2024
• End date: November 30, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: José Ribamar dos Santos Ferreira Júnior
• Grantee: Esther Dantas Batista
• Host Institution: Escola de Artes, Ciências e Humanidades (EACH). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Oleaginous yeasts are microorganisms capable of producing and storing more than 20% of their biomass in the form of lipids. Yarrowia lipolytica, an oleaginous yeast, stands out for its ability to store more than 40% lipids in its dry biomass in the form of lipid particles (LDs) and convert various carbon sources into triacylglycerols (TAGs) and sterol esters (SEs) during the stationary phase of growth. Due to these characteristics, this yeast is highly promising for the production of biofuels, in which the biosynthesis of fatty acids and the formation of TAGs and SEs are essential processes. On the other hand, ubiquinone (CoQ) is a fundamental coenzyme of the respiratory chain in eukaryotes, and its biosynthesis is operated by proteins of the Coq1p-Coq11p family. Previous work by our research group has shown that the Coq3p protein, involved in two O-methylation steps, shows structural differences between oleaginous and non oleaginous yeasts, which corroborates a probable connection between these structural variations and lipid production. It is speculated that the insertion of the ±3 and ±4 helices of Coq3p characteristic of Saccharomyces cerevisiae into Yarrowia lipolytica could potentially reduce lipid production. The aim of this project is therefore to use techniques of gene recombination, construction of mutants and quantification of lipid content by fluorescence microscopy in order to investigate whether a mutant with the insertion of the reported structure has a phenotype of lower production of lipid particles.