Acronym: EvoSCO
Project code: 162/31.07.2023. Funding contract no. 760238 / dated 28.12.2023
Implementation period: February 2024 − June 2026
Total project value: RON 6,513,908.24, equivalent to EUR 1,206,715.20, of which RON 5,999,998.14 is funded through PNRR
Title
Adaptive laboratory evolution strategies to increase the lipid accumulation process in oleaginous microorganisms and microbial oil production using agroindustrial residues as substrates
Abstract
Yarrowia lipolytica is a species considered a model for the study of oleaginous yeasts and a source of neutral intracellular lipids. Currently, at an industrial scale, only polyunsaturated fatty acids of microbial origin are produced using genetically modified Y. lipolytica strains, and their production cost remains high due to the low accumulation rate.
Aim
The aim of this project is to develop adaptive laboratory evolution (ALE) strategies for generating hyperproductive oleaginous strains. The methodology will be developed and initially applied to Y. lipolytica strains and then extended to other oleaginous organisms of industrial interest, providing significant insights into the molecular evolutionary mechanisms associated with lipogenesis.
Objectives
- Y. lipolytica, Rhodosporidium toruloides and Cunninghamella echinulata will be evolved under alternating nitrogen-limiting conditions, which favor lipid accumulation, and carbon-limiting conditions, promoting proliferation of high lipid content cells.
- After multiple rounds of evolution, enrichment of the population with oleaginous genotypes is expected. Selection of clones with enhanced oleaginous traits will be performed by cultivation on solid medium without a carbon source, where clones with an improved lipogenic genotype will form the largest colonies. These clones will be used as inoculum for the next evolutionary round and will be biochemically, genotypically, and transcriptionally characterized.
- Genes differentially expressed compared to parental strains will be analyzed and annotated using the Gene Ontology database to determine their potential role in lipogenesis. Similar adaptive laboratory evolution (ALE) strategies will then be applied to other oleaginous microorganisms. To facilitate the transition to industrial-scale microbial oil production, the evolved strains will be cultivated in lab-scale bioreactors using selected agro-industrial residues as carbon sources.
The EvoSCO brochure can be consulted at the following link: https://evosco.unibuc.ro/wp-content/uploads/2026/01/EvoSCO_brochure.pdf