Strengthening Yarrowia lipolytica as an Industrial Chassis through Metabolic Engineering for Pretreated Lignocellulosic Residue Utilization and a Library of Organelle Targeting Peptides

Abstract

This thesis advances Yarrowia lipolytica as a more versatile platform for biomanufacturing. We demonstrated robust growth and lipid production in pretreated lignocellulosic hydrolysates, supporting the use of low-cost renewable feedstocks. We then identified key constraints in xylose utilization and developed strategies based on adaptive evolution, heterologous transporters, and an alternative metabolic pathway to improve its conversion. Finally, we established a workflow for developing an organelle-targeting toolkit, enabling more precise localization of enzymes within the cell. Together, these advances improve feedstock flexibility, carbon utilization, and spatial control of metabolism, expanding the engineering potential of Y. lipolytica.