Metabolic engineering of Yarrowia lipolytica for production of triacetic acid lactone

Climate change and global warming is driving development toward use of more sustainable production processes such as microbial manufacturing of renewable compounds. [i]Yarrowia lipolytica[/i] is a non-conventional, oleaginous yeast that can accumulate high amounts of lipids in response to stress conditions, and is therefore of special interest in the biotechnological industry. The goal with this study was to harness the inherent oleaginous nature of [i]Y. lipolytica[/i] for production of triacetic acid lactone (TAL), a potential biorenewable platform chemical that is similar to fatty acids in structure and synthesis. Several metabolic engineering approaches were taken to improve TAL titer and understand mechanisms driving TAL accumulation, including introduction of a mutant enzyme in the glycolysis pathway, disruption of an enzyme involved in precursor-production for lipids, and disruption of peroxisome biogenesis. Furthermore, optimal nitrogen and carbon concentrations for accumulation of TAL in [i]Y. lipolytica[/i] were examined. This study also presents a new method for elucidating gene essentiality using CRISPR-Cas9.