Abolishing storage lipids induces protein misfolding and stress responses in Yarrowia lipolytica

Yarrowia lipolytica naturally saves carbon excess as storage lipids. Engineering efforts allow redirecting the high precursor flux required for lipid synthesis towards added-value chemicals such as polyketides, flavonoids, and terpenoids. To redirect precursor flux from storage lipids to other products, four genes involved in triacylglycerol and sterol ester synthesis (DGA1, DGA2, LRO1, ARE1) can be deleted. To elucidate the effect of the deletions on cell physiology and regulation, we performed chemostat cultivations under carbon and nitrogen limitation, followed by transcriptome analysis. We found that storage lipid-free cells show an enrichment of the unfolded protein response, and several biological processes related to protein refolding and degradation are enriched. Additionally, storage lipid-free cells show an altered lipid class distribution with an abundance of potentially cytotoxic free fatty acids under nitrogen limitation. Our findings not only highlight the importance of lipid metabolism on cell physiology and proteostasis, but can also aid the development of improved chassy strains of Y. lipolytica for commodity chemical production. Highlights Physiological and transcriptomic characterization of storage lipid free (Q4) strain Storage lipid free strain shows an increased free fatty acid fraction on nitrogen limitation Storage lipid free strain is more sensitive towards fatty acid supplementation Unfolded protein response, chaperones, and ubiquitin are enriched in the storage lipid free strain

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