Mitochondrial Engineering of Yarrowia lipolytica for Sustainable Production of a-Bisabolene from Waste Cooking Oil
ACS SUSTAINABLE CHEMISTRY & ENGINEERING(2022)
Abstract
Metabolic engineering of yeasts for terpenoid production has mostly focused on the cytoplasm, whereas harnessing their organelles as subcellular factories has been overlooked. Herein, the farnesyl diphosphate synthetic pathway and alpha-bisabolene synthase were compartmentalized into the oleaginous yeast Yarrowia lipolytica's mitochondria to enable high-level alpha-bisabolene production. Through comprehensive metabolic engineering approaches, we exploited the potential and capability of the mitochondria as a subcellular factory to achieve 257.4 mg/L of alpha-bisabolene production from glucose. By combining mitochondrial and cytoplasmic engineering, we further boosted the alpha-bisabolene titer to 765.1 mg/L by utilizing waste cooking oil as the sole carbon source. Finally, the alpha-bisabolene titer of the resulting strain reached 1058.1 mg/L in a 5 L bioreactor, which is the highest titer in the engineered Y. lipolytica cell factory reported to date. Overall, our study has provided valuable insights into the mitochondrial engineering of Y. lipolytica for sustainable and green production of valuable compounds.
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Key words
a-bisabolene,mitochondrial engineering,Yarrowia lipolytica,subcellular factory,waste cooling oil
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