Engineering of a high lipid producing Yarrowia lipolytica strain
Author(s)Friedlander, Jonathan; Tsakraklides, Vasiliki; Kamineni, Annapurna; Greenhagen, Emily H.; Consiglio, Andrew L.; MacEwen, Kyle; Crabtree, Donald V.; Afshar, Jonathan; Nugent, Rebecca L.; Hamilton, Maureen A.; Shaw, A. Joe; South, Colin R.; Stephanopoulos, Gregory; Brevnova, Elena E.; ... Show more Show less
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Background: Microbial lipids are produced by many oleaginous organisms including the well-characterized yeast Yarrowia lipolytica, which can be engineered for increased lipid yield by up-regulation of the lipid biosynthetic pathway and down-regulation or deletion of competing pathways. Results: We describe a strain engineering strategy centered on diacylglycerol acyltransferase (DGA) gene overexpression that applied combinatorial screening of overexpression and deletion genetic targets to construct a high lipid producing yeast biocatalyst. The resulting strain, NS432, combines overexpression of a heterologous DGA1 enzyme from Rhodosporidium toruloides, a heterlogous DGA2 enzyme from Claviceps purpurea, and deletion of the native TGL3 lipase regulator. These three genetic modifications, selected for their effect on lipid production, enabled a 77 % lipid content and 0.21 g lipid per g glucose yield in batch fermentation. In fed-batch glucose fermentation NS432 produced 85 g/L lipid at a productivity of 0.73 g/L/h. Conclusions: The yields, productivities, and titers reported in this study may further support the applied goal of cost effective, large -scale microbial lipid production for use as biofuels and biochemicals. Keywords: Yarrowia lipolytica, Lipid accumulation, Oleaginous yeast, Metabolic engineering
DepartmentMassachusetts Institute of Technology. Department of Chemical Engineering
Biotechnology for Biofuels
Friedlander, Jonathan, Vasiliki Tsakraklides, Annapurna Kamineni et al. "Engineering of a high lipid producing Yarrowia lipolytica strain" Biotechnology for Biofuels, 9,77 (March 2016): p. 1-12. © 2016 Friedlander et al.
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