Engineering Yarrowia lipolytica as a platform for synthesis of drop-in transportation fuels and oleochemicals.
Harnessing lipogenic pathways and rewiring acyl-CoA and acyl-ACP (acyl carrier protein) metabolism in Yarrowia lipolytica hold great potential for cost-efficient production of diesel, gasoline-like fuels, and oleochemicals. Here we assessed various pathway engineering strategies in Y. lipolytica toward developing a yeast biorefinery platform for sustainable production of fuel-like molecules and oleochemicals. Specifically, acyl-CoA/acyl-ACP processing enzymes were targeted to the cytoplasm, peroxisome, or endoplasmic reticulum to generate fatty acid ethyl esters and fatty alkanes with tailored chain length. Activation of endogenous free fatty acids and the subsequent reduction of fatty acyl-CoAs enabled the efficient synthesis of fatty alcohols. Engineering a hybrid fatty acid synthase shifted the free fatty acids to a medium chain-length scale. Manipulation of alternative cytosolic acetyl-CoA pathways partially decoupled lipogenesis from nitrogen starvation and unleashed the lipogenic potential of Y. lipolytica Taken together, the strategies reported here represent promising steps to develop a yeast biorefinery platform that potentially upgrades low-value carbons to high-value fuels and oleochemicals in a sustainable and environmentally friendly manner.
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
Peng
P
Xu
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
Kangjian
K
Qiao
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
Woo Suk
WS
Ahn
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139 gregstep@mit.edu.
Gregory
G
Stephanopoulos
D000213:Acyl Carrier Protein / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000378:metabolism; D000473:Alkanes / Q000378:metabolism; D056804:Biofuels / Q000032:analysis*; D003600:Cytosol / Q000378:metabolism; D004952:Esters / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D005233:Fatty Alcohols / Q000378:metabolism; D060847:Metabolic Engineering; D013347:Subcellular Fractions / Q000378:metabolism; D014186:Transportation; D025062:Yarrowia / Q000378:metabolism*
D000213:Acyl Carrier Protein; D000214:Acyl Coenzyme A; D000473:Alkanes; D056804:Biofuels; D004952:Esters; D005227:Fatty Acids; D005233:Fatty Alcohols
advanced biofuels; metabolic engineering; oleaginous yeast; oleochemicals; synthetic biology