title: Engineering of a high lipid producing Yarrowia lipolytica strain.
abstract: 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.
authors.affiliation: Total New Energies, 5858 Horton Street, Emeryville, CA 94610 USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Friedlander
authors.affiliation: Total New Energies, 5858 Horton Street, Emeryville, CA 94610 USA.
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Tsakraklides
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Annapurna
authors.initials: A
authors.lastname: Kamineni
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Emily H
authors.initials: EH
authors.lastname: Greenhagen
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Consiglio
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Kyle
authors.initials: K
authors.lastname: MacEwen
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Donald V
authors.initials: DV
authors.lastname: Crabtree
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Afshar
authors.affiliation: Total New Energies, 5858 Horton Street, Emeryville, CA 94610 USA ; Twist Bioscience, 455 Mission Bay Blvd South, San Francisco, CA 94158 USA.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Nugent
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Maureen A
authors.initials: MA
authors.lastname: Hamilton
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: A
authors.initials: A
authors.lastname: Joe Shaw
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA.
authors.forename: Colin R
authors.initials: CR
authors.lastname: South
authors.affiliation: Novogy, Inc., 85 Bolton Street, Cambridge, MA 02140 USA ; Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139 USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: Stephanopoulos
authors.affiliation: Total New Energies, 5858 Horton Street, Emeryville, CA 94610 USA ; Evelo Therapeutics, 620 Memorial Dr., Cambridge, MA 02139 USA.
authors.forename: Elena E
authors.initials: EE
authors.lastname: Brevnova
keywords: Lipid accumulation; Metabolic engineering; Oleaginous yeast; Yarrowia lipolytica