Lipid accumulation from glucose and xylose in an engineered, naturally oleaginous strain of Saccharomyces cerevisiae

Saccharomyces cerevisiae, a well-known industrial yeast for alcoholic fermentation, is not historically known to accumulate lipids. Four S. cerevisiae strains used in industrial applications were screened for their ability to accumulate neutral lipids. Only one, D5A, was found to accumulate up to 20% dry cell weight (dcw) lipids. This strain was further engineered by knocking out ADP-activated serine/threonine kinase (SNF1) which increased lipid accumulation to 35% dcw lipids. In addition, we engineered D5A to utilize xylose and found that D5A accumulates up to 37% dcw lipids from xylose as the sole carbon source. Further we over-expressed different diacylglycerol acyltransferase (DGA1) genes and boosted lipid accumulation to 50%. Fatty acid speciation showed that 94% of the extracted lipids consisted of 5 fatty acid species, C16:0 (palmitic), C16:1n7 (palmitoleic), C18:0 (stearic), C18:1n7 (vaccenic), and C18:1n9 (oleic), while the relative distributions changed depending on growth conditions. In addition, this strain accumulated lipids concurrently with ethanol production.