Background: Yarrowia lipolytica is a common biotechnological chassis for the production of lipids, which are the pre‑
ferred feedstock for the production of fuels and chemicals. To reduce the cost of microbial lipid production, inexpen‑
sive carbon sources must be used, such as lignocellulosic hydrolysates. Unfortunately, lignocellulosic materials often
contain toxic compounds and a large amount of xylose, which cannot be used by Y. lipolytica.
Results: In this work, we engineered this yeast to efciently use xylose as a carbon source for the production
of lipids by overexpressing native genes. We further increased the lipid content by overexpressing heterologous
genes to facilitate the conversion of xylose-derived metabolites into lipid precursors. Finally, we showed that these
engineered strains were able to grow and produce lipids in a very high yield (lipid content = 67%, titer = 16.5 g/L,
yield = 3.44 g/g sugars, productivity 1.85 g/L/h) on a xylose-rich agave bagasse hydrolysate in spite of toxic
compounds.
Conclusions: This work demonstrates the potential of metabolic engineering to reduce the costs of lipid production
from inexpensive substrates as source of fuels and chemicals.