(472c) Engineering Oleaginous Yeast Debaryomyces Hansenii Cbs 767 for Production of Fatty Acid-Derived Biofuels and Chemicals

The nonconventional yeast D. hansenii CBS 767 has attractive catabolic, anabolic, and tolerance phenotypes. It is osmotolerant and halotolerant, which confers resistance to harsh fermentation conditions and enables growth in saltwater. It naturally grows on the major monosaccharides of lignocellulosic biomass, specifically glucose, xylose, and arabinose. It exhibits a propensity for high density cell growth coupled with significant lipid accumulation. Here, we report the metabolic engineering of the CTG clade yeast D. hansenii for producing fatty acid-derived chemicals including alkanes and hexadecanol. Our developed yeasts modular cloning systems enables us to perform combinatorial metabolic pathway engineering of the D. hansenii fatty acid pathway. We targeted both acetyl-CoA overproduction and heterologous pathways to produce fatty-acid derived molecules. We report production of these molecules on the major constituents of lignocellulosic biomass – glucose, xylose, and arabinose. We obtain engineered D. hansenii produces heptadecane at the titer of 38.31 mg/L and hexadecanol at the titer of 102.06 mg/L by only overexpressing the short heterologous pathway. Thus, these strains have potential as cell factories for commodity chemicals from lignocellulose. In summary, this work highlights the potential for oleaginous yeasts D. hansenii in the biobased production of fatty acid-derived chemicals.