Yarrowia lipolytica as a Platform for Punicic Acid Production

Punicic acid (PuA) is a polyunsaturated fatty acid with significant medical, biological, and nutraceutical properties. The primary source of punicic acid is the pomegranate seed oil obtained from fruits of trees that are mainly cultivated in subtropical and tropical climates. To establish sustainable production of PuA, various recombinant microorganisms and plants have been explored as platforms with limited efficiencies. In this study, the oleaginous yeast Yarrowia lipolytica was employed as a host for PuA production. First, growth and lipid accumulation of Y. lipolytica were evaluated in medium supplemented with pomegranate seed oil, resulting in the accumulation of lipids up to 31.2%, consisting of 22% PuA esterified in the fraction of glycerolipids. In addition, lipidengineered Y. lipolytica strains, transformed with the bifunctional fatty acid conjugase/desaturase from Punica granatum (PgFADX), showed the ability to accumulate PuA de novo. PuA was detected in both polar and neutral lipid fractions, especially in phosphatidylcholine and triacylglycerols. Promoter optimization for PgFADX expression resulted in improved accumulation of PuA from 0.9 to 1.8 mg/g of dry cell weight. The best-producing strain expressing PgFADX under the control of a strong erythritol-inducible promoter produced 36.6 mg/L PuA. These results demonstrate that the yeast Y. lipolytica is a promising host for PuA production.

Automated summary

In this study, the oleaginous yeast Yarrowia lipolytica was used as a host for the production of Punicic acid (PuA). By supplementing the medium with pomegranate seed oil, lipids containing PuA esters were accumulated up to 31.2% in Y. lipolytica. Strategies involving the expression of the bifunctional fatty acid conjugase/desaturase gene from Punica granatum (PgFADX) allowed for de novo production of PuA in Y. lipolytica. The optimized promoter for PgFADX expression resulted in a PuA production of 36.6 mg/L. These findings highlight the potential of Y. lipolytica as a promising host for PuA production.