Improved de novo production of prostaglandin F2α from glucose with engineered Yarrowia lipolytica strains

Prostaglandin F_2α (PGF_2α) is an important lipid mediator with diverse physiological activities, but its reliance on chemical synthesis presents challenges and limitations, driving the need for efficient microbial fermentation. This study engineered the oleaginous yeast Yarrowia lipolytica to establish a microbial platform for the de novo biosynthesis of PGF_2α from glucose. Firstly, in vitro biotransformation of arachidonic acid (ARA) was achieved using engineered Y. lipolytica expressing prostaglandin H synthase and prostaglandin F synthase. Subsequently, the complete biosynthetic pathway was reconstituted in vivo. To overcome the rate-limiting Δ-8 desaturation step, a high-efficiency Δ-8 desaturase from I. galbana was selected, and a Δ9-elongase/Δ8-desaturase fusion protein with a GGGGS linker was constructed. Further systematic metabolic engineering including dual organelle engineering targeting the endoplasmic reticulum and lipid droplets, and enhanced acetyl-CoA supply via carnitine O-acetyltransferase (Cat2) overexpression - a strategy that proved more effective than regulating NADPH generation. Finally, fermentation optimization with the optimized YAO medium significantly boosted PGF_2α production. These combined metabolic engineering strategies enabled Y. lipolytica de novo production of PGF_2α from glucose, reaching a titer of 32.63 μg/L (nearly 3.58 μg/g DCW). Overall, this research offers a promising platform for advancing the green biosynthesis of PGF_2α and other prostaglandins.