Efficient Production\nof the Colorless Carotenoid Phytoene\nin Yarrowia lipolytica through Metabolic\nEngineering

Phytoene, a colorless carotenoid with unique ultraviolet\n(UV)–B\nabsorption properties, offers potential for applications in functional\nfood, cosmetics, and therapeutics. However, their low natural yield\nposes a challenge for large-scale production. This study aims to\nenhance phytoene production in the oleaginous yeast Yarrowia lipolytica by introducing a heterologous\nphytoene synthase gene combined with metabolic engineering approaches.\nWe enhanced phytoene synthesis by overexpressing key genes in the\nmevalonate pathway and compartmentalizing the biosynthetic pathway\nwithin peroxisomes. Moreover, we inhibited the glyoxylate cycle to\nincrease the accumulation of peroxisomal acetyl-CoA available for\nphytoene production. Our engineered strains demonstrated a significant\nincrease in phytoene production, reaching up to 1.34 g/L titer and\n58.74 mg/gDCW yield in the flask-scale fed-batch culture, which are\nthe highest levels reported to date. These results underscore the\npotential of Y. lipolytica as a robust\nplatform for producing phytoenes and other terpenoids on an industrial\nscale, offering valuable insights for future efforts in metabolic\nengineering.