Multimodule Synthetic\nRedesign of Intracellular Metabolisms\nfor the High-Titer de Novo Production of Sakuranetin in Yarrowia lipolytica

Sakuranetin, a flavonoid phytoalexin, has demonstrated\nneuroprotective\nproperties and exhibits tyrosinase inhibitory activities, making it\nhighly valuable in the cosmetics and pharmaceutical industries. In\nthis study, we engineered a Yarrowia lipolytica strain for the high-titer de novo production of sakuranetin using\nglucose as a substrate. To effectively enhance sakuranetin production,\nwe implemented a multimodule engineering strategy that included optimizing\nthe sakuranetin synthesis pathway, designing a regeneration system\nfor the methyl donor S-adenosyl methionine, increasing the malonyl-CoA\nprecursor supplement, and constructing the feedback inhibition-relieved\nshikimate pathway. Moreover, a transcriptomic analysis was conducted\nto identify potential targets for further improving sakuranetin synthesis.\nAs a result, the titer of de novo synthesized sakuranetin reached\n344.0 mg/L from glucose in a 5 L bioreactor. These achievements hold\nsignificant promise for the sustainable and large-scale production\nof sakuranetin through industrial biomanufacturing.