Long-Term Yeast Cultivation Coupled with In Situ Extraction for High Triterpenoid Production

Ginsenosides are a class of triterpenoids from the ginseng genus, with many medicinal properties. Traditionally, ginsenosides are extracted from ginseng plants to satisfy market demand; however, this approach requires substantial plant biomass and a lengthy six-year growth period before harvest. The advancement of synthetic biology allows the production of ginsenosides by engineered yeast. In this study, we combined our previously reported cultivation method with in situ extraction to enhance the production and exportation of intracellular ginsenosides by the engineered Saccharomyces cerevisiae. Remarkably, ginsenoside production reached as high as 3.4 g/L in a single shake flask, with almost 100% of ginsenosides in the organic phase. The "empty yeasts" were successfully reused 10 times in sequential cultivations. These findings are discussed in the context of cultivation intensification for natural product synthesis. Increasing the level of triterpenoid synthesis facilitates rapid development and supports the industrialization of this intriguing group of natural products.