Utilization of microbial hosts to produce natural plant products is regarded as a promising and sustainable approach. However, achieving highly efficient production of terpenoids using microorganisms remains a significant challenge. Here, mevalonate, a building block of terpenoids, was used as a demo product to explore the potential metabolic constraints for terpenoid biosynthesis in Yarrowia lipolytica. First, by regulation of the expression of ERG12 and HMGR, the mevalonate titer was improved by 7660%. Subsequently, the native mevalonate pathway (MVA pathway) was enhanced, and the production of mevalonate increased to 4.16 g/L. To ensure a sufficient supply of acetyl-CoA, the citrate route and TCA cycle were simultaneously engineered, and the mevalonate titer was further improved to 5.25 g/L in shake flasks. Ultimately, the citrate overflow metabolism of Y. lipolytica was eliminated by deleting CEX1, resulting in the highest mevalonate titer of 101 g/L with a yield of 0.255 g/g of glucose in eukaryotes. These insights could be applied to the effective production of terpenoids and biochemicals derived from central carbon metabolic pathways.

中文翻译：

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重编程酵母的碳代谢以高生成甲羟戊酸盐，甲羟戊酸盐是萜类化合物骨架的构建前体


利用微生物宿主生产天然植物产品被认为是一种有前途且可持续的方法。然而，利用微生物实现高效生产萜类化合物仍然是一项重大挑战。在这里，甲羟戊酸钠是萜类化合物的组成部分，被用作演示产品，以探索解散亚蓍草中萜类化合物生物合成的潜在代谢限制。首先，通过调节 ERG12 和 HMGR 的表达，甲羟戊酸滴度提高了 7660%。随后，天然甲羟戊酸通路 （MVA 通路） 增强，甲羟戊酸的产量增加到 4.16 g/L。为确保乙酰辅酶 A 的充足供应，同时设计了柠檬酸盐路线和 TCA 循环，并在摇瓶中将甲羟戊酸滴度进一步提高至 5.25 g/L。最终，通过删除 CEX1 消除了 Y. lipolytica 的柠檬酸盐溢出代谢，导致真核生物中甲羟戊酸滴度最高，为 101 g/L，葡萄糖产量为 0.255 g/g。这些见解可以应用于有效生产源自中央碳代谢途径的萜类化合物和生化物质。