Zhang Ge, Ma Yurui, Huang Meina, Jia Kaizhi, Ma Ting, Dai Zongjie, Wang Qinhong
Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
National Center of Technology Innovation for Synthetic Biology, Tianjin 300308, China.
J Agric Food Chem. 2025 Jan 8;73(1):606-616. doi: 10.1021/acs.jafc.4c09874. Epub 2024 Dec 17.
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 . First, by regulation of the expression of and , 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 was eliminated by deleting , 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.
利用微生物宿主生产天然植物产品被认为是一种有前景的可持续方法。然而，利用微生物高效生产萜类化合物仍然是一项重大挑战。在此，萜类化合物的一个构建模块甲羟戊酸被用作示范产品，以探索酿酒酵母中萜类生物合成的潜在代谢限制。首先，通过调节ERG10和ERG13的表达，甲羟戊酸滴度提高了7660%。随后，增强了天然甲羟戊酸途径（MVA途径），甲羟戊酸产量增加到4.16 g/L。为确保乙酰辅酶A的充足供应，同时对柠檬酸途径和三羧酸循环进行工程改造，在摇瓶中甲羟戊酸滴度进一步提高到5.25 g/L。最终，通过删除YAT1消除了酿酒酵母的柠檬酸溢流代谢，在真核生物中实现了101 g/L的最高甲羟戊酸滴度，葡萄糖产率为0.255 g/g。这些见解可应用于有效生产源自中心碳代谢途径的萜类化合物和生物化学品。
