Wang Guoli, Li Mingkai, Fan Bengui, Liang Xiqin, Wang Jun, Shi Yanbing, Zheng Qiusheng, Li Defang, An Tianyue
Featured Laboratory for Biosynthesis and Target Discovery of Active Components of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Binzhou Medical University, Yantai, 264003, China.
Microb Cell Fact. 2025 Apr 2;24(1):78. doi: 10.1186/s12934-025-02701-4.
The application of synthetic biology techniques has been recognized as an efficient alternative for the biosynthesis of high-value natural products, and various metabolic engineering strategies have been employed to develop microbial cell factories. However, exploration of more efficient metabolic pathway optimization strategies is still required to further improve the producing potential of microbial cell factories to meet the industrial requirements.
In this study, we found that the introduction of human N6,2'-O-dimethyladenosine (mAm) methyltransferase PCIF1 into Saccharomyces cerevisiae significantly promoted the biosynthesis of squalene, increased by 2.3-fold. Transcriptome analysis revealed that PCIF1 upregulated genes associated with glycolysis and acetyl-CoA biosynthesis pathways, and also activated the cell wall integrity mitogen-activated protein kinase (MAPK) pathway to improve the cell wall stress response. Importantly, PCIF1 expression notably enhanced squalene and sesquiterpenoid longifolene production in engineered yeast strains, with 2.3-fold and 1.4-fold higher increase, respectively.
This study presents a PCIF1-based metabolic engineering strategy that could serve as an effective approach for the optimization of terpene biosynthesis in yeast cell factories.
合成生物学技术的应用已被公认为是高价值天然产物生物合成的一种有效替代方法，并且已经采用了各种代谢工程策略来开发微生物细胞工厂。然而，仍需要探索更有效的代谢途径优化策略，以进一步提高微生物细胞工厂的生产潜力，满足工业需求。
在本研究中，我们发现将人N6,2'-O-二甲基腺苷（mAm）甲基转移酶PCIF1引入酿酒酵母中可显著促进角鲨烯的生物合成，产量提高了2.3倍。转录组分析表明，PCIF1上调了与糖酵解和乙酰辅酶A生物合成途径相关的基因，还激活了细胞壁完整性丝裂原活化蛋白激酶（MAPK）途径，以改善细胞壁应激反应。重要的是，PCIF1的表达显著提高了工程酵母菌株中角鲨烯和倍半萜长叶烯的产量，分别提高了2.3倍和1.4倍。
本研究提出了一种基于PCIF1的代谢工程策略，可作为优化酵母细胞工厂中萜类生物合成的有效方法。
