Jiang Fenglin, Zhou Chen, Li Yan, Deng Haidong, Gong Ting, Chen Jingjing, Chen Tianjiao, Yang Jinling, Zhu Ping
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, NHC Key Laboratory of Biosynthesis of Natural Products, CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Acta Pharm Sin B. 2022 Jul;12(7):3167-3176. doi: 10.1016/j.apsb.2022.04.012. Epub 2022 Apr 27.
Both natural ginsenoside F2 and unnatural ginsenoside 3,20-Di--Glc-DM were reported to exhibit anti-tumor activity. Traditional approaches for producing them rely on direct extraction from , enzymatic catalysis or chemical synthesis, all of which result in low yield and high cost. Metabolic engineering of microbes has been recognized as a green and sustainable biotechnology to produce natural and unnatural products. Hence we engineered the complete biosynthetic pathways of F2 and 3,20-Di--Glc-DM in the CRISPR/Cas9 system. The titers of F2 and 3,20-Di--Glc-DM were increased from 1.2 to 21.0 mg/L and from 82.0 to 346.1 mg/L at shake flask level, respectively, by multistep metabolic engineering strategies. Additionally, pharmacological evaluation showed that both F2 and 3,20-Di--Glc-DM exhibited anti-pancreatic cancer activity and the activity of 3,20-Di--Glc-DM was even better. Furthermore, the titer of 3,20-Di--Glc-DM reached 2.6 g/L by fed-batch fermentation in a 3 L bioreactor. To our knowledge, this is the first report on demonstrating the anti-pancreatic cancer activity of F2 and 3,20-Di--Glc-DM, and achieving their biosynthesis by the engineered yeasts. Our work presents an alternative approach to produce F2 and 3,20-Di--Glc-DM from renewable biomass, which lays a foundation for drug research and development.
据报道，天然人参皂苷F2和非天然人参皂苷3,20 - 二 - β - 葡萄糖 - DM都具有抗肿瘤活性。传统的生产方法依赖于从植物中直接提取、酶催化或化学合成，所有这些方法都导致产量低和成本高。微生物代谢工程已被认为是一种生产天然和非天然产物的绿色可持续生物技术。因此，我们在CRISPR/Cas9系统中构建了F2和3,20 - 二 - β - 葡萄糖 - DM的完整生物合成途径。通过多步代谢工程策略，在摇瓶水平上，F2和3,20 - 二 - β - 葡萄糖 - DM的产量分别从1.2毫克/升提高到21.0毫克/升和从82.0毫克/升提高到346.1毫克/升。此外，药理学评价表明，F2和3,20 - 二 - β - 葡萄糖 - DM都具有抗胰腺癌活性，且3,20 - 二 - β - 葡萄糖 - DM的活性更好。此外，在3升生物反应器中通过补料分批发酵，3,20 - 二 - β - 葡萄糖 - DM的产量达到了2.6克/升。据我们所知，这是首次报道证明F2和3,20 - 二 - β - 葡萄糖 - DM的抗胰腺癌活性，并通过工程酵母实现其生物合成。我们的工作提出了一种从可再生生物质生产F2和3,20 - 二 - β - 葡萄糖 - DM的替代方法，为药物研发奠定了基础。
