作为微生物细胞工厂设计的大肠杆菌用于De Novo 6'-Sialyllactose生物合成
Yifan Liu1,2, Xiangsong Chen1, Xinyang Lv1,2
1Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China.
Journal of agricultural and food chemistry
|June 10, 2025
概括
研究人员对大肠杆菌进行了基因工程,以产生6'-sialyllactose (6'-SL),这是一种关键的人类牛奶寡糖. 这种代谢工程方法实现了显著的生产量,为6'-SL.的工业规模生物合成建立了强大的平台.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 微生物生物技术 微生物生物技术
背景情况:
- 6 -乳糖 (6 -SL) 是母乳的关键成分,对婴儿发育至关重要.
- 工业规模的6 -SL生产具有挑战性,但对于各种应用非常理想.
- 微生物细胞工厂为复杂碳水化合物的可持续和高效生物合成提供了一个有希望的途径.
研究的目的:
- 为了建立一个新的生物合成途径为6 -SL在埃舍里奇亚大肠杆菌.
- 通过代谢工程优化微生物菌株,以提高6 -SL的产生.
- 为6 -SL.开发一个可扩展的生物制造平台.
主要方法:
- 关键合成基因 (neuB,neuC,neuA,α2,6-SiaT) 在单种等离子体系统中的共同表达.
- 基因无活化 (nanA,nanK,nanE,nanT) 创建一个高性能的大肠杆菌底盘.
- 系统评估和选择最优的α2,6-sialyltransferase变体.
- 代谢工程策略包括促进物优化和核糖体结合部位工程.
- 在生物反应器中进行大规模发酵.
主要成果:
- 工程化大肠杆菌菌株DAT01在摇瓶中实现了1.36g/L的6 -SL.
- 鉴定bst*为最佳的α2,6-sialyltransferase导致DAT03菌株的生产率为1.67g/L.
- 在促进剂和RBS优化后,DAT07菌株在摇瓶中达到3.42g/L的6 -SL.
- 规模化的发酵在85小时内产生了30.18 g/L的6 -SL (0.36 g/L/h的体积生产率).
结论:
- 一个模块化代谢工程战略有效地为6 -SL生物生产建立了一个强大的平台.
- 改造的大肠杆菌菌株显示出6 -SL的工业规模生物合成的巨大潜力.
- 这项工作为开发微生物细胞工厂用于复杂碳水化合物生产提供了经过验证的框架.
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