探索De NovoNMN生物合成作为增强NMN生产的替代途径
Pengju Wang1, Yidan Ma2,1, Ju Li3
1Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
ACS synthetic biology
|July 18, 2024
概括
这项研究利用大肠杆菌 (Escherichia coli) 来增强尼古丁胺单核酸 (NMN) 的产生,达到3000微米以上. 这项研究探索了一种新的NMN生物合成路径,显著提高了潜在治疗应用的产量.
科学领域:
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
- 生物技术是生物技术.
背景情况:
- 尼古丁胺胺单核酸 (NMN) 是一种重要的NAD+前体,具有潜在的健康益处.
- 现有的NMN生物合成研究主要集中在尼古丁胺酸转移酶 (NadV) 途径上.
- 探索替代NMN生物合成路径对于提高生产效率至关重要.
研究的目的:
- 为了研究和优化一种新的NMN生物合成途径在大肠杆菌中.
- 为高产量NMN生产设计一个高效的大肠杆菌细胞工厂.
- 了解NAD+救援途径和能量代谢.
主要方法:
- 对大肠杆菌进行系统的基因工程,以提高NMN的合成和积累.
- 查参与新型NMN生物合成的酶,识别MazG.
- 优化和整合de novo和NadV介导的NMN通路.
- 引入载体以改善前体的吸收和产品的分泌.
- 用PRPP合成酶进行工程,以进一步提高产量.
主要成果:
- 通过初始工程实现了NMN产量的100倍以上的增加.
- 确定了MazG作为NAD+分裂成NMN的关键酶.
- 通过整合途径达到844.10±17.40微米的细胞内NMN度.
- 增强NMN产量至1293.73±61.38微米,并引入载体.
- 通过优化PRPP合成酶,实现了3067.98 ± 27.25微M的NMN生产记录.
结论:
- 成功建造了一家高效的大肠杆菌细胞工厂,用于高水平的NMN生产.
- 证明了 de novo NMN 生物合成路径的可行性和有效性.
- 提供了对NAD+代谢及其在大肠杆菌中的调节的新见解.
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