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蛋白质工程和双模块优化,以在E.E.中有效生产NMN. 大肠杆菌
Xu Ma1, Qiang Wang1, Kewei Chen2
1Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
Journal of agricultural and food chemistry
|April 2, 2025
概括
研究人员开发了一种新型的双模块系统,用于高效的尼古丁胺胺单核酸 (NMN) 生物合成. 这种工程途径通过尿素和尼古丁胺胺增强NMN生产,为NAD+补充剂开发提供了一个有前途的途径.
科学领域:
- 生物技术是生物技术.
- 酶合成酶的合成
- 代谢工程是代谢工程.
背景情况:
- 尼古丁胺胺 mononucleotide (NMN) 是一个重要的NAD+前体,作为补充剂具有显著的兴趣.
- 现有的NMN合成方法在效率和可扩展性方面面临挑战.
研究的目的:
- 为NMN生物合成设计一种新的,高效的双模块酶系统.
- 提高NMN生产中关键酶的稳定性和活性.
- 通过使用易于获得的基板,优化系统以实现高转换率.
主要方法:
- 从尿素和尼古丁胺胺合成NMN的两模块酶反应系统的构建.
- 通过结构和进化分析,设计一种更稳定的尼古丁胺里博酸激酶 (NRK) 突变物 (KlmNRK_M4).
- 在大肠杆菌中淘汰内源性降解酶基因以改善基质和中间体转化.
- 对双模块系统的反应条件的优化.
主要成果:
- 在模块1中使用NRK和ATP再生系统实现了从尼古丁胺 рибоoside (NR) 合成NMN的高效合成.
- 在模块2中,通过NRK和胺核酸酸化酶 (PyNP) 的协同催化,从尿素和尼古丁胺胺成功合成了NMN.
- 获得了一个高度稳定的KlmNRK_M4突变体,具有改善的酶特性.
- 在优化条件下使用300毫米的尿素和尼古丁胺胺,达到81.1%的高NMN转化率.
结论:
- 开发的双模块重组系统为NMN生物合成提供了一条新且高效的途径.
- 酶工程和代谢途径优化显著提高NMN生产效率.
- 这项研究为NMN的工业生产提供了一个可扩展和具有成本效益的方法.
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