工程大肠杆菌 (Escherichia coli) 酸盐代谢产生非正规的减速功率
Derek Aspacio1, Emma Luu2, Suphanida Worakaensai1
1Department of Chemical and Biomolecular Engineering, University of California, Irvine, Irvine, California 92697-3900, United States.
概括
研究人员设计了大肠杆菌中的一个关键酶,以使用尼古丁胺胺 mononucleotide (NMN+) 作为替代的氧化还原辅因子. 这种创新克服了生物制造中的资源竞争,使新的工业应用成为可能.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 生物催化剂是一种生物催化剂.
背景情况:
- 生物制造依赖于对生物系统进行重新连接,以实现可持续的化学生产.
- 目前的生物制造受限于对原生氧化还原因子 (NAD(P) +) 的依赖,导致资源竞争.
- 非正规的氧化还原共因子,如尼古丁胺胺单核酸 (NMN+),为专用电子转移提供了解决方案.
研究的目的:
- 为了设计大肠杆菌的酸盐脱酶复合体 (PDHc),以利用NMN+作为氧化还原辅因子.
- 克服生物制造过程中本地辅助因子竞争的局限性.
- 为了扩大生物制造的工具包与一个绝缘的NMN+依赖的电子源.
主要方法:
- 对PDHc.的Lpd E3亚单元的理性和计算性酶设计.
- 定向进化和蛋白质工程来增强NMN+亲和力和特异性.
- 分子模拟来追踪辅因子结合的演变.
- 在大肠杆菌中工程 PDHc 的功能性表征.
主要成果:
- 发现了一种共因子乱交变体 (Lpd Penta),其NMN+周转率提高了2500倍.
- 设计了一个NMN+特定变体 (Lpd Ortho),对NMN+的特异性增加了2.4×10^5倍.
- 在大肠杆菌中表现出功能性NMN+依赖的PDHc活性,维持必要的pyruvate代谢.
结论:
- 成功设计了一种仅使用NMN+的PDHc,创建了一个绝缘电子通路.
- 这项工作扩展了NMN+辅助因子工具包用于生物制造应用.
- 设计的NMN+特定的PDHc为工业生物技术提供了高流量,不可逆转的电子源.
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