工程恩布登-迈耶霍夫-帕纳斯糖解产生非正规的减速功率
Edward King1, Youtian Cui2, Derek Aspacio3
1Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California 92697-3900, United States.
概括
研究人员在Embden-Meyerhof-Parnas (EMP) 途径中设计了一个关键酶,以使用尼古丁胺胺单核酸 (NMN+) 而不是NADP+. 这就产生了NMN+依赖的生物制造和葡萄糖代谢的正交控制的降解功率.
科学领域:
- 生物技术和代谢工程 生物技术和代谢工程
- 酶工程是什么? 酶工程是什么?
- 合成生物学 合成生物学
背景情况:
- 像尼古丁胺胺单核酸 (NMN+) 这样的非正规辅助因子在生物制造中为天然辅助因子 (NAD(P) +) 提供了具有成本效益的替代品.
- 工程酶利用NMN+使代谢通路的正交控制成为可能,这对于先进的应用至关重要.
研究的目的:
- 从Streptococcus mutans中重新设计糖-3-酸脱酶 (GapN),以有效地利用NMN+.
- 为生物制造应用建立一个直角的Embden-Meyerhof-Parnas (EMP) 路径.
主要方法:
- 代理性设计和*Sm*GapN的突变发生,以改变辅因子的特异性.
- 生物化学测试以测量酶活性和辅因子偏好.
- 分子动力学模拟和残留网络分析以了解突变效应.
主要成果:
- 发现了具有高NMN+依赖活性的GapN Penta变体.
- 设计的GapN Ortho变体具有从NADP+到NMN+的辅因子特异性中的约3.4 × 10^6倍切换.
- 在大肠杆菌中证明了GapN Ortho的正交功能,使NMN+依赖的葡萄糖利用成为可能.
结论:
- 在改变辅因子特异性时,协调突变对于保持酶灵活性和催化活性至关重要.
- 提供了一个设计NMN+依赖酶的策略.
- 为生物制造开发直角EMP路径铺平了道路.
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