一个非正规的核友解锁了一个新的机械路径在一个设计的酶
Amy E Hutton1, Jake Foster1, Rebecca Crawshaw1
1Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester, Manchester, UK.
Nature communications
|March 4, 2024
概括
设计酶的定向进化揭示了莫里塔-贝利斯-希尔曼 (MBH) 反应的新途径. 用Nδ-methylhistidine取代histidine,通过启用一种新的质子转移机制,创造了一个高度活性的酶.
科学领域:
- 蛋白质工程是一种蛋白质工程.
- 生物催化剂是一种生物催化剂.
- 酶进化 酶进化的过程
背景情况:
- 计算机设计的酶提供了关于催化部位出现的见解.
- 在此之前,丁核和氨酸加速了莫里塔-贝利斯-希尔曼 (MBH) 反应.
- 莫里塔-贝利斯-希尔曼反应是一个关键的碳-碳键形成反应.
研究的目的:
- 研究用Nδ-methylhistidine替换催化性histidine对酶进化和活性的影响.
- 了解由酶活性位点微妙变化引起的机械变化.
- 为了探索设计酶的新进化途径.
主要方法:
- 计算设计的酶的定向进化.
- 局部定向的突变发生引入Nδ-甲基胺.
- 酶活性测定用于测量催化效率.
- 机械学研究以阐明质子转移步骤.
主要成果:
- 用Nδ-methylhistidine替换histidine导致了一个新的催化残留物 (Glu26) 的出现,并放弃了 arginine.
- 由此产生的酶 (BHMeHis1.8) 在MBH反应中表现出一个数量级更高的活性.
- 在进化的早期阶段,Nδ-methylhistidine核对解锁新的机械路径至关重要.
- 一种含有histidine的BHMeHis1.8变种保留了显著的MBH生物催化剂活性.
结论:
- 设计的酶活性位点的微妙修改可以大大改变进化轨迹.
- 酶进化可以为复杂的化学转换提供新的机械解决方案.
- 在酶工程中,Nδ-methylhistidine可以作为有效的核,使新的催化途径成为可能.
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