由含有Ir (Me) -PIX辅因子的P450细胞染色体催化的化学选择性酶性C-H键氨基化
Paweł Dydio1,2, Hanna M Key1,2, Hiroki Hayashi1,2
1Department of Chemistry, University of California , Berkeley, California 94720, United States.
Journal of the American Chemical Society
|January 13, 2017
概括
具有共因子的工程P450酶在C-H键氨基化中获得高产量和选择性,优于基于铁的系统. 这一突破通过具有挑战性的C-H功能化实现了复杂分子的有效催化合成.
科学领域:
- 生物催化
- 有机金属化学
- 酶工程
背景情况:
- 细胞染色体P450酶被设计用于非生物学C-H键氨基化.
- 低化学选择性和基质范围限制了当前的P450催化氨化反应.
- 亚基质的竞争性降解为硫胺是一种重大挑战.
研究的目的:
- 通过改进的化学选择性和更广泛的C-H氨基化基体范围来设计P450酶.
- 在P450催化氨基化中研究甲辅因子的使用.
- 开发有效的方法来识别改进的酶变体.
主要方法:
- 来自热友生物的工程P450与甲辅因子 (Ir(Me) -PIX.
- 催化硫酸的反选择性分子内C-H键氨基化.
- 在细胞溶解物中快速选含有Ir (Me) - PIX的CYP119突变物.
主要成果:
- 在C-H氨基化比亚化还要高的化学选择性 (> 20: 1) 得到.
- 在高达98%的产量下形成的产品,营业额 () 约为300.
- 已显示出高达95:5的抗选择性和异位选择性.
- 与含铁酶相比,基质范围更广泛.
结论:
- 与基于血红素的酶相比,含有P450的Ir(Me) -PIX在C-H氨基化方面具有更高的性能.
- 在细胞溶解物中的快速选加速了高活性酶变体的发现.
- 这项工作为发现具有挑战性的C-H氨化反应的酶提供了一个平台.
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