一种进化的人造基循环酶可以通过H原子转移途径构建双循环类基架
Dongping Chen1,2,3, Xiang Zhang1,2,3, Anastassia Andreevna Vorobieva4,5
1Department of Chemistry, University of Basel, Basel, Switzerland.
Nature chemistry
|July 19, 2024
概括
研究人员使用斯特雷普塔维丁中的辅因子设计了一种人工基循环酶 (ARCase). 这种新型酶通过激素循环化有效地创建复杂的双循环类结构,具有高反体过量.
科学领域:
- 生物化学 生物化学
- 有机化学 有机化学
- 生物技术是生物技术.
背景情况:
- 天然的特类循环酶主要利用阴离子机制.
- 化物合成的激进循环化途径仍未得到充分探索.
- 金属催化H原子转移 (M-HAT) 反应为烯的水功能化提供了潜力.
研究的目的:
- 为了设计一个人造的金属酶,能够激进循环化为类合成.
- 通过将M-HAT反应性集成到蛋白质支架中来开发一个人工基循环酶 (ARCase).
主要方法:
- 在一个工程化仿制性链杆菌素中定一个生物化[Co ((Schiff-base) ]辅因子.
- 采用两轮的定向进化来优化酶活性.
- 使用X射线晶体学来确认辅因子结合和结构特征.
主要成果:
- 一个双重突变的ARCase成功催化了基结循环形成双循环产物.
- 对于cis-5-6融合环结构,ARCase实现了高反体过剩 (高达97%).
- 结晶学证实了胺与辅因子的结合.
- 时间过程实验揭示了一种级联反应,涉及激素循环和并联减少.
- 工程酶表现出基质对二烯变异的耐受性.
结论:
- 开发的ARCase为复杂的类脚手架提供了一条新的生物催化途径.
- 人工金属酶可以被设计成执行具有挑战性的激进反应.
- 这种方法扩大了生物催化剂的合成实用性,以获取各种化学结构.
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