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具有非正规的有机催化机制的酶的设计和演变
Ashleigh J Burke1, Sarah L Lovelock1, Amina Frese1
1Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, Manchester, UK.
Nature
|May 28, 2019
概括
研究人员使用计算设计和实验室进化来设计了一种新型酶. 这种酶利用Nδ-甲基胺,一种改性氨基酸,有效地催化水解,克服了标准遗传密码的局限性.
科学领域:
- 生物催化和酶工程
- 合成生物学
- 计算化学
背景情况:
- 遗传密码的局限性限制了新型催化功能的酶设计.
- 在酶活性位点中的正规核友可以形成不反应的中间体,阻碍催化效率.
- 小分子有机催化提供了酶设计的机制策略.
研究的目的:
- 开发一种使用非正规氨基酸作为催化核友的水解酶.
- 通过使用Nδ-methylhistidine来克服酶设计中的正规核友的局限性.
- 通过遗传密码扩展来扩大酶催化反应的范围.
主要方法:
- 计算的酶设计与实验室进化相结合.
- 纳入Nδ-methylhistidine作为一个催化核友.
- 针对扩大基因编码的指导进化协议.
- 晶体分析以阐明催化机制.
主要成果:
- 使用Nδ-methylhistidine生成一个水解酶.
- 与自由的Nδ-甲基胺相比,酸的催化效率提高了9000倍.
- 通过定向进化过程中的晶体学研究确定了关键的催化适应.
- 已证明Nδ-methylhistidine是二甲胺的基因编码替代品.
结论:
- 通过防止乙酶中间体的形成,Nδ-甲基胺可实现有效的催化.
- 这种方法扩大了对有价值的化学转换的酶设计范围.
- 基因编码的Nδ-methylhistidine为设计新型生物催化剂提供了一个多功能平台.
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