酶设计中的二次氨基催化:通过遗传密码扩展扩大蛋白质模板多样性
Thomas L Williams1, Irshad M Taily1, Lewis Hatton1
1School of Chemistry and Cardiff Catalysis Institute, Cardiff University, Main Building, Park Place, Cardiff, CF10 3AT, United Kingdom.
Angewandte Chemie (International ed. in English)
|March 28, 2024
概括
遗传密码的扩展使得二次胺能够被纳入各种蛋白质中,从而产生新的人工酶. 这一突破扩大了生物催化剂和进化研究的酶设计可能性.
科学领域:
- 生物化学 生化学
- 合成生物学 合成生物学
- 酶工程是什么? 酶工程是什么?
背景情况:
- 二次胺是有反应性的,可以制造人工酶.
- 目前用于将二次胺纳入蛋白质的方法范围有限.
- 遗传密码扩展提供了一种克服这些局限性的方法.
研究的目的:
- 开发一种方法,利用遗传密码扩展将二次胺纳入蛋白质的所需位置.
- 创建具有转移化活性的新型人工酶.
- 探索这些工程酶在生物催化剂中的潜力.
主要方法:
- 利用遗传密码扩展将二次氨基酸与二次氨基酸相似物纳入蛋白质中.
- 工程超级绿色光蛋白 (sfGFP),LmrR和二叶酸还原酶 (DHFR) 的变体.
- 评估工程蛋白质的蛋白质溶解稳定性和催化活性.
主要成果:
- 成功地将二次胺纳入 sfGFP,LmrR 和 DHFR.
- 工程设计的LmrR和DHFR变种表现出转移化活性.
- 一种含有D-proline的模拟物显示出蛋白质溶解稳定性和催化功能.
- DHFR变体显示使用NADPH作为化物源的立体选择性反应.
- 基于DHFR的催化被整合到一个酶循环计划中.
结论:
- 遗传密码扩展提供了一个多功能平台,用于用二次胺来设计人工酶.
- 蛋白质模板选择影响核选择和催化效率.
- 开发的方法在酶设计,进化和生物催化剂开发方面具有广泛的应用.
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