发现具有Huisgenase活动的生物分子:设计重复蛋白作为 (3 + 2) 循环添加物的生物催化剂
Iván Rivilla1, Mikel Odriozola-Gimeno1, Antonio Aires2
1Department of Organic Chemistry I, Centro de Innovación en Química Avanzada (ORFEO-CINQA) , Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU) and Donostia International Physics Center (DIPC) , P° Manuel Lardizabal 3 , E-20018 Donostia/San Sebastián , Spain.
Journal of the American Chemical Society
|December 13, 2019
概括
工程蛋白质,称为Huisgenases,催化不自然的1,3-二极反应,创建新的尼特烯. 这些蛋白质提供了对反应结果的精确控制,证明了生物催化物的新前沿.
科学领域:
- 生物催化和蛋白质工程
- 有机合成和反应机制
- 计算化学
背景情况:
- 酶通常催化特定的自然反应.
- 1,3-双极循环添加是一种基本的有机反应.
- 在蛋白质中设计新的催化活动是一个重大挑战.
研究的目的:
- 设计和描述能够催化 imines 和双极蛋白之间的 1,3-双极反应的新型蛋白质.
- 研究这些工程蛋白质的催化机制和立体化学控制.
- 通过生物催化剂探索非自然的尼特罗普林的形成.
主要方法:
- 蛋白质的设计和重复蛋白质的构造.
- 核磁共振 (NMR) 光谱用于反应监测和结构分析.
- 位点定向的突变生成以探测催化残留物.
- 量子力学/分子力学 (QM/MM) 计算用于机械洞察.
主要成果:
- 设计的重复蛋白质成功催化了溶液中的1,3-双极循环添加反应.
- 酸性和基本性氨基酸残留物被确定为关键的催化成分.
- 蛋白质的灵活性和氨酸残留的精确排列影响了立体控制.
- 工程化蛋白质产生了不寻常的二聚体循环载荷管,形成了烯酸.
结论:
- 工程蛋白质可以被设计成催化不存在于自然界的反应,例如1,3-双极循环添加.
- 催化活性和立体选择性由特定的氨基酸残留物和蛋白质结构控制.
- 这项工作建立了一个新的生物催化剂 ("Huisgenases") 类,用于合成非自然化合物.
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