一个单点突变在的过氧化酶显著增强了对抗选择性
Eugene Antipov1, Art E Cho, Alexander M Klibanov
1Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Journal of the American Chemical Society
|July 21, 2009
概括
大多数胡卜过氧化酶 (HRP) 突变体表现出反抗选择性,与野生类型不同. 在Arg178Glu变体中,表现出显著的25倍偏好于奇拉类的 (S) - 反体.
科学领域:
- 生物催化剂是一种生物催化剂.
- 酵素工程是什么? 酶工程是什么
- 有机化学 有机化学
背景情况:
- 马过氧化酶 (HRP) 是一个广泛研究的酶.
- 在酶反应中的酶选择性对于合成性化合物至关重要.
- 修改酶活性部位可以改变基质特异性和酶选择性.
研究的目的:
- 为了研究HRP在位置178的突变对其反抗选择性的影响.
- 为了识别HRP变体,增强对奇拉性醇的酶选择性.
- 阐明观测到的对抗选择性变化背后的机制.
主要方法:
- 用局部定向突变生成来创建具有178.8位置突变的HRP变体.
- 酶氧化试验是使用各种性基质进行的.
- 动态分析和分子建模被用来研究酶基质相互作用和过渡状态.
主要成果:
- 大多数HRP突变体表现出酶选择性,与野生类型酶不同.
- 该Arg178Glu变体显示了最高的反抗选择性,对 (S) - 反抗体的偏好是25倍.
- 分子建模表明,Glu178和 (R) -enantiomer的过渡状态之间的静电排斥有助于对enantioselectivity.
结论:
- 在HRP的位置178的突变可以显著增强对奇拉性醇的酶选择性.
- Arg178Glu突变代表了对不对称合成的一个有希望的变体.
- 了解静电相互作用的作用为合理的酶设计提供了洞察力.
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