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通过立体化学探针研究脱酶的基质范围和反选择性

Jian-Bo Wang^1,2, Adriana Ilie^1,2, Shuguang Yuan³

¹Department of Chemistry, Philipps-University , Hans-Meerwein-Strasse 4, 35032 Marburg, Germany.

Journal of the American Chemical Society

|July 26, 2017

概括

酸脱酶FAcD (RPA1163) 使用经典的S_N2机制,而不是双瓦尔登反转. 这种酶对 (S) 基质具有很高的选择性,在动态分辨率方面具有合成价值.

科学领域:

生物化学
酵素学
有机化学

背景情况:

酸脱酶 (FAcD) 是一种在生物催化中具有潜在应用的酶.
了解FAcD的精确反应机制对于优化其使用至关重要.

研究的目的:

研究酸脱酶FAcD (RPA1163) 的立体化学机制.
要确定FAcD是否通过双瓦尔登倒置或经典S_N2机制运行.

主要方法:

通过FAcD催化的一种立体化学探针,rac-2-fluoro-2-phenyl乙酸进行脱过程.
采用包括分子建模 (MM) 和分子动力学 (MD) 在内的计算方法来分析基质接受和立体选择性.

主要成果:

这项研究证明了配置的反转,表明了涉及Asp110的经典S_N2机制.
观察到极高的选择性因子 (E > 500),有利于 (S) 基质.
该酶表现出广泛的基质范围,甚至有效地处理像rac-2-fluoro-2-benzyl乙酸这样的硬质要求很高的基质.

结论:

FAcD (RPA1163) 是通过经典的S_N2机制进行的,而不是双反转.
该酶对 (S) 基质的高选择性使其在合成应用中具有价值.
计算分析提供了关于酸本身的基质结合和催化机制的见解.

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