Engineering an efficient and enantioselective enzyme for the Morita-Baylis-Hillman reaction.
Engineering an efficient and enantioselective enzyme for the Morita-Baylis-Hillman reaction.
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DOI：
10.1038/s41557-021-00833-9
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发表时间：
2022-03
期刊：
影响因子：
21.8
通讯作者：
Green AP
中科院分区：
文献类型：
--
作者：
Crawshaw R;Crossley AE;Johannissen L;Burke AJ;Hay S;Levy C;Baker D;Lovelock SL;Green AP
计算设计和定向进化的结合可以为创造具有新功能的酶提供一种通用策略。迄今为止，这种方法已经为一些模型反应提供了酶。在这里，我们展示了新的催化机制可以被设计成蛋白质来加速更具挑战性的化学转化。原始设计的进化优化为Morita-Baylis-Hillman （MBH）反应提供了高效的对映选择性酶（BH32.14）。BH32.14适用于制备规模转化，接受广泛的醛和烯酮偶联伙伴，并能够促进二醛的选择性单官能化。晶体学、生化和计算研究表明BH32.14通过一个复杂的催化机制起作用，该机制包括一个His23亲核试剂与一个合适位置的Arg124配对。这种催化精氨酸在构象状态之间穿梭，以稳定多个氧阴离子中间体，并作为一种遗传编码的双齿氢键催化剂（如硫脲）的替代品。这项研究表明，精细的催化装置可以从零开始构建，以促进在自然界中未观察到的苛刻的多步骤过程。
The combination of computational design and directed evolution could offer a general strategy to create enzymes with new functions. To date, this approach has delivered enzymes for a handful of model reactions. Here we show that new catalytic mechanisms can be engineered into proteins to accelerate more challenging chemical transformations. Evolutionary optimization of a primitive design afforded an efficient and enantioselective enzyme (BH32.14) for the Morita-Baylis-Hillman (MBH) reaction. BH32.14 is suitable for preparative scale transformations, accepts a broad range of aldehyde and enone coupling partners, and is able to promote selective mono-functionalizations of dialdehydes. Crystallographic, biochemical and computational studies reveal that BH32.14 operates via a sophisticated catalytic mechanism comprising a His23 nucleophile paired with a judiciously positioned Arg124. This catalytic arginine shuttles between conformational states to stabilize multiple oxyanion intermediates and serves as a genetically encoded surrogate of privileged bidentate hydrogen bonding catalysts (e.g. thioureas). This study demonstrates that elaborate catalytic devices can be built from scratch to promote demanding multi-step processes not observed in Nature.
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