Developments in computational chemistry, bioinformatics, and laboratory evolution have facilitated the de novo design and catalytic optimization of enzymes. Besides creating useful catalysts, the generation and iterative improvement of designed enzymes can provide valuable insight into the interplay between the many phenomena that have been suggested to contribute to catalysis. In this work, we follow changes in conformational sampling, electrostatic preorganization, and quantum tunneling along the evolutionary trajectory of a designed Kemp eliminase. We observe that in the Kemp Eliminase KE07, instability of the designed active site leads to the emergence of two additional active site configurations. Evolutionary conformational selection then gradually stabilizes the most efficient configuration, leading to an improved enzyme. This work exemplifies the link between conformational plasticity and evolvability and demonstrates that residues remote from the active sites of enzymes play crucial roles in controlling and shaping the active site for efficient catalysis.

中文翻译：

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设计酶中多个活性位点配置的演变。

计算化学、生物信息学和实验室进化的发展促进了酶的从头设计和催化优化。除了创造有用的催化剂外，设计酶的产生和迭代改进可以提供有价值的洞察力，以了解许多被认为有助于催化的现象之间的相互作用。在这项工作中，我们沿着设计的 Kemp 消除酶的进化轨迹跟踪构象采样、静电预组织和量子隧穿的变化。我们观察到在 Kemp Eliminase KE07 中，设计的活性位点的不稳定性导致出现两个额外的活性位点配置。然后进化构象选择逐渐稳定最有效的构型，从而产生改进的酶。