How directed evolution reshapes the energy landscape in an enzyme to boost catalysis.
How directed evolution reshapes the energy landscape in an enzyme to boost catalysis.
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进化如何将能量景观重新形成酶以增强催化作用。
DOI：
10.1126/science.abd3623
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发表时间：
2020-12-18
期刊：
影响因子：
--
通讯作者：
Kern D
中科院分区：
文献类型：
--
作者：
Otten R;Pádua RAP;Bunzel HA;Nguyen V;Pitsawong W;Patterson M;Sui S;Perry SL;Cohen AE;Hilvert D;Kern D
通过计算设计和实验室进化优化的生物催化剂的出现为探索增强催化功能的分子策略提供了机会。通过将一套核磁共振、晶体学和停流技术应用于为基本质子转移反应设计的酶，我们展示了定向进化如何逐渐改变蛋白质支架的构象整体，以填充狭窄的、高活性的构象整体，并实现近十亿倍的速率加速。优化过程中获得的突变使全局构象发生变化，包括高能主链重排，协同组织催化碱基和氧阴离子稳定剂，从而完善过渡态稳定性。在设计过程中对构象亚状态进行明确采样，特别是稳定所有非生产性构象的生产性，可以加速许多化学转化的蛋白质催化剂的开发。在多个时间尺度上改变构象亚状态的采样对于设计酶的优化至关重要。
The advent of biocatalysts designed computationally and optimized by laboratory evolution provides an opportunity to explore molecular strategies for augmenting catalytic function. Applying a suite of NMR, crystallographic, and stopped-flow techniques to an enzyme designed for an elementary proton transfer reaction, we show how directed evolution gradually altered the conformational ensemble of the protein scaffold to populate a narrow, highly active conformational ensemble and achieve a nearly billionfold rate acceleration. Mutations acquired during optimization enabled global conformational changes, including high-energy backbone rearrangements, that cooperatively organized the catalytic base and oxyanion stabilizer, thus perfecting transition-state stabilization. Explicit sampling of conformational sub-states during design, and specifically stabilizing productive over all unproductive conformations, could speed up the development of protein catalysts for many chemical transformations. Altered sampling of conformational sub-states on multiple time scales was critical for optimization of a designer enzyme.
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影响因子：
18.3
作者：
Kohen A
通讯作者：
Kohen A
DOI：
10.1039/c8cc02426j
发表时间：
2018-06-19
期刊：
Chemical communications (Cambridge, England)
影响因子：
--
作者：
Maria-Solano MA ;Serrano-Hervás E ;Romero-Rivera A ;Iglesias-Fernández J ;Osuna S
通讯作者：
Osuna S
影响因子：
7.8
作者：
Nashine, Vishal C.;Hammes-Schiffer, Sharon;Benkovic, Stephen J.
通讯作者：
Benkovic, Stephen J.
影响因子：
6.8
作者：
Campbell, Eleanor C.;Correy, Galen J.;Jackson, Colin J.
通讯作者：
Jackson, Colin J.
影响因子：
12.9
作者：
Kries, Hajo;Bloch, Joel S.;Hilvert, Donald
通讯作者：
Hilvert, Donald