How directed evolution reshapes the energy landscape in an enzyme to boost catalysis

doi:10.1126/science.abd3623

GSTDTAP > 气候变化

DOI	10.1126/science.abd3623
	How directed evolution reshapes the energy landscape in an enzyme to boost catalysis
	Renee Otten; Ricardo A. P. Pádua; H. Adrian Bunzel; Vy Nguyen; Warintra Pitsawong; MacKenzie Patterson; Shuo Sui; Sarah L. Perry; Aina E. Cohen; Donald Hilvert; Dorothee Kern
	2020-12-18
发表期刊	Science
出版年	2020
英文摘要	Whether designed computationally or uncovered in activity screening, enzymes repurposed for biocatalysis rarely start at the peak of proficiency. However, directed evolution can in some cases increase catalytic efficiency of a poor enzyme by many orders of magnitude. Otten et al. used a suite of biochemical techniques to investigate the origins of rate enhancement in a previously evolved model enzyme. Two conformational states are present in the initial, computationally designed enzyme, but only one is active. Shifting the population toward the active state is one factor in increasing catalytic efficiency during evolution. Single mutations do not greatly increase activity, but the synergistic combination of just two out of 17 substitutions can provide most of the rate enhancement seen in the final, evolved enzyme. Science , this issue p. [1442][1] 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 nuclear magnetic resonance, crystallography, 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 accelerate this transformation by nearly nine orders of magnitude. 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. The development of protein catalysts for many chemical transformations could be facilitated by explicitly sampling conformational substates during design and specifically stabilizing productive substates over all unproductive conformations. [1]: /lookup/doi/10.1126/science.abd3623
领域	气候变化 ; 资源环境
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文献类型	期刊论文
条目标识符	http://119.78.100.173/C666/handle/2XK7JSWQ/308362
专题	气候变化资源环境科学
推荐引用方式 GB/T 7714	Renee Otten,Ricardo A. P. Pádua,H. Adrian Bunzel,et al. How directed evolution reshapes the energy landscape in an enzyme to boost catalysis[J]. Science,2020.
APA	Renee Otten.,Ricardo A. P. Pádua.,H. Adrian Bunzel.,Vy Nguyen.,Warintra Pitsawong.,...&Dorothee Kern.(2020).How directed evolution reshapes the energy landscape in an enzyme to boost catalysis.Science.
MLA	Renee Otten,et al."How directed evolution reshapes the energy landscape in an enzyme to boost catalysis".Science (2020).