气候变化领域集成服务门户(CCPortal): How directed evolution reshapes the energy landscape in an enzyme to boost catalysis

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DOI	10.1126/science.abd3623
	How directed evolution reshapes the energy landscape in an enzyme to boost catalysis
	Otten R.; Pádua R.A.P.; Bunze H.A.; Nguyen V.; Pitsawong W.; Patterson M.; Sui S.; Perry S.L.; Cohen A.E.; Hilvert D.; Kern D.
发表日期	2020
ISSN	0036-8075
起始页码	1442
结束页码	1446
卷号	370 期号:6523
英文摘要	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 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. © 2020 American Association for the Advancement of Science. All rights reserved.
英文关键词	enzyme; oxyanion stabilizer; protein; stabilizing agent; unclassified drug; enzyme; catalysis; energy efficiency; enzyme; evolution; mutation; optimization; Article; biocatalyst; catalysis; complex formation; conformational transition; crystal structure; crystallization; energy; enzyme conformation; evolution; nuclear magnetic resonance; priority journal; process optimization; protein stability; structure analysis; biocatalysis; chemistry; computer aided design; directed molecular evolution; enzyme active site; genetics; protein conformation; Biocatalysis; Catalytic Domain; Computer-Aided Design; Directed Molecular Evolution; Enzymes; Nuclear Magnetic Resonance, Biomolecular; Protein Conformation; Proteins
语种	英语
来源期刊	Science
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/244962
作者单位	Howard Hughes Medical Institute and Department of Biochemistry, Brandeis University, Waltham, MA 02454, United States; Laboratory of Organic Chemistry, Eth Zürich, Zürich, 8093, Switzerland; Department of Chemical Engineering, Institute of Applied Life Sciences, University of Massachusetts, Amherst, MA 01003, United States; Stanford Synchrotron Radiation Lightsource, Menlo Park, CA 94025, United States
推荐引用方式 GB/T 7714	Otten R.,Pádua R.A.P.,Bunze H.A.,et al. How directed evolution reshapes the energy landscape in an enzyme to boost catalysis[J],2020,370(6523).
APA	Otten R..,Pádua R.A.P..,Bunze H.A..,Nguyen V..,Pitsawong W..,...&Kern D..(2020).How directed evolution reshapes the energy landscape in an enzyme to boost catalysis.Science,370(6523).
MLA	Otten R.,et al."How directed evolution reshapes the energy landscape in an enzyme to boost catalysis".Science 370.6523(2020).