Construction and in vivo assembly of a catalytically proficient and hyperthermostable de novo enzyme.
Construction and in vivo assembly of a catalytically proficient and hyperthermostable de novo enzyme.
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催化熟练且可超过的从头酶的结构和体内组装。
DOI：
10.1038/s41467-017-00541-4
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发表时间：
2017-08-25
影响因子：
16.6
通讯作者：
Anderson JLR
中科院分区：
文献类型：
--
作者：
Watkins DW;Jenkins JMX;Grayson KJ;Wood N;Steventon JW;Le Vay KK;Goodwin MI;Mullen AS;Bailey HJ;Crump MP;MacMillan F;Mulholland AJ;Cameron G;Sessions RB;Mann S;Anderson JLR
尽管天然酶的催化机制已经被很好地理解，但在重新设计的天然蛋白质中实现反应化学的多样性已被证明具有挑战性。天然酶的大规模修饰可能会受到其内在复杂性的影响，这往往掩盖了生物催化效率的基本原则。模型方法可以通过将稳健的从头设计的底盘与避免天然蛋白质的原子模拟的设计过程相结合来规避这种复杂性。在这里，我们应用这种方法来构建一个高效的，混杂的，和热稳定的人工酶，催化各种各样的底物氧化耦合到H2O2的还原。该模型表现出的动力学，匹配，甚至超过某些天然过氧化物酶，保持其活性，在高温下，在有机溶剂的存在下，并提供了一个简单的平台，询问催化中间体常见的天然血红素含酶。酶的催化机制已经很好地理解，但是在重新工程化的蛋白质中实现不同的反应化学可能是困难的。在这里，作者展示了一种高效且热稳定的人工酶，该酶催化与H2O2还原偶联的多种底物氧化。
Although catalytic mechanisms in natural enzymes are well understood, achieving the diverse palette of reaction chemistries in re-engineered native proteins has proved challenging. Wholesale modification of natural enzymes is potentially compromised by their intrinsic complexity, which often obscures the underlying principles governing biocatalytic efficiency. The maquette approach can circumvent this complexity by combining a robust de novo designed chassis with a design process that avoids atomistic mimicry of natural proteins. Here, we apply this method to the construction of a highly efficient, promiscuous, and thermostable artificial enzyme that catalyzes a diverse array of substrate oxidations coupled to the reduction of H2O2. The maquette exhibits kinetics that match and even surpass those of certain natural peroxidases, retains its activity at elevated temperature and in the presence of organic solvents, and provides a simple platform for interrogating catalytic intermediates common to natural heme-containing enzymes. Catalytic mechanisms of enzymes are well understood, but achieving diverse reaction chemistries in re-engineered proteins can be difficult. Here the authors show a highly efficient and thermostable artificial enzyme that catalyzes a diverse array of substrate oxidations coupled to the reduction of H2O2.
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影响因子：
8.4
作者：
Anderson JLR;Armstrong CT;Kodali G;Lichtenstein BR;Watkins DW;Mancini JA;Boyle AL;Farid TA;Crump MP;Moser CC;Dutton PL
通讯作者：
Dutton PL
DOI：
10.1016/j.bbabio.2015.09.008
发表时间：
2016-05
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Fry BA;Solomon LA;Leslie Dutton P;Moser CC
通讯作者：
Moser CC
影响因子：
3
作者：
Harvey, Jeremy N.;Bathelt, Christine M.;Mulholland, Adrian J.
通讯作者：
Mulholland, Adrian J.
影响因子：
14.8
作者：
Farid, Tammer A.;Kodali, Goutham;Solomon, Lee A.;Lichtenstein, Bruce R.;Sheehan, Molly M.;Fry, Bryan A.;Bialas, Chris;Ennist, Nathan M.;Siedlecki, Jessica A.;Zhao, Zhenyu;Stetz, Matthew A.;Valentine, Kathleen G.;Anderson, J. L. Ross;Wand, A. Joshua;Discher, Bohdana M.;Moser, Christopher C.;Dutton, P. Leslie
通讯作者：
Dutton, P. Leslie
影响因子：
2.9
作者：
BERRY, EA;TRUMPOWER, BL
通讯作者：
TRUMPOWER, BL