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On roads not taken in the evolution of protein catalysts: Antibody steroid isomerases that use an enamine mechanism

机译:在蛋白质催化剂尚未开发的道路上:使用烯胺机制的抗体类固醇异构酶

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摘要

Reactive immunization has emerged as a new tool for the study of biological catalysis. A powerful applica- tion resulted in catalytic antibodies that use an enamine mechanism akin to that used by the class I aldolases. With regard to the evolution of enzyme mechanisms, we investigated the utility of an enamine pathway for the allylic rearrangement exemplified by △~5-3-ketosteroid isomerase (KSI; EC 5.3.3.1). Our aldolase antibodies were found to catalyze the isomer- ization of both steroid model compounds and steroids. The kinetic and chemical studies showed that the antibodies afforded rate accelerations up to a factor of 10~4 by means of an enamine mechanism in which imine formation was the rate-determining step.
机译:反应性免疫已成为研究生物催化的新工具。强有力的应用导致了催化抗体使用了类似于I类醛缩酶的烯胺机制。关于酶机制的演变,我们研究了烯胺途径对△〜5-3-酮固醇异构酶(KSI; EC 5.3.3.1)例示的烯丙基重排的实用性。我们的醛缩酶抗体被发现可催化类固醇模型化合物和类固醇的异构化。动力学和化学研究表明,抗体通过烯胺机制使速率加速高达10〜4倍,其中亚胺的形成是速率决定步骤。

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