Jestin Jean-Luc, Kaminski Pierre Alexandre
Département de Biologie Structurale et Chimie, Unité de Chimie Organique URA 2128 CNRS, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris 15, France.
J Biotechnol. 2004 Sep 30;113(1-3):85-103. doi: 10.1016/j.jbiotec.2004.03.032.
Enzyme engineering by molecular modelling and site-directed mutagenesis can be remarkably efficient. Directed enzyme evolution appears as a more general strategy for the isolation of catalysts as it can be applied to most chemical reactions in aqueous solutions. Selections, as opposed to screening, allow the simultaneous analysis of protein properties for sets of up to about 10(14) different proteins. These approaches for the parallel processing of molecular information 'Is the protein a catalyst?' are reviewed here in the case of selections based on the formation of a specific reaction product. Several questions are addressed about in vivo and in vitro selections for catalysis reported in the literature. Can the selection system be extended to other types of enzymes? Does the selection control regio- and stereo-selectivity? Does the selection allow the isolation of enzymes with an efficient turnover? How should substrates be substituted or mimicked for the design of efficient selections while minimising the number of chemical synthesis steps? Engineering sections provide also some clues to design selections or to circumvent selection biases. A special emphasis is put on the comparison of in vivo and in vitro selections for catalysis.
通过分子建模和定点诱变进行酶工程可能非常高效。定向酶进化似乎是一种更通用的分离催化剂的策略，因为它可应用于水溶液中的大多数化学反应。与筛选不同，选择允许同时分析多达约10¹⁴种不同蛋白质的蛋白质特性。本文针对基于特定反应产物形成的选择情况，综述了这些用于并行处理分子信息“该蛋白质是催化剂吗？”的方法。文中讨论了文献中报道的关于体内和体外催化选择的几个问题。选择系统能否扩展到其他类型的酶？选择能否控制区域选择性和立体选择性？选择能否分离出具有高效周转率的酶？在设计高效选择时，应如何取代或模拟底物，同时尽量减少化学合成步骤的数量？工程部分也为设计选择或规避选择偏差提供了一些线索。特别强调了体内和体外催化选择的比较。
