酶定向进化的策略与成功。

Strategy and success for the directed evolution of enzymes.

机构信息

Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK.

出版信息

Curr Opin Struct Biol. 2011 Aug;21(4):473-80. doi: 10.1016/j.sbi.2011.05.003. Epub 2011 Jun 16.

DOI:10.1016/j.sbi.2011.05.003

Abstract

Directed evolution is widely used to improve enzymes, particularly for industrial biocatalytic processes. Molecular biology advances present many new strategies for directed evolution. Commonly used techniques have led to many successful examples of enzyme improvement, yet there is still a need to improve both the efficiency and capability of directed evolution. Recent strategies aimed at making directed evolution faster and more efficient take better advantage of available structural and sequence information. The underlying principles that lead to early dead-ends for directed evolution experiments are also discussed along with recent strategies designed to by-pass them. Several emerging methods for creating novel enzymes are also discussed including examples of catalytic activity for which there is no precedent in nature. Finally, the combined use of several strategies is likely to be required in practice to improve multiple target properties of an enzyme, as successfully shown by a recent industrial example.

摘要

定向进化被广泛用于改良酶，特别是用于工业生物催化过程。分子生物学的进步为定向进化提供了许多新的策略。常用的技术已经带来了许多成功的酶改良实例，但仍需要提高定向进化的效率和能力。最近的策略旨在使定向进化更快、更高效，从而更好地利用可用的结构和序列信息。本文还讨论了导致定向进化实验早期陷入死胡同的基本原理，以及最近为绕过这些原理而设计的策略。还讨论了几种新兴的创造新酶的方法，包括一些在自然界中没有先例的催化活性的例子。最后，如最近的一个工业实例所示，在实践中，很可能需要结合使用几种策略来改善酶的多个目标性质。