Laboratory of Organic Chemistry, ETH Zürich, CH-8093 Zürich, Switzerland; email:
Annu Rev Biochem. 2018 Jun 20;87:131-157. doi: 10.1146/annurev-biochem-062917-012034. Epub 2018 Mar 1.
Directed evolution is a powerful technique for generating tailor-made enzymes for a wide range of biocatalytic applications. Following the principles of natural evolution, iterative cycles of mutagenesis and screening or selection are applied to modify protein properties, enhance catalytic activities, or develop completely new protein catalysts for non-natural chemical transformations. This review briefly surveys the experimental methods used to generate genetic diversity and screen or select for improved enzyme variants. Emphasis is placed on a key challenge, namely how to generate novel catalytic activities that expand the scope of natural reactions. Two particularly effective strategies, exploiting catalytic promiscuity and rational design, are illustrated by representative examples of successfully evolved enzymes. Opportunities for extending these approaches to more complex biocatalytic systems are also considered.
定向进化是一种强大的技术，可用于生成广泛的生物催化应用的定制酶。遵循自然进化的原则，通过迭代的突变和筛选或选择循环来修饰蛋白质性质，提高催化活性，或开发用于非天然化学转化的全新蛋白质催化剂。本文简要综述了用于产生遗传多样性并筛选或选择改良酶变体的实验方法。重点介绍了一个关键挑战，即如何产生新的催化活性以扩大自然反应的范围。通过成功进化的酶的代表性实例说明了利用催化多功能性和合理设计的两种特别有效的策略。还考虑了将这些方法扩展到更复杂的生物催化系统的机会。
