Department of Biochemical Engineering, University College London, London, Gower Street WC1E 6BT, U.K.
Biosci Rep. 2022 Aug 31;42(8). doi: 10.1042/BSR20220168.
In enzyme engineering, the main targets for enhancing properties are enzyme activity, stereoselective specificity, stability, substrate range, and the development of unique functions. With the advent of genetic code extension technology, non-natural amino acids (nnAAs) are able to be incorporated into proteins in a site-specific or residue-specific manner, which breaks the limit of 20 natural amino acids for protein engineering. Benefitting from this approach, numerous enzymes have been engineered with nnAAs for improved properties or extended functionality. In the present review, we focus on applications and strategies for using nnAAs in enzyme engineering. Notably, approaches to computational modelling of enzymes with nnAAs are also addressed. Finally, we discuss the bottlenecks that currently need to be addressed in order to realise the broader prospects of this genetic code extension technique.
在酶工程中，提高性能的主要目标是酶活性、立体选择性特异性、稳定性、底物范围和独特功能的开发。随着遗传密码扩展技术的出现，非天然氨基酸（nnAAs）能够以特定位置或残基特异性的方式掺入蛋白质中，从而打破了蛋白质工程中 20 种天然氨基酸的限制。受益于这种方法，许多酶已经通过 nnAAs 进行了工程改造，以提高性能或扩展功能。在本综述中，我们重点介绍了 nnAAs 在酶工程中的应用和策略。值得注意的是，我们还讨论了使用 nnAAs 进行酶的计算建模的方法。最后，我们讨论了目前需要解决的瓶颈问题，以实现这项遗传密码扩展技术更广阔的前景。
