Department of Chemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA.
Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA.
Org Biomol Chem. 2022 Aug 3;20(30):5891-5906. doi: 10.1039/d2ob00443g.
Directed evolution has been remarkably successful in identifying enzyme variants with new or improved properties, such as altered substrate scope or novel reactivity. Genetically encodable biosensors (GEBs), which convert the concentration of a small molecule ligand into an easily detectable output signal, have seen increasing application to enzyme directed evolution in the last decade. GEBs enable the use of high-throughput methods to assess enzyme activity of very large libraries, which can accelerate the search for variants with desirable activity. Here, we review different classes of GEBs and their properties in the context of enzyme evolution, how GEBs have been integrated into directed evolution workflows, and recent examples of enzyme evolution efforts utilizing GEBs. Finally, we discuss the advantages, challenges, and opportunities for using GEBs in the directed evolution of enzymes.
定向进化在鉴定具有新的或改进的性质的酶变体方面取得了显著的成功，例如改变的底物范围或新颖的反应性。遗传可编码的生物传感器（GEBs）将小分子配体的浓度转化为易于检测的输出信号，在过去十年中越来越多地应用于酶定向进化。GEBs 使得可以使用高通量方法评估非常大的文库中的酶活性，从而加速寻找具有理想活性的变体。在这里，我们将回顾不同类别的 GEBs 及其在酶进化背景下的特性，GEBs 如何集成到定向进化工作流程中，以及利用 GEBs 进行酶进化的最新示例。最后，我们讨论了在酶的定向进化中使用 GEBs 的优点、挑战和机会。
