Department of Chemical and Systems Biology, Stanford University, United States.
Department of Bioengineering, Stanford University, United States.
Curr Opin Biotechnol. 2017 Dec;48:196-202. doi: 10.1016/j.copbio.2017.05.012. Epub 2017 Jun 15.
Emerging technologies are enabling ultra-high-throughput screening of combinatorial enzyme libraries to identify variants with improved properties such as increased activity, altered substrate specificity, and increased stability. Each of these enzyme engineering platforms relies on compartmentalization of reaction components, similar to microtiter plate-based assays which have been commonly used for testing the activity of enzyme variants. The technologies can be broadly divided into three categories according to their spatial segregation strategy: (1) cells as reaction compartments, (2) in vitro compartmentalization via synthetic droplets, and (3) microchambers. Here, we discuss these emerging platforms, which in some cases enable the screening of greater than 10 million enzyme variants, and highlight benefits and limitations of each technology.
新兴技术正在实现组合酶文库的超高通量筛选，以鉴定具有改良性质的变体，如提高的活性、改变的底物特异性和提高的稳定性。这些酶工程平台中的每一个都依赖于反应成分的分隔，类似于已广泛用于测试酶变体活性的微孔板测定法。这些技术可以根据其空间分离策略大致分为三类：（1）细胞作为反应室，（2）通过合成液滴进行体外分隔，以及（3）微腔室。在这里，我们讨论这些新兴平台，在某些情况下，这些平台能够筛选超过 1000 万个酶变体，并突出每种技术的优势和局限性。
