Institute of Environmental and Energy Technology, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 37673, Republic of Korea.
Department of Chemical Engineering, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 37673, Republic of Korea.
Biotechnol Adv. 2023 Dec;69:108251. doi: 10.1016/j.biotechadv.2023.108251. Epub 2023 Sep 9.
A variety of chemicals have been produced through metabolic engineering approaches, and enhancing biosynthesis performance can be achieved by using enzymes with high catalytic efficiency. Accordingly, a number of efforts have been made to discover enzymes in nature for various applications. In addition, enzyme engineering approaches have been attempted to suit specific industrial purposes. However, a significant challenge in enzyme discovery and engineering is the efficient screening of enzymes with the desired phenotype from extensive enzyme libraries. To overcome this bottleneck, genetically encoded biosensors have been developed to specifically detect target molecules produced by enzyme activity at the intracellular level. Especially, the biosensors facilitate high-throughput screening (HTS) of targeted enzymes, expanding enzyme discovery and engineering strategies with advances in systems and synthetic biology. This review examines biosensor-guided HTS systems and highlights studies that have utilized these tools to discover enzymes in diverse areas and engineer enzymes to enhance their properties, such as catalytic efficiency, specificity, and stability.
通过代谢工程方法已经产生了多种化学物质，并且可以通过使用具有高催化效率的酶来提高生物合成性能。因此，已经做出了许多努力来在自然界中发现用于各种应用的酶。此外，还尝试了酶工程方法来适应特定的工业用途。然而，酶的发现和工程中的一个重大挑战是从广泛的酶库中有效地筛选具有所需表型的酶。为了克服这一瓶颈，已经开发了遗传编码的生物传感器来专门在细胞内水平检测由酶活性产生的目标分子。特别是，生物传感器有助于针对酶的高通量筛选（HTS），通过系统和合成生物学的进步扩展了酶的发现和工程策略。本综述考察了生物传感器指导的 HTS 系统，并强调了利用这些工具在不同领域发现酶以及工程酶以提高其性能（如催化效率、特异性和稳定性）的研究。
