Department of Chemical & Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, Nebraska 68588, United States.
The Nebraska Center for Integrated Biomolecular Communication (NCIBC), University of Nebraska-Lincoln, Lincoln, Nebraska 68588, United States.
Chem Rev. 2024 Sep 25;124(18):10577-10617. doi: 10.1021/acs.chemrev.3c00938. Epub 2024 Aug 29.
Over the past two decades, genetic code expansion (GCE)-enabled methods for incorporating noncanonical amino acids (ncAAs) into proteins have significantly advanced the field of synthetic biology while also reaping substantial benefits from it. On one hand, they provide synthetic biologists with a powerful toolkit to enhance and diversify biological designs beyond natural constraints. Conversely, synthetic biology has not only propelled the development of ncAA incorporation through sophisticated tools and innovative strategies but also broadened its potential applications across various fields. This Review delves into the methodological advancements and primary applications of site-specific cellular incorporation of ncAAs in synthetic biology. The topics encompass expanding the genetic code through noncanonical codon addition, creating semiautonomous and autonomous organisms, designing regulatory elements, and manipulating and extending peptide natural product biosynthetic pathways. The Review concludes by examining the ongoing challenges and future prospects of GCE-enabled ncAA incorporation in synthetic biology and highlighting opportunities for further advancements in this rapidly evolving field.
在过去的二十年中，遗传密码扩展（GCE）使非天然氨基酸（ncAA）掺入蛋白质的方法在推进合成生物学领域的发展的同时，也从中获得了巨大的收益。一方面，它们为合成生物学家提供了一个强大的工具包，使他们能够超越自然限制，增强和多样化生物设计。另一方面，合成生物学不仅通过复杂的工具和创新策略推动了 ncAA 掺入的发展，而且还拓宽了其在各个领域的潜在应用。本文深入探讨了在合成生物学中通过定点细胞掺入非天然氨基酸的方法学进展和主要应用。这些主题包括通过非典型密码子添加扩展遗传密码、创建半自主和自主生物体、设计调控元件以及操纵和扩展肽天然产物生物合成途径。本文最后通过检查 GCE 使能 ncAA 掺入在合成生物学中面临的持续挑战和未来展望，并强调了在这个快速发展的领域进一步发展的机会。
