Over 20 years ago, the pyrrolysine encoding translation system was discovered in specific archaea. Our Review provides an overview of how the once obscure pyrrolysyl-tRNA synthetase (PylRS) tRNA pair, originally responsible for accurately translating enzymes crucial in methanogenic metabolic pathways, laid the foundation for the burgeoning field of genetic code expansion. Our primary focus is the discussion of how to successfully engineer the PylRS to recognize new substrates and exhibit higher in vivo activity. We have compiled a comprehensive list of ncAAs incorporable with the PylRS system. Additionally, we also summarize recent successful applications of the PylRS system in creating innovative therapeutic solutions, such as new antibody–drug conjugates, advancements in vaccine modalities, and the potential production of new antimicrobials.

中文翻译：

---

吡咯赖氨酰-tRNA 合成酶的进化：从产甲烷到遗传密码扩展


20多年前，在特定的古细菌中发现了吡咯赖氨酸编码翻译系统。我们的综述概述了曾经默默无闻的吡咯赖氨酰-tRNA 合成酶 (PylRS) tRNA 对，最初负责准确翻译在产甲烷代谢途径中至关重要的酶，如何为新兴的遗传密码扩展领域奠定了基础。我们的主要重点是讨论如何成功设计 PylRS 以识别新底物并表现出更高的体内活性。我们编制了一份可与 PylRS 系统合并的 ncAA 的综合列表。此外，我们还总结了 PylRS 系统最近在创建创新治疗解决方案方面的成功应用，例如新的抗体药物偶联物、疫苗模式的进步以及新抗菌药物的潜在生产。