College of Biological and Food Engineering, Changshu Institute of Technology, Changshu, 215500, China.
College of Food Science and Light Industry, Nanjing Tech University, Nanjing, 211816, China.
Appl Microbiol Biotechnol. 2023 Jul;107(13):4187-4197. doi: 10.1007/s00253-023-12585-2. Epub 2023 May 26.
Non-ribosomal peptide synthetases are mega-enzyme assembly lines that synthesize many clinically useful compounds. As a gatekeeper, they have an adenylation (A)-domain that controls substrate specificity and plays an important role in product structural diversity. This review summarizes the natural distribution, catalytic mechanism, substrate prediction methods, and in vitro biochemical analysis of the A-domain. Taking genome mining of polyamino acid synthetases as an example, we introduce research on mining non-ribosomal peptides based on A-domains. We discuss how non-ribosomal peptide synthetases can be engineered based on the A-domain to obtain novel non-ribosomal peptides. This work provides guidance for screening non-ribosomal peptide-producing strains, offers a method to discover and identify A-domain functions, and will accelerate the engineering and genome mining of non-ribosomal peptide synthetases. KEY POINTS: • Introducing adenylation domain structure, substrate prediction, and biochemical analysis methods • Advances in mining homo polyamino acids based on adenylation domain analysis • Creating new non-ribosomal peptides by engineering adenylation domains.
非核糖体肽合成酶是合成许多临床有用化合物的巨型酶组装线。作为守门员，它们具有氨酰化（A）结构域，该结构域控制底物特异性并在产物结构多样性中发挥重要作用。本综述总结了 A 结构域的自然分布、催化机制、底物预测方法和体外生化分析。以多聚氨基酸合成酶的基因组挖掘为例，我们介绍了基于 A 结构域挖掘非核糖体肽的研究。我们讨论了如何基于 A 结构域对非核糖体肽合成酶进行工程改造，以获得新型非核糖体肽。这项工作为筛选非核糖体肽产生菌株提供了指导，提供了一种发现和鉴定 A 结构域功能的方法，并将加速非核糖体肽合成酶的工程改造和基因组挖掘。
介绍氨酰化结构域结构、底物预测和生化分析方法
基于氨酰化结构域分析挖掘同聚多氨基酸的进展
通过工程化氨酰化结构域创造新的非核糖体肽。
