Kirkpatrick John P, Karanth Megha N, Carlomagno Teresa
School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Birmingham, B15 2TT, UK.
Institute of Organic Chemistry and Centre of Biomolecular Drug Research, Leibniz University of Hannover, Schneiderberg 38, 30167, Hannover, Germany.
Biomol NMR Assign. 2025 Jun;19(1):153-164. doi: 10.1007/s12104-025-10231-8. Epub 2025 Apr 25.
Non-ribosomal peptide synthetases (NRPSs) are macromolecular enzymatic complexes responsible for the biosynthesis of an array of microbial natural products, many of which have important applications for human health. The nature of the NRPS machinery, which has been likened to an assembly line, should be amenable to bio-engineering efforts directed towards efficient synthesis of novel and tailored molecules. However, the success of such endeavours depends on a detailed understanding of the mechanistic principles governing the various steps in the peptide assembly-line. Here, we report the near-complete assignment of the Ile, Met, Leu and Val methyl-groups of the 59-kDa adaptor-condensation di-domain (BN-BC) from the Tomaymycin NRPS. These assignments will provide the foundation for future NMR studies of the complex dynamic behaviour of the condensation domain both in isolation and in the context of the enzymatic cycle, which will themselves form the basis for developing a complete mechanistic description of the central condensation reaction in this prototypical NRPS.
非核糖体肽合成酶（NRPSs）是一种大分子酶复合物，负责合成一系列微生物天然产物，其中许多对人类健康具有重要应用价值。NRPS机制的本质类似于一条装配线，应该适合进行旨在高效合成新型定制分子的生物工程研究。然而，此类努力的成功取决于对肽装配线各个步骤所遵循的机制原理的详细理解。在此，我们报告了来自托马霉素NRPS的59 kDa衔接子-缩合双结构域（BN-BC）中异亮氨酸、甲硫氨酸、亮氨酸和缬氨酸甲基基团的近乎完整归属。这些归属将为未来对缩合结构域在分离状态下以及酶促循环背景下的复杂动态行为进行核磁共振研究奠定基础，而这些研究本身将为全面描述这种典型NRPS中的核心缩合反应的机制提供依据。
