State Key Laboratory of Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
Key Laboratory of Extreme Environmental Microbial Resources and Engineering of Gansu Province, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China.
Int J Mol Sci. 2024 Aug 29;25(17):9350. doi: 10.3390/ijms25179350.
Borosins are a class of ribosomally synthesized and post-translationally modified peptides (RiPPs) with α-N-methylated backbones. Although the first mature compound of borosin was reported in 1997, the biosynthetic pathway was elucidated 20 years later. Until this work, borosins have been able to be categorized into 11 types based on the features of their protein structure and core peptides. Type III borosins were reported only in fungi initially. In order to explore the sources and potential of type III borosins, a precise genome mining work of type III borosins was conducted in bacteria and KchMA's self-methylation activity was validated by biochemical experiment. Furthermore, a commercial protease and AI-assisted rational design was employed to engineer KchMA for the capacity to produce various N-methylated peptides. Our work demonstrates that type III borosins are abundant not only in eukaryotes but also in bacteria and have immense potential as a tool for synthetic biology.
博来霉素是一类核糖体合成和翻译后修饰的肽（RiPPs），具有α-N-甲基化的骨架。尽管博来霉素的第一个成熟化合物于 1997 年被报道，但直到 20 年后才阐明了其生物合成途径。在这项工作之前，根据其蛋白质结构和核心肽的特征，博来霉素已被分为 11 种类型。最初，仅在真菌中报道了 III 型博来霉素。为了探索 III 型博来霉素的来源和潜力，我们在细菌中进行了 III 型博来霉素的精确基因组挖掘工作，并通过生化实验验证了 KchMA 的自我甲基化活性。此外，我们还采用了商业蛋白酶和 AI 辅助的合理设计来对 KchMA 进行工程改造，使其具有生产各种 N-甲基化肽的能力。我们的工作表明，III 型博来霉素不仅在真核生物中丰富，而且在细菌中也大量存在，具有作为合成生物学工具的巨大潜力。
