近日，日本北海道大学Wakimoto, Toshiyuki团队研究了非核糖体肽环化酶定向化学酶合成套索形脂肽。这一研究成果于2025年11月4日发表在《自然-化学》杂志上。

套索形脂肽是一类重要的抗菌剂，但其复杂结构导致合成困难，阻碍了高效结构多样化研究。

研究组报道了一种新型化学酶法策略，可便捷制备套索形大环化合物。通过利用多功能非核糖体肽环化酶，对携带多重亲核位点（包括天然氨基末端与拟氨基末端）的未保护支链肽进行位点选择性环化，成功构建了具有多样序列与环尺寸的套索肽库。

该策略的普适性经两种青霉素结合蛋白型硫酯酶（SurE与WolJ）及一种I型硫酯酶（TycC硫酯酶）验证。此外，未参与环化过程的剩余亲核位点可作为反应柄，通过位点选择性酰化反应（即丝氨酸/苏氨酸连接）实现后续功能化。这种环化-酰基化串联策略实现了模块化一锅法合成携带不同酰基的套索形脂肽。生物活性筛选表明，位点选择性酰化赋予大环支架抗分枝杆菌活性，从中鉴定出在8-16 µg·ml^-1浓度下可抑制50%细菌生长的脂肽化合物。

附：英文原文

Title: Non-ribosomal peptide cyclase-directed chemoenzymatic synthesis of lariat lipopeptides

Author: Kobayashi, Masakazu, Matsuda, Kenichi, Yamada, Yuito, Ichihara, Rintaro, Onozawa, Naho, Fukano, Hanako, Hoshino, Yoshihiko, Hirabayashi, Aki, Suzuki, Masato, Katsuyama, Akira, Ichikawa, Satoshi, Wakimoto, Toshiyuki

Issue&Volume: 2025-11-04

Abstract: Lariat-shaped lipopeptides are important antimicrobial agents; however, their complex structures pose synthetic challenges that hamper efficient structural diversification. Here we report a new chemoenzymatic approach that facilitates access to lariat-shaped macrocycles. Unprotected, branched peptides bearing multiple nucleophiles, including a native amino terminus and a pseudo-amino terminus, were site-selectively cyclized using versatile non-ribosomal peptide cyclases, generating an array of lariat peptides with diverse sequences and ring sizes. The generality of this strategy was demonstrated using two penicillin-binding protein-type thioesterases, SurE and WolJ, as well as one type-I thioesterase, TycC thioesterase. Furthermore, the remaining nucleophile, which was not involved in the cyclization process, was exploited as a reactive handle for subsequent diversification via a site-selective acylation reaction (that is, Ser/Thr ligation). The tandem cyclization–acylation strategy enabled the one-pot, modular synthesis of lariat-shaped lipopeptides equipped with various acyl groups. Biological screening revealed that the site-selective acylation endowed the macrocyclic scaffolds with antimycobacterial activity and led to the identification of lipopeptides that inhibit 50% of growth at concentrations of 8–16μgml1.

DOI: 10.1038/s41557-025-01979-6

Source: https://www.nature.com/articles/s41557-025-01979-6