: Furan‐functionalized peptides are of significant pharmacological interest due to their pronounced bioactivities and unique potential for orthogonal bioconjugation and derivatization. However, naturally occurring peptides with furyl side chains are exceedingly rare. This study presents a streamlined method to predict and assess the microbial production of peptides incorporating 3‐furylalanine (Fua) moieties. The approach integrates genome mining and the reversible, chemoselective tagging of furyl residues, utilizing their unique Diels‐Alder reactivity, for mass spectrometry‐guided identification of candidate compounds. By employing the rhizonin Fua synthase as a bioinformatic handle and through heterologous reconstitution of Fua biosynthesis, we identified previously unknown Fua biosynthetic pathways in diverse bacterial phyla, including actinomycetes, cyanobacteria, actinobacteria, and g‐proteobacteria, suggesting that Fua‐containing peptides are remarkably widely distributed. Metabolic profiling by reversible tagging facilitated the detection of native Fua‐containing metabolites. The successful adaptation of this method for solid support enabled the direct enrichment of furyl‐substituted peptides from complex mixtures. This multi‐pronged approach enabled the discovery and characterization of two novel families of Fua cyclopeptides (rubriamides and typhamides) with potent antiproliferative effects against human tumor cells and nematodes. The innovative catch‐and‐release strategy, in conjunction with genome mining, represents a valuable tool for the discovery of new furan‐substituted natural products.

中文翻译：

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基因组学驱动发现抗增殖呋喃官能化肽的捕获-释放策略


： 呋喃官能化肽由于其显着的生物活性和正交生物偶联和衍生化的独特潜力而具有重要的药理学意义。然而，天然存在的带有呋喃侧链的肽极为罕见。本研究提出了一种简化的方法来预测和评估含有 3-呋喃丙氨酸 （Fua） 部分的肽的微生物生产。该方法结合了基因组挖掘和呋喃残基的可逆化学选择性标记，利用其独特的 Diels-Alder 反应性，用于质谱指导下对候选化合物的鉴定。通过使用根黄素 Fua 合酶作为生物信息学手柄，并通过 Fua 生物合成的异源重建，我们在不同的细菌门中鉴定了以前未知的 Fua 生物合成途径，包括放线菌、蓝细菌、放线菌门和 g-变形菌门，表明含有 Fua 的肽分布非常广泛。通过可逆标记进行的代谢分析有助于检测含有天然 Fua 的代谢物。该方法成功应用于固体载体，能够从复杂混合物中直接富集呋喃取代肽。这种多管齐下的方法能够发现和表征两个新的福阿环肽家族（红酰胺和鼠伤寒），对人类肿瘤细胞和线虫具有强大的抗增殖作用。创新的捕获和释放策略与基因组挖掘相结合，是发现新的呋喃替代天然产物的宝贵工具。