Department of Chemistry, University of Illinois at Urbana-Champaign , 600 South Mathews Avenue, Urbana, Illinois 61801, United States.
Department of Chemistry and Warwick Integrative Synthetic Biology Center, University of Warwick , Coventry CV4 7AL, United Kingdom.
J Am Chem Soc. 2017 Dec 20;139(50):18154-18157. doi: 10.1021/jacs.7b09899. Epub 2017 Dec 4.
The bottromycins belong to the ribosomally synthesized and posttranslationally modified peptide (RiPP) family of natural products. Bottromycins exhibit unique structural features, including a hallmark macrolactamidine ring and thiazole heterocycle for which divergent members of the YcaO superfamily have been biosynthetically implicated. Here we report the in vitro reconstitution of two YcaO proteins, BmbD and BmbE, responsible for the ATP-dependent cyclodehydration reactions that yield thiazoline- and macrolactamidine-functionalized products, respectively. We also establish the substrate tolerance for BmbD and BmbE and systematically dissect the role of the follower peptide, which we show serves a purpose similar to canonical leader peptides in directing the biosynthetic enzymes to the substrate. Lastly, we leverage the expanded capabilities of YcaO proteins to conduct an extensive bioinformatic survey to classify known YcaO chemistry. This analysis predicts new functions remain to be uncovered within the superfamily.
博特霉素属于核糖体合成和翻译后修饰肽 (RiPP) 天然产物家族。博特霉素具有独特的结构特征，包括一个标志性的大环内酰胺环和噻唑杂环，其中 YcaO 超家族的不同成员被认为具有生物合成作用。在这里，我们报告了两种 YcaO 蛋白（BmbD 和 BmbE）的体外重组，它们负责 ATP 依赖性环脱水反应，分别产生噻唑啉和大环内酰胺功能化产物。我们还确定了 BmbD 和 BmbE 的底物耐受性，并系统地剖析了尾随肽的作用，我们发现尾随肽在引导生物合成酶到达底物方面的作用类似于经典的前导肽。最后，我们利用 YcaO 蛋白的扩展功能进行了广泛的生物信息学调查，以对已知的 YcaO 化学进行分类。该分析预测，该超家族仍有新的功能有待发现。
