PDF(Pubmed)
Abstract：
Although sulfonation plays crucial roles in various biological processes and is frequently utilized in medicinal chemistry to improve water solubility and chemical diversity of drug leads, it is rare and underexplored in ribosomally synthesized and post-translationally modified peptides (RiPPs). Biosynthesis of RiPPs typically entails modification of hydrophilic residues, which substantially increases their chemical stability and bioactivity, albeit at the expense of reducing water solubility. To explore sulfonated RiPPs that may have improved solubility, we conducted co-occurrence analysis of RiPP class-defining enzymes and sulfotransferase (ST), and discovered two distinctive biosynthetic gene clusters (BGCs) encoding both lanthipeptide synthetase (LanM) and ST. Upon expressing these BGCs, we characterized the structures of novel sulfonated lanthipeptides and determined the catalytic details of LanM and ST. We demonstrate that SslST-catalyzed sulfonation is leader-independent but relies on the presence of A ring formed by LanM. Both LanM and ST are promiscuous towards residues in the A ring, but ST displays strict regioselectivity toward Tyr5. The recognition of cyclic peptide by ST was further discussed. Bioactivity evaluation underscores the significance of the ST-catalyzed sulfonation. This study sets up the starting point to engineering the novel lanthipeptide STs as biocatalysts for hydrophobic lanthipeptides improvement.
摘要：
尽管磺化在各种生物过程中起着至关重要的作用，并且经常在药物化学中用于改善水溶性和药物铅的化学多样性，在核糖体合成和翻译后修饰的肽（RiPP）中，它是罕见且未充分开发的。RiPP的生物合成通常需要亲水残基的修饰，大大提高了它们的化学稳定性和生物活性，尽管以降低水溶性为代价。为了探索可能具有改善的溶解度的磺化RiPP，我们进行了RiPP类定义酶和磺基转移酶（ST）的共现分析，并发现了两个独特的生物合成基因簇（BGC），它们同时编码羊毛肽合成酶（LanM）和ST。在表达这些BGC后，我们表征了新型磺化羊毛硫肽的结构，并确定了LanM和ST的催化细节。我们证明了SslST催化的磺化是不依赖于前导的，但依赖于由LanM形成的A环的存在。LanM和ST对A环中的残留物都是混杂的，但ST对Tyr5表现出严格的区域选择性。进一步讨论了ST对环肽的识别。生物活性评估强调了ST催化磺化的重要性。这项研究为工程化新型羊毛硫肽STs作为疏水性羊毛硫肽改进的生物催化剂奠定了起点。
