One, Two, and Three Rieske Routes for Catalyzing Site-Specific Oxygenations

摘要 Rieske加氧酶利用过渡金属的反应性来执行强大，高效和位点特异性 传统惰性键的转变。这些酶将[2Fe-2S]簇与非血红素偶联， 铁位点利用分子氧（O2）作为生物合成和降解途径中共底物。在这些 在反应中，O2的动力学稳定性通过使用非血红素铁位点来克服，所述非血红素铁位点结合O2， 通过形成活性氧中间体促进其裂解。这种活性物质用于 从基质中提取氢原子并引发一系列具有挑战性的转变。里斯克 已知加氧酶作为双加氧酶或单加氧酶起作用，并且甚至已经显示催化 顺序单氧化反应。正如许多生物合成途径所证明的那样， 具有抗生素、抗真菌、抗癌或麻醉活性的天然产物，以及 降解环境污染物，这些酶表现出精致的控制，在区分这些 反应类型，以确保只有预期的转化被催化。因此，这些酶代表了 酶策略的有价值来源，用于工业生产药物和商品化学品，或 促进生物修复工作。然而，严重缺乏关于这些问题如何解决的信息。 酶能够使用一组共同的双中心催化位点特异性反应， 成果。因此，在这项工作中，我们将揭示大自然用来调整 Rieske加氧酶的选择性和催化库。这些知识将提供预测 致力于重新利用Rieske加氧酶来催化定制反应，并将支持利用 他们的化学为各种各样的生物技术的努力。

Abstract Rieske oxygenases harness the reactivity of transition metals to perform powerful, efficient, and site-specific transformations of traditionally inert bonds. These enzymes, which couple a [2Fe-2S] cluster with a non-heme iron site, exploit molecular oxygen (O2) as a co-substrate in biosynthetic and degradative pathways. In these reactions, the kinetic stability of O2 is overcome by the use of the non-heme iron site, which binds O2 and promotes its cleavage via the formation of an activated oxygen intermediate. This reactive species is used to abstract a hydrogen atom from a substrate and initiate an array of challenging transformations. Rieske oxygenases are known to function as dioxygenases or monooxygenases, and have even been shown to catalyze sequential monooxygenation reactions. As demonstrated in a number of biosynthetic pathways that produce natural products with antibiotic, antifungal, anticancer, or anesthetic activities, as well as in pathways that degrade environmental pollutants, these enzymes demonstrate exquisite control in differentiating between these reaction types to ensure that only the intended transformation is catalyzed. Thus, these enzymes represent a valuable source of enzymatic strategies to industrially produce pharmaceuticals and commodity chemicals, or facilitate bioremediation efforts. However, there is a critical lack of information available about how these enzymes are able to use a common set of metallocenters to catalyze site-specific reactions with diverse outcomes. Therefore, in this work, we will uncover the architectural strategies that Nature uses to tune the selectivity and catalytic repertoire of the Rieske oxygenase enzymes. This knowledge will provide predictive power towards repurposing Rieske oxygenases to catalyze custom reactions, and will support efforts to exploit their chemistry for a wide variety of biotechnological endeavors.