Discovery and Application of New Halogenases

项目摘要 分子多样性的快速和模块化生成是寻找新的化学功能的关键。 一个特别有用的官能团是卤素(X=氯、溴、碘)，它使许多选择性和 创造结构复杂性的有效下游战略。在这方面，卤素酶有 为区域和立体选择性合成催化剂提供了一条重要的补充途径 在复合支架上引入卤素取代基。虽然存在许多卤代酶家族，但 自由基卤化酶提供了反应多样性的最大潜力，因为它们有能力取代 与亲电或亲核的C-H键不同，未活化的C-H键与卤素不同 机械装置。然而，到目前为止，这些酶的底物范围仅限于以下两种蛋白质之一： 结合底物或大型后期天然产品中间体。我们小组发现了一个新的支系 能够与小分子底物反应的自由基卤化酶。我们现在试图利用 以开发体外和体内合成的新工具。这项提案的具体目标 包括：(I)阐明这些新的自由基卤化酶的结构和机制，这将提供 对其工程的重要洞察，(Ii)卤代酶的研究和工程选择性。

Project Summary The rapid and modular generation of molecular diversity is key to the search for new chemical functions. One particularly useful functional group is the halogen (X = Cl, Br, I), which enables many selective and effective downstream strategies for creating structural complexity. In this regard, halogenase enzymes have provided an important and complementary approach to synthetic catalysts for regio- and stereoselective introduction of a halogen substituent on a complex scaffold. While many families of halogenases exist, the radical halogenases provide the greatest potential for reaction diversity, as they are competent to replace unactivated C-H bonds with a halogen unlike those that operate by electrophilic or nucleophilic mechanisms. However, the substrate scope of these enzymes has been limited to date to either protein- bound substrates or large late-stage natural product intermediates. Our group has discovered a new clade of radical halogenases capable of reacting with small molecule substrates. We now seek to take advantage of this discovery to develop new tools for in vitro and in vivo synthesis. Specific aims of this proposal include: (i) elucidating the structure and mechanism of these new radical halogenases, which will provide important insight into their engineering, (ii) investigating and engineering selectivity in halogenases.