Heterocycles in natural products: Enzymes, pathways, and chemistry

从活着的有机体中分离出来的分子是迄今为止分离出来的一些化学上最复杂、药用上最相关的化合物。这些天然产物分子扮演着抗生素和其他药物的关键角色。我的研究小组对了解自然如何构建天然产品感兴趣，我们的长期目标是利用这一知识开发新的分子，并了解新型酶是如何发挥作用的。在这个项目中，我们将使用吲哚霉素，一种独特的抗生素，具有翻译潜力，作为对抗耐甲氧西林金黄色葡萄球菌(MRSA)的试剂，作为阐明基础生物有机化学的起点。我的团队之前的工作确定了吲哚霉素基因簇以及几种独特的酶在构建这种独特结构中的作用。这些先前的结果为阐明新的酶机制和研究从基因组挖掘中识别的相关系统奠定了基础。此外，我们现在可以通过结合有机化学、酶学和基因工程的技术来开发新的分子。 在这个项目中，我们将追求三个相关的目标：第一，我们将使用X射线结晶学来解析吲哚霉素生物合成酶的结构。几种吲哚霉素酶催化了前所未有的反应，我们想了解它们是如何工作的。通过为这些酶生成分子“蓝图”，我们将了解这些蛋白质的结构，并为设计它们进行新的反应奠定基础。其次，我们将阐明与吲哚霉素相关的三个新途径。利用生物信息学工具，我们发现了三个基因簇，它们具有与吲哚霉素基因相关的基因。我们的目标是了解这些新基因簇编码的酶，并阐明这些新途径的产物。第三，我们将结合化学和生物学的技术，建立一个吲哚霉素衍生物库。这项工作将利用并改造吲哚霉素基因簇来制造我们可以测试其抗生素活性的分子。总体而言，我们的工作将提供对酶学、生物合成和生物有机化学的基本见解，并提供对新的吲哚霉素类分子文库的可持续访问，以使下游的翻译工作能够开发新的抗生素。

Molecules isolated from living organisms are some of the most chemically complex and medicinally relevant compounds ever isolated. These natural product molecules play critical roles as antibiotics and other drugs. My research group is interested in understanding how nature constructs natural products, and our long-term goal is to harness this knowledge to develop new molecules and to understand how novel enzymes function. In this program, we will use indolmycin, a unique antibiotic, with translational potential as an agent against methicillin-resistant Staphylococcus aureus (MRSA), as a starting point to elucidate fundamental bioorganic chemistry. Previous work in my group identified the indolmycin gene cluster and the roles of several unique enzymes in building this unique structure. These prior results set the stage to elucidate new enzyme mechanisms and investigate related systems identified from genomic mining. Furthermore, we are now positioned to develop new molecules by combining techniques from organic chemistry, enzymology, and genetic engineering. In this program, we will pursue three related objectives: first, we will use X-ray crystallography to solve the structures of indolmycin biosynthetic enzymes. Several of the indolmycin enzymes catalyze reactions that are unprecedented, and we want to understand how they work. By generating molecular “blueprints” for these enzymes, we will understand the architecture of these proteins and set the stage for engineering them to do new reactions. Second, we will elucidate three new pathways related to indolmycin. Using bioinformatics tools, we have discovered three gene clusters that have genes related to the indolmycin genes. Our goals are to understand the enzymes encoded by these new gene clusters and to elucidate the products of these new pathways. Third, we will generate a library of indolmycin derivatives by combining techniques from chemistry and biology. This work will harness and engineer the indolmycin gene cluster to make molecules that we can test for their antibiotic activity. Overall, our work will provide fundamental insights into enzymology, biosynthesis, and bioorganic chemistry and provide sustainable access to library of new indolmycin-like molecules to enable downstream translational efforts to develop new antibiotics.