Expanding the synthetic utility of natural product biosynthetic enzymes - Diversity Supplement

项目摘要 氧化转化是最常用的反应类型之一，在合成 用于研究生物系统的药剂和小分子。虽然取得了巨大的进步， 在开发强有力的氧化反应方法方面已经取得了进展，但执行这些方法仍然具有挑战性 在复杂分子上具有高水平的化学、位置和立体选择性的转化。与之形成鲜明对比的是 来自天然产物生物合成途径的小分子催化剂和试剂、酶已经进化到 进行高选择性的氧化反应。氧化酶反应的发现和 这些催化剂用途的开发有可能使合成策略成为可能，并使我们 获得具有强大生物活性的新分子。本建议书描述了以下几种策略 开发强大的酶介导的氧化反应并利用这些工具来简化 合成具有药用潜力的分子。 这项工作利用了大自然在其合成的化合物中进化出的强大的反应性和选择性 通向复杂的次生代谢物的路线。从自然提供的这个起点出发，我们(1)描述了 每种酶的功能，并确定每种酶的底物灵活性，(2)使用结构导向蛋白 改变给定催化剂的天然位置和立体选择性的工程，(3)开发化学仿生 生物催化反应和(4)利用策略扩大给定酶的底物范围。一起， 这些方法提供了一套通用的催化剂，将应用于生物活性的合成。 目标分子。这些分子的生物学特性将通过直接合作进行评估。 密歇根大学医学院和化学基因组学中心的研究人员 密歇根大学生命科学研究所的高通量筛选设施。 综上所述，本提案描述了化学、位置和立体选择性氧化的发展 由生物合成酶介导的转化。这些方法将直接使合成 与人类健康相关的复杂生物活性分子。

Project Summary Oxidative transformations are one of the most utilized classes of reactions, indispensible in the synthesis of pharmaceutical agents and small molecules used to study biological systems. While significant strides have been made in developing powerful methods for oxidative reactions, it remains challenging to carry out these transformations with high levels of chemo-, site- and stereoselectivity on complex molecules. In contrast to small molecule catalysts and reagents, enzymes from natural product biosynthetic pathways have evolved to carry out oxidation reactions with high levels of selectivity. The discovery of oxidative enzymatic reactions and development of the utility of these catalysts has the potential to enable to synthetic strategies and grant us access to new molecules with potent biological activity. This proposal describes several strategies for developing robust enzyme-mediated oxidation reactions and leveraging these tools for the streamlined synthesis of molecules with pharmaceutical potential. This work takes advantage of the powerful reactivity and selectivity that Nature has evolved within its synthetic routes to complex secondary metabolites. From this starting point provided by Nature, we (1) characterize the function of each enzyme and define the substrate flexibility of each enzyme, (2) use structure-guided protein engineering to alter the innate site- and stereoselectivity of a given catalysts, (3) develop chemomimetic biocatalytic reactions and (4) utilize strategies to expand the substrate scope of a given enzyme. Together, these approaches provide a suite of versatile catalysts that will be applied to the synthesis of biologically active target molecules. The biological properties of these molecules will be evaluated through direct collaborations with investigators at the University of Michigan Medical School as well as the Center for Chemical Genomics High Throughput Screening facility at the University of Michigan Life Sciences Institute. In summary, this proposal describes the development of chemo-, site- and stereoselective oxidative transformations mediated by biosynthetic enzymes. These methods will directly enable the synthesis of complex biologically active molecules relevant to human health.