Expanding the synthetic utility of natural product biosynthetic enzymes

项目摘要 氧化转化是最常用的反应类别之一，在合成 用于研究生物系统的药剂和小分子。虽然取得了重大进展， 尽管在开发用于氧化反应的强大方法方面取得了进展，但实施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选择性、位点选择性和立体选择性的转化。相比 小分子催化剂和试剂、来自天然产物生物合成途径的酶已经发展到 以高选择性进行氧化反应。氧化酶反应的发现， 开发这些催化剂的效用有可能使合成策略，并赋予我们 获得具有强大生物活性的新分子。该提案描述了几种战略， 开发强大的酶介导的氧化反应，并利用这些工具， 具有药物潜力的分子的合成。 这项工作利用了自然界在其合成过程中进化出的强大的反应性和选择性。 复杂次级代谢产物的途径。从自然界提供的这个出发点，我们（1）描述了 每种酶的功能，并定义每种酶的底物灵活性，（2）使用结构指导蛋白质 工程化以改变给定催化剂的固有位点和立体选择性，（3）开发化学模拟物 生物催化反应和（4）利用策略来扩大给定酶的底物范围。在一起， 这些方法提供了一套通用的催化剂， 靶分子。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。 总之，该提案描述了化学选择性、位点选择性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.