Development of Selective Oxidative Biocatalytic Methods

选择性氧化生物催化方法的发展

• 批准号: 10606798
• 负责人: Sarah Elizabeth Champagne
• 金额: $ 4.03万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10606798
• 关键词: 3-Dimensional; Acceleration; Active Sites; Address; Affinity; Amino Acids; Antimicrobial Resistance; Binding; Biological; Breathing; COVID-19; Chemicals; Communicable Diseases; Complex; Data; Databases; Derivation procedure; Development; Disease; Docking; Drug resistance; Ebola; Electronics; Emerging Communicable Diseases; Enzymes; Family; Flavins; Health; Human; Hydroxylation; Indoles; Investigation; Left; Libraries; Logic; Mediating; Metabolic; Methodology; Methods; Mixed Function Oxygenases; Natural Products; Nature; Oral; Oxidants; Pharmaceutical Preparations; Pharmacologic Substance; Phenols; Positioning Attribute; Property; Reaction; Research; Route; Sequence Alignment; Sequence Analysis; Site; Solubility; Substrate Interaction; Techniques; Visualization; catalyst; computerized tools; improved; method development; next generation; non-Native; novel; novel therapeutics; oxidation; predictive modeling; pyridine; scaffold; screening; small molecule; tool; trend

Project Summary The ability to access complex target molecules with sustainable methodology is imperative to the development of novel drugs, which will be necessary to treat antimicrobial resistance and emerging infectious diseases.1 Site- selective oxidation reactions are fundamental to the current synthetic logic towards complex scaffolds and the decoration of these cores, which is key to the development of novel drugs.2,3 Nature can use enzymes to impart exquisite site-selectivity in oxidations based on the three-dimensional control exerted by the active site while bringing a substrate and oxidant together. Taking inspiration from Nature, I plan to utilize flavin-dependent monooxygenases to site-selectively oxidize a variety of non-native arenes and heteroarenes substrates (Aim 1), which will allow access to novel building blocks, pharmaceuticals, and natural products and their derivatives. The sequence space of flavin-dependent monooxygenases is vast, therefore, logically picking biocatalysts that have improved, site-selectivity, yield, or substrate scope, requires a strategy. I plan to use sequence similarity networks, (to visualize how similar sequences are), sequence alignments, (to probe amino acid changes proximal to active sites), three-dimensional active site analysis, (to identify key amino acids and size of active site), and docking studies, (to understand how a substrate interacts with an active site) (Aim 2). These techniques can be used together to explain and predict trends in substrate scope, and site-selectivity. Ultimately, these techniques will help me to logically choose biocatalysts that have improved properties such as expanded substrate scope, divergent site-selectivity, or improved yield to strengthen the proposed oxidative method development. Together these aims will develop site-selective biocatalytic oxidation methods applicable to a variety of arene and heteroarene substrates. This research will broadly impact biocatalysis investigations by developing a high throughput platform for nonnative substrate screening across a family of enzymes, and informing what tools are important for rationally choosing existing biocatalysts. Finally, the proposed site-selective oxidative method will improve the ease and efficiency of synthesis and derivatization of molecules that are important to human health.

项目摘要 以可持续的方法获得复杂目标分子的能力对于开发 新的药物，这将是必要的，以治疗抗生素耐药性和新出现的传染病。 选择性氧化反应是目前复杂支架合成逻辑的基础， 这些核心的装饰，这是开发新药的关键。2，3自然可以使用酶来赋予 基于活性位点所施加的三维控制的氧化中的精确位点选择性， 将底物和氧化剂结合在一起。从大自然中获得灵感，我计划利用黄素依赖 单加氧酶位点选择性氧化多种非天然芳烃和杂芳烃底物（Aim 1）， 这将允许获得新的结构单元、药物和天然产品及其衍生物。 黄素依赖性单加氧酶的序列空间是巨大的，因此，从逻辑上挑选生物催化剂， 有改进，位点选择性，产量，或底物范围，需要一个策略。我计划用序列相似性 网络（可视化序列的相似程度）、序列比对（探测氨基酸变化 活性位点近端）、三维活性位点分析（以鉴定活性位点的关键氨基酸和大小）， 位点）和对接研究（以了解底物如何与活性位点相互作用）（目标2）。这些技术 可以一起用来解释和预测底物范围和位点选择性的趋势。最终，这些 技术将帮助我合理地选择具有改进性能的生物催化剂，如扩展 底物范围、不同的位点选择性或提高的产率，以加强所提出的氧化方法 发展 这些目标将共同发展适用于各种芳烃的定点生物催化氧化方法 和杂芳烃底物。这项研究将广泛影响生物催化研究，通过开发一个高 通量平台，用于跨酶家族的非天然底物筛选，并告知 对于合理选择现有的生物催化剂很重要。最后，所提出的位点选择性氧化方法将 改善对人类健康重要的分子的合成和衍生化的容易性和效率。