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 使用可持续方法访问复杂的目标分子的能力对于发展至关重要 新型药物，这对于治疗抗菌素耐药性和新兴的传染病是必不可少的。1部位 - 选择性氧化反应是当前合成逻辑对复杂支架和 这些核心的装饰，这是新药开发的关键。2,3自然可以使用酶赋予 基于活性部位施加的三维控制，氧化中精致的位点选择性 将底物和氧化剂融合在一起。从大自然中汲取灵感，我计划利用黄素依赖 单加氧化酶以定位氧化各种非本地领域和杂种底物（AIM 1）， 这将允许使用新颖的构件，药品和天然产品及其衍生产品。 因此，在逻辑上选择生物催化剂，依赖黄素依赖性单加氧酶的序列空间很大 改善了现场选择性，产量或底物范围，需要策略。我计划使用序列相似性 网络，（可视化序列的相似序列），序列比对，（探测氨基酸变化 三维主动位点分析（识别关键氨基酸和活性的大小） 站点）和对接研究（了解底物如何与活动地点相互作用）（AIM 2）。这些技术 可以一起解释和预测底物范围的趋势和现场选择性。最终，这些 技术将帮助我从逻辑上选择具有改进属性的生物催化剂，例如扩展 底物范围，不同的位点选择性或提高产率以增强提出的氧化方法 发展。 这些目标共同开发出适用于各种Arene的现场选择性生物催化氧化方法 和异源底物。这项研究将通过发展高 跨酶家族的非新型底物筛查的吞吐量平台，并告知哪些工具是什么工具 对于合理选择现有的生物催化剂的重要性。最后，提出的现场选择性氧化方法将 提高对人类健康很重要的分子的合成和衍生化的易度性和效率。