Discovery and Characterization of Natural Product Systems

天然产物系统的发现和表征

• 批准号: 9277486
• 负责人: DAVID H SHERMAN
• 金额: $ 16.73万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9277486
• 关键词: Area; Biochemical; Bioinformatics; Biological; Carrier Proteins; Characteristics; Clinical; Complement; Complex; Coupling; Cytochrome P450; Data Set; Engineering; Enzymes; Erythromycin; Focus Groups; Gene Cluster; Genomics; Health; Human; Knowledge; Lead; Metabolism; Metagenomics; Methods; Mixed Function Oxygenases; Modification; National Institute of General Medical Sciences; Natural Products; Pathway interactions; Pharmaceutical Preparations; Population; Process; Property; Reaction; Research; System; Technology; Therapeutic Agents; Tylosin; Ursidae Family; Work; base; bryostatin; design; fascinate; functional group; improved; interdisciplinary approach; microbial host; microbiome; novel; oxidation; peptide synthase; picromycin; polyketide synthase; programs; public health relevance; success; synthetic biology

 DESCRIPTION (provided by applicant): This proposed MIRA project employs a range of multi-disciplinary approaches toward the discovery and analysis of natural products and the biosynthetic pathways that assemble and modify complex metabolites. The proposal covers three areas that have been supported by NIGMS during the past 20 years. Each has been articulated as a Grand Challenge designed to complement our accomplishments and continue to push forward vigorously to discover new knowledge and offer solutions with high potential for improving human health. Grand Challenge I of this MIRA application is based on the exciting momentum of a highly productive and collaborative program lead by my group that focuses on the pikromycin (Pik), erythromycin (DEBS), tylosin (Tyl), curacin (Cur) and bryostatin (Bry) pathways whose detailed analysis has been further developed during the previous cycle of support. These systems each bear fascinating biochemical features that will expand our understanding of substrate selectivity, and structural characteristics that enable functional activity within and between native and engineered polyketide synthase/non-ribosomal peptide synthetase modules. Grand Challenge II of this proposal focuses on studies relating to natural product pathway tailoring enzymes. A fundamental aspect of structural diversification in secondary metabolism involves oxidative processes that contribute significantly to biological activity. This can be readily appreciated in a number of important molecules that are clinical therapeutic agents, or show significant potential as drug leads. Based on the important successes in our research relating to P450 substrate and enzyme engineering over the past four years, we have been emboldened to expand our work in exciting new directions. This includes plans to investigate a range of P450 monoxygenases that catalyze iterative oxidative processes. We will also investigate monooxygenases that catalyze C-C coupling involving substrates in both inter- and intramolecular oxidation reactions, including aromatic, alkyl and alkenyl functional groups. One of the most underexplored, yet very important classes of tailoring enzyme includes the acyl/peptidyl carrier protein dependent monooxygenases, and we propose to explore mechanisms of selectivity and proceed with efforts to expand their substrate recognition and biocatalytic properties. Grand Challenge III focuses on natural product discovery and pathway engineering. We have established the technologies and bioinformatics capabilities to readily assemble and mine genomic, and metagenomic datasets from diverse microbiome populations toward natural product gene cluster discovery, which is now poised for heterologous expression in amenable microbial hosts. The next wave of progress will rely on ready identification of the most novel pathways, and our ability to express them using facile synthetic biology methods. We plan to attack these problems with utmost energy and determination to gain access to important compounds with valuable medicinal properties.

 描述（由申请人提供）：这个拟议的MIRA项目采用了一系列多学科的方法来发现和分析天然产物以及组装和修饰复杂代谢物的生物合成途径。该提案涵盖了过去20年来得到国家地理定位系统支持的三个领域。每一项都被视为一项重大挑战，旨在补充我们的成就，并继续大力推进，以发现新知识，并提供具有改善人类健康潜力的解决方案。该MIRA应用程序的大挑战I是基于由我的团队领导的高效协作计划的令人兴奋的势头，该计划专注于吡克霉素（Pik），红霉素（DEBS），泰乐菌素（Tyl），Curacin（Cur）和苔藓抑素（Bry）途径，其详细分析已在上一个支持周期中得到进一步发展。这些系统各自具有迷人的生物化学特征，这将扩大我们对底物选择性的理解，以及使天然和工程化聚酮化合物合酶/非核糖体肽合成酶模块内和之间的功能活性成为可能的结构特征。该提案的大挑战II侧重于与天然产物途径剪裁酶相关的研究。次级代谢结构多样化的一个基本方面涉及对生物活性有重大贡献的氧化过程。这可以很容易地在许多重要的分子中被理解，这些分子是临床治疗剂，或者显示出作为药物先导的显著潜力。基于我们在过去四年中与P450底物和酶工程相关的研究取得的重要成功，我们有勇气在令人兴奋的新方向上扩大我们的工作。这包括研究一系列催化迭代氧化过程的P450单加氧酶的计划。我们还将研究单加氧酶，催化C-C耦合涉及底物在分子间和分子内氧化反应，包括芳香族，烷基和烯基官能团。其中一个最underexplored，但非常重要的一类裁剪酶包括酰基/肽基载体蛋白依赖性单加氧酶，我们建议探索机制的选择性和继续努力扩大其底物识别和生物催化性能。Grand Challenge III专注于天然产物发现和途径工程。我们已经建立了技术和生物信息学能力，可以轻松地从不同的微生物组群体中组装和挖掘基因组和宏基因组数据集，以发现天然产物基因簇，该基因簇现已准备在顺从的微生物宿主中进行异源表达。下一波进展将依赖于对最新途径的快速识别，以及我们使用简单的合成生物学方法表达它们的能力。我们计划以最大的精力和决心来解决这些问题，以获得具有宝贵药用价值的重要化合物。