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（由适用提供）：该提出的MIRA项目采用了一系列多学科方法来发现和分析天然产物以及组装和修改复杂代谢物的生物合成途径。该提案涵盖了过去20年中奈IMS支持的三个领域。每个人都被阐明为旨在完成我们的成就的巨大挑战，并继续向前迈进，以发现新知识，并提供具有更大潜力改善人类健康的解决方案。此MIRA应用程序的盛大挑战I是基于我的小组领导的高产和协作计划的激动人心的时刻，该计划的重点是pikroycin（Pik）（Pik），红霉素（DEBS），Tylosin（Tyl），curacin（Cur）（CUR）（CUR）和Bryostatin（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）（Bry）途径在上一个周期的详细分析中得到了进一步的支持。这些系统具有引人入胜的生化特征，这些特征将扩大我们对底物选择性的理解，以及在天然和工程聚酮化合物合成酶/非核糖体肽合成酶模块内以及在本地和工程聚酮化合酶内和之间实现功能活性的结构特征。该提案的大挑战II着重于与自然产品途径调整酶有关的研究。次生代谢中结构多样化的一个基本方面涉及对生物活性产生重大贡献的氧化过程。这可以在许多是临床治疗剂的重要分子中很容易理解的，或者表现出像药物铅一样重要的潜力。基于过去四年中与P450底物和酶工程有关的研究的重要成功，我们一直在勇敢地将工作扩展到令人兴奋的新方向上。这包括研究一系列催化迭代氧化物过程的P450一氧合酶的计划。我们还将研究单加氧酶，以催化涉及氧化物间和分子内氧化物反应（包括芳族，烷基和烷烯基官能团）的C-C耦合。最不受欢迎但非常重要的剪裁酶之一包括酰基/肽基载体蛋白依赖性单加掺杂酶，我们建议探索选择性的机制，并继续努力扩大其底物识别和生物催化特性。大挑战III专注于自然产品发现和途径工程。我们已经建立了很容易组装和矿山基因组的技术和生物信息学能力，以及从潜水员微生物组人群到天然产物基因群集发现的宏基因组数据集，现在在可机动微生物宿主中有异源表达中毒。下一步的进步将依赖于最新颖的途径的现成识别，以及我们使用轻松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攻击这些问题，以获取具有宝贵医疗特性的重要化合物。