Biosynthetic Analysis of Marine Cyanobacterial Pathways

海洋蓝藻途径的生物合成分析

• 批准号: 8106995
• 负责人: William Henry Gerwick
• 金额: $ 35.78万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8106995
• 关键词: Ally; Antineoplastic Agents; Bacterial Infections; Biochemical; Biological; Biological Assay; Biological Factors; Cells; Chemicals; Complex; Cryptophycin; Cyanobacterium; Development; Engineering; Enzymes; Genetic; Genomics; Grant; In Vitro; Inflammatory Response; Insecta; Institution; Kinetics; Knowledge; Laboratories; Laboratory culture; Lead; Malignant Neoplasms; Marines; Metabolic; Metabolic Diseases; Methodology; Methods; Michigan; Molecular; Oceanography; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Process; Production; Property; Proteins; Research; Research Institute; Sequence Analysis; Series; Site-Directed Mutagenesis; Source; Structure; Substrate Specificity; System; Technology; Universities; analog; base; design; drug development; human disease; in vivo; killings; microbial; microorganism; novel; pathogen; peptide synthase; polyketide synthase; programs; structural biology

DESCRIPTION (provided by applicant): Marine cyanobacteria are extraordinarily rich in their production of biologically active and structurally unique natural products. A number of these secondary metabolites or their derivatives are lead compounds in drug development programs aimed at providing new therapies to treat cancer, bacterial infections, inflammatory responses and in crop protection to kill harmful microbial pathogens and insects. Isolation and structural analysis of marine and terrestrial cyanobacterial natural products has provided access to an unusually large number of mixed non-ribosomal peptide synthetase/polyketide synthase (NRPS/PKS) systems. The corresponding metabolic systems are comprised of an intriguing set of complex multifunctional proteins that along with allied enzymes generate structurally complex molecules via a modular multi-step process. Over the past several years the Sherman, Gerwick and Smith laboratories have developed a complementary program to clone and characterize the biosynthetic pathways of novel cyanobacterial secondary metabolites that possess significant potential for biotechnological applications. Despite considerable progress, a full understanding of the molecular mechanisms, catalytic activities, kinetic properties, and substrate specificities within cyanobacterial biosynthetic pathways is just beginning to unfold. The proposed research will build upon our accomplishments on the curacin, jamaicamide and cryptophycin/ arenastatin metabolic systems, three robust pathways that have been a rich source of new information. The expected metabolic, biochemical and structural understanding will facilitate the design of new biosynthetic systems that harness the growing potential of cyanobacterial natural product pathways. The full promise of cyanobacterial natural products to yield new lead compounds for development as useful pharmaceuticals will only be realized by closing a series of key gaps in knowledge and technology. Solving these challenges will require development and optimization of genetic and biochemical methods that allow us to 1) manipulate cyanobacterial natural product metabolic systems to produce analog structures, 2) utilize unique secondary metabolite enzymes for creation of novel bioactive molecules and, 3) screen new compounds and analogs to identify promising new anticancer compounds for further development. The specific aims are: 1. To harness the inherent versatility of cyanobacterial natural product systems to create new anticancer lead compounds. Sub-aims include: a. Investigate ability of cyanobacterial biosynthetic pathways to generate novel analogs using unique laboratory culture and mutasynthesis methodologies. b. Investigate the unique enzymatic capabilities of marine cyanobacterial pathways to engineer new metabolic systems and tailoring processes to generate new bioactive compounds. c. Employ structural biology and site-directed mutagenesis approaches to understand the precise biochemical mechanisms of unique biosynthetic enzymes. d. Develop new chemoenzymatic, in vivo, and in vitro pathways to create new anticancer agents with enhanced medicinal properties 2. Perform bioassays on new compounds resulting from Specific Aim 1. a. New compounds derived from the proposed research will be transferred to Eisai Research Institute and University of Michigan Center for Chemical Genomics for analysis of biological activity using a series of biochemical and cell based assays relevant to cancer. PUBLIC HEALTH RELEVANCE: The proposed research will focus on elucidating the detailed function and mechanistic basis of complex biosynthetic pathways from marine cyanobacteria that create chemically diverse natural products with anti-cancer activity. The ability to understand and subsequently engineer these remarkable biochemical systems will create new opportunities to discover and develop effective drugs for the treatment of human diseases, particularly cancer and related metabolic disorders.

描述（由申请人提供）：海洋蓝细菌在生物学活性和结构独特的天然产物中的生产非常丰富。这些二级代谢物或其衍生物中的许多是药物开发计划中的铅化合物，旨在提供新的疗法来治疗癌症，细菌感染，炎症反应和作物保护，以杀死有害的微生物病原体和昆虫。海洋和陆生蓝细菌天然产物的隔离和结构分析已为异常数量的混合非核糖体肽合成酶/聚酮化酶/聚酮化合酶（NRPS/PKS）系统提供了访问。相应的代谢系统由一组有趣的复杂多功能蛋白组成，这些蛋白质与盟军一起通过模块化的多步骤生成结构上复杂的分子。在过去的几年中，谢尔曼（Sherman），格里克（Gerwick）和史密斯（Smith）实验室制定了一个互补程序，以克隆和表征新型蓝细菌次生代谢产物的生物合成途径，这些代谢物具有生物技术应用的重要潜力。 尽管取得了很大的进步，但对蓝细菌生物合成途径内的分子机制，催化活性，动力学特性和底物特异性有充分的了解刚刚开始展开。拟议的研究将基于我们在素素，牙买酰胺和隐霉素/竞技替汀代谢系统上的成就，这是三种强大的途径，这些途径已成为新信息的丰富来源。预期的代谢，生化和结构理解将促进新的生物合成系统的设计，从而利用蓝细菌天然产物途径的潜力越来越大。 蓝细菌天然产品的全部希望只能通过缩小知识和技术的一系列关键差距来实现新的铅化合物进行开发。解决这些挑战将需要开发和优化遗传和生化方法，使我们能够到达1）操纵氰基细菌天然产物代谢系统以生成模拟结构，2）利用独特的二级代谢物酶来创建新型的生物活性分子，以及筛选新的化合物和类似物的新型化合物，以实现良好的新混合物，以实现新的化合物，以实现新颖的新混合物。具体目的是：1。利用蓝细菌天然产品系统的固有多功能性来创建新的抗癌铅化合物。子iams包括：研究了使用独特的实验室培养和突变合成方法来研究蓝细菌生物合成途径的能力。 b。研究海洋蓝细菌途径的独特酶促能力，以设计新的代谢系统和剪裁过程，以生成新的生物活性化合物。 c。采用结构生物学和定向诱变方法来了解独特的生物合成酶的精确生化机制。 d。开发新的化学酶，体内和体外途径，以创建具有增强药用特性的新抗癌剂2。从拟议的研究中得出的新化合物将转移到EISAI研究所和密歇根大学化学基因组中心，以使用与癌症相关的一系列生化和基于细胞的测定法分析生物学活性。 公共卫生相关性：拟议的研究将着重于阐明从海洋蓝细菌中复杂的生物合成途径的详细功能和机械基础，这些途径从海洋蓝细菌中创造出具有抗癌活性的化学多样性天然产物。理解和随后设计这些非凡的生化系统的能力将创造新的机会，以发现和开发有效的药物来治疗人类疾病，尤其是癌症和相关代谢疾病。