Evaluation of Novel Antimicrobial Agents and a Novel Shuttle Vector

新型抗菌剂和新型穿梭载体的评价

• 批准号: 7708548
• 负责人: MARK R LILES
• 金额: $ 23.01万
• 依托单位: AUBURN UNIVERSITY AT AUBURN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7708548
• 关键词: Anabolism; Antibiotic Resistance; Antibiotics; Bacterial Artificial Chromosomes; Biochemical; Bioinformatics; Biological Assay; Biological Factors; Chemical Structure; Chemicals; Chemistry; Civilization; Cloning; Cloning Vectors; Collection; Communicable Diseases; Communities; Complex; DNA Sequence; Development; Drug Industry; Engineering; Escherichia coli; Eukaryotic Cell; Evaluation; Funding; Future; Genes; Genetic; Genomic Library; Genomics; Goals; Gram-Negative Bacteria; Health; Human; Incidence; Infectious Agent; Lead; Libraries; Liquid Chromatography; Metagenomics; Molecular Weight; Pathway interactions; Pharmacologic Substance; Phylogenetic Analysis; Plague; Process; Production; Property; Recombinants; Relative (related person); Research; Resources; Shuttle Vectors; Soil; Structure; Therapeutic; Therapeutic Agents; Toxic effect; Type I Polyketide Synthase; antimicrobial; antimicrobial drug; combat; design; expression cloning; genetic analysis; microbial community; microorganism; new technology; next generation; novel; novel therapeutics; pathogen; pathogenic bacteria; polyketide synthase; public health relevance; technology development; vector

DESCRIPTION (provided by applicant): Antibiotics are our first line of defense against bacterial pathogens that have historically been a plague on human civilization. Increasingly, human pathogens that had been regarded as treatable infectious agents are becoming serious problems as a result of antibiotic resistance. There is an urgent need for novel antibiotics in the pharmaceutical pipeline, yet for many reasons the pharmaceutical industry has not invested heavily in the antibiotic discovery process. The best possible resource for antibiotic discovery is to explore microbial communities, where complex chemistries have naturally evolved. To access the complex chemistries from diverse microorganisms, this proposal uses a community genomic approach in which biosynthetic genes from diverse microorganisms are directly recovered from soils and expressed in E. coli or other bacterial hosts. A collection of fourty-four clones that produce antibiotic activity has already been isolated from these community genomic libraries. The proposed research will determine the chemical structure of antibiotic compounds from this collection, and determine the potency and any toxicity associated with these compounds. In addition to clones that express antibiotic activity, another collection of clones has been isolated that contain the genes for a specific class of natural product, the polyketides. Polyketide compounds have diverse structures, and diverse activities, such as antimicrobial and anticancer properties. A genetic analysis of polyketide biosynthesis pathways discovered from a soil microbial community library indicates that the 34 polyketide-containing clones are very different from known polyketide pathways. Our research efforts will transfer these polyketide pathways into a novel cloning vector, designed to allow transfer of genetic pathways between Gram-negative bacteria, and then genetic and bioassay screens will determine whether these polyketide pathways are expressed in different bacterial species. The results of this research will be a collection of lead candidate compounds with antimicrobial activity, novel polyketide biosynthetic pathways useful in engineering novel polyketide structures with therapeutic properties, an evaluation of a novel Gram- negative shuttle vector, and the next generation of microbial community genomic library for future development of this technology. PUBLIC HEALTH RELEVANCE: Antibiotics are critical to our ability to combat infectious disease, and especially in an era in which the incidence of antibiotic resistance is on the rise there is a need for discovery and development of clinically useful antibiotics. Using new technology for cloning and expressing genetic pathways from diverse microorganisms, a collection of clones that either produce an antimicrobial activity or encode a biosynthetic pathway have been isolated. The proposed research would evaluate this collection of clones at a genetic and biochemical level to determine which lead candidates have the best potential as novel therapeutic agents.

描述（由申请人提供）：抗生素是我们对抗历史上一直困扰人类文明的细菌病原体的第一道防线。由于抗生素耐药性，曾经被视为可治疗的传染源的人类病原体正日益成为严重的问题。制药管道中迫切需要新型抗生素，但由于多种原因，制药行业并未在抗生素发现过程中投入大量资金。抗生素发现的最佳资源是探索微生物群落，其中复杂的化学物质自然进化。为了获取来自不同微生物的复杂化学物质，该提案采用了群落基因组方法，其中直接从土壤中回收来自不同微生物的生物合成基因并在大肠杆菌或其他细菌宿主中表达。已经从这些群落基因组文库中分离出了产生抗生素活性的四十四个克隆的集合。拟议的研究将确定该集合中抗生素化合物的化学结构，并确定与这些化合物相关的效力和毒性。除了表达抗生素活性的克隆之外，还分离出另一组克隆，其中含有特定类别的天然产物聚酮化合物的基因。聚酮化合物具有多种结构和多种活性，例如抗菌和抗癌特性。对土壤微生物群落文库中发现的聚酮化合物生物合成途径的遗传分析表明，34 个含聚酮化合物的克隆与已知的聚酮化合物途径有很大不同。我们的研究工作将把这些聚酮化合物途径转移到一种新型克隆载体中，该载体旨在允许革兰氏阴性细菌之间的遗传途径转移，然后遗传和生物测定筛选将确定这些聚酮化合物途径是否在不同细菌物种中表达。这项研究的结果将是一系列具有抗菌活性的主要候选化合物、可用于设计具有治疗特性的新型聚酮化合物结构的新型聚酮化合物生物合成途径、对新型革兰氏阴性穿梭载体的评估以及用于该技术未来发展的下一代微生物群落基因组文库。公共卫生相关性：抗生素对于我们对抗传染病的能力至关重要，尤其是在抗生素耐药性发生率上升的时代，需要发现和开发临床上有用的抗生素。使用克隆和表达来自不同微生物的遗传途径的新技术，已经分离出一组产生抗菌活性或编码生物合成途径的克隆。拟议的研究将在遗传和生化水平上评估这组克隆，以确定哪些主要候选药物最有可能成为新型治疗剂。