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

基本信息

批准号：
7849914
负责人：
MARK R LILES
金额：
$ 18.08万
依托单位：
AUBURN UNIVERSITY AT AUBURN
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7849914
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

项目摘要

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）：抗生素是我们针对细菌病原体的第一道防线，这些病原体一直是人类文明的瘟疫。由于抗生素耐药性，越来越多地认为被视为可治疗感染剂的人正在成为严重的问题。在药品管道中，迫切需要新型的抗生素，但由于许多原因，制药行业尚未在抗生素发现过程中投入大量投资。抗生素发现的最佳资源是探索复杂化学自然发展的微生物群落。为了从多种微生物中获取复杂的化学物质，该提案采用了一种社区基因组方法，其中从土壤中直接回收了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。已经从这些社区基因组库中隔离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任何毒性。除了表达抗生素活性的克隆外，还分离了另一个集合的克隆，该克隆包含特定类别的天然产物Polyketides的基因。聚酮化合物具有多种结构和不同的活性，例如抗菌和抗癌特性。从土壤微生物社区文库中发现的聚酮化合物生物合成途径的遗传分析表明，含34个聚酮化合物的克隆与已知的聚酮化合物途径大不相同。我们的研究工作将将这些聚酮途径转移到新的克隆载体中，旨在允许在革兰氏阴性细菌之间转移遗传途径，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的细菌物种中表达。这项研究的结果将是具有抗菌活性的铅候选化合物的集合，新型的Polyketide Biosynthetic途径可用于具有治疗特性的工程新型Polyketide结构，对新型的革兰氏阴性快车载体的评估以及下一代Microbial社区基因组库，以实现这项技术的未来开发。公共卫生相关性：抗生素对我们打击传染病的能力至关重要，尤其是在一个抗生素耐药性发病率上升的时代，需要发现和开发临床上有用的抗生素。已经分离了新技术来克隆和表达各种微生物的遗传途径，已经分离出产生抗菌活性或编码生物合成途径的克隆集合。拟议的研究将在遗传和生化水平上评估该集合的克隆，以确定哪些主要候选者作为新型治疗剂具有最佳潜力。