Genomic approaches of discovery broad-spectrum antimicrobial agents

发现广谱抗菌药物的基因组方法

• 批准号: 7669772
• 负责人: Jon Clardy
• 金额: $ 34.21万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-25 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7669772
• 关键词: Actinobacteria class; Actinomyces; Actinomycetales; Antibiotics; Bacteria; Bioinformatics; Biological Factors; Chemistry; Culture Media; Ecology; Elements; Environment; Funding; Gene Cluster; Genetic; Genome; Genomics; Goals; Grant; Institutes; Methodology; Microbe; National Institute of Allergy and Infectious Disease; New England; Nutrient; Pilot Projects; Production; Resistance; Screening procedure; Simulate; antimicrobial; antimicrobial drug; base; biodefense; combat; microbial; novel; pathogen; programs; promoter; scaffold; small molecule; tool

Most clinically-used antibiotics are bacterially-produced small molecules known as natural products, or their derivatives. However, natural products are not being featured in antibiotic discovery programs due to high rates of rediscovery and low rates of new molecule discovery. This proposal describes a new genomicsbased approach to natural product discovery that employs a combination of bioinformatics, microbial ecology, genetics, and chemistry to identify cryptic biosynthetic loci and stimulate them to produce their encoded natural products. All molecules identified will be characterized for antimicrobial activity. Specific Aim 1. Use bioinformatics to identify biosynthetic gene clusters and predict their products. New bioinformatic tools will be developed to identify biosynthetic gene clusters in the genomes of actinomycetes and other microbes, and to predict structural elements of their small molecule products. These predictions will be leveraged toward antibiotic discovery by using them as the basis for efforts to stimulate the production of cryptic natural products, as described in Specific Aims 2 and 3: Specific Aim 2. Stimulate the production of cryptic metabolites by simulating a multispecies environment. Most screens for new natural products have been performed with strains grown as pure cultures in nutrientrich growth media. A new screening format has recently been developed in which strains are grown as microcolonies on nutrient-poor growth media. This microcolony screening methodology will be used to identify cryptic natural products with antimicrobial activity. Specific Aim 3. Stimulate the production of cryptic metabolites by genetically manipulating producers. Most natural product-encoding gene clusters are thought to be repressed under standard culture conditions. Using actinomycete strains whose biosynthetic gene clusters have been identified by the efforts described in Specific Aim 1, endogenous gene cluster promoters will be systematically replaced with a strong, inducible promoter, enabling the controlled synthesis and isolation of their small molecule products and the subsequent characterization of their antimicrobial activity. .

大多数临床使用的抗生素是细菌产生的小分子，称为天然产物，或其