A Genome-Mining Strategy for identification of new antibiotics produced by short LanB enzymes

用于鉴定由短 LanB 酶产生的新抗生素的基因组挖掘策略

基本信息

批准号：
9767527
负责人：
Chi Pan Ting
金额：
$ 4.72万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-16 至 2020-06-02
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9767527
关键词：
Amino Acids Anabolism Antibiotics Antifungal Agents Bacterial Antibiotic Resistance Bioinformatics Biological C-terminal Carbon Chemicals Complex Cyclization Cysteine Dehydration Development Enzymes Escherichia coli Excision Exhibits Family Food Preservatives Gene Cluster Genes Genetic Genome Gram-Negative Bacteria Health Human Hydro-Lyases Infection Inherited Investigation Lead Light Logic Mass Spectrum Analysis Methods Mining Modeling Modification Natural Products Nisin Organism Pathogenicity Penicillins Peptides Production Pseudomonas syringae Reaction Recording of previous events Resistance Ribosomes Roentgen Rays Role Serine Societies Source Structure System Testing Therapeutic Threonine Time Transfer RNA antimicrobial bacterial resistance base combat dehydroalanine dehydrobutyrine enzyme activity enzyme biosynthesis glutamic acid-tRNA homologous recombination in vivo novel pathogenic bacteria scaffold

项目摘要

Project Summary/Abstract Development of antibiotic resistance by bacterial pathogens poses an inherit need for the identification of new chemical entities that will serve as new classes of antibiotics. Lanthipeptides are a family of ribosomally synthesized post-translationally modified peptides (RiPPs) consisting of structurally diverse natural products many of which are antibiotics. In this proposal, a genome mining strategy for the identification of new natural product RiPPs will be explored. The recent discovery of a new class of truncated LanB enzymes, herein termed short LanBs (sLanB), has prompted investigation of the natural products they produce. Given the rich history of lanthipeptides, formerly known as lantibiotics, as antimicrobial compounds, the natural products derived from sLanB enzymes are expected to possess similar therapeutic value and structural diversity. The first aim of this proposal involves the isolation of a natural product from a model gene cluster in Pseudomonas syringae containing a single sLanB gene. The enzymes encoded by the gene cluster will be studied by heterologous expression in E. coli. After determining the natural product structure, isolation from the native organism will be attempted by chemical derivatization. Chromosomal disruption by homologous recombinations represents an alternative method for natural product determination by isolation of advanced biosynthetic intermediates. Building upon our initial studies, the second aim of this proposal involves identification of a natural product from a complex gene cluster in Desmospora sp. 8437, containing a total of seven sLanB enzymes. The enzymes encoded by the gene cluster will be heterologous expressed in E. coli. In vivo co-expression of all seven sLanBs with tagged precursor peptide will allow for rapid determination of enzyme activity. The modifications by the sLanB enzymes will be analyzed by mass spectrometry. This overall strategy is expected to allow for rapid isolation of natural products derived from multiple sLanB enzymes. Natural products isolated from these two gene clusters will be evaluated for their antimicrobial activity as potential therapeutics.