Biosynthesis of antifungal nucleoside antibiotics

抗真菌核苷抗生素的生物合成

基本信息

批准号：
10264167
负责人：
Kenichi Yokoyama
金额：
$ 32.86万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10264167
关键词：
Acids Amides Anabolism Antibiotics Antifungal Agents Bacteria Carbohydrates Cell Wall Cellular Structures Chemicals Chitin Clinical Communicable Diseases Data Development Drug resistance Engineering Enzymatic Biochemistry Enzymes Excision Exhibits Foundations Funding Fungal Components Future Gene Cluster Gene Combinations Generations Genetic Genome Genomics Goals Growth Human Immunocompromised Host Knowledge Ligase Ligation Mediating Mining Mycoses Natural Products Nucleosides Oxygenases Pathway interactions Phosphorylation Phosphotransferases Physiological Polysaccharides Preparation Protein Dephosphorylation Public Health Reaction Reporting Research Resistance Specificity Streptomyces Structure Structure-Activity Relationship Substrate Specificity Testing Toxic effect Travel alpha ketoglutarate analog base carbohydrate structure clinical development combat desert fever experimental study fungus guided inquiry in vivo inorganic phosphate nikkomycin novel novel therapeutics nucleoside analog nucleoside triphosphatase pathogenic fungus reconstitution sugar text searching

项目摘要

Project Summary/Abstract Peptidyl nucleosides (PNs) are naturally occurring antifungal agents active against multiple pathogenic fungi such as the causal agent of “Valley Fever”. They also exhibit potent synergistic effects with clinically approved antifungal drugs. Our long-term goal is to provide a comprehensive understanding of both the biosynthesis of PNs and their mode of action. The current application focuses on the biosynthesis of PNs. Understanding PN biosynthetic pathways will provide a basis for creating structurally diverse PN analogs through engineered biosynthesis, semi synthesis and genome mining. In the previous funding cycle, we found that PNs are biosynthesized through cryptic phosphorylation and carbohydrate tailoring by oxidative C-C bond cleavage. On the basis of these findings, in this application, we will perform functional and mechanistic characterization of the biosynthetic enzymes to provide the foundation for genome mining discovery of novel nucleoside natural products and chemoenzymatic synthesis of unnatural PNs. In Aim 1, the mechanism of removal of cryptic phosphorylation and the generality of cryptic phosphorylation in other nucleoside biosynthesis pathways will be investigated. In Aim 2, the radical mediated divergent biosynthesis of nucleoside natural products will be investigated by studying the mechanism of oxidative C-C bond cleavage and genome mining characterization of homologous enzymes. In Aim 3, the mechanism of amide ligation by NikS/PolG enzymes will be characterized, and their potentials for use in the chemoenzymatic preparation of PNs will be investigated. The proposed research is significant because it will provide a basis for the future biosynthetic and chemoenzymatic generation of novel therapeutic PNs as well as genome mining discovery of novel antifungal nucleoside natural products.

项目摘要/摘要肽基核酶（PNS）是天然存在的抗真菌剂，对多种致病真菌进行活性例如“山谷热”的因果因素。他们还通过临床批准，暴露了有效的协同作用抗真菌药物。我们的长期目标是对同时的生物合成提供全面的理解 PNS及其作用方式。当前的应用集中于PNS的生物合成。了解PN 生物合成途径将为通过工程设计而创建结构上的PN类似物提供基础生物合成，半合成和基因组开采。在上一个资金周期中，我们发现PN是通过氧化C-C键裂解量身定制的加密磷酸化和碳水化的生物合成。在这些发现的基础，在此应用中，我们将执行功能和机械表征生物合成酶为新型核球自然的基因组开采发现基础非天然PNS的产物和化学酶合成。在AIM 1中，去除加密的机制其他核苷生物合成途径中隐磷酸化的磷酸化和一般性将是调查。在AIM 2中，核外侧天然产物的自由基介导的发散生物合成将是通过研究氧化C-C键裂解的机制和基因组挖掘表征同源酶。在AIM 3中，将表征NIKS/POLG酶的酰胺连接机理，他们将研究它们在PNS化学酶制剂中使用的潜力。提议研究很重要，因为它将为未来的生物合成和化学酶产生提供基础新型治疗PNS以及新型抗真菌核核苷天然产物的基因组开采发现。