Comprehensive analysis of NRPS-derived metabolomes of three Aspergillus species

三种曲霉属 NRPS 衍生代谢组的综合分析

基本信息

批准号：
8798807
负责人：
NANCY P KELLER
金额：
$ 29.26万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-12-15 至 2018-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8798807
关键词：
Agriculture Anabolism Antibiotics Aspergillus Aspergillus nidulans Bacterial Artificial Chromosomes Biological Biological Models Biology Chemicals Communities Complement Complex Coupled Drug Design Engineering Evaluation Fungal Genome Future Gene Cluster Genes Genetic Genetic Models Genetic Recombination Genome Goals Grant Immunosuppressive Agents Knowledge Laboratories Lipids Methodology Modeling Molds NMR Spectroscopy Orphan Pathway interactions Peptides Pharmaceutical Preparations Pharmacologic Substance Play Production Property Research Resolution Resources Role Source Sterigmatocystin Structure Substrate Domain Surveys Techniques Virulence Factors Work Yeasts antimicrobial base comparative drug discovery fungus genome sequencing innovation interest member metabolomics overexpression pathogen peptide synthase preference promoter public health relevance small molecule tool vector

项目摘要

DESCRIPTION (provided by applicant): Filamentous fungi produce a vast universe of secondary metabolites (SM), which display a broad range of useful activities for pharmaceutical and agricultural purposes, e.g. antibiotic, immunosuppressant, lipid lowering, or antimicrobial properties. One group of metabolites of particular interest are non-ribosomal peptide synthetase- (NRPS-) derived metabolites which have provided important leads for drug design. However, production of NRPS-based pathways is often very low (or undetectable) under laboratory conditions, in large part due to complex endogenous regulatory networks that control the biosyntheses of these metabolites, which remain, at best, poorly understood. As a result, many gene clusters containing NRPS or NRPS-like genes have no known metabolites ("orphan gene clusters"), even in well-studied species such as the genetic model Aspergillus nidulans. This R01 proposes in Aim 1 to identify metabolites associated with all remaining orphan NRPS and NRPS-like gene clusters in A. nidulans and the opportunistic pathogen A. fumigatus using an inducible promoter activation strategy coupled with comparative metabolomics based on high-resolution MS and 2D NMR spectroscopy. In Aim 2 we will utilize a Bacterial Artificial Chromosome (BAC)-based strategy to identify metabolites for all NRPS and NRPS-like gene clusters of A. terreus, an industrially important fungal species. A final goal of this aim is to examine the hypothesis that specific genes contained within a fungal NRPS/NRPS-like cluster may be predictive of the biological activity - and drug discovery potential - of the SM products. Overall, this parallel study of different species and comparison of different expression approaches will significantly advance understanding of NRPS-pathways and the scope of heterologous expression. Moreover, this work will provide advanced fungal lines for the community for SM cluster expression, a comprehensive assessment of A-domain substrate preference for fungal NRPS and an innovative methodology to identify new metabolites for biological evaluation.

描述（由申请人提供）：丝状真菌产生了广阔的二级代谢产物（SM），这些宇宙显示出用于药物和农业用途的广泛有用活动，例如抗生素，免疫抑制剂，脂质降低或抗菌特性。一组特别感兴趣的代谢产物是非核糖体肽合成酶（NRPS-）衍生的代谢产物，这些代谢物为药物设计提供了重要的铅。然而，在实验室条件下，基于NRPS的途径的产生通常非常低（或无法检测到），这在很大程度上是由于控制这些代谢物的生物合成的复杂内源性调节网络，这些网络充其量仍然是尚不清楚的。结果，即使在诸如遗传模型曲霉曲霉（Aspergillus nidulans）等精心研究的物种中，许多含有NRP或类似NRP的基因的基因簇也没有已知的代谢产物（“孤儿基因簇”）。该R01在AIM 1中提出，旨在鉴定与A. nidulans中所有剩余的孤儿NRP和类似NRPS的基因簇相关的代谢产物，以及使用诱导启动子激活策略与基于高分辨率NMR和2D NMR光谱的比较代谢组学结合使用诱导型启动子激活策略。在AIM 2中，我们将利用基于细菌的人工染色体（BAC）策略来鉴定所有NRP和NRPS样基因簇的代谢物，A. terreus是一种工业重要的真菌物种。该目的的最终目标是检验以下假设：真菌NRPS/NRPS样簇中包含的特定基因可能可以预测SM产物的生物学活性和药物发现潜力。总体而言，这项对不同物种的平行研究以及对不同表达方法的比较将显着提高对NRPS轨迹和异源表达范围的理解。此外，这项工作将为社区提供SM簇表达的高级真菌线，对A域基板对真菌NRP的偏好进行了全面评估，以及一种创新方法，以识别用于生物学评估的新代谢物。