A defined minimal medium for systems analyses of reveals strain-specific metabolic requirements.

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ID: 45938
2019
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Abstract
is a Gram-positive pathogenic bacterium that colonizes an estimated one-third of the human population and can cause a wide spectrum of disease, ranging from superficial skin infections to life-threatening sepsis. The adaptive mechanisms that contribute to the success of this pathogen remain obscure partially due to a lack of knowledge of its metabolic requirements. Systems biology approaches can be extremely useful in predicting and interpreting metabolic phenotypes; however, such approaches rely on a chemically defined minimal medium as a basis to investigate the requirements of the cell. In this work, a chemically defined minimal medium formulation, termed synthetic minimal medium (SMM), was investigated and validated to support growth of three strains: LAC and TCH1516 (USA300 lineage), and D592 (USA100 lineage). The formulated SMM was used in an adaptive laboratory evolution (ALE) experiment to probe the various mutational trajectories of all three strains leading to optimized growth capabilities. The evolved strains were phenotypically characterized for their growth rate and antimicrobial susceptibility. Strains were also re-sequenced to examine the genetic basis for observed changes in phenotype and to design follow-up metabolite supplementation assays. Our results reveal evolutionary trajectories that arose from strain-specific metabolic requirements. SMM and the evolved strains can also serve as important tools to study antibiotic resistance phenotypes of As researchers try to understand and combat the development of antibiotic resistance in pathogens, there is a growing need to thoroughly understand the physiology and metabolism of the microbes. is a threatening pathogen with increased antibiotic resistance and well-studied virulence mechanisms. However, knowledge regarding the adaptive mechanisms used by this pathogen to survive environmental stresses remain unclear, mostly due to the lack of information about its metabolic requirements. Defining the minimal metabolic requirements for growth is a first step towards unraveling the mechanisms by which it adapts to metabolic stresses. Here we present the development of a chemically defined minimal medium supporting growth of three strains, and reveal key genetic mutations contributing to improved growth in minimal medium.
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machado2019aapplied Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Machado, Henrique;Weng, Liam L;Dillon, Nicholas;Seif, Yara;Holland, Michelle;Pekar, Jonathan E;Monk, Jonathan M;Nizet, Victor;Palsson, Bernhard O;Feist, Adam M;
Journal Applied and environmental microbiology
Year 2019
DOI
AEM.01773-19
URL
Keywords
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