A comprehensive method to elucidate pyoverdines produced by fluorescent Pseudomonas spp. by UHPLC-HR-MS/MS
| العنوان: | A comprehensive method to elucidate pyoverdines produced by fluorescent Pseudomonas spp. by UHPLC-HR-MS/MS |
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| المؤلفون: | Rehm, Karoline, Vollenweider, Vera, Kümmerli, Rolf, Bigler, Laurent |
| المساهمون: | University of Zurich |
| المصدر: | Analytical and Bioanalytical Chemistry. 414:2671-2685 |
| بيانات النشر: | Springer Science and Business Media LLC, 2022. |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | 10120 Department of Chemistry, Tandem Mass Spectrometry, Pseudomonas, 540 Chemistry, Siderophores, Oligopeptides, 11493 Department of Quantitative Biomedicine, Biochemistry, Chromatography, High Pressure Liquid, Analytical Chemistry |
| الوصف: | Microbial secondary metabolites represent a rich source for drug discovery, plant protective agents, and biotechnologically relevant compounds. Among them are siderophores, iron-chelating molecules, that show a great influence on bacterial community assembly and the potential to control pathogen invasions. One of such a siderophore is pyoverdine that is produced by fluorescent Pseudomonas members and consists of different peptide chains specific to each bacterial species. The identification and structural elucidation of such suites of siderophores remain widely underexplored as general high-throughput analytical protocols are missing. Therefore, a dedicated method was established allowing a rapid localization and structural elucidation of pyoverdines. Liquid bacterial culture samples were purified by an easy small-scale solid-phase extraction (SPE). Ultra-high-performance liquid chromatography high-resolution tandem mass spectrometry (UHPLC-HR-MS/MS) separated highly polar pyoverdines and their derivatives. All ion fragmentation (AIF) generated mass spectra containing the characteristic fragments of the biological precursor of pyoverdine, ferribactin. This led to the revelation of the mass of secreted pyoverdines. Targeted MS/MS experiments at multiple collision energies accomplished the full structure elucidation of the pyoverdine peptide chain. A mass calculator and a fragmentation predictor facilitated greatly the interpretation of MS/MS spectra by providing accurate masses for a straightforward comparison of measured and theoretical values. The method was successfully validated using four well-known pyoverdines with various peptide chains. Finally, the applicability was proven by the analysis of 13 unknown pyoverdines secreted by sampled bacterial cultures. Among these, 4 novel pyoverdine peptide chains were discovered and are herein reported for the first time. Graphical abstract |
| وصف الملف: | 2022_Article_AComprehensiveMethodToElucidat.pdf - application/pdf |
| تدمد: | 1618-2650 1618-2642 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::36d445891d9e23340ab71eed74b964eaTest https://doi.org/10.1007/s00216-022-03907-wTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....36d445891d9e23340ab71eed74b964ea |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 16182650 16182642 |
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