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Analysis of Gram-negative Bacteria Peptidoglycan by Ultra-performance Liquid Chromatography
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).ORCID iD: 0000-0003-2429-7542
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).ORCID iD: 0000-0001-5995-718x
2020 (English)In: Bio-protocol, E-ISSN 2331-8325, Vol. 10, no 19, article id e3780Article in journal (Refereed) Published
Abstract [en]

Bacteria are surrounded by a protective peptidoglycan cell wall. Provided that this structure and the enzymes involved are the preferred target for our most successful antibiotics, determining its structural and chemical complexity is of the highest interest. Traditionally, high-performance liquid chromatography (HPLC) analyses have been performed, but these methods are very time consuming in terms of sample preparation and chromatographic separation. Here we describe an optimized method for preparation of Gram-negative bacteria peptidoglycan and its subsequent analysis by ultra-performance liquid chromatography (UPLC). The use of UPLC in peptidoglycan analyses provides a dramatic reduction of the sample volume and hands-on time required and, furthermore, permits in-line mass spectrometry (MS) of the UPLC resolved muropeptides, thus facilitating their identification. This method improves our capability to perform high throughput analysis to better understand the cell- wall biology.

Place, publisher, year, edition, pages
Bio-protocol , 2020. Vol. 10, no 19, article id e3780
Keywords [en]
UPLC, Cell Wall, Peptidoglycan, Muropeptide, Muramidase, Crosslink
National Category
Microbiology Analytical Chemistry
Identifiers
URN: urn:nbn:se:umu:diva-176864DOI: 10.21769/BioProtoc.3780ISI: 000583376800012OAI: oai:DiVA.org:umu-176864DiVA, id: diva2:1502472
Funder
Knut and Alice Wallenberg FoundationSwedish Research CouncilThe Kempe FoundationsAvailable from: 2020-11-20 Created: 2020-11-20 Last updated: 2022-09-15Bibliographically approved

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Alvarez, LauraCava, Felipe

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