title: Revisiting Protocols for the NMR Analysis of Bacterial Metabolomes.
abstract: Over the past decade, metabolomics has emerged as an important technique for systems biology. Measuring all the metabolites in a biological system provides an invaluable source of information to explore various cellular processes, and to investigate the impact of environmental factors and genetic modifications. Nuclear magnetic resonance (NMR) spectroscopy is an important method routinely employed in metabolomics. NMR provides comprehensive structural and quantitative information useful for metabolomics fingerprinting, chemometric analysis, metabolite identification and metabolic pathway construction. A successful metabolomics study relies on proper experimental protocols for the collection, handling, processing and analysis of metabolomics data. Critically, these protocols should eliminate or avoid biologically-irrelevant changes to the metabolome. We provide a comprehensive description of our NMR-based metabolomics procedures optimized for the analysis of bacterial metabolomes. The technical details described within this manuscript should provide a useful guide to reliably apply our NMR-based metabolomics methodology to systems biology studies.
authors.affiliation: Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0304.
authors.forename: Steven
authors.initials: S
authors.lastname: Halouska
authors.affiliation: Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0304.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68588-0905.
authors.forename: Rosmarie
authors.initials: R
authors.lastname: Gaupp
authors.affiliation: Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0304.
authors.forename: Shulei
authors.initials: S
authors.lastname: Lei
authors.affiliation: Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0304.
authors.forename: Emily
authors.initials: E
authors.lastname: Snell
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68588-0905.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Fenton
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68588-0905.
authors.forename: Raul G
authors.initials: RG
authors.lastname: Barletta
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE 68588-0905.
authors.forename: Greg A
authors.initials: GA
authors.lastname: Somerville
authors.affiliation: Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0304.
authors.forename: Robert
authors.initials: R
authors.lastname: Powers
keywords: Mycobacterium; NMR; Staphylococcus; bacteria; chemometrics; metabolomics