ArticleMicrobial cell factories2015
Metabolic fingerprinting of Lactobacillus paracasei: the optimal quenching strategy.
Article in Microbial cell factories, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 7 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
7 citing papers in PubMed, 13 citations in OpenAlex.
- Bacterial Metabolomics: Sample Preparation Methods.Biochemistry research international · 2022Review
- Improved Sample Preparation for Untargeted Metabolomics Profiling of Escherichia coli.Microbiology spectrum · 2021Article
- Review of recent developments in GC-MS approaches to metabolomics-based research.Metabolomics : Official journal of the Metabolomic Society · 2018Review
- Analysis of Intracellular Metabolites from Microorganisms: Quenching and Extraction Protocols.Metabolites · 2017Review
- Extracellular Microbial Metabolomics: The State of the Art.Metabolites · 2017Review
- Targeted redox and energy cofactor metabolomics inBiotechnology for biofuels · 2017Article
- Erratum to: Metabolic fingerprinting of Lactobacillus paracasei: the optimal quenching strategy.Microbial cell factories · 2016Article
Corrections and comments
- Erratum issued
Authors and funding
3 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundQuenching in cold buffered methanol at -40 °C has long been the preferred method for sub-second inactivation of cell metabolism during metabolic fingerprinting. However, methanol is known to cause intracellular metabolite leakage of microbial cells, making the distinction between intra- and extracellular metabolites in microbial systems challenging. In this paper, we tested three quenching protocols proposed for microbial cultures: fast filtration, cold buffered methanol and cold glycerol saline.
resultsOur results clearly showed that cold glycerol saline quenching resulted in the best recovery of intracellular metabolites in Lactobacillus paracasei subsp. paracasei (L. paracasei). Membrane integrity assayed by propidium iodide revealed that approximately 100 % [Corrected] of the L. paracasei cell membranes were damaged by contact with the cold buffered methanol solution, whilst cold glycerol saline quenching led to minimal cell damage. Due to the nature of the L. paracasei culture, fast filtration took several minutes, which is far from ideal for metabolites with high intracellular turnover rates.
conclusionThe implementation of a reliable, reproducible quenching method is essential within the metabolomics community. Cold glycerol saline prevented leakage of intracellular metabolites, and, thus, allowed more accurate determinations of intracellular metabolite levels.
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