Co-Culture of Gut Bacteria and Metabolite Extraction Using Fast Vacuum Filtration and Centrifugation
Journal article
Authors | Guraka, A., Duff, R., Waldron, J., Tripathi, G. and Kermanizadeh, A. |
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Abstract | This protocol describes a robust method for the extraction of intra and extracellular metabolites of gut bacterial mono and co-cultures. In recent years, the co-culture techniques employed in the field of microbiology have demonstrated significant importance in regard to understanding cell–cell interactions, cross-feeding, and the metabolic interactions between different bacteria, fungi, and microbial consortia which enable the mimicking of complex co-habitant conditions. This protocol highlights a robust reproducible physiologically relevant culture and extraction protocol for the co-culture of gut bacterium. The novel extraction steps are conducted without using quenching and cell disruption through bead-cell methods, freeze–thaw cycles, and sonication, which tend to affect the physical and biochemical properties of intracellular metabolites and secretome. The extraction procedure of inoculated bacterial co-cultures and monocultures use fast vacuum filtration and centrifugation. The extraction methodology is fast, effective, and robust, requiring 4 h to complete. |
Keywords | gut bacteria; co-culture; Bacteriodes thetaiotomicron; Limosilactobacillus fermentum; bacterial metabolites; short-chain fatty acids |
Year | 2024 |
Journal | Methods and Protocols |
Journal citation | 7 (74), pp. 1-13 |
Publisher | MPDI |
ISSN | 2409-9279 |
Digital Object Identifier (DOI) | https://doi.org/10.3390/mps7050074 |
Web address (URL) | https://www.mdpi.com/2409-9279/7/5/74 |
Publisher's version | License File Access Level Open |
Output status | Published |
Publication dates | |
Online | 19 Sep 2024 |
Publication process dates | |
Deposited | 25 Sep 2024 |
https://repository.derby.ac.uk/item/q9341/co-culture-of-gut-bacteria-and-metabolite-extraction-using-fast-vacuum-filtration-and-centrifugation
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