Growth-phase dependence of bacterial membrane lipid profile and labeling for in-cell solid-state NMR applications.
Biochim Biophys Acta Biomembr
; 1864(2): 183819, 2022 02 01.
Article
in En
| MEDLINE
| ID: mdl-34800428
ABSTRACT
Cell labeling is a preliminary step in multiple biophysical approaches, including the solid-state nuclear magnetic resonance (NMR) study of bacteria in vivo. Deuterium solid-state NMR has been used in the past years to probe bacterial membranes and their interactions with antimicrobial peptides, following a standard labeling protocol. Recent results from our laboratory on a slow-growing bacterium has shown the need to optimize this protocol, especially the bacterial growth time before harvest and the concentration of exogenous labeled fatty acids to be used for both Escherichia coli and Bacillus subtilis. It is also essential for the protocol to remain harmless to cells while providing optimal labeling. We have therefore developed a fast and facile approach to monitor the lipid composition of bacterial membranes under various growth conditions, combining solution 31P NMR and GCMS. Using this approach, the optimized labeling conditions of Escherichia coli and Bacillus subtilis with deuterated palmitic acid were determined. Our results show a modification of B. subtilis phospholipid profile as a function of the growth stage, as opposed to E. coli. Our protocol recommends low concentrations of exogenous palmitic acid in the growth medium, and bacteria harvest after the exponential phase.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Bacillus subtilis
/
Magnetic Resonance Spectroscopy
/
Cell Membrane
/
Palmitic Acid
/
Escherichia coli
/
Lipid Bilayers
/
Membrane Lipids
Language:
En
Journal:
Biochim Biophys Acta Biomembr
Year:
2022
Document type:
Article
Affiliation country:
Canada