Journal
JACS AU
Volume 3, Issue 3, Pages 929-942Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jacsau.3c00025
Keywords
Methylobacterium extorquens; lipopolysaccharide (LPS); lipid bilayers; NMR spectroscopy; ESI FT-ICR mass spectrometry; lipid A MALDI MS; electron paramagnetic resonance (EPR); small-angle neutron scattering (SANS)
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The ability of Methylobacterium extorquens to grow on methanol as the sole carbon and energy source is influenced by the composition and function of the lipopolysaccharide (LPS) in the outer membrane. The LPS of M. extorquens is unique, with non-phosphorylated and heavily O-methylated core oligosaccharides, and a distinctive lipid A structure. These chemical features impact the resistance of the membrane to methanol, regulating membrane ordering and dynamics.
The ability of Methylobacterium extorquens to grow on methanol as the sole carbon and energy source has been the object of intense research activity. Unquestionably, the bacterial cell envelope serves as a defensive barrier against such an environmental stressor, with a decisive role played by the membrane lipidome, which is crucial for stress resistance. However, the chemistry and the function of the main constituent of the M. extorquens outer membrane, the lipopolysaccharide (LPS), is still undefined. Here, we show that M. extorquens produces a rough-type LPS with an uncommon, non-phosphory-lated, and extensively O-methylated core oligosaccharide, densely substituted with negatively charged residues in the inner region, including novel monosaccharide derivatives such as O-methylated Kdo/Ko units. Lipid A is composed of a non-phosphorylated trisaccharide backbone with a distinctive, low acylation pattern; indeed, the sugar skeleton was decorated with three acyl moieties and a secondary very long chain fatty acid, in turn substituted by a 3 -O-acetyl-butyrate residue. Spectroscopic, conformational, and biophysical analyses on M. extorquens LPS highlighted how structural and tridimensional features impact the molecular organization of the outer membrane. Furthermore, these chemical features also impacted and improved membrane resistance in the presence of methanol, thus regulating membrane ordering and dynamics.
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