ELECTROSPRAY MASS-SPECTROMETRY FOR CHARACTERIZATION OF LIPID-A FROM ENTEROBACTER-AGGLOMERANS

Negative ion and positive ion electrospray mass spectrometry have been employed to characterize the lipid A mixture produced by hydrolysis of lipopolysaccharides from Enterobacter agglomerans, a Gram-negative bacterium commonly found in field cotton Neutral monophosphoryl lipid A molecules form stable anions vis deprotonation, but adduct formation via cation attachment occurs in low yield. Dephosphorylated lipid A molecules, on the other hand, readily form adducts with Na+, whereas deprotonation occurs in low yield. The mass spectra of lipid A produced by E. agglomerans reveal the presence of lipid A ions which differ in the nature of attached fatty acid side chains. At least two heptaacyl forms of lipid A are present, one of which has a structure which appears to be the same as the structure of heptaacyl lipid A produced by Salmonella minnesota. The second structure differs only by the nature of the side chain at position 3' of the disaccharide backbone where a hydroxymyristoyloxymyristoyl group replaces the myristoyloxymyristoyl substituent. Collisionally activated dissociations prior to mass analysis enable the identification of fragment ions which can be distinguished from at least eight intact deprotonated molecules present in crude lipid A.