Purification and characterisation of lipoglycan macroamphiphiles from Propionibacterium acnes.
Lipidated macroamphiphiles such as the lipoteichoic acids and mycobacterial lipoarabinomannans are cell envelope components of Gram-positive bacteria that have been extensively associated with the pathogenesis of disease. In order to study such associations, purification of these macroamphiphiles is essential for resolving their structures and diverse biological effects. We describe herein a method for purification of lipoglycan components from Propionibacterium acnes. This method uses the existing phenol-water extraction, followed by hydrophobic interaction chromatography and an additional purification step that utilises preparative electrophoresis for the separation of two lipoglycan components. Analysis of these lipoglycans revealed evidence for a lipid anchor based on fatty acids whilst the polysaccharide moiety contained significant amounts of mannose, glucose and galactose, together with an amino sugar suspected of being a diaminohexuronic acid. These latter components have been previously identified as components of the P. acnes cell wall polysaccharide. Consequently, it is proposed that there may be a relationship between the structures of these distinctive cell envelope polymers.
Whale
Gary A
GA
School of Life Science, The Robert Gordon University, Aberdeen, AB25 1HG, UK. g.whale@microscience.com
Sutcliffe
Iain C
IC
Morrisson
Andrew R
AR
Pretswell
Emma L
EL
Emmison
Neil
N
D002845:Chromatography / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000032:analysis / Q000302:isolation & purification; D005690:Galactose / Q000032:analysis / Q000302:isolation & purification; D005947:Glucose / Q000032:analysis / Q000302:isolation & purification; D006603:Hexuronic Acids / Q000032:analysis / Q000302:isolation & purification; D057927:Hydrophobic and Hydrophilic Interactions; D008070:Lipopolysaccharides / Q000737:chemistry* / Q000302:isolation & purification*; D008358:Mannose / Q000032:analysis / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000032:analysis / Q000302:isolation & purification; D011425:Propionibacterium acnes / Q000737:chemistry* / Q000378:metabolism
D005227:Fatty Acids; D006603:Hexuronic Acids; D008070:Lipopolysaccharides; D011135:Polysaccharides, Bacterial; D005947:Glucose; D008358:Mannose; D005690:Galactose