Purification and characterisation of lipoglycan macroamphiphiles from Propionibacterium acnes

• Published in: Antonie van Leeuwenhoek 2004-08, Vol.86 (1), p.77-85
• Main Authors: WHALE, Gary A, SUTCLIFFE, Iain C, MORRISSON, Andrew R, PRETSWELL, Emma L, EMMISON, Neil
• Format: Article
• Language: English
• Subjects: Bacteria; Bacteriology; Biogenesis of cell structures, supramolecular organization; Biological and medical sciences; Biological effects; Chromatography - methods; Electrophoresis, Polyacrylamide Gel; Fatty Acids - analysis; Fatty Acids - isolation & purification; Fundamental and applied biological sciences. Psychology; Galactose - analysis; Galactose - isolation & purification; Glucose - analysis; Glucose - isolation & purification; Hexuronic Acids - analysis; Hexuronic Acids - isolation & purification; Hydrophobic and Hydrophilic Interactions; Lipopolysaccharides - chemistry; Lipopolysaccharides - isolation & purification; Mannose - analysis; Mannose - isolation & purification; Microbiology; Phenols; Polymers; Polysaccharides, Bacterial - analysis; Polysaccharides, Bacterial - isolation & purification; Propionibacterium acnes; Propionibacterium acnes - chemistry; Propionibacterium acnes - metabolism
• ISSN: 0003-6072; 1572-9699
• DOI: 10.1023/B:ANTO.0000024911.67625.27

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.