Simple Protocol to Purify Cell Wall Polysaccharide from Gram-Positive Bacteria and Assess Its Structural Integrity
Cell wall polysaccharides (CWPS), which are usually covalently bound to the peptidoglycan and are closely associated with the cell wall, are considered as ubiquitous components of the cell envelope of gram-positive bacteria and play an important role as mediators of bacterial interactions with the environment. Here, we describe a simple method for purifying CWPS by extraction of bacterial cells with consecutive acid treatments. Purified CWPS are obtained by gel-filtration chromatography following treatment with HF. We also provide the methodology to easily assess the integrity of CWPS using high-resolution magic-angle spinning (HR-MAS) NMR.
Key wordsCell wall polysaccharide Purification HR-MAS NMR
We acknowledge Douwe VanSideren (School of Microbiology, University College Cork, Ireland) and Marie-Pierre Chapot Chartier (Micalis Institute, INRA, France) for the use of Lactococcus lactis MG1363 and SK11. This work was supported by French ANR project “Lactophages” (ANR-11-BSV8-004-01).