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6.9 Techniques for Analysis of Peptidoglycans

Methods in Microbiology, ISSN: 0580-9517, Vol: 27, Issue: C, Page: 277-286
1998
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This chapter discusses the techniques for analysis of peptidoglycans. The backbone of the eubacterial cell wall is peptidoglycan, a structural macromolecule that gives the wall its strength and integrity, and serves as the infrastructure to which a variety of proteins and lipids may be anchored. The basic molecular structure of peptidoglycan is simple and well established. The definitive identification of the structure of each of muropeptides requires mass spectrometry (MS). Further, the chapter discusses peptidoglycan isolation. A typical protocol for isolation of peptidoglycan from Gram-negative bacteria is discussed in the chapter. To isolate peptidoglycan from Gram positive and acid-fast bacteria, the procedure must be varied to inhibit degradation by endogenous autolytic enzymes and to overcome the stability imparted to the wall by accessory proteins. In addition, the chapter explains reverse-phase high-pressure liquid chromatography (RP-H PLC).

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