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Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography
Published on: January 15, 2014
A toolbox to measure changes in the cell wall glycopolymer composition during differentiation of Streptomyces
Steffen Sigle1, Nadja Steblau1, Wolfgang Wohlleben1
1Interfakultäres Institut für Mikrobiologie und Infektionsmedizin Tübingen IMIT, Mikrobiologie/Biotechnologie, Eberhard Karls Universität Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany.
Abstract:
Cell wall glycopolymers (CWG) represent an important component of the Gram-positive cell envelope with many biological functions. The mycelial soil bacterium Streptomyces coelicolor A3(2) incorporates two distinct CWGs, polydiglycosylphosphate (PDP) and teichulosonic acid, into the cell wall of its vegetative mycelium but only little is known about their role in the complex life cycle of this microorganism. In this study we established assays to measure the total amount of CWGs in mycelial cell walls and spore walls, to quantify the individual CWGs and to determine the length of PDP. By applying these assays, we discovered that the relative amount of CWGs, especially of PDP, is reduced in spores compared to vegetative mycelium. Furthermore we found that PDP extracted from mycelial cell walls consisted of at least 19 repeating units, whereas spore walls contained substantially longer PDP polymers.

