The diacetamidodideoxyuronic-acid-containing glycan chain of Bacillus stearothermophilus NRS 2004/3a represents the secondary cell-wall polymer of wild-type B. stearothermophilus strains.
The diacetamidodideoxymannuronic-acid-containing glycan of Bacillus stearothermophilus NRS 2004/3a with the repeating unit structure [-->4)-beta-D-ManpA2,3(NAc)2-(1-->6)-alpha-D-Glcp-(1-->4)-beta-D-+ ++ManpA2,3 (NAc)2-(1-->3)-alpha-D-GlcpNAc-(1-->], was examined to identify its linkage to the bacterial cell wall. In a previous paper it was suggested that this glycan is covalently linked to the surface layer (S-layer) glycoprotein of that organism. By improved chromatographic techniques (gel permeation over Sephacryl S-1000 SF; C4 reversed-phase HPLC) the diacetamidodideoxyuronic-acid-containing material was completely separated from the S-layer glycoprotein. This implicates only low, if any, specific affinity between these cell-wall components. To obtain sufficient amounts for the chemical characterization of its linkage region, the identical diacetamidodideoxyuronic-acid-containing material was isolated from sonicated cells of that organism by a purification procedure different to that for preparation of S-layers. This method allowed collection of the intact molecule including its linkage region. From the combined results of the chemical characterization and 600 MHz NMR spectroscopy it is proposed that the diacetamidodideoxyuronic-acid-containing glycan chain, consisting of approximately six tetrasaccharide repeating units, is directly linked via a pyrophosphate bridge to carbon 6 of muramic acid residues of the peptidoglycan sacculus. About 20-25% of the muramic acid residues are substituted with these polysaccharide chains. Thus, the diacetamidodideoxyuronic-acid-containing glycan represents a secondary cell-wall polymer of B. stearothermophilus NRS 2004/3a.
Schäffer
Christina
C
Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Molekulare Nanotechnologie, Universität für Bodenkultur, Gregor-Mendel-Str. 33, A-1180 Wien, Austria.
Kählig
Hanspeter
H
Institut für Organische Chemie, Universität Wien, A-1090 Wien, Austria.
Christian
Rudolf
R
Scientific Software Company, A-1140 Wien, Austria.
Schulz
Gerhard
G
Novartis Forschungsinstitut Wien, A-1235 Wien, Austria.
Zayni
Sonja
S
Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Molekulare Nanotechnologie, Universität für Bodenkultur, Gregor-Mendel-Str. 33, A-1180 Wien, Austria.
Messner
Paul
P
Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Molekulare Nanotechnologie, Universität für Bodenkultur, Gregor-Mendel-Str. 33, A-1180 Wien, Austria.
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002473:Cell Wall / Q000737:chemistry*; D001411:Geobacillus stearothermophilus / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism; D005965:Glucuronates / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D011135:Polysaccharides, Bacterial / Q000737:chemistry
D001426:Bacterial Proteins; D005965:Glucuronates; D008562:Membrane Glycoproteins; D010457:Peptidoglycan; D011108:Polymers; D011135:Polysaccharides, Bacterial; C055764:S-layer proteins; C031522:2,3-diacetamido-2,3-dideoxy-glucuronic acid