| RRC ID |
15840
|
| Author |
Kondo K, Takeda M, Ejima W, Kawasaki Y, Umezu T, Yamada M, Koizumi J, Mashima T, Katahira M.
|
| Title |
Study of a novel glycoconjugate, thiopeptidoglycan, and a novel polysaccharide lyase, thiopeptidoglycan lyase.
|
| Journal |
Int J Biol Macromol
|
| Abstract |
A typical filamentous bacterium, Sphaerotilus natans, secretes a thiolic glycoconjugate which is assembled into a microtube, so called sheath. The glycoconjugate is known to consist of a pentasaccharide-dipeptide repeating unit, but its chemical structure has not been completely elucidated. In order to determine its chemical structure, the sheath was broken down by performic acid oxidation. The released sulfonated derivative was water soluble which was suitable for detailed NMR analysis. The data exhibited the presence of two stoichiometric and one substoichiometric (relative abundance was about 0.5) acetylations, suggesting that the glycoconjugate is composed of two equimolar pentasaccharide-dipeptide repeating units each having either two or three acetyl groups. However, the position of substoichiometric acetylation could not be defined. To determine the position, the sheath was derivatized with a thiol selective fluorescent reagent followed by digestion with a specific polysaccharide lyase prepared from a sheath-degrading bacterium, Paenibacillus koleovorans. As expected, two fluorescent digests were recovered by reverse-phase HPLC and were subjected to NMR analysis. The data revealed that both digests are pentasaccharide-dipeptides which have unsaturated glucuronic acid and galactosamine residues at their reducing and non-reducing ends, respectively. It was also confirmed that one digest has 3-O-acetylated glucose residue while the other has non-derivatized glucose residue. The substoichiometric acetylation was thus identified with the 3-O-acetylation, and structural determination of the thiolic glycoconjugate was completed. By virtue of the clarification of the two digests' structures, the cleavage site was specified as (1→4)-α-galactosaminic bond to glucuronic acid. Based on the present and earlier findings, we propose a novel glycoconjugate category named thiopeptidoglycan and a novel polysaccharide lyase named thiopeptidoglycan lyase.
|
| Volume |
48(2)
|
| Pages |
256-62
|
| Published |
2011-3-1
|
| DOI |
10.1016/j.ijbiomac.2010.11.009
|
| PII |
S0141-8130(10)00339-9
|
| PMID |
21095202
|
| MeSH |
Carbohydrate Conformation
Carbohydrate Sequence
Chromatography, High Pressure Liquid
Glycopeptides / analysis*
Glycopeptides / chemistry
Magnetic Resonance Spectroscopy
Molecular Sequence Data
Paenibacillus / enzymology
Peptidoglycan / analysis*
Peptidoglycan / chemistry
Polysaccharide-Lyases / isolation & purification
Polysaccharide-Lyases / metabolism*
Polysaccharides, Bacterial / analysis*
Polysaccharides, Bacterial / chemistry
Solubility
Sphaerotilus / chemistry
|
| IF |
5.162
|
| Times Cited |
10
|
|
WOS Category
|
POLYMER SCIENCE
CHEMISTRY, APPLIED
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| General Microbes |
JCM 11186
JCM 20382 |
