Glucosamine Research - Chondroitin Sulfate, Uses, Effects, Benefits, Arthritis

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Structural analysis of the sheath of a sheathed bacterium, Leptothrix cholodnii.

Takeda M, Makita H, Ohno K, Nakahara Y, Koizumi J

Division of Materials Science and Chemical Engineering, Faculty of Engineering, Yokohama National University, Tokiwadai 79-5, Hodogaya, Yokohama 240-8501, Japan. mtake@ynu.ac.jp

Leptothrix cholodnii is an aerobic sheath-forming bacterium often found in oligotrophic and metal-rich aquatic environments. The sheath of this bacterium was isolated by selectively lysing the cells. Glycine and cysteine were the major amino acids of the sheath. The sheath was readily dissolved in hydrazine, and a polysaccharide substituted with cysteine was recovered from the solution. Galactosamine, glucosamine and galacturonic acid were detected in the hydrazinolysate by gas liquid chromatography analysis. FAB-MS analysis of the hydrazinolysate suggested a sugar sequence of HexN-GalA-HexN-HexN. Methylation linkage analysis revealed the presence of 4-linked GalA, 3-linked HexN and 4-linked HexN. The sulfhydryl groups of the sheath were used for labeling with the fluorogenic reagent, 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F). The labeled sheath (ABD-sheath) was partially hydrolyzed and three fluorescent fragments were purified by HPLC. One of them was identified as ABD-cysteine. The second one was found to be the ABD-cysteine tetramer. Another fragment was indicated to be a pentasaccharide substituted with ABD-cysteine by nuclear magnetic resonance (NMR) analysis. It can be assumed that the polysaccharide and peptide moieties of the sheath are connected by a cysteine residue. NMR analysis of the hydrazinolysate revealed that the polysaccharide moiety of the sheath was constructed from a pentasaccharide repeating unit containing 2-amino-2-deoxygalacturonic acid (GalNA), as shown below. -->4)-alpha-GalNA-(1-->4)-alpha-D-GalN(p)-(1-->4)-alpha-D-GalA(p)-(1-->4)-beta-D-GlcN(p)-(1-->3)-beta-D-GalN(p)-(1-->.

Published 31 October 2005 in Int J Biol Macromol, 37(1): 92-8.
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Glucosamine Books

User's Guide to Glucosamine and Chondroitin: Don't Be a Dummy: Become an Expert on What Glucosamine & Choneroitin Can Do (Basic Health Publications User's Guide)

User's Guide to Glucosamine and Chondroitin: Don't Be a Dummy: Become an Expert on What Glucosamine & Choneroitin Can Do (Basic Health Publications User's Guide)