TAILIEUCHUNG - Báo cáo Y học: Structural study on lipid A and the O-specific polysaccharide of the lipopolysaccharide from a clinical isolate of Bacteroides vulgatus from a patient with Crohn’s disease

Bacteroides vulgatus has been shown to be involved in the aggravation of colitis. Previously, we separated two potent virulence factors, capsular polysaccharide (CPS) and lipopolysaccharide (LPS), from a clinical isolate of B. vulgatus and characterized the structure of CPS. In this study, we elucidated the structures of O-antigen polysaccharide (OPS) and lipid A in the LPS. LPS was subjected to weak acid hydrolysis to produce the lipid A fraction and polysaccharide fraction. Lipid A was isolated by preparative TLC, and its structure determined by MS and NMR to be similar to that of Bacteroides fragilis except for the number of. | Eur. J. Biochem. 269 3715-3721 2002 FEBS 2002 doi Structural study on lipid A and the O-specific polysaccharide of the lipopolysaccharide from a clinical isolate of Bacteroides vulgatus from a patient with Crohn s disease Masahito Hashimoto1 2 Fumiko Kirikae1. Taeko Dohi1. Seizi Adachi3. Shoichi Kusumoto3. Yasuo Suda3 4 Tsuyoshi Fujita5 Hideo Naoki5 and Teruo Kirikae1 1 Research Institute International Medical Center of Japan department of Oral Microbiology Asahi University School of Dentistry Japan Graduate School of Science Osaka University Japan department of Bacteriology Hyogo College of Medicine Japan 5Suntory Institute for Bioorganic Research Japan Bacteroides vulgatus has been shown to be involved in the aggravation of colitis. Previously we separated two potent virulence factors capsular polysaccharide CPS and i po--polysaccharide LPS from a clinical isolate of B. vulgatus and characterized the structure of CPS. In this study we elucidated the structures of O-antigen polysaccharide OPS and lipid A in the LPS. LPS was subjected to weak acid hydrolysis to produce the lipid A fraction and polysaccharide fraction. Lipid A was isolated by preparative TLC and its structure determined by MS and NMR to be similar to that of Bacteroides fragilis except for the number of fatty acids. The polysaccharide fraction was subjected to gelfiltration chromatography to give an OPS-rich fraction. The structure of OPS was determined by chemical analysis and NMR spectroscopy to be a polysaccharide composed of the following repeating unit fi4 a-L-Rhap 1fi3 b-D-Manp 1fi . Keywords Bacteroides vulgatus fast-atom-bombardment tandem mass spectrometry lipopolysaccharide MALDI-TOF-MS NMR. Commensal flora are thought to be significantly involved in the pathogenesis of inflammatory bowel diseases Crohn s disease and ulcerative colitis reviewed in 1 . As chronic intestinal inflammation in several rodent models is prevented in a germ-free environment 2 .

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