tailieunhanh - Báo cáo Y học: Trehalose-based oligosaccharides isolated from the cytoplasm of Mycobacterium smegmatis Relation to trehalose-based oligosaccharides attached to lipid

A series of trehalose-based oligosaccharides were isolated from the cytoplasmic fraction of Mycobacterium smegmatis and purified by gel-filtration and paper chromatography and TLC. Their structures were determined by HPLC and GLC to determine sugar composition and ratios, MALDI-TOF MS to measure molecular mass, methylation analysis to determine linkages, 1H-NMR to obtain anomeric configurations of glycosidic linkages, and exoglycosidase digestions followed by TLC to determine sequences and anomeric configurations of the monosaccharides | Eur. J. Biochem. 269 3142-3149 2002 FEBS 2002 doi Trehalose-based oligosaccharides isolated from the cytoplasm of Mycobacterium smegmatis Relation to trehalose-based oligosaccharides attached to lipid Masaya Ohta1 Y. T. Pan2 Roger A. Laine3 and Alan D. Elbein2 1 Department of Biochemistry Fukuyuma University Japan department of Biochemistry and Molecular Biology University of Arkansas for Medical Sciences Little Rock AR 72205 USA 3Departments of Biological Sciences and Chemistry Louisiana State University Baton Rouge LA USA A series of trehalose-based oligosaccharides were isolated from the cytoplasmic fraction of Mycobacterium smegmatis and purified by gel-filtration and paper chromatography and TLC. Their structures were determined by HPLC and GLC to determine sugar composition and ratios MALDI-TOF MS to measure molecular mass methylation analysis to determine linkages H-NMR to obtain anomeric configurations of glycosidic linkages and exoglycosidase digestions followed by TLC to determine sequences and anomeric configurations of the monosaccharides. Six different oligosaccharides were identified all with trehalose as the basic structure and additional glucose or galactose residues attached in various linkages. One of these oligosaccharides is the disaccharide trehalose Glca1-1aGlc which is present in substantial amounts in these cells and also in other mycobacteria. Two other oligosaccharides the tetrasaccharides Glca1-4Glca1-1aGlc6-1aGal and Gala1-6Gala1-6Glca1-1aGlc have not previously been isolated from natural sources or synthesized chemically. The fourth oligosaccharide Glcp1-6Glcp1-6Glca1-1aGlc has been isolated from corynebacteria but not reported in other organisms. Two other oligosaccharides Glca1-4Glca1-1aGlc which has been synthesized chemically and isolated from insects but not previously reported in mycobacteria and Glcb1-6Glca1-1aGlc which was previously isolated from Mycobacterium fortuitum and yeast were also .