从表达衣原体脂多糖属特异性表位的明尼苏达沙门氏菌和大肠杆菌重组菌株的脂多糖中获得的寡糖单磷酸酯的制备及结构分析
Preparation and structural analysis of oligosaccharide monophosphates obtained from the lipopolysaccharide of recombinant strains of Salmonella minnesota and Escherichia coli expressing the genus-specific epitope of Chlamydia lipopolysaccharide.
作者信息
Holst O, Thomas-Oates J E, Brade H
机构信息
Division of Biochemical Microbiology, Forschungsinstitut Borstel, Germany.
出版信息
Eur J Biochem. 1994 May 15;222(1):183-94. doi: 10.1111/j.1432-1033.1994.tb18856.x.
The lipopolysaccharide of the recombinant strain Salmonella minnesota r595-207 expressing the genus-specific epitope of Chlamydia lipopolysaccharide [Holst, O., Brade, L., Kosma, P. and Brade, H. (1991) J. Bacteriol, 173, 1862-1866] was sequentially de-O- and de-N-acylated by mild hydrazinolysis and treatment with 4 M KOH, respectively. The resulting mixture of compounds was separated by high-performance anion-exchange chromatography and gel-permeation chromatography, yielding four oligosaccharide phosphates two of which were readily identified by their 1H-NMR- and 13C-NMR spectra as alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha-D-Glcp N 1,4'-bisphosphate (tetrasaccharide bisphosphate; Kdo = 3-deoxy-D-manno-octulopyranosonic acid) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha-D- GlcpN 1,4'-bisphosphate (pentasaccharide bisphosphate) [Holst, O., Broer, W., Thomas-Oates, J.E., Mamat, U. and Brade, H. (1993) Eur. J. Biochem. 214, 703-710]. The structures of the other two compounds were established by chemical analysis, NMR spectroscopy, and fast-atom-bombardment mass spectrometry as alpha-Kdo- (2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha-D-GlcpN 1-phosphate (tetrasaccharide 1-phosphate) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha-D- GlcpN 1-phosphate (pentasaccharide 1-phosphate). alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha/beta- D-GlcpN 4'-phosphate (tetrasaccharide 4'-phosphate) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha/beta-D-GlcpN 4'-phosphate (pentasaccharide 4'-phosphate) were prepared from the 1,4'-bisphosphates isolated from the recombinant strain Escherichia coli F515-207 by treatment with alkaline phosphatase and purification by high-performance anion-exchange chromatography and gel-permeation chromatography. Their structures were characterised by chemical analysis, NMR spectroscopy, and fast-bombardment mass spectrometry.
表达衣原体脂多糖属特异性表位的重组菌株明尼苏达沙门氏菌r595 - 207的脂多糖，分别通过温和的肼解和用4M氢氧化钾处理依次进行脱O - 酰化和脱N - 酰化。所得化合物混合物通过高效阴离子交换色谱和凝胶渗透色谱进行分离，得到四种寡糖磷酸酯，其中两种通过其1H - NMR和13C - NMR光谱很容易鉴定为α - Kdo - (2→4) - α - Kdo - (2→6) - β - D - GlcpN - (1→6) - α - D - GlcpN 1,4'-双磷酸酯（四糖双磷酸酯；Kdo = 3 - 脱氧 - D - 甘露糖 - 辛吡喃糖酸）和α - Kdo - (2→8) - α - Kdo - (2→4) - α - Kdo - (2→6) - β - D - GlcpN - (1→6) - α - D - GlcpN 1,4'-双磷酸酯（五糖双磷酸酯）[霍尔斯特·O、布勒·W、托马斯 - 奥茨·J.E、马马特·U和布勒·H（1993年）欧洲生物化学杂志214卷，703 - 710页]。另外两种化合物的结构通过化学分析、核磁共振光谱和快原子轰击质谱确定为α - Kdo - (2→4) - α - Kdo - (2→6) - β - D - GlcpN - (1→6) - α - D - GlcpN 1 - 磷酸酯（四糖1 - 磷酸酯）和α - Kdo - (2→8) - α - Kdo - (2→4) - α - Kdo - (2→6) - β - D - GlcpN - (1→6) - α - D - GlcpN 1 - 磷酸酯（五糖1 - 磷酸酯）。α - Kdo - (2→4) - α - Kdo - (2→6) - β - D - GlcpN - (1→6) - α/β - D - GlcpN 4'-磷酸酯（四糖4'-磷酸酯）和α - Kdo - (2→8) - α - Kdo - (2→4) - α - Kdo - (2→6) - β - D - GlcpN - (1→6) - α/β - D - GlcpN 4'-磷酸酯（五糖4'-磷酸酯）是通过用碱性磷酸酶处理从重组菌株大肠杆菌F515 - 207中分离得到的1,4'-双磷酸酯，并通过高效阴离子交换色谱和凝胶渗透色谱纯化而制备的。它们的结构通过化学分析、核磁共振光谱和快轰击质谱进行了表征。
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