Dmitriev B A, Knirel Y A, Kocharova N A, Kochetkov N K, Stanislavsky E S, Mashilova G M
Eur J Biochem. 1980 May;106(2):643-51. doi: 10.1111/j.1432-1033.1980.tb04612.x.
A loosely bound lipopolysaccharide-protein complex was extracted from cells of Pseudomonas aeruginosa strain 170015 (O:7ab; Lanyi classification) by saline solution and purified from contaminant nucleic acid by Cetavlon treatment followed by precipitation in an ultracentrifuge. The saline-treated cells were re-extracted with hot aqueous phenol to give firmly bound lipopolysaccharide which was isolated from the phenol layer and purified by ultracentrifugaiton. The identity of both lipopolysaccharide preparations was proved by serological and chemical evidence. Mild acid degradation of the lipopolysaccharide resulted in the splitting off of a lipid component and led to polysaccharide which was purified by gel-filtration on a Sephadex G-50 column. The polysaccharide consisted of N-acetyl-D-fucosamine, N-acetyl-L-fucosamine and D-glucose in the ratio 1:1:1. On the basis of nuclear magnetic resonance spectra, results of methylation analysis and two sequential Smith degradations, the following structure can be assigned to the repeating unit of the polysaccharide: -3)LFucNAc(alpha 1-3)DFucNAc(beta 1-2)DGlc(beta 1-. The polysaccharide did not show serological activity whereas alkali-treated lipopolysaccharide readily sensitised sheep erythrocytes and inhibited the passive haemagglutination reaction with anti-(O:7a,b)serum. Evidence is presented that the oligosaccharide repeating units of the polysaccharide and alkali-treated lipopolysaccharide are indistinguishable. Ps. aeruginosa strain 170016 (O:7a,c) was shown to have the O-specific lipopolysaccharide identical with that from strain 170015. The presented data show that subfactors 7b and 7c in the Lanyi classification of Ps. aeruginosa O-antigens seem to relate to components of the bacterial surface other than lipopolysaccharides.
用盐溶液从铜绿假单胞菌170015菌株(O:7ab;兰伊分类)的细胞中提取一种松散结合的脂多糖-蛋白质复合物,通过西他氯铵处理从污染的核酸中纯化,然后在超速离心机中沉淀。用热的苯酚水溶液对经盐处理的细胞进行再提取,得到紧密结合的脂多糖,将其从苯酚层中分离出来并通过超速离心进行纯化。两种脂多糖制剂的同一性通过血清学和化学证据得以证明。脂多糖的温和酸降解导致一种脂质成分的裂解,并产生多糖,该多糖通过在葡聚糖G-50柱上进行凝胶过滤进行纯化。该多糖由N-乙酰-D-岩藻糖胺、N-乙酰-L-岩藻糖胺和D-葡萄糖以1:1:1的比例组成。基于核磁共振光谱、甲基化分析结果和两次连续的史密斯降解,可将以下结构赋予多糖的重复单元:-3)LFucNAc(α1-3)DFucNAc(β
1-2)DGlc(β1-。该多糖未显示血清学活性,而经碱处理的脂多糖很容易使绵羊红细胞致敏,并抑制与抗-(O:7a,b)血清的被动血凝反应。有证据表明,该多糖和经碱处理的脂多糖的寡糖重复单元无法区分。已证明铜绿假单胞菌170016菌株(O:7a,c)具有与170015菌株相同的O-特异性脂多糖。所呈现的数据表明,在铜绿假单胞菌O抗原的兰伊分类中,亚因子7b和7c似乎与细菌表面除脂多糖以外的成分有关。