Ziółkowski A, Swierzko A S, Cedzyński M
Pracownia Struktur Komórkowych Centrum Mikrobiologii i Wirusologii PAN w Lodzi.
Med Dosw Mikrobiol. 1998;50(3-4):215-22.
In the paper, we propose a method for estimation of the mean molecular weight of lipopolysaccharide, which is important for accuracy of endotoxin activity investigation. In our study, it was assumed that lipid A portion in Enterobacterial lipopolysaccharide is substituted by four 3-hydroxytetradecanoic acid residues. Lipopolysaccharides of S, Ra, Rc and Re chemotypes being laboratory preparations as well as purchased from Sigma were investigated. Fatty acids were determined by of gas chromatography as methyl esters according to the procedure described by Wollenweber and Rietschel. Mean molecular weight was calculated by the formula: MMW = [formula: see text]. A high agreement between the estimated and the theoretical molecular weight values was demonstrated in the case of Salmonella minnesota R595 (Re) LPS preparation. As expected, LPS heterogeneity increase together with enlargement of polysaccharide chain length which is visible in electrophoregrams also. Except for LPS mean molecular weight estimation, the method allows its detection in various preparations and samples, distinguishing of R and S LPS forms as well as the determination of mean length of O-specific chain in lipopolysaccharides which structures are known.
在本文中，我们提出了一种估算脂多糖平均分子量的方法，这对于内毒素活性研究的准确性很重要。在我们的研究中，假定肠杆菌脂多糖中的类脂A部分被四个3-羟基十四烷酸残基取代。对S、Ra、Rc和Re化学型的脂多糖进行了研究，这些脂多糖既有实验室制备的，也有从Sigma购买的。按照Wollenweber和Rietschel所述的方法，通过气相色谱法将脂肪酸测定为甲酯。平均分子量通过以下公式计算：MMW = [公式：见正文]。在明尼苏达沙门氏菌R595（Re）脂多糖制剂的情况下，估算的分子量值与理论分子量值之间显示出高度一致性。正如预期的那样，脂多糖的异质性随着多糖链长度的增加而增加，这在电泳图中也很明显。除了估算脂多糖的平均分子量外，该方法还能在各种制剂和样品中检测脂多糖，区分R型和S型脂多糖形式，以及测定已知结构的脂多糖中O-特异性链的平均长度。
