Separation of lipopolysaccharides containing different fatty acid chains using hydrophobic interaction chromatography

Separation of lipopolysaccharides containing different fatty acid chains using hydrophobic interaction chromatography
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使用疏水相互作用色谱法分离含有不同脂肪酸链的脂多糖

DOI:
10.1039/c2ay05663a
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发表时间:
2012-03
期刊:
影响因子:
3.1
通讯作者:
Wang, Xiaoyuan
Wang, Xiaoyuan
中科院分区:
化学3区
文献类型:
--
作者:
Ren, Ke;Li, Ye;Shi, Feng;Wang, Xiaoyuan

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内毒素是大多数革兰氏阴性细菌外膜的主要成分。由于它们的两亲性,很难分离内毒素。本研究采用疏水相互作用层析技术分离含有不同数量脂肪酸链的脂多糖。从大肠杆菌W3110、MLK1067和MKV15b中分别提取了3种脂多糖。研究了它们与丁基琼脂糖、苯基琼脂糖和辛基琼脂糖三种疏水基质的相互作用。用几种有机溶剂从不同的基质中洗脱结合的脂多糖。以正丁基琼脂糖凝胶、丙醇和0.8mol L−-1乙酸钠(pH4.5)为负载缓冲液,对含有不同脂肪酸链的脂多糖进行了分离。该体系可用于从大肠杆菌产生的蛋白中去除受污染的脂多糖,或分离不同种类的脂多糖。
Lipopolysaccharides, known as endotoxins, are major components in the outer membrane of most Gram-negative bacteria. Because of their amphipathic property, it is difficult to separate lipopolysaccharides. In this study, hydrophobic interaction chromatography was used for the separation of lipopolysaccharides containing different numbers of fatty acid chains. Three species of lipopolysaccharides were extracted from Escherichia coli W3110, MLK1067, and MKV15b, respectively. Their interactions with three types of hydrophobic matrixes Butyl-Sepharose, Phenyl-Sepharose and Octyl-Sepharose were investigated. Several organic solvents were used to elute the bound lipopolysaccharides from different matrixes. Butyl-Sepharose, propyl alcohol and 0.8 mol l−1 sodium acetate (pH 4.5) loading buffer were chosen for the best separation system, and lipopolysaccharides containing different fatty acid chains were separated. This system might be useful for removing the contaminated lipopolysaccharides from proteins produced by E. coli, or separating different species of lipopolysaccharides.
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