IMPROVED PREPARATION OF LIPOTEICHOIC ACIDS
IMPROVED PREPARATION OF LIPOTEICHOIC ACIDS
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DOI：
10.1111/j.1432-1033.1983.tb07495.x
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发表时间：
1983-01-01
期刊：
影响因子：
--
通讯作者：
HAAS, R
中科院分区：
文献类型：
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作者：
FISCHER, W;KOCH, HU;HAAS, R
描述了一种从革兰氏阳性细菌中提取脂磷壁酸的绝对值的测量方法。如果细胞被破坏，则可以用热酚/水从各种细菌中几乎完全提取。提取完整的和脱脂的细胞以及膜部分的产率相当低。从被破坏的细胞中共同提取的大部分核酸都是通过核酸酶处理来去除的。通过DEAE-Sephacel阴离子交换层析或辛基-Sepharose疏水作用层析，从脂磷壁酸中分离出抗核酸酶的核酸、蛋白质、多糖和磷壁酸。纯化的制剂基本上没有聚合物污染物，保留了它们的丙氨酸酯取代，并以钠盐的形式存在。疏水相互作用层析还可以识别脱酰化和溶血型脂磷壁酸的污染，以及区分含有2和3个或2和4个酰基的分子物种。
A procedure is described for measuring the extraction of lipoteichoic acids from gram-positive bacteria in absolute terms. Virtually complete extraction was achieved from various bacteria by hot phenol/water if the cells were disrupted. Extraction of whole and delipidated cells and of the membrane fraction gave considerably lower yields. Most of the nucleic acids co-extracted from disrupted cells was removed by treatment with nucleases. Nuclease-resistant nucleic acid, protein, polysaccharide and teichoic acid were separated from lipoteichoic acid by anion-exchange chromatography on DEAE-Sephacel or hydrophobic interaction chromatography on octyl-Sepharose. Purified preparations were essentially free of polymeric contaminants, retained their alanine ester substitution and were in the Na salt form. Hydrophobic interaction chromatography also made it possible to recognize contamination of lipoteichoic acid with its deacylated and lyso-form and to discriminate molecular species containing 2 and 3, or 2 and 4 acyl groups.