The use of acid alumina and sephadex LH-20 for the separation and characterization of ethanol-soluble peptides produced by Bacillus brevis.
The use of acid alumina and sephadex LH-20 for the separation and characterization of ethanol-soluble peptides produced by Bacillus brevis.
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DOI：
10.1042/bj1270489
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发表时间：
1972-04
期刊：
影响因子：
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通讯作者：
I. Bartley;B. Hodgson;J. S. Walker;G. Holme
中科院分区：
文献类型：
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作者：
I. Bartley;B. Hodgson;J. S. Walker;G. Holme
从短芽孢杆菌（A.T.C.C. 10068），将某些（14）C-标记的氨基酸（其通常是短杆菌素的组分）掺入到通过乙醇从通过加入酸形成的沉淀物中提取的材料中。当这种物质通过纸和硅胶薄层色谱和纸电泳分离时（14）C位于也含有短杆菌肽和短杆菌肽的区域。根据对负责掺入的系统的性质的研究，推断存在非短杆菌素材料。结果表明，通常用于表征短杆菌肽的方法不能充分区分短杆菌肽和非短杆菌肽材料。然而，设计了一种分离这些材料的方法。这涉及用乙醇从酸性氧化铝柱上洗脱，然后用二甲基甲酰胺-水溶剂在Sephadex LH-20上凝胶过滤。研究了短杆菌肽和短杆菌肽在SephadexLH-20柱上的行为，得出的结论是，分离不是简单地由未缔合分子的凝胶过滤引起的。此外，具有不同氨基酸组成的酪氨酸分子似乎对Sephadex LH-20柱具有不同的亲和力。
Cell extracts from Bacillus brevis (A.T.C.C. 10068), grown with various media, incorporated certain (14)C-labelled amino acids that are normally components of tyrothricin into material that was extracted by ethanol from the precipitate formed by adding acid. When this material was separated by paper and silica-gel thin-layer chromatography and paper electrophoresis (14)C was located in those regions that also contained gramicidin and tyrocidine. From a study of the properties of the system responsible for the incorporation it was deduced that non-tyrothricin materials were present. It was shown that the methods normally used to characterize tyrothricin do not adequately distinguish between tyrothricin and non-tyrothricin materials. However, a method for separating these materials was devised. This involved elution with ethanol from columns of acid alumina followed by gel filtration on Sephadex LH-20 with dimethylformamide-water solvent. The behaviour of gramicidin and tyrocidine on the Sephadex LH-20 column was examined, and it was concluded that the separation was not caused simply by gel filtration of unassociated molecules. Also, tyrocidine molecules with different amino acid compositions seemed to have different affinities for the Sephadex LH-20 column.