EXTRACTION AND COMPOSITION OF POLAR LIPIDS FROM THE ARCHAEBACTERIUM, METHANOBACTERIUM-THERMOAUTOTROPHICUM - EFFECTIVE EXTRACTION OF TETRAETHER LIPIDS BY AN ACIDIFIED SOLVENT
EXTRACTION AND COMPOSITION OF POLAR LIPIDS FROM THE ARCHAEBACTERIUM, METHANOBACTERIUM-THERMOAUTOTROPHICUM - EFFECTIVE EXTRACTION OF TETRAETHER LIPIDS BY AN ACIDIFIED SOLVENT
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DOI：
10.1093/oxfordjournals.jbchem.a121969
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发表时间：
1987-04-01
影响因子：
2.7
通讯作者：
KOGA, Y
中科院分区：
文献类型：
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作者：
NISHIHARA, M;KOGA, Y
通常的Bligh和Dyer法只能提取热自养甲烷杆菌的一小部分脂质。当溶剂中的水被5%的三氯乙酸取代时，脂质回收率达到最高水平，比前一种方法提高了6倍。使用盐酸（2m）或破坏细胞也是有效的，但用含盐酸的溶剂长时间提取会导致一些磷酸糖脂的降解。在总脂质薄层色谱上检测到23个极性脂质点。10种磷脂（18%）、6种氨基磷脂（17%）、3种氨基磷脂糖脂（15%）、2种磷酸糖脂（31%）和2种糖脂（19%）。主要极性脂质为高极性磷酸糖脂（PGL1, 30%）和糖脂（GL1a, 16%）。其他主要脂质包括一种氨基磷脂（PNL1a, 9%）和一种氨基磷酸糖脂（PNGL1, 7%）。本文描述了PNL1a、GL1a和PNGL1的完整结构测定。为了使极性头基团完全裂解，需要对总脂质进行乙酰解，然后进行酸甲醇解，释放出二phytanyl glycerol diether （C20 diether）和二biphytanyl digglycerol tetraether （C40 tetraether）的核心残基。薄层色谱密度分析表明C20二醚与C40四醚的比例为1:14。对BCl3处理后的总脂质制备的烷基氯化物进行GLC分析，结果表明，植烷基（C20）、联植烷基（C40）和未识别的烷基链分别占总烷基链的10%、83%和7mol %。脂质提取后得到的大分子残留物被强酸水解得到大量的C40四醚，这些四醚可能与细胞内的其他物质共价结合。
The usual Bligh and Dyer method could extract only a small part of the lipids of Methanobacterium thermoautotrophicum. When the water in the solvent was replaced by 5% trichloroacetic acid, the lipid recovery reached the maximum level, which was 6 times higher than that by the former method. The use of HCl (2 M) or disruption of cells was also effective but prolonged extraction with the HCl-containing solvent caused degradation of some phosphoglycolipids. Twenty-three spots of polar lipids were detected on a thin-layer chromatogram of the total lipid. These were 10 phospholipids (18%), 6 aminophospholipids (17%), 3 aminophosphoglycolipids (15%), 2 phosphoglycolipids (31%), and 2 glycolipids (19%). The predominant polar lipids were a highly polar phosphoglycolipid (PGL1, 30%) and a glycolipid (GL1a, 16%). The other major lipids included an aminophospholipid (PNL1a, 9%), and an aminophosphoglycolipid (PNGL1, 7%). The complete structure determination of PNL1a, GL1a, and PNGL1 is described in the accompanying paper. Acetolysis of the total lipids followed by acid methanolysis was required for the complete cleavage of polar head groups, releasing core residues of diphytanyl glycerol diether (C20 diether) and dibiphytanyl diglycerol tetraether (C40 tetraether). A densitometric assay of a thin-layer chromatogram showed that the ratio of C20 diether and C40 tetraether was 1:14. GLC analysis of alkyl chlorides prepared from the total lipid by BCl3 treatment showed that phytanyl (C20), biphytanyl (C40), and unidentified alkyl chains accounted for 10, 83, and 7 mol% of the total alkyl chains, respectively. Strong acid hydrolysis of the macromolecular residue obtained after lipid extraction gave a significant amount of C40 tetraether, which had probably been bound covalently to other substances in the cells.