Řezanka Pavel, Řezanka Michal, Kyselová Lucie, Řezanka Tomáš
Department of Analytical Chemistry, Faculty of Chemical Engineering, University of Chemistry and Technology, Prague, 166 28, Prague 6, Czech Republic.
Institute for Nanomaterials, Advanced Technologies and Innovation, Technical University of Liberec, Studentská 1402/2, 461 17, Liberec 1, Czech Republic.
Folia Microbiol (Praha). 2025 Feb;70(1):225-233. doi: 10.1007/s12223-024-01235-3. Epub 2024 Dec 17.
Lipids from microorganisms, and especially lipids from Archaea, are used as taxonomic markers. Unfortunately, knowledge is very limited due to the uncultivability of most Archaea, which greatly reduces the importance of the diversity of lipids and their ecological role. One possible solution is to use lipidomic analysis. Six radioactive sources were investigated, two of which are surface (Wettinquelle and Radonka) and four deep from the Svornost mine (Agricola, Behounek, C1, and Curie). A total of 15 core lipids and 82 intact polar lipids were identified from the membranes of microorganisms in six radioactive springs. Using shotgun lipidomics, typical Archaea lipids were identified in spring water, namely dialkyl glycerol tetraethers, archaeol, hydroxyarchaeol and dihydroxyarchaeol. Diverse groups of polar heads were formed in archaeal IPLs, whose polar heads are formed mainly by hexose, deoxyhexose, and phosphoglycerol. The analysis was performed using shotgun lipidomics and the structure of all molecular species was confirmed by tandem mass spectrometry. After acid hydrolysis, a mixture of polar compounds was obtained from the polar head. Further analysis by GC-MS confirmed that the carbohydrates were glucose and rhamnose. Analysis by HPLC-MS of diastereoisomers of 2-(polyhydroxyalkyl)-3-(O-tolylthiocarbamoyl)thiazolidine-4(R)-carboxylates revealed that both L-rhamnose and D-glucose are present in spring samples only in varying amounts. The glycoside composition depends on the type of spring, that is, Wettinquelle and Radonka springs are basically shallow groundwater, while the samples from the Svornost mine are deep groundwater and do not contain glycosides with rhamnose. This method enables quick screening for characteristic Archaea lipids, allowing decisions on whether to pursue further analyses, such as metagenomic analysis, to directly confirm the presence of Archaea.
来自微生物的脂质，尤其是古菌的脂质，被用作分类标记。不幸的是，由于大多数古菌无法培养，相关知识非常有限，这大大降低了脂质多样性及其生态作用的重要性。一种可能的解决方案是使用脂质组学分析。研究了六个放射性源，其中两个是地表的（韦廷泉和拉东卡），四个是来自斯沃尔诺斯特矿深部的（阿格里科拉、贝霍内克、C1和居里）。从六个放射性泉水中微生物的膜中总共鉴定出15种核心脂质和82种完整的极性脂质。使用鸟枪法脂质组学，在泉水中鉴定出了典型的古菌脂质，即二烷基甘油四醚、古菌醇、羟基古菌醇和二羟基古菌醇。古菌完整极性脂质中形成了不同的极性头部基团群体，其极性头部主要由己糖、脱氧己糖和磷酸甘油形成。使用鸟枪法脂质组学进行分析，并通过串联质谱确认了所有分子物种的结构。酸水解后，从极性头部获得了极性化合物混合物。通过气相色谱-质谱联用进一步分析证实，碳水化合物为葡萄糖和鼠李糖。通过高效液相色谱-质谱联用对2-(多羟基烷基)-3-(O-甲苯基硫代氨基甲酰基)噻唑烷-4(R)-羧酸盐的非对映异构体进行分析表明，L-鼠李糖和D-葡萄糖在泉水样品中仅以不同含量存在。糖苷组成取决于泉水类型，也就是说，韦廷泉和拉东卡泉基本上是浅层地下水，而来自斯沃尔诺斯特矿的样品是深层地下水，不含含鼠李糖糖苷。这种方法能够快速筛选出特征性的古菌脂质，从而决定是否进行进一步分析，如宏基因组分析，以直接确认古菌的存在。
