In quantitative metabolomics studies, the most crucial step was arresting snapshots of all interesting metabolites. However, the procedure customized for Streptomyces was so rare that most studies consulted the procedure from other bacteria even yeast, leading to inaccurate and unreliable metabolomics analysis. In this study, a base solution (acetone: ethanol = 1:1, mol/mol) was added to a quenching solution to keep the integrity of the cell membrane. Based on the molar transition energy (E_T) of the organic solvents, five solutions were used to carry out the quenching procedures. These were acetone, isoamylol, propanol, methanol, and 60% (v/v) methanol. To the best of our knowledge, this is the first report which has utilized a quenching solution with E_T values. Three procedures were also adopted for extraction. These were boiling, freezing–thawing, and grinding ethanol. Following the analysis of the mass balance, amino acids, organic acids, phosphate sugars, and sugar alcohols were measured using gas chromatography with an isotope dilution mass spectrometry. It was found that using isoamylol with a base solution (5:1, v/v) as a quenching solution and that freezing–thawing in liquid nitrogen within 50% (v/v) methanol as an extracting procedure were the best pairing for the quantitative metabolomics of Streptomyces ZYJ-6, and resulted in average recoveries of close to 100%. The concentration of intracellular metabolites obtained from this new quenching solution was between two and ten times higher than that from 60% (v/v) methanol, which until now has been the most commonly used solution. Our findings are the first systematic quantitative metabolomics tools for Streptomyces ZYJ-6 and, therefore, will be important references for research in fields such as ¹³C based metabolic flux analysis, multi-omic research and genome-scale metabolic model establishment, as well as for other Streptomyces.

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链霉菌中基于质谱的代谢组学的优化采样方案

在定量代谢组学研究中，最关键的步骤是捕获所有有趣代谢物的快照。但是，为链霉菌定制的程序非常罕见，以至于大多数研究都从其他细菌甚至酵母中咨询了该程序，从而导致代谢组学分析不准确且不可靠。在这项研究中，将基本溶液（丙酮：乙醇= 1：1，mol / mol）添加到淬灭溶液中，以保持细胞膜的完整性。基于有机溶剂的摩尔转变能（E _T），使用了五种溶液进行淬灭程序。这些是丙酮，异戊醇，丙醇，甲醇和60％（v / v）甲醇。据我们所知，这是第一份使用E _T淬火溶液的报告。价值观。还采用了三种程序进行提取。这些是沸腾的，冻融的和研磨的乙醇。在质量平衡分析之后，使用具有同位素稀释质谱法的气相色谱法测量氨基酸，有机酸，磷酸糖和糖醇。研究发现，使用异戊醇与碱溶液（5：1，v / v）作为淬灭溶液，在液氮中于50％（v / v）甲醇中冷冻解冻作为萃取步骤是最佳的配对方法。链霉菌的定量代谢组学ZYJ-6，平均回收率接近100％。从这种新的淬灭溶液中获得的细胞内代谢物浓度比从60％（v / v）甲醇获得的浓度高2到10倍，而甲醇是迄今为止最常用的溶液。我们的发现是链霉菌ZYJ-6的第一个系统的定量代谢组学工具，因此，将为基于¹³ C的代谢通量分析，多组学研究和基因组规模的代谢模型建立以及其他领域的研究提供重要参考。对于其他链霉菌。