Spura Jana, Reimer Lorenz Christian, Wieloch Patricia, Schreiber Kerstin, Buchinger Sebastian, Schomburg Dietmar
Department of Bioinformatics and Biochemistry, Technische Universität Braunschweig, 38106 Braunschweig, Germany.
Anal Biochem. 2009 Nov 15;394(2):192-201. doi: 10.1016/j.ab.2009.07.016. Epub 2009 Jul 15.
Although microbial metabolome analysis has now become a widely used method, no generally applicable quenching method has been published so far. Either the methods were established for only one defined organism or the metabolite coverage was quite low. In the current work, a novel, reliable, and robust quenching method for different types of organisms is described. Compared with the commonly used quenching procedure with 60% methanol (-50 degrees C), we obtained improved results for three examined organisms with different cell wall and membrane structures using a 40% ethanol/0.8% sodium chloride solution (-20 degrees C). Increased metabolite levels were achieved for 60-80% of all identified compounds. Moreover, the estimated standard error of the relative concentrations of 120-160 different substances was only 14+/-4% compared with 17+/-3% in unquenched samples and 24+/-7% in samples quenched with methanol for the different tested organisms.
尽管微生物代谢组分析现已成为一种广泛使用的方法，但迄今为止尚未发表普遍适用的淬灭方法。这些方法要么仅针对一种特定的生物体建立，要么代谢物覆盖范围相当低。在当前的工作中，描述了一种针对不同类型生物体的新型、可靠且稳健的淬灭方法。与常用的60%甲醇（-50℃）淬灭程序相比，使用40%乙醇/0.8%氯化钠溶液（-20℃），我们在三种具有不同细胞壁和膜结构的受试生物体中获得了更好的结果。所有已鉴定化合物中有60-80%的代谢物水平有所提高。此外，对于不同测试生物体，120-160种不同物质相对浓度的估计标准误差仅为14±4%，而未淬灭样品为17±3%，甲醇淬灭样品为24±7%。
