Villas-Bôas Silas G, Noel Samantha, Lane Geoffrey A, Attwood Graeme, Cookson Adrian
AgResearch Limited, Grasslands Research Centre, Private Bag 11008, Palmerston North 5301, New Zealand.
Anal Biochem. 2006 Feb 15;349(2):297-305. doi: 10.1016/j.ab.2005.11.019. Epub 2005 Dec 1.
This work reports the implementation and optimization of a method for high-throughput analysis of metabolites produced by the breakdown of natural polysaccharides by microorganisms. Our simple protocol enables simultaneous separation and quantification of more than 40 different sugars and sugar derivatives, in addition to several organic acids in complex media, using 50-mul samples and a standard gas chromatography-mass spectrometry platform that was fully optimized for this purpose. As an implementation proof-of-concept, we assayed extracellular metabolite levels of three bacterial strains cultivated on complex medium rich in polysaccharides and under identical growth conditions. We demonstrate that the metabolic footprinting profile data distinguish among sample types such as typical metabolomics data. Moreover, we demonstrate that the differential metabolite-level data provide insight on specific fibrolytic activity of the different microbial strains and lay the groundwork for integrated proteome-metabolome studies of fiber-degrading microorganisms.
这项工作报告了一种用于高通量分析微生物分解天然多糖产生的代谢物的方法的实施与优化。我们的简单方案能够使用50微升样品以及为此目的而完全优化的标准气相色谱-质谱平台，在复杂培养基中同时分离和定量40多种不同的糖和糖衍生物以及几种有机酸。作为概念验证的实施，我们测定了在富含多糖的复杂培养基上且在相同生长条件下培养的三种细菌菌株的细胞外代谢物水平。我们证明，代谢足迹图谱数据能够像典型的代谢组学数据一样区分不同的样品类型。此外，我们证明，差异代谢物水平数据为不同微生物菌株的特定纤维分解活性提供了见解，并为纤维降解微生物的蛋白质组-代谢组综合研究奠定了基础。
