液相质谱联用的代谢组方法优化及其在蓝细菌分析中的应用

Optimization and application of targeted LC-MS metabolomic analyses in photosynthetic cyanobacteria

摘要:

为了准确鉴定光合蓝细菌中的各种代谢物，需要对基于液相色谱–质谱联用仪 (LC-MS) 的代谢组学分析方法进行有针对性的优化。本研究选取了24种涉及中心碳代谢和能量代谢的代谢物作为LC-MS的检测目标，获得了每个代谢物的最适色谱分离条件和质谱参数；同时以光合蓝细菌Synechocystis sp. PCC6803为主要对象，针对性地优化了样品前处理条件，结果显示适当延长梯度洗脱顺序表的时间并将流速设为0.2 mL/min可以得到最佳的分离效果，同时选择80% (V/V) 甲醇 (?80 ?C) 作为代谢物萃取剂。分析结果证明这一代谢组分析技术可以成功地应用到光合蓝细菌的研究中。

Abstract:

To accurately analyze metabolites in industry-important photosynthetic microbes, LC-MS based metabolomics protocol needs to be optimized specifically for individual species. In this study, an LC-MS based metabolomics method was optimized for cyanobacterium Synechocystis sp. PCC 6803. With the optimized extraction, liquid chromatographic and mass spectral parameters, the method was capable of detecting 24 important metabolites related to central carbohydrate and energy metabolism in Synechocystis sp. PCC 6803. The study laid an important foundation for the metabolomics analysis of cyanobacteria.