高山被孢霉淬灭方法及胞内代谢产物的气相色谱-质谱分析比较  

Comparison of sampling method of intracellular metabolites in Mortierella alpina and the analysis of the metabolites by GC-MS

作　　者：刘欣[1] 傅宁华[1] 张红漫[2] 纪晓俊[1] 张鑫[2] 杨健健[1] 黄和[1] 

机构地区：[1]南京工业大学生物与制药工程学院材料化学工程国家重点实验室,南京210009 [2]南京工业大学理学院,南京210009

出　　处：《生物加工过程》2013年第6期63-67,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金(21176124);国家自然科学基金重点项目(20936002);国家科技支撑计划(2011BAD23B03);江苏省高校优势学科建设工程

摘　　要：在花生四烯酸生产菌高山被孢霉代谢组学研究中,需利用胞内代谢物的提取手段并基于气相色谱-质谱(GC-MS)分析方法对其进行检测。比较了3种胞内代谢物提取方法及不同色谱柱条件下GC-MS分析结果。研究表明:采用冷甲醇淬灭分别较液氮直接淬灭及真空过滤后,减少了胞内代谢物的泄露并更好地实现了胞外及胞内代谢物的分离。在对代谢物分析的比较中,极性色谱柱(DB-FFAP)检出的代谢物仅为11种,主要为有机酸、醛类;而代谢物经衍生化后采用非极性色谱柱(DB-5)共检出32种化合物,主要为糖、糖苷及醇类。In the metabolomics study on arachidonic acid production by Mortierella alpina, sampling method of intracellular metabolites and their detection based on GC-MS analysis are needed. The comparisons of three sampling methods and GC-MS analysis results in two chromatographic columns were carried out. The results showed that cold methanol quenching caused less metabolites leakage and more effective separation of extra-and intracellular metabolites than direct quenching in liquid nitrogen and vacuum filtration followed by quenching in liquid nitrogen. In the comparison of analysis of metabolites, only 11 metabolites were determined by polar column （DB-FFAP）, including organic acids and aldehydes. However, 32 metabolites were determined by non polar column （DB-5）, including sugars, glycosides, and alcohols.

关 键 词：花生四烯酸 高山被孢霉 代谢组学 代谢产物 取样 GC-MS分析 

分 类 号：S182[农业科学—农业基础科学]