【摘要】 近年来微生物代谢组学逐渐成为一种新兴的分析手段.微生物虽然有着结构简单、营养需求单一的特点,但也存在自身种类繁多、细胞内代谢物质理化性质各不相同的弊端.因此,在前期样品处理方面仍存在很大的不足.基于GC-MS的分析方法,建立了一种用于酵母菌S288C的代谢组学分析方法,通过考察甲醇(-20℃)、V(乙腈):V(水)=1:1(-20℃)、V(乙腈):V(甲醇):V(水)=2:2:1(-20℃)、V(水):V(异丙醇):V(甲醇)=2:2:5(-20℃)、75%乙醇(煮沸):5种溶剂对胞内物提取的影响,确定最佳的提取溶剂为V(水):V(异丙醇):V(甲醇)=2:2:5(-20℃).该方法可检测348种代谢物,通过数据库检索可定性出142种物质,包括46种氨基酸类代谢物、9种糖类代谢物、30种有机酸代谢物、17种醇类代谢物以及23种酯类代谢物.更多还原

【Abstract】 In recent years,microbial metabolomics has gradually become a new analysis method.Although microorganisms have the characteristics of simple structure and single nutritional requirement,they also have many kinds of their own.The physical and chemical properties of intracellular metabolites are different.Therefore,there are still a lot of deficiencies in the pre-sample treatment.Based on GC-MS,a metabolomics analysis method for yeast S288 C has been established in this paper.The effects of five solvents,including methanol(-20 ℃),V(acetonitrile)■V(water)=1■1(-20 ℃),V(acetonitrile)■V(methanol/water)=2■2■1(-20 ℃),V(water)■V(isopropanol/methanol)=2■2■5(-20 ℃) and 75% ethanol(boiling) on the extraction of intracellular substances are investigated.The optimum solvent is V(water)■V(isopropanol)■V(methanol)=2■2■5(-20 ℃).The method can detect 348 metabolites,and 142 metabolites are identified by NIST library searching,including 46 amino acids,9 carbohydrates,30 organic acids,17 alcohols and 23 esters.更多还原