基于GC-MS的棉子糖肠球菌代谢组学样品前处理方法的优化  

GC-MS-Based Optimization of Pretreatment of Enterococcus raffinosus Samples for Metabonomics

作　　者：杨新达 牛肖凡 刘仲浩 张健[1] 张巧珍[2] 高强[1] YANG Xin-da;NIU Xiao-fan;LIU Zhong-hao;ZHANG Jian;ZHANG Qiao-zhen;GAO Qiang(Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education,Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control,College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China;College of Sciences,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区：[1]工业发酵微生物教育部重点实验室,天津市微生物代谢与发酵过程控制技术工程中心,天津科技大学生物工程学院,天津300457 [2]天津科技大学理学院,天津300457

出　　处：《食品研究与开发》2022年第18期130-136,共7页Food Research and Development

基　　金：国家自然科学基金项目(31902193、61603273);天津自然科学基金重点项目(20JCZDJC00140)。

摘　　要：探究不同的样品前处理方法对气相色谱-质谱联用(gas chromatography-mass spectrometry,GC-MS)法检测棉子糖肠球菌(Enterococcus raffinosus)胞内小分子代谢物种类和数量的影响。通过GC-MS分析测定不同的菌体破碎方式、菌体清洗次数以及冻融时间处理后的棉子糖肠球菌样品的胞内代谢物的种类。结果显示,液氮研磨较超声破碎更有利于胞内物质的释放;清洗3次既可清洗掉菌体表面的杂质,也能够防止胞内小分子代谢物外泄;冻融3 min更利于胞内代谢物的提取。通过优化后的前处理方法处理的样品,共检测出253种胞内小分子物质,经NIST 17.0数据库可精确匹配到8类共47种物质,其中酯类5种、烯烃类1种、烷烃类2种、醇类5种、酸类30种、糖类1种、苯环类2种、其他1种。The small molecular intracellular metabolites of Enterococcus raffinosus samples pretreated with different methods were detected by gas chromatography-mass spectrometry(GC-MS).The effects of different cell crashing methods,washing times,and freeze-thaw time on the detection results were studied.The results showed that liquid nitrogen grinding was more favorable for the release of intracellular substances than ultrasonic crushing.Washing the cells for 3 times not only cleaned the impurities on the cell surface but also prevented the release of small molecular metabolites.Freeze-thaw for 3 min was most favorable for the extraction of intracellular metabolites.After optimization of the pretreatment method,253 small molecular metabolites were detected,in which 47 metabolites were accurately matched with those in NIST 17.0,including 5 esters,1 alkene,2 alkanes,5 alcohols,30 acids,1 sugar,2 benzenes,and 1 other.

关 键 词：棉子糖肠球菌 代谢组学 样品前处理方法 气相色谱-质谱法(GC-MS) 胞内代谢物 

分 类 号：O657.63[理学—分析化学] TS201.3[理学—化学]