大肠杆菌9种有机酸代谢产物的高效液相检测条件优化  

Optimization of HPLC determination Conditions for 9 Organic Acids Produced by E.coli

作　　者：马建刚[1] 刘娇[2] 曾祥峰[2] 郑平[2] 贾士儒[1] 孙际宾[2] 

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]中国科学院天津工业生物技术研究所,天津300308

出　　处：《现代食品科技》2011年第5期591-594,519,共5页Modern Food Science and Technology

基　　金：中国科学院知识创新工程重要方向项目(KSCX2-YW-G-030;G-070);天津市科技支撑计划重点项目(09ZCKFSH00700)

摘　　要：以高交联度磺化苯乙烯-二乙烯基苯共聚物多孔微球作为固定相的Aminex HPX-87H离子色谱柱为分析柱,通过优化条件对E.coli MG1655及其突变株厌氧发酵过程中九种有机酸（琥珀酸、甲酸、乙酸、乳酸、草酰乙酸、丙酮酸、富马酸、乙醛酸、柠檬酸）进行了有效的分离,尤其做到了对富马酸与其他有机酸的较好分离,为代谢过程监测及后期菌株改造提供了重要支持。优化后条件为：流动相2.75 mM H2SO4、流速0.6 mL/min、柱温50℃,本方法的回收率在97.9%~106.7%;RSD为0.8%~3.3%Ion chromatographic column Aminex HPX-87H filled with highly cross-linked sulfonated polystyrene divinylbenzene porous microspheres was used to determine 9 kinds of organic acids in the anaerobic fermentation broth of E.coli MG1655 and its mutant.After optimization of HPLC conditions,nine kinds of organic acids（succinic acid,formic acid,acetic acid,lactic acid,oxaloacetate,pyruvate,fumaric acid,glyoxylic acid,citric acid） were successfully separated.In particular,fumaric acid was well separated from the other organic acids,which provides important support for metabolic process monitoring and subsequent strain development.The optimal determination conditions were： mobile phase 2.75 mM H2SO4,flow rate 0.6 mL/min,and column temperature 50 ℃.The recovery of this method was 97.9%~106.7% and RSD was 0.8%~3.3%.

关 键 词：大肠杆菌 有机酸 代谢产物 

分 类 号：TQ921[轻工技术与工程—发酵工程]