重组大肠杆菌YK537/pSB┐TK高密度培养过程中有机酸的RP┐HPLC快速分析

RP HPLC Rapid Analysis of Organic Acids during High Cell Density Culture of Recombinant E.coli YK537/pSB TK

采用高压液相色谱法（ＨＰＬＣ），利用Ｃ１８反相柱快速检测重组大肠杆菌ＹＫ５３７／ｐＳＢ－ＴＫ高密度发酵过程中细菌代谢产生的各种有机酸。分析表明，经乙醚和水两次萃取处理后能去除样品中大部分的干扰峰，可获得理想的分离效果，并能定量测出发酵过程中各种有机酸的积累。不同培养方式的结果表明用甘油代替葡萄糖作为细菌生长的碳源可减少乙酸的积累，降低其对细菌生长和外源蛋白表达的抑制作用，从而更易实现重组大肠杆菌的高表达、高密度培养。

Various organic acids produced by bacteria during high cell density culture of recombinant E.coli YK537/pSB TK were rapidly analysed by HPLC with reversed phase colmn C 18 .Ideal separation and quantitative analysis of organic acids were achieved by ether and water extractio,which could remove most of interferential peaks in the sample.Results indicated that acetic acid decreased using glycerol as carbon resource for bacteria growth in place of glucose,while the inhibition of cell growth and hetero...