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《China Biotechnology》 2009-07
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Analysis of Mass Isotopomer Distributions of Proteingenic Amino Acids Used GC-MS during ~(13)CMFA

WANG Jian WANG Ze-jian HUANG Ming-zhi QIAN Jiang-chao CHU Ju ZHANG Si-liang(The State Key Laboratory of Bioreactor Engineering of East China University of Science and Technology/The National Engineering and Technology Research Center for Biotechnology,Shanghai 200237,China)  
13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.
【CateGory Index】: Q936
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Chinese Journal Full-text Database 1 Hits
1 Li XiaojingChen TaoChen XunLi XianggaoZhao Xueming~(**)(Department of Biochemical Engineering,School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China);Advances in ~(13)C Metabolic Flux Analysis[J];Progress in Chemistry;2006-Z2
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