Analysis of Mass Isotopomer Distributions of Proteingenic Amino Acids Used GC-MS during ~(13)CMFA
- VernacularTitle:~(13)CMFA过程中GC-MS分析菌体蛋白氨基酸的~(13)C标记丰度
- Author:
Jian WANG
;
Ze-Jian WANG
;
Ming-Zhi HUANG
;
Jiang-Chao QIAN
;
Ju CHU
;
Si-Liang ZHANG
;
- Publication Type:Journal Article
- Keywords:
13C metabolic flux analysis Proteinogenic amino acids Amino acids derivatized GC-MS 13C labelling pattern
- From:
China Biotechnology
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
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.
