期刊
JOURNAL OF BIOTECHNOLOGY
卷 150, 期 4, 页码 497-508出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2010.09.944
关键词
Metabolic flux analysis; Mammalian cells; Metabolic networks; Underdetermined systems; Convex analysis
资金
- Interuniversity Attraction Poles Program
this article the metabolic flux analysis of growing CHO-320 cells is performed for a detailed metabolic network which involves 100 reactions and embraces all the significant pathways describing the metabolism of CHO cells. The purpose is to investigate the efficiency of the flux analysis when it is based on a relatively small set of extracellular measurements that can be easily achieved in most laboratories. In this case the flux analysis problem leads to a generally underdetermined mass balance system, as data are not sufficient to uniquely define the metabolic fluxes. Our main contribution is to show that, provided the system of mass balance equations is well-posed, although it is underdetermined, very narrow intervals may be found for most fluxes. The importance of checking the well-posedness of the problem is emphasized and the influence of the number of available measurements on the accuracy of the metabolic flux intervals is systematically investigated. In all cases the computed flux intervals are bounded and a single well defined value is obtained for the formation rates of the cellular macromolecules (proteins, DNA. RNA, lipids) that are not measured. The potential gain of a simple theoretical assumption regarding the metabolism of Threonine is also discussed and compared with an optimal solution calculated by maximizing the biomass formation rate. Alternative network structures obtained by inverting the direction of reversible reactions are also considered. Finally, the results of the metabolic flux analysis are exploited to estimate the total energy production resulting from the metabolism of growing CHO-320 cells. (C) 2010 Elsevier By. All rights reserved.
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