期刊
BIOTECHNOLOGY JOURNAL
卷 6, 期 11, 页码 1377-1387出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201000446
关键词
Escherichia coli; Free amino acids; Metabolic engineering; Metabolic flux analysis
资金
- Osaka University under the Ministry of Education, Culture, Sports, Science and Technology, Japan
Metabolic flux analysis using C-13 enrichment data of intracellular free amino acids (FAAs) can improve the time resolution of flux estimation compared to analysis of proteinogenic amino acid data owing to the faster turnover times of FAAs. The nature of the C-13 enrichment dynamics of FAAs remains obscure, however, especially with regard to its dependence on culture conditions, even though an understanding of dynamic behavior is important for precise metabolic flux estimation. In this study, we analyzed the C-13 enrichment dynamics of free and proteinogenic amino acids in a series of continuous culture experiments with Escherichia coli. The results indicated that the effect of protein degradation on the C-13 enrichment of FAAs was negligible under cellular growth conditions. Furthermore, they showed that the time scale necessary for C-13 enrichment dynamics of FAAs to reach a steady state depends on culture conditions such as oxygen uptake rate, which was likely due to different pool sizes of intracellular metabolites. The results demonstrate the importance of analyzing C-13 enrichment dynamics for the precise estimation of metabolic fluxes using FAA data.
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