Isotopically non-stationary metabolic flux analysis: complex yet highly informative
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- 作者：Wolfgang Wiechert ; Katharina N枚h
- 刊名：Current Opinion in Biotechnology
- 出版年：December, 2013
- 年：2013
- 卷：24
- 期：6
- 页码：979-986
- 全文大小：1333 K
文摘
Metabolic flux analysis (MFA) using isotopic tracers aims at the experimental determination of in vivo reaction rates (fluxes). In recent years, the well-established 13C-MFA method based on metabolic and isotopic steady state was extended to INST-MFA (isotopically non-stationary MFA), which is performed in a transient labeling state. INST-MFA offers short-time experiments with a maximal information gain, and can moreover be applied to a wider range of growth conditions or organisms. Some of these conditions are not accessible by conventional methods. This comes at the price of significant methodological complexity involving high-frequency sampling and quenching, precise analysis of many samples and an extraordinary computational effort. This review gives a brief overview of basic principles, experimental workflows, and recent progress in this field. Special emphasis is laid on the trade-off between total effort and information gain, particularly on the suitability of INST-MFA for certain types of biological questions. In order to integrate INST-MFA as a viable method into the toolbox of MFA, some major challenges must be addressed in the coming years. These are discussed in the outlook.