基于气相色谱-质谱联用的¹³C代谢通量分析。

GC-MS-based ¹³C metabolic flux analysis.

作者信息

Becker Judith, Wittmann Christoph

机构信息

Institute of Systems Biotechnology, Saarland University, Campus A 1 5, 66123, Saarbrücken, Germany.

出版信息

Methods Mol Biol. 2014;1191:165-74. doi: 10.1007/978-1-4939-1170-7_10.

DOI:10.1007/978-1-4939-1170-7_10

PMID:25178790

Abstract

The in vivo analysis of metabolic fluxes has become a valuable method for the investigation of microorganisms. It turned out especially useful in industrial biotechnology for the prediction of beneficial genetic targets for rational strain optimization. Here, we describe in detail the procedure for the state-of-the-art approach of (13)C metabolic flux analysis comprising steady-state cultivations in a mineral salt medium with (13)C-labeled substrates, GC-MS measurement for labeling analysis, as well as metabolic modeling using the open-source software OpenFlux.

摘要

代谢通量的体内分析已成为研究微生物的一种有价值的方法。事实证明，它在工业生物技术中对于合理菌株优化的有益遗传靶点预测特别有用。在此，我们详细描述了采用先进的（13）C代谢通量分析方法的步骤，包括在含有（13）C标记底物的矿物盐培养基中进行稳态培养、用于标记分析的气相色谱-质谱联用（GC-MS）测量以及使用开源软件OpenFlux进行代谢建模。