期刊
METABOLIC ENGINEERING
卷 12, 期 4, 页码 392-400出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2010.02.001
关键词
Metabolic flux analysis; GC-C-IRMS; C-13-labeling; Proteinogenic amino acid; Corynebacterium glutamicum
资金
- German Research Foundation DFG [GRK-532]
C-13-based metabolic flux analysis ((13)CMFA) is limited to smaller scale experiments due to very high costs of labeled substrates. We measured C-13 enrichment in proteinogenic aminio acid hydrolyzates using gas chromotography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) from a series of parallel batch cultivations of Corynebacterium glutamicum utilizing mixtures of natural glucose and [1-C-13] glucose, containing 0%, 0.5%, 1%, 2% and 10% [1-C-13] glucose. Decreasing the [1-C-13] glucose content, kinetic isotope effects played an increasing role but could be corrected. From the corrected C-13 enrichments in vivo fluxes in the central metabolism were determined by numerical optimization. The obtained flux distribution was very similar to those obtained from parallel labeling experiments using conventional high labeling GC-MS method and to published results. The GC-C-IRMS-based method involving low labeling degree of expensive tracer substrate, e.g. 1%, is well suited for larger laboratory and industrial pilot scale fermentations. (C) 2010 Elsevier Inc. All rights reserved.
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