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13C metabolic flux analysis for larger scale cultivation using gas chromatography-combustion-isotope ratio mass spectrometry

Paper ID Volume ID Publish Year Pages File Format Full-Text
31730 44833 2010 9 PDF Available
Title
13C metabolic flux analysis for larger scale cultivation using gas chromatography-combustion-isotope ratio mass spectrometry
Abstract

13C-based metabolic flux analysis (13CMFA) is limited to smaller scale experiments due to very high costs of labeled substrates. We measured 13C enrichment in proteinogenic amino acid hydrolyzates using gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) from a series of parallel batch cultivations of Corynebacterium glutamicum utilizing mixtures of natural glucose and [1-13C] glucose, containing 0%, 0.5%, 1%, 2%, and 10% [1-13C] glucose. Decreasing the [1-13C] glucose content, kinetic isotope effects played an increasing role but could be corrected. From the corrected 13C 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.

Keywords
Metabolic flux analysis; GC-C-IRMS; 13C-labeling; Proteinogenic amino acid; Corynebacterium glutamicum
First Page Preview
13C metabolic flux analysis for larger scale cultivation using gas chromatography-combustion-isotope ratio mass spectrometry
Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 12, Issue 4, July 2010, Pages 392–400
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering