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Stationary versus non-stationary 13C-MFA: A comparison using a consistent dataset

Paper ID Volume ID Publish Year Pages File Format Full-Text
23956 43484 2011 12 PDF Available
Title
Stationary versus non-stationary 13C-MFA: A comparison using a consistent dataset
Abstract

Besides the well-established 13C-metabolic flux analysis (13C-MFA) which characterizes a cell's fluxome in a metabolic and isotopic stationary state a current area of research is isotopically non-stationary MFA. Non-stationary 13C-MFA uses short-time isotopic transient data instead of long-time isotopic equilibrium data and thus is capable to resolve fluxes within much shorter labeling experiments. However, a comparison of both methods with data from one single experiment has not been made so far. In order to create a consistent database for directly comparing both methods a 13C-labeling experiment in a fed-batch cultivation with a Corynebacterium glutamicum lysine producer was carried out. During the experiment the substrate glucose was switched from unlabeled to a specifically labeled glucose mixture which was immediately traced by fast sampling and metabolite quenching. The time course of labeling enrichments in intracellular metabolites until isotopic stationarity was monitored by LC–MS/MS. The resulting dataset was evaluated using the classical as well as the isotopic non-stationary MFA approach.The results show that not only the obtained relative data, i.e. intracellular flux distributions, but also the more informative quantitative fluxome data significantly depend on the combination of the measurements and the underlying modeling approach used for data integration. Taking further criteria on the experimental and computational part into consideration, the current limitations of both methods are demonstrated and possible pitfalls are concluded.

Keywords
Metabolic flux analysis; Isotopic labeling; 13C-MFA
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Stationary versus non-stationary 13C-MFA: A comparison using a consistent dataset
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 154, Issues 2–3, 10 July 2011, Pages 179–190
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us