Response of fluxome and metabolome to temperature-induced recombinant protein synthesis in Escherichia coli

Wittmann, Christoph, Weber, Jan, Betiku, Eriola, Kromer, Jens, Böhm, Daniela and Rinas, Ursula (2007) Response of fluxome and metabolome to temperature-induced recombinant protein synthesis in Escherichia coli. Journal of Biotechnology, 132 4: 375-384. doi:10.1016/j.jbiotec.2007.07.495


Author Wittmann, Christoph
Weber, Jan
Betiku, Eriola
Kromer, Jens
Böhm, Daniela
Rinas, Ursula
Title Response of fluxome and metabolome to temperature-induced recombinant protein synthesis in Escherichia coli
Formatted title
Response of fluxome and metabolome to temperature-induced recombinant protein synthesis in Escherichia coli
Journal name Journal of Biotechnology   Check publisher's open access policy
ISSN 0168-1656
Publication date 2007-12-01
Sub-type Article (original research)
DOI 10.1016/j.jbiotec.2007.07.495
Volume 132
Issue 4
Start page 375
End page 384
Total pages 10
Place of publication Amsterdam
Publisher Elsevier Science.
Language eng
Subject 060101 Analytical Biochemistry
060104 Cell Metabolism
060114 Systems Biology
Abstract The response of the central carbon metabolism of Escherichia coli to temperature-induced recombinant production of human fibroblast growth factor was studied on the level of metabolic fluxes and intracellular metabolite levels. During production, E. coli TG1:pFGFB, carrying a plasmid encoded gene for the recombinant product, revealed stress related characteristics such as decreased growth rate and biomass yield and enhanced by-product excretion (acetate, pyruvate, lactate). With the onset of production, the adenylate energy charge dropped from 0.85 to 0.60, indicating the occurrence of a severe energy limitation. This triggered an increase of the glycolytic flux which, however, was not sufficient to compensate for the increased ATP demand. The activation of the glycolytic flux was also indicated by the readjustment of glycolytic pool sizes leading to an increased driving force for the reaction catalyzed by phosphofructokinase. Moreover, fluxes through the TCA cycle, into the pentose phosphate pathway and into anabolic pathways decreased significantly. The strong increase of flux into overflow pathways, especially towards acetate was most likely caused by a flux redirection from pyruvate dehydrogenase to pyruvate oxidase. The glyoxylate shunt, not active during growth, was the dominating anaplerotic pathway during production. Together with pyruvate oxidase and acetyl CoA synthase this pathway could function as a metabolic by-pass to overcome the limitation in the junction between glycolysis and TCA cycle and partly recycle the acetate formed back into the metabolism.
Keyword Human basic fibroblast growth factor
Adenylate energy charge
Cellular stress
¹³C metabolic flux
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Thu, 12 Feb 2009, 15:18:08 EST by Alexandra Cooney on behalf of Aust Institute for Bioengineering & Nanotechnology