Recombinant protein production-associated metabolic burden reflects anabolic constraints and reveals similarities to a carbon overfeeding response

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Autoren

  • Zhaopeng Li
  • Ursula Rinas

Organisationseinheiten

Externe Organisationen

  • Helmholtz-Zentrum für Infektionsforschung GmbH (HZI)
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Details

OriginalspracheEnglisch
Seiten (von - bis)94-105
Seitenumfang12
FachzeitschriftBiotechnology and Bioengineering
Jahrgang118
Ausgabenummer1
Frühes Online-Datum3 Sept. 2020
PublikationsstatusVeröffentlicht - 31 Dez. 2020

Abstract

A comparison of the metabolic response of Escherichia coli BL21 (DE3) towards the production of human basic fibroblast growth factor (hFGF-2) or towards carbon overfeeding revealed similarities which point to constraints in anabolic pathways. Contrary to expectations, neither energy generation (e.g., ATP) nor provision of precursor molecules for nucleotides (e.g., uracil) and amino acids (e.g., pyruvate, glutamate) limit host cell and plasmid-encoded functions. Growth inhibition is assumed to occur when hampered anabolic capacities do not match with the ongoing and overwhelming carbon catabolism. Excessive carbon uptake leads to by-product secretion, for example, pyruvate, acetate, glutamate, and energy spillage, for example, accumulation and degradation of adenine nucleotides with concomitant accumulation of extracellular hypoxanthine. The cellular response towards compromised anabolic capacities involves downregulation of cAMP formation, presumably responsible for subsequently better-controlled glucose uptake and resultant accumulation of glucose in the culture medium. Growth inhibition is neglectable under conditions of reduced carbon availability when hampered anabolic capacities also match with catabolic carbon processing. The growth inhibitory effect with accompanying energy spillage, respectively, hypoxanthine secretion and cessation of cAMP formation is not unique to the production of hFGF-2 but observed during the production of other proteins and also during overexpression of genes without transcript translation.

ASJC Scopus Sachgebiete

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Recombinant protein production-associated metabolic burden reflects anabolic constraints and reveals similarities to a carbon overfeeding response. / Li, Zhaopeng; Rinas, Ursula.
in: Biotechnology and Bioengineering, Jahrgang 118, Nr. 1, 31.12.2020, S. 94-105.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

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