Details zur Publikation

Referenztyp Zeitschriften
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Creative Commons Lizenz creative commons licence
Titel (primär) The metabolic flux probe (MFP)—Secreted protein as a non-disruptive information carrier for 13C-based metabolic flux analysis
Autor Dusny, C.; Schmid, A.
Journal / Serie International Journal of Molecular Sciences
Erscheinungsjahr 2021
Department SOMA
Band/Volume 22
Heft 17
Seite von art. 9438
Sprache englisch
Topic T7 Bioeconomy
Supplements https://www.mdpi.com/1422-0067/22/17/9438/s1
Keywords 13C-labeling; metabolic flux analysis; metabolic flux probe; isotope mapping
Abstract Novel cultivation technologies demand the adaptation of existing analytical concepts. Metabolic flux analysis (MFA) requires stable-isotope labeling of biomass-bound protein as the primary information source. Obtaining the required protein in cultivation set-ups where biomass is inaccessible due to low cell densities and cell immobilization is difficult to date. We developed a non-disruptive analytical concept for 13C-based metabolic flux analysis based on secreted protein as an information carrier for isotope mapping in the protein-bound amino acids. This “metabolic flux probe” (MFP) concept was investigated in different cultivation set-ups with a recombinant, protein-secreting yeast strain. The obtained results grant insight into intracellular protein turnover dynamics. Experiments under metabolic but isotopically nonstationary conditions in continuous glucose-limited chemostats at high dilution rates demonstrated faster incorporation of isotope information from labeled glucose into the recombinant reporter protein than in biomass-bound protein. Our results suggest that the reporter protein was polymerized from intracellular amino acid pools with higher turnover rates than biomass-bound protein. The latter aspect might be vital for 13C-flux analyses under isotopically nonstationary conditions for analyzing fast metabolic dynamics.
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=25062
Dusny, C., Schmid, A. (2021):
The metabolic flux probe (MFP)—Secreted protein as a non-disruptive information carrier for 13C-based metabolic flux analysis
Int. J. Mol. Sci. 22 (17), art. 9438