Publication Details |
| Category | Text Publication |
| Reference Category | Journals |
| DOI | 10.3390/ijms22179438 |
Licence  |
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| Title (Primary) | The metabolic flux probe (MFP)—Secreted protein as a non-disruptive information carrier for 13C-based metabolic flux analysis |
| Author | Dusny, C.; Schmid, A. |
| Source Titel | International Journal of Molecular Sciences |
| Year | 2021 |
| Department | SOMA |
| Volume | 22 |
| Issue | 17 |
| Page From | art. 9438 |
| Language | englisch |
| Topic | T7 Bioeconomy |
| Supplements | Supplement 1 |
| 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. |
| Persistent UFZ Identifier | 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 10.3390/ijms22179438 |
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