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Article

The Metabolic Flux Probe (MFP)—Secreted Protein as a Non-Disruptive Information Carrier for 13C-Based Metabolic Flux Analysis

Department Solar Materials, Helmholtz Centre for Environmental Research (UFZ), Permoser Str. 15, D-04318 Leipzig, Germany
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Author to whom correspondence should be addressed.
Academic Editor: Thomas Kietzmann
Int. J. Mol. Sci. 2021, 22(17), 9438; https://doi.org/10.3390/ijms22179438
Received: 15 July 2021 / Revised: 26 August 2021 / Accepted: 27 August 2021 / Published: 30 August 2021
(This article belongs to the Section Biochemistry)
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. View Full-Text
Keywords: 13C-labeling; metabolic flux analysis; metabolic flux probe; isotope mapping 13C-labeling; metabolic flux analysis; metabolic flux probe; isotope mapping
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MDPI and ACS Style

Dusny, C.; Schmid, A. The Metabolic Flux Probe (MFP)—Secreted Protein as a Non-Disruptive Information Carrier for 13C-Based Metabolic Flux Analysis. Int. J. Mol. Sci. 2021, 22, 9438. https://doi.org/10.3390/ijms22179438

AMA Style

Dusny C, Schmid A. The Metabolic Flux Probe (MFP)—Secreted Protein as a Non-Disruptive Information Carrier for 13C-Based Metabolic Flux Analysis. International Journal of Molecular Sciences. 2021; 22(17):9438. https://doi.org/10.3390/ijms22179438

Chicago/Turabian Style

Dusny, Christian, and Andreas Schmid. 2021. "The Metabolic Flux Probe (MFP)—Secreted Protein as a Non-Disruptive Information Carrier for 13C-Based Metabolic Flux Analysis" International Journal of Molecular Sciences 22, no. 17: 9438. https://doi.org/10.3390/ijms22179438

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