A novel platform for automated high-throughput fluxome profiling of metabolic variants

Stephanie Heux (1) , Juliette Poinot (1) , Stéphane Massou (1) , Serguei Sokol (1) , Jean-Charles Portais (1)

1 LISBP - Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés

Advances in metabolic engineering are enabling the creation of a large number of cell factories. However, high throughput platforms do not yet exist for rapidly analyzing the metabolic network of the engineered cells. To fill the gap, we developed an integrated solution for fluxome profiling of large sets of biological systems and conditions. This platform combines a robotic system for C-13-labelling experiments and sampling of labelled material with NMR-based isotopic fingerprinting and automated data interpretation. As a proof-of-concept, this workflow was applied to discriminate between Escherichia coli mutants with gradual expression of the glucose-6-phosphate dehydrogenase. Metabolic variants were clearly discriminated while pathways that support metabolic flexibility towards modulation of a single enzyme were elucidating. By directly connecting the data flow between cell cultivation and flux quantification, considerable advances in throughput, robustness, release of resources and screening capacity were achieved. This will undoubtedly facilitate the development of efficient cell factories. (C) 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Mots clés: High-throughput; Fluxomics; Automation; NMR; Statistical analysis; Flux calculation; ISOTOPE LABELING EXPERIMENTS; CENTRAL CARBON METABOLISM; MAMMALIAN-CELL CULTURES; QUANTITATIVE 2D NMR; ZWF GENE-KNOCKOUT; ESCHERICHIA-COLI; CORYNEBACTERIUM-GLUTAMICUM; CLOSTRIDIUM-ACETOBUTYLICUM; MICROBIAL CULTIVATIONS; INFORMATION-CONTENT

Domaines: Sciences du Vivant [q-bio]

Stephanie Heux, Juliette Poinot, Stéphane Massou, Serguei Sokol, Jean-Charles Portais. A novel platform for automated high-throughput fluxome profiling of metabolic variants. Metabolic Engineering, 2014, 25, pp.8 - 19. ⟨10.1016/j.ymben.2014.06.001⟩. ⟨hal-01268982⟩