An energetic profile of Corynebacterium glutamicum underpinned by measured biomass yield on ATP

• Published in: Metabolic engineering Vol. 65; pp. 66 - 78
• Main Authors: Zelle, E., Pfelzer, N., Oldiges, M., Koch-Koerfges, A., Bott, M., Nöh, K., Wiechert, W.
• Format: Journal Article
• Language: English
• Published: Belgium Elsevier Inc 01-05-2021
• Subjects: ATP yield; Branched respiratory chain; Constraint based methods; Energy metabolism; Metabolic flux analysis; ATP yield; Constraint based methods; Branched respiratory chain; Energy metabolism; Metabolic flux analysis
• ISSN: 1096-7176; 1096-7184
• DOI: 10.1016/j.ymben.2021.03.006

The supply and usage of energetic cofactors in metabolism is a central concern for systems metabolic engineering, particularly in case of energy intensive products. One of the most important parameters for systems wide balancing of energetic cofactors is the ATP requirement for biomass formation YATP/Biomass. Despite its fundamental importance, YATP/Biomass values for non-fermentative organisms are still rough estimates deduced from theoretical considerations. For the first time, we present an approach for the experimental determination of YATP/Biomass using comparative 13C metabolic flux analysis (13C MFA) of a wild type strain and an ATP synthase knockout mutant. We show that the energetic profile of a cell can then be deduced from a genome wide stoichiometric model and experimental maintenance data. Particularly, the contributions of substrate level phosphorylation (SLP) and electron transport phosphorylation (ETP) to ATP generation become available which enables the overall energetic efficiency of a cell to be characterized. As a model organism, the industrial platform organism Corynebacterium glutamicum is used. C. glutamicum uses a respiratory type of energy metabolism, implying that ATP can be synthesized either by SLP or by ETP with the membrane-bound F1FO-ATP synthase using the proton motive force (pmf) as driving force. The presence of two terminal oxidases, which differ in their proton translocation efficiency by a factor of three, further complicates energy balancing for this organism. By integration of experimental data and network models, we show that in the wild type SLP and ETP contribute equally to ATP generation. Thus, the role of ETP in respiring bacteria may have been overrated in the past. Remarkably, in the genome wide setting 65% of the pmf is actually not used for ATP synthesis. However, it turns out that, compared to other organisms C. glutamicum still uses its energy budget rather efficiently.
•Comparative 13CMFA shows that 30 g ATP are needed for synthesis of 1 g cells.•Only 45% of ATP is produced by ATP synthase.•Overall efficiency of substrate usage is 27% of the theoretical maximal ATP yield.•Genome-wide model predictes in vivo growth rate for supercomplex usage higher 43%.•65% of the pmf is used for cellular processes other than ATP synthesis.