Design and thermodynamic analysis of a pathway enabling anaerobic production of poly-3-hydroxybutyrate in Escherichia coli
Olavarria K, Becker MV, Sousa DZ, van Loosdrecht MC, Wahl SA (2023)
Publication Type: Journal article
Publication year: 2023
Journal
Book Volume: 8
Pages Range: 629-639
Journal Issue: 4
DOI: 10.1016/j.synbio.2023.09.005
Abstract
Utilizing anaerobic metabolisms for the production of biotechnologically relevant products presents potential advantages, such as increased yields and reduced energy dissipation. However, lower energy dissipation may indicate that certain reactions are operating closer to their thermodynamic equilibrium. While stoichiometric analyses and genetic modifications are frequently employed in metabolic engineering, the use of thermodynamic tools to evaluate the feasibility of planned interventions is less documented. In this study, we propose a novel metabolic engineering strategy to achieve an efficient anaerobic production of poly-(R)-3-hydroxybutyrate (PHB) in the model organism Escherichia coli. Our approach involves re-routing of two-thirds of the glycolytic flux through non-oxidative glycolysis and coupling PHB synthesis with NADH re-oxidation. We complemented our stoichiometric analysis with various thermodynamic approaches to assess the feasibility and the bottlenecks in the proposed engineered pathway. According to our calculations, the main thermodynamic bottleneck are the reactions catalyzed by the acetoacetyl-CoA β-ketothiolase (EC 2.3.1.9) and the acetoacetyl-CoA reductase (EC 1.1.1.36). Furthermore, we calculated thermodynamically consistent sets of kinetic parameters to determine the enzyme amounts required for sustaining the conversion fluxes. In the case of the engineered conversion route, the protein pool necessary to sustain the desired fluxes could account for 20% of the whole cell dry weight.
Authors with CRIS profile
Involved external institutions
Wageningen University & Research
Netherlands (NL)
Delft University of Technology (TU Delft)
Netherlands (NL)
Netherlands (NL)
Delft University of Technology (TU Delft)
Netherlands (NL)
How to cite
APA:
Olavarria, K., Becker, M.V., Sousa, D.Z., van Loosdrecht, M.C., & Wahl, S.A. (2023). Design and thermodynamic analysis of a pathway enabling anaerobic production of poly-3-hydroxybutyrate in Escherichia coli. Synthetic and Systems Biotechnology, 8(4), 629-639. https://dx.doi.org/10.1016/j.synbio.2023.09.005
MLA:
Olavarria, Karel, et al. "Design and thermodynamic analysis of a pathway enabling anaerobic production of poly-3-hydroxybutyrate in Escherichia coli." Synthetic and Systems Biotechnology 8.4 (2023): 629-639.
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