Abstract
Cupriavidus necator DSM 545 can utilise glycerol to synthesise poly(3-hydroxybutyric acid) under unbalanced growth conditions, i.e., nitrogen limitation. To improve poly(3-hydroxybutyric acid) (PHB) batch production by C. necator through model-guided bioprocessing or genetic engineering, insights into the dynamic effect of the fermentation conditions on cell metabolism are crucial. In this work, we have used dynamic flux balance analysis (DFBA), a constrained-based stoichiometric modelling approach, to study the metabolic change associated with PHB synthesis during batch cultivation. The model employs the ‘minimisation of all fluxes’ as cellular objectives and measured extracellular fluxes as additional constraints. The mass balance constraints are further adjusted based on thermodynamic considerations. The resultant flux distribution profiles characterise the evolution of metabolic states due to adaptation to dynamic extracellular conditions and provide further insights towards improvements that can be implemented to enhance PHB productivity.
Highlights
In recent decades, biopolymers produced naturally have gained much attention due to rising concerns about environmental issues caused by fossil fuel-based plastics along with a growing strategic interest in bioeconomy
We focused on analysing the dynamics of metabolism of C. necator DSM 545 under different batch conditions using dynamic flux balance analysis (DFBA)
We have applied a dynamic flux balance analysis (DFBA) framework to examine the shift of metabolic states during bacterial fermentation
Summary
Biopolymers produced naturally have gained much attention due to rising concerns about environmental issues caused by fossil fuel-based plastics along with a growing strategic interest in bioeconomy. Poly(3-hydroxybutyric acid) (PHB) is the most commonly produced type of natural polyhydroxyalkanoates (PHAs), which features biodegradability and biocompatibility [1]. PHB can be synthesised as an intracellular carbon and energy storage compound in the form of cytoplasmic inclusion when bacteria grow under unbalanced nutrient conditions, typically when essential growth nutrients, such as N and P, are limiting [2,3,4]. The choice of carbon sources remains a major limiting factor in the development of economically viable, large-scale and technologically feasible biopolymer processes. The use of low-cost substrates derived from sustainable sources is desirable and has been intensively investigated [11,12,13,14]
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