4 years ago

Metabolic modeling of energy balances in Mycoplasma hyopneumoniae shows that pyruvate addition increases growth rate

Metabolic modeling of energy balances in Mycoplasma hyopneumoniae shows that pyruvate addition increases growth rate
Simen-Jan Slagman, Vitor A. P. Martins dos Santos, Peter J. Schaap, Maria Suarez-Diez, Jetta J. E. Bijlsma, Tjerko Kamminga
Mycoplasma hyopneumoniae is cultured on large-scale to produce antigen for inactivated whole-cell vaccines against respiratory disease in pigs. However, the fastidious nutrient requirements of this minimal bacterium and the low growth rate make it challenging to reach sufficient biomass yield for antigen production. In this study, we sequenced the genome of M. hyopneumoniae strain 11 and constructed a high quality constraint-based genome-scale metabolic model of 284 chemical reactions and 298 metabolites. We validated the model with time-series data of duplicate fermentation cultures to aim for an integrated model describing the dynamic profiles measured in fermentations. The model predicted that 84% of cellular energy in a standard M. hyopneumoniae cultivation was used for non-growth associated maintenance and only 16% of cellular energy was used for growth and growth associated maintenance. Following a cycle of model-driven experimentation in dedicated fermentation experiments, we were able to increase the fraction of cellular energy used for growth through pyruvate addition to the medium. This increase in turn led to an increase in growth rate and a 2.3 times increase in the total biomass concentration reached after 3–4 days of fermentation, enhancing the productivity of the overall process. The model presented provides a solid basis to understand and further improve M. hyopneumoniae fermentation processes. Biotechnol. Bioeng. 2017;9999: 1–9. © 2017 Wiley Periodicals, Inc. Production of Mycoplasma hyopneumoniae as active component in whole-cell inactivated vaccines for pigs is challenging due to the largely unknown growth requirements. In this study, we created and validated a genome-scale metabolic model and applied the model to optimize an aerobic fermentation process. We found that by adding pyruvate to the growth medium we could increase the amount of cellular energy used for growth and thereby increased the economic potential of the process by increasing growth rate and biomass yield.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/bit.26347

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