Flux measurements and maintenance energy for carbon dioxide utilization by Methanococcus maripaludis

Background: The rapidly growing mesophilic methanogen Methanococcus maripaludis S2 has a unique ability to consume both CO2 and N-2, the main components of a flue gas, and produce methane with H-2 as the electron donor. The existing literature lacks experimental measurements of CO2 and H-2 uptake rates and CH4 production rates on M. maripaludis. Furthermore, it lacks estimates of maintenance energies for use with genome-scale models. In this paper, we performed batch culture experiments on M. maripaludis S-2 using CO2 as the sole carbon substrate to quantify three key extracellular fluxes (CO2, H-2, and CH4) along with specific growth rates. For precise computation of these fluxes from experimental measurements, we developed a systematic process simulation approach. Then, using an existing genome-scale model, we proposed an optimization procedure to estimate maintenance energy parameters: growth associated maintenance (GAM) and non-growth associated maintenance (NGAM). Results: The measured extracellular fluxes for M. maripaludis showed excellent agreement with in silico predictions from a validated genome-scale model (iMM518) for NGAM = 7.836 mmol/gDCW/h and GAM = 27.14 mmol/gDCW. M. maripaludis achieved a CO2 to CH4 conversion yield of 70-95 % and a growth yield of 3.549 +/- 0.149 g DCW/mol CH4 during the exponential phase. The ATP gain of 0.35 molATP/molCH(4) for M. maripaludis, computed using NGAM, is in the acceptable range of 0.3-0.7 mol ATP/molCH(4) reported for methanogens. Interestingly, the uptake distribution of amino acids, quantified using iMM518, confirmed alanine to be the most preferred amino acids for growth and methanogenesis. Conclusions: This is the first study to report experimental gas consumption and production rates for the growth of M. maripaludis on CO2 and H-2 in minimal media. A systematic process simulation and optimization procedure was successfully developed to precisely quantify extracellular fluxes along with cell growth and maintenance energy parameters. Our growth yields, ATP gain, and energy parameters fall within acceptable ranges known in the literature for hydrogenotrophic methanogens.