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Experimental methods for screening parameters influencing the growth to product yield (Y(x/CH4)) of a biological methane production (BMP) process performed with Methanothermobacter marburgensis

1 Institute of Chemical Engineering, Research Area Biochemical Engineering, Vienna University of Technology, Gumpendorferstraße 1a, 1060 Vienna, Austria
2 Department of Ecogenomics and Systems Biology, Althanstraße 14, 1090 Vienna, Austria
3 Krajete GmbH, Scharitzerstraße 30, 4020 Linz, Austria

Special Issue: Bioconversion for Renewable Energy and Biomaterials

New generation biofuels are a suitable approach to produce energy carriers in an almost CO2 neutral way. A promising reaction is the conversion of carbon dioxide (CO2) and molecular hydrogen (H2) to methane (CH4) and water (H2O). In this contribution, this so-called Sabatier reaction was performed biologically by using hydrogenotrophic and autotrophic methanogenic microorganisms from the archaea life domain. For the development of a biological methane production (BMP) process, one key parameter is the ratio of biomass production rate (rx) to methane evolution rate (MER) reflected in the growth to product yield (Y(x/CH4)) because it represents both a physiological and a scalable entity for the bioprocesses development as it quantify the selectivity of reaction with respect to the carbon. Y(x/CH4) needs also to be held constant in order to establish an adaptable media composition for developing a scalable feeding strategy. Identification of parameters and quantification of their impact on Y(x/CH4) is a necessary prerequisite for obtaining a growth kinetic model and developing advanced process control strategies especially for dynamic operation modes. In this work, process conditions and parameters impacting Y(x/CH4) were investigated by using a combination of multivariate and univariate chemostat cultures, as well as dynamic experiments. The proposed combination of methods is a novel modular approach for the development of BMP processes. It allowed determining the effects of multiple process factors on physiology and methane productivity of Methanothermobacter marburgensis. In fact, quantitative analysis of basal medium, sulphide and ammonium dilution rates, as well as the ammonium concentration revealed that all these variables vary rx without affecting MER. Hence Y(x/CH4) can be used to identify limiting or inhibiting conditions during media development tasks as well as for tuning the carbon flux of the bioprocess in an industrial application by reducing Y(x/CH4) to improve the carbon balance of the reaction.
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Keywords Design of Experiments; chemostat; dynamic experiments; bioprocess quantification; carbon balance, biological methanogenesis, bioCH4

Citation: Sébastien Bernacchi, Simon Rittmann, Arne H. Seifert, Alexander Krajete, Christoph Herwig. Experimental methods for screening parameters influencing the growth to product yield (Y(x/CH4)) of a biological methane production (BMP) process performed with Methanothermobacter marburgensis. AIMS Bioengineering, 2014, 1(2): 72-87. doi: 10.3934/bioeng.2014.2.72


  • 1. Specht M, Brellochs J, Frick V, et al. (2010) Storage of renewable energy in the natural gas grid. Erdoel, Erdgas, Kohle 126: 342-345.
  • 2. Thauer RK, Kaster AK, Goenrich M, et al. (2010) Hydrogenases from methanogenic archaea, nickel, a novel cofactor, and H2 storage. Annu Rev Biochem 79: 507-536.    
  • 3. Liu Y, Whitman WB (2008) Metabolic, phylogenetic, and ecological diversity of the methanogenic archaea. Ann N Y Acad Sci 1125: 171-189.    
  • 4. Kaster AK, Goenrich M, Seedorf H, et al. (2011) More than 200 genes required for methane formation from H2 and CO2 and energy conservation are present in Methanothermobacter marburgensis and Methanothermobacter thermautotrophicus. Archaea ID973848: 1-23.
  • 5. Seifert AH, Rittmann S, Herwig C (2014) Analysis of process related factors to increase volumetric productivity and quality of biomethane with Methanothermobacter marburgensis Appl Energ 132: 155-162.
  • 6. Bernacchi S, Weissgram M, Wukovits W, et al. (2014) Process efficiency simulation for key process parameters in biological methanogenesis. AIMS bioengineering 1: 53-71.    
  • 7. Thauer RK, Kaster AK, Seedorf H, et al. (2008) Methanogenic archaea: ecologically relevant differences in energy conservation. Nat Rev Microbiol 6: 579-591.    
  • 8. Schill N, van Gulik WM, Voisard D, et al. (1996) Continuous cultures limited by a gaseous substrate: development of a simple, unstructure mathematical model and experimental verification with Methanobacterium thermoautotrophicum. Biotechnol Bioeng 51: 645-658.
  • 9. Jud G, Schneider K, Bachofen R (1997) The role of hydrogen mass transfer for the growth kinetics of Methanobacterium thermoautotrophicum in batch and chemostat cultures. J Ind Microbiol Biotechnol 19: 246-251.    
  • 10. Tsao JH, Kaneshiro SM, Yu SS, et al. (1994) Continuous culture of Methanococcus jannaschii, an extremely thermophilic methanogen. Biotechnol Bioeng 43: 258-261.    
  • 11. Schill N, van Gulik WM, Voisard D, et al. (1996) Continuous cultures limited by a gaseous substrate: development of a simple, unstructured mathematical model and experimental verification with Methanobacterium thermoautotrophicum. Biotechnol Bioeng 51: 645-658.
  • 12. Peillex JP, Fardeau ML, Belaich JP (1990) Growth of Methanobacterium thermoautotrophicum on hydrogen-carbon dioxide: high methane productivities in continuous culture. Biomass 21:315-321.    
  • 13. Nishimura N, Kitaura S, Mimura A, et al. (1992) Cultivation of thermophilic methanogen KN-15 on hydrogen-carbon dioxide under pressurized conditions. J Ferment Bioeng 73: 477-480.    
  • 14. Morii H, Koga Y, Nagai S (1987) Energetic analysis of the growth of Methanobrevibacter arboriphilus A2 in hydrogen-limited continuous cultures. Biotechnol Bioeng 29: 310-315.    
  • 15. de Poorter LMI, Geerts WJ, Keltjens JT (2007) Coupling of Methanothermobacter thermautotrophicus methane formation and growth in fed-batch and continuous cultures under different H2 gassing regimens. Appl Environ Microbiol 73: 740-749.    
  • 16. Schoenheit P, Moll J, Thauer RK (1980) Growth parameters (Ks, μmax, Ys) of Methanobacterium thermoautotrophicum. Arch Microbiol 127: 59-65.    
  • 17. Gerhard E, Butsch BM, Marison IW, et al. (1993) Improved growth and methane production conditions for Methanobacterium thermoautotrophicum. Appl Microbiol Biotechnol 40: 432-437.    
  • 18. Seifert AH, Rittmann S, Bernacchi S, et al. (2013) Method for assessing the impact of emission gasses on physiology and productivity in biological methanogenesis. Bioresour Technol 136:747-751.    
  • 19. Fardeau ML, Peillex JP, Belaich JP (1987) Energetics of the growth of Methanobacterium thermoautotrophicum and Methanococcus thermolithotrophicus on ammonium chloride and dinitrogen. Arch Microbiol 148: 128-131.    
  • 20. Fardeau ML, Belaich JP (1986) Energetics of the growth of Methanococcus thermolithotrophicus. Arch Microbiol 144: 381-385.    
  • 21. Morgan RM, Pihl TD, Nolling J (1997) Hydrogen regulation of growth, growth yields, and methane gene transcription in Methanobacterium thermoautotrophicum Delta H. J Bacteriol 179:889-898.
  • 22. Archer DB (1985) Uncoupling of methanogenesis from growth of Methanosarcina barkeri by phosphate limitation. Appl Environ Microbiol 50: 1233-1237.
  • 23. Rittmann S, Seifert A, Herwig C (2012) Quantitative analysis of media dilution rate effects on Methanothermobacter marburgensis grown in continuous culture on H2 and CO2. Biomass Bioenerg 36: 293-301.    
  • 24. Fuchs G, Stupperich E, Thauer RK (1978) Acetate assimilation and the synthesis of alanine, aspartate and glutamate in Methanobacterium thermoautotrophicum. Arch Microbiol 117: 61-66.    
  • 25. Schoenheit P, Moll J, Thauer RK (1979) Nickel, cobalt, and molybdenum requirement for growth of Methanobacterium thermoautotrophicum. Arch Microbiol 123: 105-107.    
  • 26. Bonacker LG, Baudner S, Thauer RK (1992) Differential expression of the two methyl-coenzyme M reductases in Methanobacterium thermoautotrophicum as determined immunochemically via isoenzyme-specific antisera. Eur J Biochem 206: 87-92.    
  • 27. Bonacker LG, Baudner S, Moerschel E, et al. (1993) Properties of the two isoenzymes of methyl-coenzyme M reductase in Methanobacterium thermoautotrophicum. Eur J Biochem 217:587-595.    
  • 28. Hallenbeck PC, Ghosh D (2009) Advances in fermentative biohydrogen production: the way forward? Trends Biotechnol 27: 287-297.    
  • 29. Wang J, Wan W (2008) Optimization of fermentative hydrogen production process by response surface methodology. Int J Hydrogen Energy 33: 6976-6984.    
  • 30. Rittmann S, Herwig C (2012) A comprehensive and quantitative review of dark fermentative biohydrogen production. Microbial Cell Factories 11:115.    
  • 31. Spadiut O, Rittmann S, Dietzsch C, et al. (2013) Dynamic process conditions in bioprocess development. Eng Life Sci 13: 88-101.    
  • 32. Rittmann S, Seifert A, Herwig C (2013) Essential prerequisites for successful bioprocess development of biological CH4 production from CO2 and H2. Crit Rev Biotechnol 1-11.
  • 33. Costa KC, Yoon SH, Pan M, et al. (2013) Effects of H2 and formate on growth yield and regulation of methanogenesis in Methanococcus maripaludis. J Bacteriol 195: 1456-1462.    
  • 34. Haydock AK, Porat I, Whitman WB, et al. (2004) Continuous culture of Methanococcus maripaludis under defined nutrient conditions. FEMS Microbiol Lett 238: 85-91.


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