AIMS Microbiology, 2016, 2(3): 262-277. doi: 10.3934/microbiol.2016.3.262

Research article

Export file:


  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text


  • Citation Only
  • Citation and Abstract

Experimental workflow for developing a feed forward strategy to control biomass growth and exploit maximum specific methane productivity of Methanothermobacter marburgensis in a biological methane production process (BMPP)

1 Institute of Chemical Engineering, Vienna University of Technology, Gumpendorferstraße 1a, 1060 Vienna, Austria
2 Krajete GmbH, Scharitzerstraße 30, 4020 Linz, Austria

Recently, interests for new biofuel generations allowing conversion of gaseous substrate(s) to gaseous product(s) arose for power to gas and waste to value applications. An example is biological methane production process (BMPP) with Methanothermobacter marburgensis. The latter, can convert carbon dioxide (CO2) and hydrogen (H2), having different origins and purities, to methane (CH4), water and biomass. However, these gas converting bioprocesses are tendentiously gas limited processes and the specific methane productivity per biomass amount (qCH4) tends to be low. Therefore, this contribution proposes a workflow for the development of a feed forward strategy to control biomass, growth (rx) and qCH4 in a continuous gas limited BMPP. The proposed workflow starts with a design of experiment (DoE) to optimize media composition and search for a liquid based limitation to control selectively growth. From the DoE it came out that controlling biomass growth was possible independently of the dilution and gassing rate applied while not affecting methane evolution rates (MERs). This was done by shifting the process from a natural gas limited state to a controlled liquid limited growth. The latter allowed exploiting the maximum biocatalytic activity for methane formation of Methanothermobacter marburgensis. An increase of qCH4 from 42 to 129 mmolCH4 g−1 h−1 was achieved by applying a liquid limitation compare with the reference state. Finally, a verification experiment was done to verify the feeding strategy transferability to a different process configuration. This evidenced the ratio of the fed KH2PO4 to rx (R(FKH2PO4/rx)) has an appropriate parameter for scaling feeds in a continuous gas limited BMPP. In the verification experiment CH4 was produced in a single bioreactor step at a methane evolution rate (MER) of   132 mmolCH4*L−1*h−1 at a CH4 purity of 93 [Vol.%].
  Article Metrics


1. Demirbas MF, Balat M (2006) Recent advances on the production and utilization trends of bio-fuels: A global perspective. Energy Convers Manag 47: 2371–2381.    

2. Demirbas A (2007) Progress and recent trends in biofuels. Prog Energy Combust Sci 33: 1–18.    

3. Pinto AC, Guarieiro LLN, Rezende MJC, et al. (2005) Biodiesel: an overview. J Braz Chem Soc 16: 1313–1330.    

4. Jajesniak P, Omar Ali HEM, Wong TS (2014) Carbon Dioxide Capture and Utilization using Biological Systems: Opportunities and Challenges. J Bioprocess Biotech 4:155.

5. Aresta M, Dibenedetto A, Angelini A (2014) Catalysis for the Valorization of Exhaust Carbon: from CO2 to Chemicals, Materials, and Fuels. Technological Use of CO2. Chem Rev 114: 1709–1742.

6. Abubackar HN, Veiga MC, Kennes C (2011) Biological conversion of carbon monoxide: rich syngas or waste gases to bioethanol. Biofuels Bioprod Biorefining 5: 93–114.    

7. Li H, Liao JC (2013) Biological conversion of carbon dioxide to photosynthetic fuels and electrofuels. Energy Environ Sci 6: 2892–2899.    

8. Demirbas A (2008) Biofuels sources, biofuel policy, biofuel economy and global biofuel projections. Energy Convers Manag 49: 2106–2116.    

9. Bernacchi S, Lorantfy B, Martinez E, et al. (2014) Restructuring renewable energy sources for more efficient biofuels production with extremophilic microorganisms. Symposium Energieinnovation, Graz, Austria.

10. Blanch HW (2012) Bioprocessing for biofuels. Curr Opin Biotechnol 23: 390–395.    

11. Ghimire A, Frunzo L, Pirozzi F, et al. (2015) A review on dark fermentative biohydrogen production from organic biomass: Process parameters and use of by-products. Appl Energy 144: 73–95.    

12 Rittmann SK-MR, Lee HS, Lim JK, et al. (2015) One-carbon substrate-based biohydrogen production: Microbes, mechanism, and productivity. Biotechnol Adv 33: 165–177.    

13. Patni N, Shah P, Agarwal S, et al. (2013) Alternate Strategies for Conversion of Waste Plastic to Fuels. ISRN Renew Energy 2013: 1–7.

14. Rodríguez Couto S (2008) Exploitation of biological wastes for the production of value‐added products under solid‐state fermentation conditions. Biotechnol J 3: 859–870.    

15. Angenent LT, Karim K, Al-Dahhan MH, et al. (2004) Production of bioenergy and biochemicals from industrial and agricultural wastewater. Trends Biotechnol 22: 477–485.    

16. van Groenestijn JW, Kraakman NJR (2005) Recent developments in biological waste gas purification in Europe. Chem Eng J 113: 85–91.    

17. Kennes C, Rene ER, Veiga MC (2009) Bioprocesses for air pollution control. J Chem Technol Biotechnol 84: 1419–1436.    

18. Choi D, Chipman DC, Bents SC, et al. (2010) A Techno-economic Analysis of Polyhydroxyalkanoate and Hydrogen Production from Syngas Fermentation of Gasified Biomass. Appl Biochem Biotechnol 160: 1032–1046.    

19. Rittmann S, Seifert A, Herwig C (2013) Essential prerequisites for successful bioprocess development of biological CH4 production from CO2 and H2. Crit Rev Biotechnol 1–12.

20. Yasin M, Jeong Y, Park S, et al. (2015) Microbial synthesis gas utilization and ways to resolve kinetic and mass-transfer limitations. Bioresour Technol 177: 361–374.    

21. Porqueras EM, Rittmann S, Herwig C (2012) Biofuels and CO 2 neutrality: an opportunity. Biofuels 3: 413–426.    

22. Bernacchi S, Weissgram M, Wukovits W, et al. (2014) Process efficiency simulation for key process parameters in biological methanogenesis. AIMS Bioeng 1: 53–71.    

23. Lehner M, Tichler R, Steinmüller H, et al. (2014) Power-to-Gas: Technology and Business Models. Springer International Publishing.

24. Götz M, Koch AM, Graf F (2014) State of the Art and Perspectives of CO2 Methanation Process Concepts for Power-to-Gas Applications. International Gas Union Research Conference, Copenhagen.

25. 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.    

26. Götz M, Lefebvre J, Mörs F, et al. (2016) Renewable Power-to-Gas: A technological and economic review. Renew Energy 85: 1371–1390.    

27. Martin MR, Fornero JJ, Stark R, et al. (2013) A Single-Culture Bioprocess of Methanothermobacter thermautotrophicus to Upgrade Digester Biogas by CO2-to-CH4 Conversion with H2. Archaea 2013: 1–11.

28. Sambusiti C, Bellucci M, Zabaniotou A, et al. (2015) Algae as promising feedstocks for fermentative biohydrogen production according to a biorefinery approach: A comprehensive review. Renew. Sustain Energy Rev 44: 20–36.    

29. Cusick RJ (1974) Space station prototype Sabatier reactor design verification testing. presented at the Intersociety Conference on Environmental Systems, Seattle, WA, US.

30. Murdoch K, Goldblatt L, Carrasquillo R, et al. (2015) Sabatier Methanation Reactor for Space Exploration.

31. Bernacchi S, Seifert AH, Rittmann S (2013) Benefits of Biological Methanation. presented at the conference: DBI-Fachforum Energiespeicher - Pilotprojekte, Berlin, Germany.

32. Li J, Wong C-F, Wong MT, et al. (2014) Modularized Evolution in Archaeal Methanogens Phylogenetic Forest. Genome Biol Evol 6: 3344–3359.    

33. 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 Bioenergy 36: 293–301.    

34. Seifert AH, Rittmann S, Herwig C (2014) Analysis of process related factors to increase volumetric productivity and quality of biomethane with Methanothermobacter marburgensis. Appl Energy 132: 155–162.    

35. Bernacchi S, Rittmann S, Seifert AH, et al. (2014) 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 Bioeng 1: 72–86.    

36. 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.

37. Archer DB (1985) Uncoupling of Methanogenesis from Growth of Methanosarcina barkeri by Phosphate Limitation. Appl Environ Microbiol 50: 1233–1237.

38. Wrede C, Walbaum U, Ducki A, et al (2913) Localization of Methyl-Coenzyme M Reductase as Metabolic Marker for Diverse Methanogenic Archaea. Archaea 2013: 1–7.

39. Bernacchi S, Seifert AH, Krajete A, et al. (2013) Method and system for producing methane using methanogenic microorganisms and applying specific nitrogen concentrations in the liquid phase. Available from:

40. Schönheit P, Moll J, Thauer RK (1980) Growth parameters (K s, μmax, Y s) of Methanobacterium thermoautotrophicum. Arch Microbiol 127: 59–65.    

41. 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.    

42. Thauer RK, Kaster A-K, Goenrich M (2010) Hydrogenases from Methanogenic Archaea, Nickel, a Novel Cofactor, and H 2 Storage. Annu Rev Biochem 79: 507–536.    

43. Duin E, Cosper N, Mahlert F (2003) Coordination and geometry of the nickel atom in active methyl-coenzyme M reductase from Methanothermobacter marburgensis as detected by X-ray absorption spectroscopy. J Biol Inorg Chem 8: 141–148.    

44. Ullmann E, Tan TC, Gundinger T, et al. (2014) A novel cytosolic NADH:quinone oxidoreductase from Methanothermobacter marburgensis. Biosci Rep 34: 893–904.    

45. Martin DD, Ciulla RA, Roberts MF (1999) Osmoadaptation in Archaea. Appl Environ Microbiol 65: 1815–1825.

46. Ciulla R, Clougherty C, Belay N (1994) Halotolerance of Methanobacterium thermoautotrophicum delta H and Marburg. J Bacteriol 176: 3177–3187.

47. Kempf B, Bremer E (1988) Stress responses ofBacillus subtilis to high osmolarity environments: Uptake and synthesis of osmoprotectants. J Biosci 23: 447–455.

48. 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.    

Copyright Info: © 2016, Christoph Herwig, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (

Download full text in PDF

Export Citation

Article outline

Show full outline
Copyright © AIMS Press All Rights Reserved