, Volume 22, Issue 4, pp 805–814 | Cite as

Thermophilic, anaerobic co-digestion of microalgal biomass and cellulose for H2 production

  • Sarah M. Carver
  • Chris J. Hulatt
  • David N. Thomas
  • Olli H. Tuovinen
Original Paper


Microalgal biomass has been a focus in the sustainable energy field, especially biodiesel production. The purpose of this study was to assess the feasibility of treating microalgal biomass and cellulose by anaerobic digestion for H2 production. A microbial consortium, TC60, known to degrade cellulose and other plant polymers, was enriched on a mixture of cellulose and green microalgal biomass of Dunaliella tertiolecta, a marine species, or Chlorella vulgaris, a freshwater species. After five enrichment steps at 60°C, hydrogen yields increased at least 10% under all conditions. Anaerobic digestion of D. tertiolecta and cellulose by TC60 produced 7.7 mmol H2/g volatile solids (VS) which were higher than the levels (2.9–4.2 mmol/g VS) obtained with cellulose and C. vulgaris biomass. Both microalgal slurries contained satellite prokaryotes. The C. vulgaris slurry, without TC60 inoculation, generated H2 levels on par with that of TC60 on cellulose alone. The biomass-fed anaerobic digestion resulted in large shifts in short chain fatty acid concentrations and increased ammonium levels. Growth and H2 production increased when TC60 was grown on a combination of D. tertiolecta and cellulose due to nutrients released from algal cells via lysis. The results indicated that satellite heterotrophs from C. vulgaris produced H2 but the Chlorella biomass was not substantially degraded by TC60. To date, this is the first study to examine H2 production by anaerobic digestion of microalgal biomass. The results indicate that H2 production is feasible but higher yields could be achieved by optimization of the bioprocess conditions including biomass pretreatment.


Dunaliella tertiolecta Chlorella vulgaris Cellulose degradation Anaerobic digestion Hydrogen production 



Partial funding for this study was received from the Finland Distinguished Professorship Program (402/2006) of the Finnish Agency for Technology and Innovation (S.M. Carver and O.H. Tuovinen), USDA National Needs Graduate Fellowship Program (S.M. Carter), Academy of Finland (D.N. Thomas), and Engineering and Physical Sciences Research Council Studentship in conjunction with RWE Power (U.K.) and Varicon Aqua Ltd. (C.J. Hulatt and D.N. Thomas).


  1. Ben-Amotz A, Polle JEW, Rao DVS (2010) The alga Dunaliella: biodiversity, physiology, genomics and biotechnology. Science, EnfieldGoogle Scholar
  2. Benemann JR (2000) Hydrogen production by microalgae. J Appl Phycol 12:291–300CrossRefGoogle Scholar
  3. Cordóba LT, Bocanegra ARD, Llorente BR, Hernández ES, Echegoyen FB, Borja R, Bejines FR, Morcillo MFC (2008) Batch culture growth of Chlorella zofingiensis on effluent derived from two-stage anaerobic digestion of two-phase olive mill solid waste. Electron J Biotechnol. doi: 10.2225/vol11-issue2-fulltext-1 11(2):1–8
  4. Department of Energy (2007) Roadmap for bioenergy and biobased products in the U.S. Biomass Research and Development, Technical Advisory Committee, Washington DC ADA494661Google Scholar
  5. Eaton AD, Clesceri LS, Rice EW, Greenberg AE, Franson MAH (2005) Standard methods for the examination of water and wastewater. American Public Health Association, New YorkGoogle Scholar
  6. Ghirardi ML, Dubini A, Yu J, Maness PC (2009) Photobiological hydrogen-producing systems. Chem Soc Rev 38:52–61PubMedCrossRefGoogle Scholar
  7. Golueke CG, Oswald WJ, Gotaas HB (1957) Anaerobic digestion of algae. Appl Microbiol 5:47–55PubMedGoogle Scholar
  8. Hernández EPS, Córdoba LT (1993) Anaerobic digestion of Chlorella vulgaris for energy production. Resour Conserv Recycl 9:127–132CrossRefGoogle Scholar
  9. Hodaifa G, Martínez ME, Sánchez S (2008) Use of industrial wastewater from olive-oil extraction for biomass production of Scenedesmus obliquus. Bioresour Technol 99:1111–1117PubMedCrossRefGoogle Scholar
  10. Holmes RM, Aminot A, Kérouel R, Hooker BA, Peterson BJ (1999) A simple and precise method for measuring ammonium in marine and freshwater ecosystems. Can J Fish Aquat Sci 56:1801–1808Google Scholar
  11. Horiuchi J, Ohba I, Tada K, Kobayashi M, Kanno T, Kishimoto M (2003) Effective cell harvesting of the halotolerant microalga Dunaliella tertiolecta with pH control. J Biosci Bioeng 95:412–415PubMedGoogle Scholar
  12. Hulatt CJ, Thomas DN (2010) Dissolved organic matter (DOM) in microalgal photobioreactors: a potential loss in solar energy conversion? Bioresour Technol 101:8690–8697PubMedCrossRefGoogle Scholar
  13. Ike A, Saimura C, Hirata K, Miyamoto K (1996) Environmentally friendly production of H2 incorporating microalgal CO2 fixation. J Mar Biotechnol 4:47–51Google Scholar
  14. Ike A, Toda N, Hirata K, Miyamoto K (1997) Hydrogen photoproduction from CO2-fixing microalgal biomass: application of lactic acid fermentation by Lactobacillus amylovorus. J Ferment Bioeng 84:428–433CrossRefGoogle Scholar
  15. Kawaguchi H, Hashimoto K, Hirata K, Miyamoto K (2001) H2 production from algal biomass by a mixed culture of Rhodobium marinum A-501 and Lactobacillus amylovorus. J Biosci Bioeng 91:277–282PubMedCrossRefGoogle Scholar
  16. Kojima H, Lee YK (2001) Photosynthetic microorganisms in environmental biotechnology. Springer, Hong KongGoogle Scholar
  17. Lardon L, Hélias A, Sialve B, Steyer J-P, Bernard O (2009) Life-cycle assessment of biodiesel production from microalgae. Environ Sci Technol 43:6475–6481PubMedCrossRefGoogle Scholar
  18. Levin DB, Pitt L, Love M (2004) Biohydrogen production: prospects and limitations to practical application. Int J Hydrogen Energy 29:173–185CrossRefGoogle Scholar
  19. Mata TM, Martins AA, Caetano NS (2010) Microalgae for biodiesel production and other applications: a review. Renew Sustain Energy Rev 14:217–232CrossRefGoogle Scholar
  20. Melis A, Happe T (2001) Hydrogen production. Green algae as a source of energy. Plant Physiol 127:740–748PubMedCrossRefGoogle Scholar
  21. Oswald WJ, Golueke CG (1960) Biological transformation of solar energy. Adv Appl Microbiol 2:223–262PubMedCrossRefGoogle Scholar
  22. Parkin GF, Owen WF (1986) Fundamentals of anaerobic digestion of wastewater sludge. J Environ Eng 112:867–920CrossRefGoogle Scholar
  23. Perlack RD, Wright LL, Turhollow AF, Graham RL, Stokes BJ, Erbach DC (2005) Biomass as feedstock for a bioenergy and bioproducts industry: the technical feasibility of a billion-ton annual supply. U.S. Department of Energy DOE/GO-102005-2135Google Scholar
  24. Peu P, Béline F, Martinez J (2004) Volatile fatty acids analysis from pig slurry using high-performance liquid chromatography. Int J Environ Anal Chem 84:1017–1022CrossRefGoogle Scholar
  25. Ren N, Wang A, Cao G, Xu J, Gao L (2009) Bioconversion of lignocellulosic biomass to hydrogen: potential and challenges. Biotechnol Adv 27:1051–1060PubMedCrossRefGoogle Scholar
  26. Sialve B, Bernet N, Bernard O (2009) Anaerobic digestion of microalgae as a necessary step to make microalgal biodiesel sustainable. Biotechnol Adv 27:409–416PubMedCrossRefGoogle Scholar
  27. Tam NFY, Wong YS (1996) Effect of ammonia concentrations on growth of Chlorella vulgaris and nitrogen removal from media. Bioresour Technol 57:45–50CrossRefGoogle Scholar
  28. Yen HW, Brune DE (2007) Anaerobic co-digestion of algal sludge and waste paper to produce methane. Bioresour Technol 98:130–134PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Sarah M. Carver
    • 1
    • 2
  • Chris J. Hulatt
    • 3
  • David N. Thomas
    • 3
    • 4
  • Olli H. Tuovinen
    • 1
    • 2
  1. 1.Department of MicrobiologyOhio State UniversityColumbusUSA
  2. 2.Department of Chemistry and BioengineeringTampere University of TechnologyTampereFinland
  3. 3.School of Ocean Sciences, College of Natural SciencesBangor UniversityAngleseyUK
  4. 4.Marine Research Centre, Finnish Environment InstituteHelsinkiFinland

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