Skip to main content

Advertisement

Log in

Non-destructive oil extraction from Botryococcus braunii (Chlorophyta)

  • Published:
Journal of Applied Phycology Aims and scope Submit manuscript

Abstract

Some of the key reasons for why the production of biofuels from microalgae have not yet succeeded as a source of sustainable transport fuel are the costs involved and the amount of energy needed to obtain the oils compared to the energy contained in the final fuel. The key energy costs are in the dewatering of biomass followed by extraction of the oil, disposal of biomass, and the energy content of the nutrient fertiliser needed for regrowing the algae. In this study, we bypass all of these barriers by using a different approach towards cutting energy and fertiliser costs in the production of biofuels from microalgae—rather than growing the algae in the presence of fertilisers such as N and P, followed by harvesting the whole algae cells, and the energetically costly drying of cells and extraction of the fuel from the cells, this process makes use of the natural tendency of the green alga, Botryococcus braunii to release oils from the cell into the extracellular matrix during and after growth. Here, we non-destructively and repeatedly harvest the external oil (hydrocarbons) from B. braunii CCAP 807/2. Extraction with several solvents showed that hexane was not compatible with B. braunii, but that heptane in contact with B. braunii for less than 20 min did not negatively affect this alga. As an alternative, solvent-free method, we tested physical methods of extracting the extracellular oil. Light and temperature did not affect the extraction of the external oil from Botryococcus, but gentle pressure (i.e. ‘blotting’) was an effective method for external oil recovery. Less than 1 h of blotting also did not affect the physiology of Botryococcus. Both the heptane extraction and the non-destructive ‘blotting’ methods had no significant effect on growth and photosynthesis (F v/F m, ETRmax) of B. braunii. Our results indicate that over a period of 6 days, we can repeatedly extract over 35 % (using heptane) and 1 % (using ‘blotting’) of the total oil, mainly in the form of external hydrocarbon in stationary phase cells without damage to the cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ackman RG (1991) Application of thin-layer chromatography separation: neutral lipids. In: Perkins EG (ed) Analyses of fats, oils and lipoproteins. American Oil Chemists Society, Champaign, Illinois, pp 60–82

    Google Scholar 

  • An JY, Sim S-J, Kim BW, Lee JS (2004) Improvement of hydrocarbon recovery by two-stage cell-recycle extraction in the cultivation of Botryococcus braunii. J Microbiol Biotech 14:932–937

    CAS  Google Scholar 

  • Blackburn KB (1936) Botryococcus and the algal coals. Part I: a reinvesigation of the alga Botryococcus braunii Kutzing. Trans R Soc Edinburgh 58:841–854

  • Borowitzka MA, Moheimani NR (2013) Sustainable biofuels from algae. Mitig Adapt Strat Glob Change 18:13–25

    Article  Google Scholar 

  • Cane RF, Albion PR (1973) The organic geochemistry of torbanite precursors. Geochim Cosmochim Acta 37:1543–1549

    Article  CAS  Google Scholar 

  • Carpenter JH (1966) New measurements of oxygen solubility in pure and natural water. Limnol Oceanogr 11:264–277

    Article  CAS  Google Scholar 

  • Cordell D, Drangert JO, White S (2009) The story of phosphorus: global food security and food for thought. Glob Environ Chang 19:292–305

    Article  Google Scholar 

  • Dayananda C, Sarada R, Srinivas P, Shamala T, Ravishankar G (2006) Presence of methyl branched fatty acids and saturated hydrocarbons in botryococcene producing strain of Botryococcus braunii. Acta Physiol Plant 28:251–256

    Article  CAS  Google Scholar 

  • de Boer K, Moheimani N, Borowitzka M, Bahri P (2012) Extraction and conversion pathways for microalgae to biodiesel: a review focused on energy consumption. J Appl Phycol 24:1681–1698

    Google Scholar 

  • Eroglu E, Melis A (2010) Extracellular terpenoid hydrocarbon extraction and quantitation from the green microalgae Botryococcus braunii var. Showa. Biores Technol 101:2359–2366

    Article  CAS  Google Scholar 

  • Fon Sing S, Isdepsky A, Borowitzka MA, Moheimani NR (2013) Production of biofuels from microalgae. Mitig Adapt Strat Glob Change 18:47–72

    Article  Google Scholar 

  • Frenz J, Largeau C, Casadevall E (1989) Hydrocarbon recovery by extraction with a biocompatible solvent from free and immobilized cultures of Botryococcus braunii. Enzym Microb Technol 11:717–724

    Article  CAS  Google Scholar 

  • Fried B, Sherma J (1982) Thin-layer chromatography: techniques and applications. Marcel Dekker, New York, pp 105–122

    Google Scholar 

  • Hejazi MA, Holwerda E, Wijffels RH (2004) Milking microalga Dunaliella salina for β-carotene production in two-phase bioreactors. Biotech Bioeng 85:475–481

    Article  CAS  Google Scholar 

  • Kanazawa Z, Fujita C, Yuhara T, Sasa T (1958) Mass culture of unicellular algae using the "open pond circulation method". J Gen Appl Microbiol 4:135–139

    Article  Google Scholar 

  • Kates M, Volcani BE (1966) Lipid components of diatoms. Biochim Biophys Acta 116:264–278

    Article  PubMed  CAS  Google Scholar 

  • Kirk JTO (1994) Light and Photosynthesis in Aquatic Ecosystems. Cambridge University Press, Cambridge

    Book  Google Scholar 

  • Kleinegris D, van Es M, Janssen M, Brandenburg W, Wijffels R (2011) Phase toxicity of dodecane on the microalga Dunaliella salina. J Appl Phycol 23:949–958

    Article  PubMed  CAS  Google Scholar 

  • Lundquist TJ, Woertz IC, Quinn NWT, Benemann JR (2010) A realistic technology and engineering assessment of algae biofuel production. California Polytechnic State University, San Luis Obispo, p 178

    Google Scholar 

  • Largeau C, Casadevall E, Berkaloff C, Dhamelincourt P (1980) Sites of accumulation and composition of hydrocarbons in Botryococcus braunii. Phytochemistry 19:1043–1051

    Article  CAS  Google Scholar 

  • Mercz TI (1994) A study of high lipid yielding microalgae with potential for large-scale production of lipids and polyunsaturated fatty acids. PhD Thesis. Murdoch University, Perth, p 278

    Google Scholar 

  • Metzger P, Largeau C (2005) Botryococcus braunii: a rich source for hydrocarbons and related ether lipids. Appl Microbiol Biotech 66:486–496

    Article  CAS  Google Scholar 

  • Moheimani NR (2013) Long term outdoor growth and lipid productivity of Tetraselmis suecica, Dunaliella tertiolecta, and Chlorella sp (Chlorophyta) in bag photobioreactors. J Appl Phycol 25:167–176

    Article  CAS  Google Scholar 

  • Moheimani N, Borowitzka M (2006) The long-term culture of the coccolithophore Pleurochrysis carterae (Haptophyta) in outdoor raceway ponds. J Appl Phycol 18:703–712

    Article  Google Scholar 

  • Moheimani NR, Borowitzka MA (2007) Limits to productivity of the alga Pleurochrysis carterae (Haptophyta) grown in outdoor raceway ponds. Biotech Bioeng 96:27–36

    Article  CAS  Google Scholar 

  • Moheimani NR, Lewis D, Borowitzka MA, Pahl S (2011) Harvesting, thickening and dewatering microalgae. In: Carioca JOB (ed) International Microalgae and Biofuels Workshop. CENER, Fortaleza, pp 267–277

    Google Scholar 

  • Moheimani NR, Isdepsky A, Fon Sing S, Borowitzka MA (2013) Standard methods for measuring growth of algae and their composition. In: Borowitzka MA, Moheimani NR (eds) Algae for biofuels and energy. Springer, Dordrecht, pp 265–285

    Chapter  Google Scholar 

  • Platt T, Gallegos CL, Harrison WG (1980) Photoinhibition of photosynthesis in natural assemblages of marine phytoplanton. J Mar Res 38:687–701

    Google Scholar 

  • Ralph PJ, Gademann R (2005) Rapid light curves: A powerful tool to assess photosynthetic activity. Aquat Bot 82:222–237

    Article  CAS  Google Scholar 

  • Ratledge C, Kristiansen B (2001) Basic Biotechnology. Cambridge University Press, Cambridge

    Google Scholar 

  • Redfield AC (1958) The biological control of chemical factors in the environment. Am Sci 46:230A–221

    Google Scholar 

  • Sauer T, Galinski EA (1998) Bacterial milking: a novel bioprocess for production of compatible solutes. Biotech Bioeng 57:306–313

    Article  CAS  Google Scholar 

  • Sheehan J, Dunahay T, Benemann J, Roessler P (1998) A look back at the U.S. department of energy's aquatic species program-biodiesel from algae. NREL Report No. TP-580-24190. 325 pp

  • Sim SJ, An JY, Kim BW (2001) Two-phase extraction culture of Botryococcus braunii producing long-chain unsaturated hydrocarbons. Biotech Lett 23:201–205

    Article  CAS  Google Scholar 

  • Sim S, Chang H (1993) Increased shikonin production by hairy roots of Lithospermum erythrorhizon in two phase bubble column reactor. Biotech Lett 15:145–150

    Article  CAS  Google Scholar 

  • Stumm W, Morgan JJ (1996) Aquatic chemistry: chemical equilibria and rates in natural waters. John Wiley & Sons, NewYork, 1022 pp

    Google Scholar 

  • Wake L, Hillen L (1981) Nature and hydrocarbon content of blooms of the alga Botryococcus braunii occurring in Australian freshwater lakes. Aust J Mar Freshwat Res 32:353–367

    Article  CAS  Google Scholar 

  • Zittelli G, Zittelli GC, Rodolfi L, Tredici MR (2003) Mass cultivation of Nannochloropsis sp. in annular reactors. J Appl Phycol 15:107–114

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study was funded by the Murdoch University Strategic Research Fund. The authors would like to thank Azadeh Mohammadbagheri, Emily Quek and Celia Smuts for their excellent technical assistance.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Navid R. Moheimani.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Moheimani, N.R., Cord-Ruwisch, R., Raes, E. et al. Non-destructive oil extraction from Botryococcus braunii (Chlorophyta). J Appl Phycol 25, 1653–1661 (2013). https://doi.org/10.1007/s10811-013-0012-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10811-013-0012-9

Keywords

Navigation