The green algae Botryococcus braunii is widely recognized as a source of oil, including hydrocarbons. However, the slow rate of growth B. braunii hampers its commercial development. This stu- dy addresses this by examining the effects of three growth media on biomass and oil production in two B. braunii Race B strains, Kossou-4 and Overjuyo-3. Growth of B. braunii strains in BG11 medium resulted in significantly higher growth (2.3 - 4.2 and 2.9 - 6.0 fold increases in Kossou-4 and Overjuyo-3 respectively) compared to the JM and BBM-3N media after 15 days. A similar trend was obtained when biomass was measured indirectly using optical density (OD) and chlorophyll fluo-rescence. Oil production was also significantly higher in BG11 whether measured as oil weight or absorbance (ODs at 680 and 750 nm). However, the presence of extracellular oil was shown to in-crease absorbance values making OD measurements less reliable than dry weight assays. Maximum recovery of oil was recorded when hexane was used as solvent compared to hexane-isopro- panol and heptane. These results suggest that BG11 is the best growth medium for these two strains under the conditions of this experiment.
References
[1]
Costa, M.H., Yanagi, S.N., Souza, P.J., Ribeiro, A. and Rocha, E.J. (2007) Climate Change in Amazonia Caused by Soybean Cropland Expansion, as Compared to Caused by Pastureland Expansion. Geophysical Research Letters, 34, 1-4. http://dx.doi.org/10.1029/2007GL029271
[2]
Cheng, J.J. and Timilsina, G.R. (2011) Status and Barriers of Advanced Biofuel Technologies: A Review. Renewable Energy, 36, 3541-3549. http://dx.doi.org/10.1016/j.renene.2011.04.031
[3]
Smith, V.H., Sturm, B.S., Denoyelles, F.J. and Billings, S.A. (2010) The Ecology of Algal Biodiesel Production. Trends in Ecology & Evolution, 25, 301-309. http://dx.doi.org/10.1016/j.tree.2009.11.007
[4]
Chisti, Y. (2007) Biodiesel from Microalgae. Biotechnology Advances, 25, 294-306.
http://dx.doi.org/10.1016/j.biotechadv.2007.02.001
[5]
Brown, A., Knights, B. and Conway, E. (1969) Hydrocarbon Content and Its Relationship to Physiological State in the Green Alga Botryococcus braunii. Phytochemistry, 8, 543-547. http://dx.doi.org/10.1016/S0031-9422(00)85397-2
[6]
Andersen, R.A. (2005) Algal Culturing Techniques. Academic Press, 578 p.
[7]
Cepák, V. and Lukavsky, J. (1994) The Effect of High Irradiances on Growth, Biosynthetic Activities and the Ultrastructure of the Green Alga Botryococcus braunii Strain Droop 1950/807-1. Algological Studies, 72, 115-131.
[8]
Croft, M.T., Lawrence, A.D., Raux-Deery, E., Warren, M.J. and Smith, A.G. (2005) Algae Acquire Vitamin B12 through a Symbiotic Relationship with Bacteria. Nature, 438, 90-93. http://dx.doi.org/10.1038/nature04056
[9]
Rani, H., Kumar, K. and Eswarappa, V. (2011) Effect of Nitrogen Source on the Growth and Lipid Production of Microalgae. Current Biotica, 4, 426-433.
[10]
Ge, Y., Liu, J. and Tian, G. (2011) Growth Characteristics of Botryococcus braunii 765 under High CO2 Concentration in Photobioreactor. Bioresource Technology, 102, 130-134. http://dx.doi.org/10.1016/j.biortech.2010.06.051
[11]
Velichkova, K., Sirakov, I. and Georgiev, G. (2012) Cultivation of Botryococcus braunii Strain in Relation of Its Use for Biodiesel Production. Journal of Bioscience and Biotechnology, 157-162.
[12]
Li, Y., Moore, R.B., Qin, J.G., Scott, A. and Ball, A.S. (2013) Extractable Liquid, Its Energy and Hydrocarbon Content in the Green Alga Botryococcus braunii. Biomass and Bioenergy, 52, 103-112.
http://dx.doi.org/10.1016/j.biombioe.2013.03.002
[13]
Dayananda, C., Sarada, R., Usha Rani, M., Shamala, T. and Ravishankar, G. (2007) Autotrophic Cultivation of Botryococcus braunii for the Production of Hydrocarbons and Exopolysaccharides in Various Media. Biomass and Bioenergy, 31, 87-93. http://dx.doi.org/10.1016/j.biombioe.2006.05.001
[14]
Ambati, R.R., Ravi, S. and Aswathanarayana, R.G. (2010) Enhancement of Carotenoids in Green Alga-Botryococcus braunii in Various Autotrophic Media under Stress Conditions. International Journal of Biomedical and Pharmaceutical Sciences, 4, 87-92.
[15]
Kalita, N., Baruah, G., Dev G.R.C., Talukdar, J. and Kalita, M.C. (2011) Ankistrodesmus falcatus: A Promising Candidate for Lipid Production, Its Biochemical Analysis and Strategies to Enhance Lipid Productivity. Journal of Microbiology and Biotechnology Research, 1, 148-157.
[16]
Sakamoto, K., Baba, M., Suzuki, I., Watanabe, M.M. and Shiraiwa, Y. (2012) Optimization of Light for Growth, Photosynthesis, and Hydrocarbon Production by the Colonial Microalg Botryococcus braunii BOT-22. Bioresource Technology, 110, 474-479. http://dx.doi.org/10.1016/j.biortech.2012.01.091
[17]
Zhang, F., Cheng, L.H., Xu, X.H., Zhang, L. and Chen, H.L. (2013) Application of Memberane Dispersion for Enhanced Lipid Milking from Botryococcus braunii FACHB 357. Journal of Biotechnology, 165, 22-29.
http://dx.doi.org/10.1016/j.jbiotec.2013.02.010
[18]
Griffiths, M.J., Garcin, C., van Hille, R.P. and Harrison, S.T. (2011) Interference by Pigment in the Estimation of Microalgal Biomass Concentration by Optical Density. Journal of Microbiological Methods, 85, 119-123.
http://dx.doi.org/10.1016/j.mimet.2011.02.005
[19]
Sánchez Mirón, A., Contreras Gómez, A., Garc??a Camacho, F., Molina Grima, E. and Chisti, Y. (1999) Comparative Evaluation of Compact Photobioreactors for Large-Scale Monoculture of Microalgae. Journal of Biotechnology, 70, 249-270. http://dx.doi.org/10.1016/S0168-1656(99)00079-6
[20]
Ryckebosch, E., Muylaert, K. and Foubert, I. (2012) Optimization of an Analytical Procedure for Extraction of Lipids from Microalgae. Journal of the American Oil Chemists’ Society, 89, 189-198.
http://dx.doi.org/10.1007/s11746-011-1903-z
[21]
Mercer, P. and Armenta, R.E. (2011) Developments in Oil Extraction from Microalgae. European Journal of Lipid Science and Technology, 113, 539-547. http://dx.doi.org/10.1002/ejlt.201000455
[22]
Eroglu, E. and Melis, A. (2010) Extracellular Terpenoid Hydrocarbon Extraction and Quantitation from the Green Microalgae Botryococcus braunii var. Showa. Bioresource Technology, 101, 2359-2366.
http://dx.doi.org/10.1016/j.biortech.2009.11.043
[23]
Metzger, P., Allard, B., Casadevall, E., Berkaloff, C. and Couté, A. (1990) Structure and Chemistry of a New Chemical Race of Botryococcus braunii (Chlorophyceae) that Produces Lycopadiene, a Tetraterpenoid Hydrocarbon. Journal of Phycology, 26, 258-266. http://dx.doi.org/10.1111/j.0022-3646.1990.00258.x
[24]
Boonma, S., Vacharapiyasophon, P., Peerapornpisal, Y., Pekkoh, J. and Pumas, C. (2014) Isolation and Cultivation of Botryococcus braunii Kutzing from Northern Thailand. Chiang Mai Journal of Science, 41, 298-306.
[25]
Mohsenpour, S.F., Richards, B. and Willoughby, N. (2012) Spectral Conversion of Light for Enhanced Microalgae Growth Rates and Photosynthetic Pigment Production. Bioresource Technology, 125, 75-81.
http://dx.doi.org/10.1016/j.biortech.2012.08.072
[26]
Zhu, C. and Lee, Y. (1997) Determination of Biomass Dry Weight of Marine Microalgae. Journal of Applied Phycology, 9, 189-194. http://dx.doi.org/10.1023/A:1007914806640
[27]
Sawayama, S., Minowa, T., Dote, Y. and Yokoyama, S. (1992) Growth of the Hydrocarbon-Rich Microalga Botryococcus braunii in Secondarily Treated Sewage. Applied Microbiology and Biotechnology, 38, 135-138.
http://dx.doi.org/10.1007/BF00169433
[28]
Eroglu, E., Okada, S. and Melis, A. (2011) Hydrocarbon Productivities in Different Botryococcus Strains: Comparative Methods in Product Quantification. Journal of Applied Phycology, 23, 763-775.
http://dx.doi.org/10.1007/s10811-010-9577-8
[29]
Moheimani, N.R., Cord-Ruwisch, R., Raes, E. and Borowitzka, M.A. (2013) Non-Destructive Oil Extraction from Botryococcus braunii (Chlorophyta). Journal of Applied Phycology, 25, 1653-1661.
http://dx.doi.org/10.1007/s10811-013-0012-9
[30]
Kalacheva, G., Zhila, N., Volova, T. and Gladyshev, M. (2002) The Effect of Temperature on the Lipid Composition of the Green Alga Botryococcus. Microbiology, 71, 286-293. http://dx.doi.org/10.1023/A:1015898426573
[31]
Padovan, A. (1992) Isolation and Culture of Five Species of Freshwater Algae from the Alligator Rivers Region, Northern Territory. Australian Government Publishing Service. http://155.187.2.69/ssd/publications/tm/pubs/tm37.pdf
[32]
Ferreira-Dias, S., Valente, D.G. and Abreu, J.M. (2003) Comparison between Ethanol and Hexane for Oil Extraction from Quercus suber L. Fruits. Grasas y Aceites, 54, 378-383. http://dx.doi.org/10.3989/gya.2003.v54.i4.225
