limitaEffect of nutrient availability on lipid productivity of Botryococcus sp. (Botryococcaceae, Chlorophyta), a newly isolated tropical microalgae strain from Puerto Rico

Authors:Catalina Dávila, Gary A. Toranzos


Microalgae are promising sources of biofuels due to the high lipid content of some species. However, growing microalgae at large scales involves high production costs, mainly associated with nutrient inputs and harvesting processes. Therefore, to be cost competitive, species to be used as a source of fuels should be capable of accumulating lipids and biomass at lower fertilizer inputs. In the present study, we isolated and identified a native microalgal strain of Botryococcus sp. which was cultured under varying nitrogen, phosphorus and carbon dioxide regimes. The effects of nutrient availability on biomass, lipid production and fatty acid profiles were examined. We observed an increase in the relative lipid content from 25.5% under nitrogen non-limiting conditions to 41.8% under nitrogen deprivation. The lipid profile induced by nitrogen limitation was found to be dominated by saturated and monounsaturated lipid classes, meeting the European Standards for biodiesel and oil suitable for biofuel production. Thus, this Botryococcus sp. has the potential to be used at large scale cultures with the purpose of producing biofuels with lower fertilizer costs.

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Banerjee, A., R. Sharma, Y. Chisti, and U. C. Banerjee, U. C. 2002. Botryococcus braunii : A Renewable Source of Hydrocarbons and Other Chemicals. Critical Reviews in Biotechnology 22: 245–279.

Beal, C. M., R. E. Hebner, M. E. Webber, R. S. Ruoff, A. F. Seibert, and C. W. King. 2012. Comprehensive Evaluation of Algal Biofuel Production: Experimental and Target Results. Energies 5: 1943–1981.

Beal, C. M., R. E. Hebner, M. E. Webber, R. S. Ruoff, and A. F. Seibert. 2012. The energy return on investment for algal biocrude: Results for a research production facility. Bioenergy Research 5: 341–362.

Becker, E. W. 1994. Microalgae: biotechnology and microbiology. Cambridge University Press.

Brennan, L. and P. Owende. 2010. Biofuels from microalgae—A review of technologies for production, processing, and extractions of biofuels and co-products. Renewable and Sustainable Energy Reviews 14: 557–577.

Chen, C.-Y., K.-L. Yeh, R. Aisyah, D.-J. Lee, and J.-S. Chang. 2011. Cultivation, photobioreactor design and harvesting of microalgae for biodiesel production: a critical review. Bioresource Technology 102: 71–81.

Chen, M., H. Tang, H. Ma, T. C. Holland, K. Y. S. Ng, and S. O. Salley. 2011. Effect of nutrients on growth and lipid accumulation in the green algae Dunaliella tertiolecta. Bioresource Technology 102: 1649–1655.

Bechemin, C., D. Grzebyk, F. Hachame, C. Hummert, and S. Y. Maestrinil. 1999. Effect of different nitrogen/phosphorus nutrient ratios on the toxin content in Alexandrium minutum. Aquatic Microbial Ecology 20: 157–165.

Chu, S. P. 1942. The Influence of the Mineral Composition of the Medium on the Growth of Planktonic Algae : Part I . Methods and Culture Media. Journal of Ecology 30: 284–325.

Converti, A., A. A. Casazza, E. Y. Ortiz, P. Perego, and M. Del Borghi. 2009. Effect of temFperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculata and Chlorella vulgaris for biodiesel production. Chemical Engineering and Processing: Process Intensification 48: 1146–1151.

Doan, T. T. Y., B. Sivaloganathan, and J. P. Obbard. 2011. Screening of marine microalgae for biodiesel feedstock. Biomass and Bioenergy 35: 2534–2544.

Elsey, D., D. Jameson, B. Raleigh, and M. J. Cooney. 2007. Fluorescent measurement of microalgal neutral lipids. Journal of Microbiological Methods 68: 639–642.

Eriksen, N. T. 2008. The technology of microalgal culturing. Biotechnology Letters 30: 1525–1536.

Fawley, M. and K. Fawley. 2004. A simple and rapid technique for the isolation of DNA from microalgae. Journal of Phycology 40: 223–225.

Fields, M. W., A. Hise, E. J. Lohman, T. Bell, R. D. Gardner, L. Corredor, K. Moll, B. M. Peyton, G. W. Characklis, and R. Gerlach. 2014. Sources and resources: importance of nutrients, resource allocation, and ecology in microalgal cultivation for lipid accumulation. Applied Microbiology and Biotechnology 98: 4805–4816.

Geider, R. J. and J. La Roche. 2002. Redfield revisited : variability of C : N : P in marine microalgae and its biochemical basis. European Journal of Phycology 37: 1–17.

Gouveia, L. and A. C. Oliveira. 2009. Microalgae as a raw material for biofuels production. Journal of Industrial Microbiology & Biotechnology 36: 269–274.

Griffiths, M. J. and S. T. L. Harrison. 2009. Lipid productivity as a key characteristic for choosing algal species for biodiesel production. Journal of Applied Phycology 21: 493–507.

Han, F., J. Huang, Y. Li, W. Wang, M. Wan, G. Shen, and J. Wang. 2013. Enhanced lipid productivity of Chlorella pyrenoidosa through the culture strategy of semi-continuous cultivation with nitrogen limitation and pH control by CO2. Bioresource Technology 136: 418–424.

Hu, Q. 2004. Environmental Effects on Cell Composition. Pp. 83–98 in Handbook of Microalgal Culture: Biotechnology and Applied Phycology (1st ed.), ed. A. Richmond. Blackwell Science Ltd.

Huang, G., F. Chen, D. Wei, X. Zhang, G. Chen. 2010. Biodiesel production by microalgal biotechnology. Applied Energy 87: 38–46.

Juneja, A., R. M. Ceballos, and G. S. Murthy. 2013. Effects of environmental factors and nutrient availability on the biochemical composition of algae for biofuels production: A review. Energies 6: 4607–4638.

Khozin-Goldberg, I. and Z.Cohen. 2006. The effect of phosphate starvation on the lipid and fatty acid composition of the fresh water eustigmatophyte Monodus subterraneus. Phytochemistry 67: 696–701.

Lee, J., C. Yoo, S. Jun, C. Ahn, and H. Oh. 2010. Comparison of several methods for effective lipid extraction from microalgae. Bioresource Technology 101: 575–577.

Lohman, E. J., R. D. Gardner, L. D. Halverson, B. M. Peyton, and R. Gerlach. 2014. Carbon partitioning in lipids synthesized by Chlamydomonas reinhardtii when cultured under three unique inorganic carbon regimes. Algal Research 5: 171–180.

Matsumoto, H., N. Shioji, A. Hamasaki, Y. Ikuta, Y. Fukuda, M. Sato, N. Endo, and T. Tsukamoto. 1995. Carbon dioxide fixation by microalgae photosynthesis using actual flue gas discharged from a boiler. Applied Biochemistry and Biotechnology 51: 681–692.

Mazzuca, T. and Y. Chisti. 2010. Potential fuel oils from the microalga Choricystis minor. Journal of Chemical Technology and Biotechnology 85: 100–108.

McMillan, J. R., I. A. Watson, M. Ali, M., and W. Jaafar. 2013. Evaluation and comparison of algal cell disruption methods: Microwave, waterbath, blender, ultrasonic and laser treatment. Applied Energy 103: 128–134.

Milano, J., H. C. Ong, H. H. Masjuki, W. T. Chong, M. K. Lam, P. K. Loh, and V. Vellayan. 2016. Microalgae biofuels as an alternative to fossil fuel for power generation. Renewable and Sustainable Energy Reviews 58: 180–197.

Msanne, J., D. Xu, A. Reddy, J. A. Casas-Mollano, T. Awada, E. B. Cahoon and H. Cerutti. 2012. Metabolic and gene expression changes triggered by nitrogen deprivation in the photoautotrophically grown microalgae Chlamydomonas reinhardtii and Coccomyxa sp. C-169. Phytochemistry 75: 50–59.

Murphy, C. F. and D. T. Allen. 2011. Energy-water nexus for mass cultivation of algae. Environmental Science and Technology 45: 5861–5868.

Qu, J., H.-Z. Mao, W. Chen, S.-Q. Gao, Y.-N. Bai, Y.-W. Sun, Y.-F. Geng, and J. Ye. 2012. Development of marker-free transgenic Jatropha plants with increased levels of seed oleic acid. Biotechnology for Biofuels, 5: article 10.

Rowley, W. M. 2010. Nitrogen and phosphorus biomass-kinetic model for Chlorella vulgaris in a biofuel production scheme. Thesis submitted to the Graduate School of Engineering and Management, Air Force Institute of Technology.

Sakthivel, R., S. Elumalai, and M. Mohommad. 2011. Microalgae lipid research, past, present: A critical review for biodiesel production, in the future. Journal of Experimental Sciences 2: 29–49.

Scarsella, M., G. Torzillo, A. Cicci, G. Belotti, P. De Filippis, and M. Bravi. 2012. Mechanical stress tolerance of two microalgae. Process Biochemistry 47: 1603–1611.

Schenk, P. M., S. R. Thomas-Hall, E. Stephens, U. C. Marx, J. H. Mussgnug, C. Posten, O. Kruse, and B. Hankamer. 2008. Second Generation Biofuels: High-Efficiency Microalgae for Biodiesel Production. BioEnergy Research 1: 20–43.

Sobczuk, T. M., F. García Camacho, E. Molina Grima, and Y. Chisti. 2006. Effects of agitation on the microalgae Phaeodactylum tricornutum and Porphyridium cruentum. Bioprocess and Biosystems Engineering 28: 243–250.

Spijkerman, E. 2007. Phosphorus acquisition by Chlamydomonas acidophila under autotrophic and osmo-mixotrophic growth conditions. Journal of Experimental Botany 58: 4195–4202.

Spijkerman E. and P. F. M. Coesel. 1996. Competition for phosphorus among planktonic desmid species in continuous-flow culture. Journal of Phycology 32: 939–948.

Suganya, T., M. Varman, H. H. Masjuki, and S. Renganathan. 2016. Macroalgae and microalgae as a potential source for commercial applications along with biofuels production: A biorefinery approach. Renewable and Sustainable Energy Reviews 55: 909–941.

Tomas, C. R. 1979. Olisthodiscus luteus (Chrysophyceae). III.Uptake and utilization of nitrogen and phosphorus.Journal of Phycology 15: 5–12.

Widjaja, A., C. Chien, and Y. Ju. 2009. Study of increasing lipid production from fresh water microalgae Chlorella vulgaris. Journal of the Taiwan Institute of Chemical Engineers 40: 13–20.

Xin, L., H. Hu, G. Ke, and Y. Sun. 2010. Effects of different nitrogen and phosphorus concentrations on the growth, nutrient uptake, and lipid accumulation of a freshwater microalga Scenedesmus sp. Bioresource Technology 101: 5494–5500.

Yamaberi, K., M. Takagi, and T. Yoshida. 1998. Nitrogen depletion for intracellular triglyceride accumulation to enhance liquefaction yield of marine microalgal cells into a fuel oil. Journal of Marine Biotechnology 6: 44–48.

Yoo, C., S. Jun, J. Lee, C. Ahn, and H. Oh. 2010. Selection of microalgae for lipid production under high levels carbon dioxide. Bioresource Technology 101: S71–S74.

Zhila, N. O., G. S. Kalacheva, and T. G. Volova. 2005. Effect of Nitrogen Limitation on the Growth and Lipid Composition of the Green Alga Botryococcus braunii Kütz IPPAS H-252. Russian Journal of Plant Physiology 52: 311–319.

Zhu, F., R. Massana, F. Not, D. Marie, and D. Vaulot. 2005. Mapping of picoeucaryotes in marine ecosystems with quantitative PCR of the 18S rRNA gene. FEMS Microbiology Ecology 52: 79–92.


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