Influence of nutrients on biomass and oil yield from microalgae Chlorella vulgaris for biodiesel production

Glacio Souza Araujo, Dilliani Naiane Mascena Lopes, Clarice da Silva Santiago, José William Alves da Silva, Fabiano André Narciso Fernandes


Microalgae are commonly used in aquiculture as feed for postlarval mollusks, fish and crustaceans because they are easy to grow, small in size, grow rapidly and have high levels of fatty acids. These microorganisms also accumulate high amounts of oil, which can be extracted and converted into biodiesel using chemical processes. In this work, freshwater microalgae Chlorella vulgaris was grown in the Live Food Production Laboratory (LABPAV/IFCE Aracati Campus), with urea (stock solution 1), triple superphosphate (stock solution 2) and vitamins (stock solution 3), in growth medium, in triplicate, using three different quantities of stock solutions 1 and 2, but with a constant amount of vitamins. The quantities of 0.5, 1 and 2 mL (T0.5, T1 and T2, respectively) were used for both stock solutions. We then monitored the growth of the microalgae, flocculated through chemical flocculation by adding a NaOH 2N solution, air-oven dried at 60 ºC for 24 hours, weighed the dried biomass on a semi-analytical balance, and extracted the oil using solvents. We thus observed that algal growth intensified and dry biomass increased as the amount of nutrients increased in the growth media; inversely, the best oil level was observed in the treatment using the lowest amount of nutrients in the growth media where the microalgae developed (20.13±0.19%). Finally, in Treatment T2, even with the lowest percentage of oil (18.95%), the amount of biomass produced compensates in the oil productivity, and using a lower amount of nutrients in the media of culture.


Biodiesel; Microalgae; Oil

Texto completo:



AMERICAN OIL CHEMISTS’ SOCIETY. Official Method Ca 5a-40. Free fatty acids. In: FIRESTONE, D. E. (ed.) Official methods and recommended practices of the AOCS. Champaign, IL: AOCS Press, 1997.

ARAUJO, G. S. et al. Bioprospecting for oil producing microalgal strains: evaluation of oil and biomass production for ten microalgal strains. Bioresource Technology, v. 102, n. 8, p. 5248-5250, 2011.

ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS. Official method of analysis of Official Analytical Chemists. 16. ed. Washington, USA: AOAC International, 1995.

AZMA, M. et al. Improvement of medium composition for heterotrophic cultivation of green microalgae, Tetraselmis suecica, using response surface methodology. Biochemical Engineering Journal, v. 53, n. 2, p. 187-195, 2011.

BLIGH, E. G.; DYER, W. J. A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology, v. 37, n. 8, p. 911-917, 1959.

CHISTI, Y. Biodiesel from microalgae beats bioethanol. Trends in Biotechnology, v. 26, n. 3, p. 126-131, 2008.

COURCHESNE, N. M. D. et al. Enhancement of lipid production using biochemical, genetic and transcription factor engineering approaches. Journal of Biotechnology, v. 141, n. 1/2, p. 31-41, 2009.

DE LA HOZ, S. et al. A rapid method of total lipid extraction and purification. Bioresource Technology, v. 102, n. 10, p. 5764-5774, 2011.

DERNER, R. B. et al. Microalgas, produtos e aplicações. Ciência Rural, v. 36, n. 6, p. 1959-1967, 2006.

DEVAPPA, K. R. et al. Quality of biodiesel prepared from phorbol ester extracted Jatropha curcas oil. Journal of the American Oil Chemists’ Society, v. 87, n. 6, p. 697-704, 2010.

DOAN, T. T. Y.; SIVALOGANATHAN, B.; PHILIP, J. Screening of marine microalgae for biodiesel feedstock. Biomass and Bioenergy, v. 35, n. 7, p. 2534-2544, 2011.

DRAGONE, G. et al. Nutrient limitation as a strategy for increasing starch accumulation in microalgae. Applied Energy, v. 88, n. 10, p. 3331-3335, 2011.

FRUMENTO, D. et al. Cultivation of Chlorella vulgaris in tubular photobioreactors: a lipid source for biodiesel production. Biochemical Engineering Journal, v. 81, p. 120-125, 2013.

GAO, C. et al. Application of sweet sorghum for biodiesel production by heterotrophic microalgae Chlorella protothecoides. Applied Energy, v. 87, n. 3, p. 756-761, 2010.

GUEDES, A. C.; AMARO, H. M.; MALCATA, F. X. Microalgae as sources of high added-value compounds-a brief review of recent work. Biotechnology Progress, v. 27, n. 3, p. 597-613, 2011.

HE, P. J. et al. Cultivation of Chlorella vulgaris on wastewater containing high levels of ammonia for biodiesel production. Bioresource Technology, v. 129, p. 177-181, 2013.

LYNCH, J. M.; BARBANO, D. M. Kjeldahl nitrogen analysis as a reference method for protein determination in dairy products. Journal of AOAC International, v. 82, p. 1389-1398, 1999.

MANDAL, S.; MALLICK, N. Microalgae Scenedesmus obliquus as a potential source for biodiesel production. Applied Microbiology and Biotechnology, v. 84, n. 2, p. 281-291, 2009.

MENG, X. et al. Biodiesel production from oleaginous microorganisms. Renew Energy, v. 34, p. 1-5, 2009.

RICHMOND, A. Handbook of microalgal culture: biotechnology and applied phycology. Oxford: Blackwell Science, 2004. 169 p.

SAFI, C. et al. Influence of microalgae cell wall characteristics on protein extractability and determination of nitrogen-to-protein conversion factors. Journal of Applied Phycology, v. 25, p. 523-529, 2013.

SÁNCHEZ, S.; MARTINEZ, M. E.; ESPINOLA, F. Biomass production and biochemical variability of the marine microalgae Isochrysis galbana in relation to culture medium. Biochemical Engineering Journal, v. 6, p. 13-18, 2000.

SHEN, Q. et al. Saline wastewater treatment by Chlorella vulgaris with simultaneous algal lipid accumulation triggered by nitrate deficiency. Bioresource Technology, v. 193, p. 68-75, 2015.

SILVA, A. F.; LOURENÇO, S. O.; CHALOUB, R. M. Effects of nitrogen starvation on the photosynthetic physiology of tropical marine microalga Rhodomonas sp. (Cryptophyceae). Aquatic Botany, v. 91, n. 4, p. 291-297, 2009.

SUBHADRA, B.; EDWARDS, M. An integrated renewable energy park approach for algal biofuel production in United States. Energy Policy, v. 38, n. 9, p. 4897-4902, 2010.

TAKAGI, M. et al. Limited feeding of potassium nitrate for intracellular lipid and triglyceride accumulation of Nannochloris sp. UTEX LB1999. Applied Microbiology and Biotechnology, v. 54, p. 112-117, 2000.

URI, P.; TATYANA, Z.; MEIRA, W. Accumulation of triglycerides in green microalgae: a potential source for biodiesel. Federation of European Biochemical Societies, v. 277, n. 1, p. 5-36, 2010.

VARGAS, R. M. Transesterificação de óleos vegetais, catalisada por bases não-iônicas, em fases homogênea e heterogênea/1996. 135 f. Tese (Doutorado em Química Orgânica) - Universidade Estadual de Campinas, Campinas, 1996.

WIDJAJA, A.; CHIEN, C. C.; JU, Y. H. Study of increasing lipid production from fresh water microalgae Chlorella vulgaris. Journal of the Taiwan Institute of Chemical Engineers, v. 40, p. 13-20, 2009.

WIJFFELS, R. H. Potential of sponges and microalgae for marine biotechnology. Trends in Biotechnology, v. 26, n. 1, 2007.

ZENG, X. et al. Microalgae bioengineering: From CO2 fixation to biofuel production. Renewable and Sustainable Energy Reviews, v. 15, n. 6, p. 3252-3260, 2011.

ZHENG, H. et al. Disruption of Chlorella vulgaris cells for the release of biodiesel-producing lipids: a comparison of grinding, ultrasonication, bead milling, enzymatic lysis, and microwaves. Applied Biochemical Biotechnology, v. 164, n. 7, p. 1215-1224, 2011.

Revista Ciência Agronômica ISSN 1806-6690 (online) 0045-6888 (impresso), Site:, e-mail: - Fone: (85) 3366.9702 - Expediente: 2ª a 6ª feira - de 7 às 17h.