Waste and Biomass Valorization

, Volume 10, Issue 7, pp 1873–1881 | Cite as

Efficacy of Enzymatic Transesterification of Chlorococcum sp. Algal Oils for Biodiesel Production

  • Pandian Prabakaran
  • Virumandi Pradeepa
  • Gopal SelvakumarEmail author
  • A. David Ravindran
Original Paper


Increasing oil prices and climate change concerns, biodiesel has gained attention as an alternative energy source. Biodiesel derived from microalgae is a potentially renewable and carbon neutral alternative to petroleum fuels. One of the most important decisions in obtaining oil from microalgae is the choice of algal species to use. Seven microalgae from a total of 18 isolated cultures were selected as based on their purity and growth rates. Seven cultures were identified as Chlorella sp., Haematococcus sp., Chlorococcum sp., Scenedesmus sp., Rivularia sp., Neochloris sp. and Nostoc sp. Based on the higher lipid production, three species such as Chlorella sp. (25.8 ± 0.15%), Chlorococcum sp. (27.0 ± 0.25%) and Scenedesmus sp. (29.7 ± 0.32%) were selected for further studies. An efficient lipase producing Pseudomonas sp. was isolated and used for biological transesterification studies for microalgal oil conversion. Immobilized lipase from Pseudomonas sp. and chemical transesterification by NaOH were tried. Fatty acid methyl esters (FAMEs) were analysed by GC-MS which showed that Chlorococcum sp. crude oil contained C35H46O8 as the major constituent. In transesterification studies, C50H82O9 was identified by chemical method whereas two compounds such as C36H56O6 and C24H28N2O5 were identified through biological transesterification. Hence the tested seven microalgal species, Chlorococcum sp. was more suitable for the production of good quality biodiesel.


Microalgae Chlorococcum sp. Biodiesel Transesterification Lipase 



The authors are thankful to the authorities of Gandhigram Rural Institute-Deemed University and Alagappa University for providing required facilities and one of the author P. Prabakaran is grateful to the SERB-DST for the N-PDF (PDF/2015/000652) for financial assistance.

Compliance with Ethical Standards

Conflict of interest

The authors declare that there is no conflict of interest regarding the publication of this paper.


  1. 1.
    Krawczyk, T.: Biodiesel: alternative fuel makes inroads but hurdles remain. Inform. 7, 801–829 (1996)Google Scholar
  2. 2.
    Von Wedel, R.: Technical Handbook for Marine Biodiesel in Recreational Boats. National Renewable Energy Laboratory, US Department of Energy 32 (1999)Google Scholar
  3. 3.
    Patil, V., Tran, K.-Q., Giselrød, H.R.: Towards sustainable production of biofuels from microalgae. Int. J. Mol. Sci. 9, 1188–1195 (2008)CrossRefGoogle Scholar
  4. 4.
    Zhang, X., Peterson, C., Reece, D., Haws, R., Möller, G.: Biodegradability of biodiesel in the aquatic environment. Am. Soc. Agric. Eng. 41(5), 1423–1430 (1998)CrossRefGoogle Scholar
  5. 5.
    Fukuda, H., Kondo, A., Noda, H.: Biodiesel fuel production by transesterification of oils. J. Biosci. Bioeng. 92, 405–416 (2001)CrossRefGoogle Scholar
  6. 6.
    Kayode, B., Hart, A.: An overview of transesterification methods for producing biodiesel from waste vegetable oils. Biofuels (2017). Google Scholar
  7. 7.
    Kato, M., Fuchimoto, J., Tanino, T., Kondo, A., Fukuda, H.: Preparation of a whole cell biocatalyst of mutated Candida Antarctica Lipase B (mCALB) by a yeast molecular display system and its practical properties. Appl. Microbiol. Biotechnol. 75, 549–555 (2007)CrossRefGoogle Scholar
  8. 8.
    Kanz, T., Bold, H.C.: Physiological Studies, Morphological and Taxonomical Investigation of Nostoc and Anabaena in Culture. University of Texas, Austin (TX), Publication No. 6924 (1969)Google Scholar
  9. 9.
    John, D.M., Whitton, B.A., Brook, A.J.: The Freshwater Algal Flora of the British Isles an Identification Guide to Freshwater and Terrestrial Algae, pp. 39–43. Cambridge University Press, Cambridge (2003)Google Scholar
  10. 10.
    Prescott, G.W.: How to Know the Fresh Water Algae, vol. 1. Cranbrook press, Michigan (1959)Google Scholar
  11. 11.
    Ehimen, E.A., Sun, Z.F., Carrington, C.G.: Variables affecting the in situ transesterification of microalgae lipids. Fuel 89(3), 677–684 (2010)CrossRefGoogle Scholar
  12. 12.
    Zhu, M., Zhou, P.P., Yu, L.J.: Extraction of lipids from Mortierella alpine and Enrichment of arachidoni acid from the fungal lipids. Bioresour. Technol. 84, 93–95 (2002)CrossRefGoogle Scholar
  13. 13.
    Bligh, E.G., Dyer, W.J.: A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37, 911–917 (1959)CrossRefGoogle Scholar
  14. 14.
    Schreiner, M.: Optimization of solvent extraction and direct transmethylation methods for the analysis of egg yolk lipids. Int. J. Food Prop. 9, 573–581 (2006)CrossRefGoogle Scholar
  15. 15.
    Xu, N., Zhang, X., Fan, X., Han, L., Zeng, C.: Effects of nitrogen source and concentration on growth rate and fatty acid composition of Elipsoidion sp. (Eustigmatophyta). J. Appl. Phycol. 13, 463–469 (2001)CrossRefGoogle Scholar
  16. 16.
    Shukla, P., Gupta, K.: Ecological screening for lipolytic molds and process optimization for lipase production from Rhizopus Orzyae KG-5. Appl. Sci. Environ. Sanit. 2, 35–42 (2007)Google Scholar
  17. 17.
    Apun, K., Jong, B.C., Salleh, M.A.: Screening and isolation of a cellulolytic and amylolytic Bacillus from Sago pith waste. J. Gen. Appl. Microbiol. 46, 263–267 (2000)CrossRefGoogle Scholar
  18. 18.
    Holt, J.G., Krieg, N.R., Sneath, P.H.A., Staley, J.T., Williams, S.T.: Bergey’s Manual of Determinative Bacteriology, 9th edn. Williams and Wilkins, Baltimore, MS (1994)Google Scholar
  19. 19.
    Sadasivam, S., Manickam, A.: Biochemical Methods for Agricultural Sciences. Wiley Eastern Limited and Tamilnadu Agricultural University, New Delhi (1991)Google Scholar
  20. 20.
    Bhushan, I., Parshad, R., Qazi, G.N., Gupta, V.K.: Immobilzation of lipase by entrapment in Ca-alginate beads. J. Bioact. Compat. Polym. 23, 552–562 (2008)CrossRefGoogle Scholar
  21. 21.
    Richmond, A.: Handbook of Microalgal Culture: Biotechnology and Applied Phycology. Blackwell Science Ltd, Oxford (2004)Google Scholar
  22. 22.
    Chisti, Y.: Biodiesel from microalgae. Biotechnol. Adv. 25(3), 294–306 (2007)CrossRefGoogle Scholar
  23. 23.
    Wang, L., Yecong, L., Chen, P., Min, M., Chen, Y., Zhu, J., et al.: Anaerobic digested dairy manure as a nutrient supplement for cultivation of oil-rich green microalgae Chlorella sp. Bioresour. Technol. 101, 2623–2628 (2010)CrossRefGoogle Scholar
  24. 24.
    Ratha, S., Babu, S., Renuka, N., Prasanna, R., Prasad, R., Saxena, A.: Exploring nutritional modes of cultivation for enhancing lipid accumulation in microalgae. J. Basic Microbiol. 53(5), 440–450 (2013)CrossRefGoogle Scholar
  25. 25.
    Sheehan, J., Dunahay, T., Benemann, J., Roessler, P.: A Look Back at the U.S. Department of Energy’s Aquatic Species Program-Biodiesel From Algae. National Renewable Energy Laboratory, Golden, CO. Report NREL/TP, 580-24190 (1998)Google Scholar
  26. 26.
    Gao, Y., Yang, M., Wang, C.: Nutrient deprivation enhances lipid content in marine microalgae. Bioresour. Technol. 147, 484–491 (2013)CrossRefGoogle Scholar
  27. 27.
    Allard, B., Templier, J.: Comparison of neutral lipid profile of various trilaminar outer cell wall (TLS)-containing microalgae with emphasis on algae occurrence. Phytochemistry 54, 369–380 (2000)CrossRefGoogle Scholar
  28. 28.
    Rodolfi, L., Zittelli, G.C., Bassi, N., Padovani, G., Biondi, N., Bonin, G., et al.: Microalgae for oil: strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor. Biotechnol. Bioeng. 102, 100–112 (2009)CrossRefGoogle Scholar
  29. 29.
    Knothe, G.: Designer biodiesel: optimizing fatty esters composition to improve fuel properties. Energy Fuel 22, 1358–1364 (2008)CrossRefGoogle Scholar
  30. 30.
    Mohan, T.S., Palavesam, A., Immanvel, G.: Isolation and characterization of lipase-producing Bacillus strains from oil mill waste. Afr. J. Biotechnol. 7, 2728–2735 (2008)Google Scholar
  31. 31.
    Bora, L., Kalita, M.C.: Production of extra cellular lipase from Bacillus sp. LBN 4 by solid state fermentation. Int. J. Microbiol. 7(2), 172–179 (2009)Google Scholar
  32. 32.
    Singh, M., Singh, S., Singh, R.S., Chisti, Y., Banerjee, U.C.: Transesterification of primary and secondary alcohols using Pseudomonas aeruginosa lipase. Bioresour. Technol. 99, 116–2120 (2008)Google Scholar
  33. 33.
    Maulidiyah, N.M., Fatma, F., Natsir, M., Wibowo, D.: Characterization of methyl ester compound of biodiesel from industrial liquid waste of crude palm oil processing. Anal. Chem. Res. 12, 1–9 (2017)CrossRefGoogle Scholar

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© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of MicrobiologyAlagappa UniversityKaraikudi, Sivagangai DistrictIndia
  2. 2.Department of ZoologySeethalakshmi Achi College for WomenPallathur, Sivagangai DistrictIndia
  3. 3.Department of BiologyGandhigram Rural Institute-Deemed UniversityGandhigram, Dindigul DistrictIndia

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