Journal of Applied Phycology

, Volume 30, Issue 6, pp 3063–3073 | Cite as

Lipid production enhancement in tropically isolated microalgae by azide and its effect on fatty acid composition

  • Nurul Ashyikin Yahya
  • Noraiza Suhaimi
  • Marshila Kaha
  • Hirofumi Hara
  • Zuriati Zakaria
  • Norio Sugiura
  • Nor ‘Azizi Othman
  • Koji IwamotoEmail author
8th Asian Pacific Phycological Forum


Microalgae, as photosynthetic microorganisms, have been referred to as the third generation of biodiesel feedstock due to their ability to accumulate high amounts of lipids in their intracellular bodies. However, the commercialization of microalgal biodiesel is currently not economical due to high production cost. Recently, a novel strategy was found to overcome this issue and, at the same time, boost the accumulation of lipids by the addition of azide to the microalgal culture. Azide was added to study its effect on the growth and accumulation of fatty acids in three strains of tropical microalgae, identified as Acutodesmus obliquus, Desmodesmus maximus, and Chlorella pyrenoidosa, which were isolated from the Hulu Langat River, Selangor, Malaysia. Lipid production of A. obliquus and D. maximus after azide treatment increased by 40 and 20%, respectively. In contrast, the induction of azide led to insignificant lipid accumulation in C. pyrenoidosa, likely because azide targeted the respiration system rather than lipid production, which consequently caused a slight growth retardation. There was a great difference in the types of fatty acids found in azide-treated cells and non-treated cells. In A. obliquus, more saturated fatty acids (SFAs) were found in azide-treated cells, while more polyunsaturated fatty acids (PUFAs) were found in non-treated cells. In C. pyrenoidosa, the induction of lipid accumulation by azide led to the formation of more SFAs and PUFAs. For D. maximus, the cells synthesized more monounsaturated fatty acids (MUFAs) in both azide-treated and untreated media but azide treatment increased an insignificant amount of PUFAs.


Azide Biodiesel Growth Lipid Microalgae Chlorophyta 


Funding information

This study was supported by a grant-in-aid from MJIIT Takasago Education and Research Grant (Vote No. PY/2015/05518) sponsored by Takasago Thermal Engineering Co. Ltd. from 2015 to 2017 and the International Grant by University of Tsukuba (Vote No. PY/2016/07077).


  1. Atabani AE, Silitonga AS, Badruddin IA, Mahlia TMI, Masjuki HH, Mekhilef S (2012) A comprehensive review on biodiesel as an alternative energy resource and its characteristics. Renew Sust Energ Rev 16:2070–2093CrossRefGoogle Scholar
  2. Basil G, Imran P, Chahana D, Kaumeel C, Chetan P, Tonmoy G, Sandhya M (2014) Effects of different media composition, light intensity and photoperiod on morphology and physiology of freshwater microalgae Ankistrodesmus falcatus—a potential strain for biofuel production. Bioresour Technol 171:367–374CrossRefGoogle Scholar
  3. Borowitzka MA (2013) Species and strain selection. In: Borowitzka MA, Moheimani NR (eds) Algae for biofuels and energy. Springer, Dordrecht, pp 77–89CrossRefGoogle Scholar
  4. Brennan L, Owende P (2010) Biofuels from microalgae—a review of technologies for production processing and extractions of biofuels and co-products. Renew Sust Energ Rev 14:557–577CrossRefGoogle Scholar
  5. Chisti Y (2007) Biodiesel from microalgae. Biotechnol Adv 25:294–306CrossRefGoogle Scholar
  6. Converti A, Casazza AA, Ortiz EY, Perego P, Borghi MD (2009) Effect of temperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculata and Chlorella vulgaris for biodiesel production. Chem Eng Process 48:1146–1151CrossRefGoogle Scholar
  7. Corro G, Tellez N, Jimenez T, Tapia A, Banuelos F, Vazquez-Cuchillo O (2011) Biodiesel from waste frying oil: two steps process using acidified SiO2 for esterification step. Catal Today 166:116–122CrossRefGoogle Scholar
  8. Damiani MC, Popovich CA, Constenla D, Leonardi PI (2010) Lipid analysis in Haematococcus pluvialis to assess its potential use as a biodiesel feedstock. Bioresour Technol 101:3801–3807CrossRefGoogle Scholar
  9. Folch J, Lees M, Stanley GHS (1957) A simple method for the isolation and purification of total lipid from animal tissues. J Biol Chem 226:497–509PubMedGoogle Scholar
  10. Griffiths MJ, Harrison STL (2009) Lipid productivity as a key characteristic for choosing algal species for biodiesel production. J Appl Phycol 21:493–507CrossRefGoogle Scholar
  11. Jin L, Junchao H, Zheng S, Yujuan Z, Yue J, Feng C (2011) Differential lipid and fatty acid profiles of photoautotrophic and heterotrophic Chlorella zofingiensis: assessment of algal oils for biodiesel production. Bioresour Technol 102:106–110CrossRefGoogle Scholar
  12. Kawachi M, Ishimoto M, Mori F, Yumoto K, Sato M, Noël MH (2013) MCC-NIES list of strains: microalgae, endangered macroalgae and protists, 9th edn. Microbial culture collection at National Institute for Environmental Studies, TsukubaGoogle Scholar
  13. Keilin D (1936) The action of sodium azide on cellular respiration and on some catalytic oxidation reactions. Proc R Soc Lond B Biol Sci 121:165–173CrossRefGoogle Scholar
  14. Knothe G (2013) Production and properties of biodiesel from algal oils. In: Borowitzka MA, Moheimani NR (eds) Algae for biofuels and energy. Springer, Dordrecht, pp 207–219CrossRefGoogle Scholar
  15. Kotajima T, Shiraiwa Y, Suzuki I (2014) Functional screening of a novel Δ15 fatty acid desaturase from the coccolithophorid Emiliania huxleyi. Biochim Biophys Acta 1841:1451–1458CrossRefGoogle Scholar
  16. Laurens LML, Quinn M, Wychen SV, Templeton DW, Wolfrum EJ (2012) Accurate and reliable quantification of total microalgal fuel potential as fatty acid methyl esters by in situ transesterification. Anal Bioanal Chem 403:167–178CrossRefGoogle Scholar
  17. Lim S, Lee KT (2010) Recent trends, opportunities, and challenges of biodiesel in Malaysia: an overview. Renew Sust Energ Rev 14:938–954CrossRefGoogle Scholar
  18. Lim DKY, Garg S, Timmins M, Zhang ESB, Thomas-Hall SR, Schuhmann H, Li Y, Schenk PM (2012) Isolation and evaluation of oil-producing microalgae from subtropical coastal and brackish waters. PLoS One 7:e40751CrossRefGoogle Scholar
  19. Pedersen TC, Gardner RD, Gerlach R, Peyton BM (2018) Assessment of Nannochloropsis gaditana growth and lipid accumulation with increased inorganic carbon delivery. J Appl Phycol 30:2155–2166CrossRefGoogle Scholar
  20. Rashid N, Rehman MSU, Sadeq M, Mahmood T, Han J-I (2014) Current status, issues and developments in microalgae derived biodiesel production. Renew Sust Energ Rev 40:760–778CrossRefGoogle Scholar
  21. Rodolfi L, Zittelli GC, Bassi N, Padovani G, Biondi N, Bonini G, Tredici MR (2008) Microalgae for oil: strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor. Biotechnol Bioeng 102:100–112CrossRefGoogle Scholar
  22. Sekimoto S, Rochon D, Long JE, Dee JM, Berbee ML (2011) A multiple phylogeny of Olpidium and its implications for early fungal evolution. BMC Evol Biol 11:331CrossRefGoogle Scholar
  23. Singh P, Kumari S, Gulde A, Misra R, Rawat I, Bux F (2016) Trends and novel strategies for enhancing lipid accumulation and quality in microalgae. Renew Sust Energ Rev 55:1–16CrossRefGoogle Scholar
  24. Stansell GR, Gray VM, Sym SD (2012) Microbial fatty acid composition: implications for biodiesel quality. J Appl Phycol 24:791–801CrossRefGoogle Scholar
  25. Taleb A, Legrand J, Takache H, Taha S, Pruvost J (2018) Investigation of lipid production by nitrogen-starved Parachlorella kessleri under continuous illumination and day/night cycles for biodiesel application. J Appl Phycol 30:761–772CrossRefGoogle Scholar
  26. Throndsen J (1973) Special method—micromanipulators. In: Stein JR (ed) Handbook of phycological methods: culture methods and growth measurements. Cambridge University Press, Cambridge, pp 139–144Google Scholar
  27. Wychen SV, Laurens LML (2013) Determination of total lipids as fatty acid methyl esters (FAME) by in situ transesterification. National Renewable Energy Laboratory. NREL/TP-5100-60958, pp 1–12Google Scholar
  28. Yeesang C, Cheirsilp B (2011) Effect of nitrogen, salt and iron content in the growth medium and light intensity on lipid production by microalgae isolated from freshwater sources in Thailand. Bioresour Technol 102:3034–3040CrossRefGoogle Scholar
  29. Zalogin TR, Pick U (2014a) Azide improves triglycerides yield in microalgae. Algal Res 3:8–16CrossRefGoogle Scholar
  30. Zalogin TR, Pick U (2014b) Inhibition of nitrate reductase by azide in microalgae results in triglycerides accumulation. Algal Res 3:17–23CrossRefGoogle Scholar
  31. Zhou W (2014) Potential applications of microalgae in wastewater treatments. In: Liu L, Sun Z, Gerken H (eds) Recent advances in microalgal biotechnology. OMICS Group eBooks, USA, pp 1–8Google Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Environmental and Green Technology, Malaysia-Japan International Institute of TechnologyUniversiti Teknologi MalaysiaKuala LumpurMalaysia
  2. 2.Department of Chemical Process Engineering, Malaysia-Japan International Institute of TechnologyUniversiti Teknologi MalaysiaKuala LumpurMalaysia
  3. 3.Department of Mechanical Precision Engineering, Malaysia-Japan International Institute of TechnologyUniversiti Teknologi MalaysiaKuala LumpurMalaysia

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