Algae-Based Wastewater Treatment for Biofuel Production: Processes, Species, and Extraction Methods

  • Stephen R. Lyon
  • Hossein AhmadzadehEmail author
  • Marcia A. Murry
Part of the Biofuel and Biorefinery Technologies book series (BBT, volume 2)


This chapter develops the principles and rationale for an algae-based biofuel production coupled to bioremediation of municipal and agricultural wastewaters . A synergistic model for algal wastewater treatment is proposed, which addresses several economic bottlenecks to earlier algal systems and promotes value-added products, including a high-quality effluent in addition to biodiesel to improve the economic feasibility of algal biofuels . Finally, we review candidate species for full-scale algae production ponds based on algal structure, physiology and ecology , and methods for extraction of algal oils for biodiesel production and coproducts. The dominant strains of algae that are commonly found in wastewater ponds, including  Euglenia , Scenedesmus , Selenastrum , Chlorella, and  Actinastrum , are suggested as candidates for large-scale culturing based on their ability to strip nutrients and organic matter from wastewater , grow rapidly, and produce a significant level of algal oil. Oil extraction by supercritical fluid extraction (SFE) is discussed as an efficient means of isolating algal oil and other commercially important high-value compounds from algal biomass . Together with water and CO2 reclamation, such products may shift the economics of algal biomass production to allow production of low-value commodities including biodiesel and biogas.


Biochemical Oxygen Demand Supercritical Fluid Extraction Phaeodactylum Tricornutum Tertiary Treatment Euglena Gracilis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



H. Ahmadzadeh thanks ATF Committee for the financial support.


  1. Adams C, Godfrey V, Wahlen B, Seefeldt L, Bugbee B (2013) Understanding precision nitrogen stress to optimize the growth and lipid content tradeoff in oleaginous green microalgae. Bioresour Technol 131:188–194CrossRefGoogle Scholar
  2. Anderson RA (2004) Biology and systematics of heterokont and haptophyte algae. Am J Bot 91:1508–1522CrossRefGoogle Scholar
  3. Becker EW (1994) Microalgae: biotechnology and microbiology. Cambridge University Press, CambridgeGoogle Scholar
  4. Benemann JR, Hallenbeck PC, Weissman JC, Murry M, Oswald WJ (1977) Fertilizer production with nitrogen-fixing heterocystous blue-green algae. Final Report to the National Science Foundation, Sanitary Engineering Research Laboratory, University California BerkeleyGoogle Scholar
  5. Benemann JR, Oswald WJ (1996) Systems and economic analysis of microalgae ponds for conversion of CO2 to biomass, final report, Pittsburgh Energy Technology CenterGoogle Scholar
  6. Bligh EG, Dyer WJ (1959) A rapid method for total lipid extraction and purification. Can J Biochem Physiol 37(8):911–917CrossRefGoogle Scholar
  7. Borowitzka MA (1997) Microalgae for aquaculture: opportunities and constraints. J Appl Phycol 9(5):393–401CrossRefGoogle Scholar
  8. Borowitzka MA, Moheimani NR (2013) Sustainable biofuels from algae. Mitig Adapt Strat Glob Change 18:13–25CrossRefGoogle Scholar
  9. Brussaard CP (2004) Viral control of phytoplankton populations—a review. J Eukaryot Microbiol 51(2):125–138CrossRefGoogle Scholar
  10. Cavalier-Smith T (2007) A revised six-kingdom system of life. Biol Rev 73(3):203–266CrossRefGoogle Scholar
  11. Chisti Y (2007) Biodiesel from microalgae. Biotechnol Adv 25(3):294–306CrossRefGoogle Scholar
  12. Choi KJ, Nakhost Z, Krukonis VJ, Karel M (1987) Supercritical fluid extraction and characterization of lipids from algae Scenedesmus obliquus. Food Biotechnol 1(2):263–281CrossRefGoogle Scholar
  13. Colla LM, Bertolina TE, Costa JAV (2004) Fatty acids profile of Spirulina platensis grown under different temperatures and nitrogen concentrations. Zeitschrift Fur Naturforschung C 59(1/2):55–59Google Scholar
  14. Couto RM, Simões PC, Reis A, da Silva TL, Martins VH, Sanchez-Vincente Y (2010) Supercritical fluid extraction of lipids from the heterotrophic microalga Crypthecodinium cohnii. Eng Life Sci 10(2):158–164Google Scholar
  15. Craggs RJ, Lundquist TJ, Benemann JR (2012) Wastewater treatment pond algal production for biofuel. In: Gordon R, Seckbach J (eds) The science of algal fuels: phycology, geology, biophotonics, genomics and nanotechnology. Springer, DordrechtGoogle Scholar
  16. Das P, Lei W, Aziz SS, Obbard JP (2011) Enhanced algae growth in both phototrophic and mixotrophic culture under blue light. Bioresour Technol 102(4):3883–3887CrossRefGoogle Scholar
  17. Davis R, Aden A (2011) Techno-economic analysis of autotrophic microalgae for fuel production. Appl Energy 88:3524–3531CrossRefGoogle Scholar
  18. Davis SC, Kucharik CJ, Fazio S, Monti A (2013) Environmental sustainability of advanced biofuels. Biofuels Bioprod Biorefin 7:638–646CrossRefGoogle Scholar
  19. De Pauw N, Van Vaerenbergh E (1981) Microalgae wastewater treatment systems: potentials and limits. In: Conference phytodepuration and employment of the biomass produced, Parma, Italy, 15–16 May 1981Google Scholar
  20. De Wever A, Leliaert F, Verleyen E, Vanormelingen P, Van der Gucht K, Hodgson DA, Sabbe K, Vyverman W (2009) Hidden levels of phylodiversity in Antarctic green algae: further evidence for the existence of glacial refugia. Proc Roy Soci B Biol Sci 276:3591–3599CrossRefGoogle Scholar
  21. Downing JB, Bracco E, Green FB, Ku AY, Lundquist TJ, Zubieta IX, Oswald WJ (2002) Low cost wastewater reclamation using the advanced integrated wastewater pond system technology and reverse osmosis. Water Sci Technol 45(1):117–125Google Scholar
  22. Fawley KP, Fawley MW (2007) Observations on the diversity and ecology of freshwater Nannochloropsis (Eustigmatophyceae), with descriptions of new Taxa. Protist 158:325–336CrossRefGoogle Scholar
  23. Frank ED, Elgowainy A, Han J, Wang Z (2013) Life cycle comparison of hydrothermal liquefaction and lipid extraction pathways to renewable diesel from algae. Mitig Adapt Strat Glob Change 18(1):137–158Google Scholar
  24. Fulton LM (2009) Nutrient removal by algae grown in CO2-enriched wastewater over a range of nitrogen-to-phosphorus ratios. Master’s thesis, civil and environmental engineering department, California Polytechnic State University, San Luis Obispo, p 63Google Scholar
  25. Garnier M, Carrier G, Rogniaux H, Nicolau E, Bougaran G, Saint-Jean B, Cadoret JP (2014) Comparative proteomics reveals proteins impacted by nitrogen deprivation in wild-type and high lipid-accumulating mutant strains of Tisochrysis lutea. J Proteomics 105:107–120 S1874-3919(14)00071-2CrossRefGoogle Scholar
  26. Gerken HG, Donohoe B, Knoshaug EP (2013) Enzymatic cell wall degradation of Chlorella vulgaris and other microalgae for biofuels production. Planta 237(1):239–253CrossRefGoogle Scholar
  27. Gladden JM, Allgaier M, Miller CS, Hazen TC, VanderGheynst JS, Hugenholtz P, Simmons BA, Singer SW (2011) Glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass. Appl Environ Microbiol 77:5804–5812CrossRefGoogle Scholar
  28. Griffiths MJ, Harrison STL (2009) Lipid productivity as a key characteristic for choosing algal species for biodiesel production. J Appl Phycol 21(5):493–507CrossRefGoogle Scholar
  29. Grobbelaar JU (2004) Algal nutrition. In: Richmond A (ed) Handbook of microalgal culture: biotechnology and applied phycology. Blackwell Publishing, Ames, pp 97–115Google Scholar
  30. Guckert JB, Cooksey KE, Jackson LL (1988) Lipid solvent systems are not equivalent for analysis of lipid classes in the microeukaryotic green algae, Chlorella. J Microbiol Methods 8:139–149CrossRefGoogle Scholar
  31. Guiry MD (2012) How many species of algae are there? J Phycol 48:1057–1061CrossRefGoogle Scholar
  32. Halim R, Gladman B, Danquah MK, Webley PA (2011) Oil extraction from microalgae for biodiesel production. Bioresour Technol 102:178–185CrossRefGoogle Scholar
  33. Herrero M, Mendiola JA, Cifuentes A, Ibáñez E (2010) Supercritical fluid extraction: recent advances and applications. J Chromatogr A 1217:2495–2511CrossRefGoogle Scholar
  34. Hillebrand H (2011) Temperature mediates competitive exclusion and diversity in benthic microalgae under different N: P stoichiometry. Ecol Res 26(3):533–539CrossRefGoogle Scholar
  35. Horner RA (2002) A taxonomic guide to some common marine phytoplankton. Biopress, Bristol, pp 25–30Google Scholar
  36. Huerlimann R, de Nys Heimann (2010) Growth, lipid content, productivity, and fatty acid composition of tropical microalgae for scale-up production. Biotechnol Bioeng 107(2):245–257CrossRefGoogle Scholar
  37. Illman AM, Scragg AH, Shales SW (2000) Increase in Chlorella strains calorific values when grown in low nitrogen medium. Enzyme Microb Technol 27:631–635CrossRefGoogle Scholar
  38. Jakobsen AN, Aasen IM, Josefsen KD, Strom AR (2008) Accumulation of docosahexaenoic acid-rich lipid in thraustochytrid Aurantiochytrium sp strain T66: effects of N and P starvation and O2 limitation. Appl Microbiol Biotechnol 80(2):297–306CrossRefGoogle Scholar
  39. Kent AD, Triplett EW (2002) Microbial communities and their interactions in soil and rhizosphere ecosystems. Annu Rev Microbiol 56:211–236CrossRefGoogle Scholar
  40. Lang W (1974) Competitive exclusion among three planktonic blue-green algal species. J Phycol 10(4):411–414Google Scholar
  41. Lewis L, Lewis P (2005) Unearthing the molecular phylodiversity of desert soil green algae (Chlorophyta). Syst Biol 54(6):936–947CrossRefGoogle Scholar
  42. Li Y, Horsman M, Wang B, Wu N, Lan CQ (2008) Effects of nitrogen sources on cell growth and lipid accumulation of green alga Neochloris oleoabundans. Appl Microbiol Biotechnol 81(4):629–636CrossRefGoogle Scholar
  43. Lucentinii J (2005) Secondary endosymbiosis exposed. Scientist 19:22–23Google Scholar
  44. Lundquist TJ, Woertz IC, Benemann JR (2011) Algal high rate ponds with CO2 addition for energy efficient nutrient recovery. In: Paper written for the Nutrient Recovery and Management conference, Water Environment Federation, Miami, Florida, p 14Google Scholar
  45. Lundquist TJ, Woertz IC, Quinn NWT, Benemann JR (2010) A realistic technological and economic assessment of algae biofuels. Report prepared for the BP Energy Biosciences Institute. Berkeley, California, p 154Google Scholar
  46. Lv JM, Cheng LH, Xu XH, Zhang L, Chen HL (2010) Enhanced lipid production of Chlorella vulgaris by adjustment of cultivation conditions. Bioresour Technol 101(17):6797–6804CrossRefGoogle Scholar
  47. Mahapatra DM, Ramachandra TV (2013) Algal biofuel: bountiful lipid from Chlorococcum sp. proliferating in municipal wastewater. Curr Sci 105(1):47–55Google Scholar
  48. Martinez ME, Sanchez S, Jimenez JM, El Yousfi F, Munoz L (2000) Nitrogen and phosphorus removal from urban wastewater by the microalga Scenedesmus obliquus. Bioresour Technol 73:263–272CrossRefGoogle Scholar
  49. McCormick PV, Cairns J (1994) Algae as indicators of environmental change. J Appl Phycol 6:509–526CrossRefGoogle Scholar
  50. McGinnis KM, Dempster TA, Sommerfeld MR (1997) Characterization of the growth and lipid content of the diatom Chaetoceros muelleri. J Appl Phycol 9:19–24CrossRefGoogle Scholar
  51. Miaoa X, Wu Q (2006) Biodiesel production from heterotrophic microalgal oil. Bioresour Technol 97:841–846CrossRefGoogle Scholar
  52. Moestrup O (2001a) Algae: phylogeny and evolution. In: Encyclopedia of Life Sciences. Wiley, ChichesterGoogle Scholar
  53. Moestrup O (2001b) Algal taxonomy: historical overview. In: eLS. Wiley, ChichesterGoogle Scholar
  54. Molina Grima EH, Belarbia FG, Acien FA, Robles M, Chisti Y (2003) Recovery of microalgal biomass and metabolites: process options and economics. Biotechnol Adv 20:491–515CrossRefGoogle Scholar
  55. Murry MA, Benemann JR (1980) Fresh and brackish water aquatic plant resources. In: Zaborsky O (ed) Handbook of Biosolar resources, vol II. CRC Press, Boca Raton, pp 407–470Google Scholar
  56. Neori A (2008) Essential role of seaweed cultivation in integrated multi-trophic aquaculture farms for global expansion of mariculture: an analysis. J Appl Phycol 20:567–570CrossRefGoogle Scholar
  57. Oilgae (2009) Oilgae guide to algae-based wastewater treatment. A Sample ReportGoogle Scholar
  58. Ojala A, Tikka M, Valkonen K (2013) Algal cultivation for lipid production in waste water. In: Arnold M (ed) Sustainable algal biomass products by cultivation in waste water flows. VTT Technical Research Centre of Finland, p 147Google Scholar
  59. Oswald WJ (1963) Light efficiency of algae grown in sewage. Trans Am Soc Civ Eng 128:47–83Google Scholar
  60. Oswald WJ (2003) My 60 years in applied algology. J Appl Phycol 15:99–106CrossRefGoogle Scholar
  61. Park JJ, Lechno-Yossef S, Wolk CP, Vieille C (2013) Cell-specific gene expression in Anabaena variabilis grown phototrophically, mixotrophically, and heterotrophically. BMC Genomic 14:759CrossRefGoogle Scholar
  62. Patil PD, Gude VG (2011) Optimization of direct conversion of wet algae to biodiesel under supercritical methanol conditions. Bioresour Technol 102(1):118–122CrossRefGoogle Scholar
  63. Pickett-Heaps J, Schmid AMM, Edgar LA (1990) The cell biology of diatom valve formation. Prog Phycol Res 7:1–168Google Scholar
  64. Ponnuswamy I, Madhavan S, Shabudeen S (2013) Isolation and characterization of green microalgae for carbon sequestration, waste water treatment and bio-fuel production. Int J Bio-Sci Bio-Technol 5(2):17–25Google Scholar
  65. Radner RJ, Parker BC (1994) Commercial applications of algae: opportunities and constraints. J Appl Phycol 6:93–98CrossRefGoogle Scholar
  66. Raeesossadati MJ, Ahmadzadeh H, McHenry MP, Moheimani NR (2014) CO2 bioremediation by microalgae in photobioreactors: impacts of biomass and CO2 concentrations, light, and temperature. Algal Res 6:78–85CrossRefGoogle Scholar
  67. Ranganathan S, Narasimhan S, Muthukumar K (2008) An overview of enzymatic production of biodiesel. Bioresour Technol 99:3975–3981CrossRefGoogle Scholar
  68. Rawson DS (1956) Algal indicators of trophic lake types. Limnol Oceanogr 1(1):18–25CrossRefMathSciNetGoogle Scholar
  69. Renewable fuels association (2012).
  70. Rodolfi LC, Zittelli G, Bassi N, Padovani G, Biondi N, Bonini G, Tredici MR (2009) Microalgae for oil: strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor. Biotechnol Bioeng 102(1):100–112CrossRefGoogle Scholar
  71. Roessler PG (1988) Changes in the activities of various lipid and carbohydrate biosynthetic enzymes in the diatom Cyclotella cryptica in response to silicon deficiency. Arch Biochem Biophys 267:521–528CrossRefGoogle Scholar
  72. Roessler PG (1990) Environmental control of glycerolipid metabolism in microalgae: commercial implications and future research directions. J Phycol 26:393–399CrossRefGoogle Scholar
  73. Rosgaard L, de Porcellinis AJ, Jacobsen JH, Frigaard NU, Sakuragi Y (2012) Bioengineering of carbon fixation, biofuels, and biochemicals in cyanobacteria and plants. J Biotechnol 162(1):134–147CrossRefGoogle Scholar
  74. Round FE, Crawford RM, Mann DG (1990) The diatoms: biology and morphology of the genera. Cambridge University Press, Cambridge, pp 1–11Google Scholar
  75. Sheehan J, Dunahay T, Benemann JR, Roessler P (1987) A look back at the U.S. Department of energy’s aquatic species program: Biodiesel from Algae. National Renewable Energy Laboratory, Golden, CO NERL/TP-580-24190Google Scholar
  76. Spolaore P, Joannis-Cassan C, Duran E, Isambert A (2006) Commercial applications of microalgae. J Biosci Bioeng 101:87–96CrossRefGoogle Scholar
  77. Takagi M, Watanabe K, Yamaberi K, Yoshida T (2000) Limited feeding of potassium nitrate for intracellular lipid and triglyceride accumulation of Nannochloris sp. UTEX LB1999. Appl Microbiol Biotechnol 54:112–117CrossRefGoogle Scholar
  78. Walker TL, Collet C, Purton S (2005) Algal transgenics in the genomic era. J Phycol 41:1077–1093CrossRefGoogle Scholar
  79. Wang W, Liu X, Lu X (2013) Engineering cyanobacteria to improve photosynthetic production of alka(e)nes. Biotechnol Biofuels 6:69CrossRefGoogle Scholar
  80. Weissman JC, Goebel RP (1987) Factors affecting the photosynthetic yield of microalgae. FY 1986 Aquatic Species Program Annual Report, Solar Energy Research Institute, Golden, Colorado, SERI/SP-231-3071, pp 139–168Google Scholar
  81. Williams PJ, Laurens LML (2010) Microalgae as biodiesel & biomass feedstocks: Review & analysis of the biochemistry, energetics & economics. Energy Environ Sci 3:554CrossRefGoogle Scholar
  82. Wisconsin department of natural resources (2010) Introduction to activated sludge study guide, bureau of science services, operator certification program, P.O. Box 7921, Madison, WI 53707.
  83. Woertz IC, Fulton L, Lundquist TJ (2009) Nutrient removal & greenhouse gas abatement with CO2-supplemented algal high rate ponds. In: Paper written for the WEFTEC annual conference, Water Environment FederationGoogle Scholar
  84. Yool A, Tyrrell T (2003) Role of diatoms in regulating the ocean’s silicon cycle. Global Biogeochem Cycles 17(4):1103–1124Google Scholar
  85. Zelitch I (1971) Photosynthesis, photorespiration and plant productivity. Academic Press, Waltham, p 275Google Scholar
  86. Zheng H, Yin J, Gao Z, Huang H, Ji X, Dou C (2011) Disruption of Chlorella vulgaris Cells for the release of biodiesel-producing lipids: a comparison of grinding, ultrasonication, bead milling, enzymatic lysis, and microwaves. Appl Biochem Biotechnol 164:1215–1224CrossRefGoogle Scholar
  87. Zhou Y, Schideman L, Yu G, Zhang Y (2013) A synergistic combination of algal wastewater treatment and hydrothermal biofuel production maximized by nutrient and carbon recycling. Energy Environ Sci 6:3765–3779CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Stephen R. Lyon
    • 1
  • Hossein Ahmadzadeh
    • 2
    Email author
  • Marcia A. Murry
    • 3
    • 4
  1. 1.AlgaXperts, LLCMilwaukeeUSA
  2. 2.Department of ChemistryFerdowsi University of MashhadMashhadIran
  3. 3.California State Polytechnic UniversityPomonaUSA
  4. 4.Sinai Technology CorporationLos AngelesUSA

Personalised recommendations