Effect of trophic conditions on microalga growth, nutrient removal, algal organic matter, and energy storage products in Scenedesmus (Acutodesmus) obliquus KGE-17 cultivation

  • Wook Jin Choi
  • A. Na Chae
  • Kyung Guen SongEmail author
  • Joonhong ParkEmail author
  • Byung Chan Lee
Research Paper


This study compared the performance of microalga growth, nutrient removal, algal organic matter, and energy storage products in mixotrophic, photoautotrophic, and heterotrophic conditions. Scenedesmus obliquus was used as a model species. Mixotrophic condition showed the highest specific growth rate of 0.96 d−1 as well as the fastest nitrogen and phosphorus removal rate of 85.17 mg-N g-cell−1 day−1 and 11.49 mg-P g-cell−1 day−1, respectively, compared with photoautotrophic and heterotrophic conditions. Mixotrophic microalgae had relatively higher carbohydrates and lipids contents (21.8 and 24.0%) than photoautotrophic and heterotrophic conditions. Meanwhile, algal organic matter (AOM) in the medium was produced at the highest level under photoautotrophic condition. Mixotrophic condition was more efficient in terms of microalga growth, nutrient removal, production of energy storage products, and suppression of AOM, and would be adaptable for wastewater treatment process.


Microalgae Heterotrophic Mixotrophic Photoautotrophic Scenedesmus obliquus 



This work was supported by a Grant (18CTAP-C116746-03) from the Technology Advancement Research Program funded by the Ministry of Land, Infrastructure and Transport of the Korean government, and the Korea Institute of Science and Technology (KIST) Institutional Program (2E29660).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

449_2019_2120_MOESM1_ESM.docx (382 kb)
Supplementary file1 (DOCX 381 kb)


  1. 1.
    Oswald WJ, Gotaas HB (1957) Photosynthesis in sewage treatment. Trans Am Soc Civ Eng 122:73–105Google Scholar
  2. 2.
    Umamaheswari J, Shanthakumar S (2016) Efficacy of microalgae for industrial wastewater treatment: a review on operating conditions, treatment efficiency and biomass productivity. Rev Environ Sci Biotechnol 15:265–284Google Scholar
  3. 3.
    Li Y, Chen YF, Chen P, Min M, Zhou W, Martinez B, Zhu J, Ruan R (2011) Characterization of a microalga Chlorella sp. well adapted to highly concentrated municipal wastewater for nutrient removal and biodiesel production. Bioresour Technol 102:5138–5144Google Scholar
  4. 4.
    Kim HC, Choi WJ, Chae AN, Park J, Kim HJ, Song KG (2016) Evaluating integrated strategies for robust treatment of high saline piggery wastewater. Water Res 89:222–231Google Scholar
  5. 5.
    Dang NM, Lee K (2018) Recent trends of using alternative nutrient sources for microalgae cultivation as a feedstock of biodiesel production. Appl Chem Eng 29(1):1–9Google Scholar
  6. 6.
    Suali E, Sarbatly R (2012) Conversion of microalgae to biofuel. Renew Sust Energ Rev 16:4316–4342Google Scholar
  7. 7.
    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 73:263–272Google Scholar
  8. 8.
    Sydney EB, da Silva TE, Tokarski A, Novak AC, de Carvalho JC, Woiciecohwski AL, Larroche C, Soccol CR (2011) Screening of microalgae with potential for biodiesel production and nutrient removal from treated domestic sewage. Appl Energy 88:3291–3294Google Scholar
  9. 9.
    Schulze PSC, Carvalho CFM, Pereira H, Gangadhar KN, Schuler LM, Santos TF, Varela JCS, Barreira L (2017) Urban wastewater treatment by Tetraselmis sp. CTP4 (Chlorophyta). Bioresour Technol 223:175–183Google Scholar
  10. 10.
    Perez-Garcia O, Escalante FM, de Bashan LE, Bashan Y (2011) Heterotrophic cultures of microalgae: metabolism and potential products. Water Res 45:11–36Google Scholar
  11. 11.
    Combres C, Laliberté G, Sevrin Reyssac J, de la Noüe J (1994) Effect of acetate on growth and ammonium uptake in the microalga Scenedesmus obliquus. Physiol Plant 91:729–734Google Scholar
  12. 12.
    Babaei A, Mehrnia MR, Shayegan J, Sarrafzadeh M-H, Amini E (2018) Evaluation of nutrient removal and biomass production through mixotrophic, heterotrophic, and photoautotrophic cultivation of chlorella in nitrate and ammonium wastewater. Int J Environ Res 12:167–178Google Scholar
  13. 13.
    Kim S, Park JE, Cho YB, Hwang SJ (2013) Growth rate, organic carbon and nutrient removal rates of Chlorella sorokiniana in autotrophic, heterotrophic and mixotrophic conditions. Bioresour Technol 144:8–13Google Scholar
  14. 14.
    Ogawa T, Aiba S (1981) Bioenergetic analysis of mixotrophic growth in Chlorella vulgaris and Scenedesmus acutus. Biotechnol Bioeng 23:1121–1132Google Scholar
  15. 15.
    Marquez FJ, Sasaki K, Kakizono T, Nishio N, Nagai S (1993) Growth characteristics of Spirulina platensis in mixotrophic and heterotrophic conditions. J Ferment Bioeng 76:408–410Google Scholar
  16. 16.
    Quinn J, de Winter L, Bradley T (2011) Microalgae bulk growth model with application to industrial scale systems. Bioresour Technol 102:5083–5092Google Scholar
  17. 17.
    Cheirsilp B, Torpee S (2012) Enhanced growth and lipid production of microalgae under mixotrophic culture condition: effect of light intensity, glucose concentration and fed-batch cultivation. Bioresour Technol 110:510–516Google Scholar
  18. 18.
    Mohammad-Mirzaie MA, Kalbasi M, Mousavi SM, Ghobadian B (2016) Investigation of mixotrophic, heterotrophic, and autotrophic growth of Chlorella vulgaris under agricultural waste medium. Prep Biochem Biotechnol 46:150–156Google Scholar
  19. 19.
    Park YT, Lee H, Yun HS, Song KG, Yeom SH, Choi J (2013) Removal of metal from acid mine drainage using a hybrid system including a pipes inserted microalgae reactor. Bioresour Technol 150:242–248Google Scholar
  20. 20.
    Hegewald EH (1997) Taxonomy and phylogeny of Scenedesmus. Algae 12:235–246Google Scholar
  21. 21.
    Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61Google Scholar
  22. 22.
    APHA, AWWA, WEF (1998) Standard methods for the examination of water and wastewater 21st edn. American Public Health Association, WashingtonGoogle Scholar
  23. 23.
    Huber SA, Balz A, Abert M, Pronk W (2011) Characterisation of aquatic humic and non-humic matter with size-exclusion chromatography—organic carbon detection—organic nitrogen detection (LC-OCD-OND). Water Res 45:879–885Google Scholar
  24. 24.
    Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Calorimetric method for determination of sugars and related substances. Anal Chem 28:350–356Google Scholar
  25. 25.
    Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem phys 37:911–917Google Scholar
  26. 26.
    Kim H-C, Lee S (2006) Pump diffusion flash mixing (PDFM) for improving coagulation process in drinking water treatment. Sep Purif Technol 52:117–125Google Scholar
  27. 27.
    Lamsal R, Walsh ME, Gagnon GA (2011) Comparison of advanced oxidation processes for the removal of natural organic matter. Water Res 45:3263–3269Google Scholar
  28. 28.
    Laliberté G, de la Noüe J (1993) Auto-, hetero-, and mixotrophic growth of Chlamydomonas humicola (Chlorophyceae) on acetate. J Phycol 29:612–620Google Scholar
  29. 29.
    Smith RT, Bangert K, Wilkinson SJ, Gilmour DJ (2015) Synergistic carbon metabolism in a fast growing mixotrophic freshwater microalgal species Micractinium inermum. Biomass Bioenerg 82:73–86Google Scholar
  30. 30.
    Ho SH, Chen CY, Chang JS (2012) Effect of light intensity and nitrogen starvation on CO2 fixation and lipid/carbohydrate production of an indigenous microalga Scenedesmus obliquus CNW-N. Bioresour Technol 113:244–252Google Scholar
  31. 31.
    Huang A, Sun L, Wu S, Liu C, Zhao P, Xie X, Wang G (2016) Utilization of glucose and acetate by Chlorella and the effect of multiple factors on cell composition. J Appl Phycol 29(1):23–33Google Scholar
  32. 32.
    Geider RJ, Osborne BA (1989) Respiration and microalgal growth: a review of the quantitative relationship between dark respiration and growth. New Phytol 112(3):327–341Google Scholar
  33. 33.
    Cardol P, Forti G, Finazzi G (2011) Regulation of electron transport in microalgae. Biochim Biophys Acta Bioenerg 1807:912–918Google Scholar
  34. 34.
    Wang ZW, Liu Y, Tay JH (2007) Biodegradability of extracellular polymeric substances produced by aerobic granules. Appl Microbiol Biotechnol 74:462–466Google Scholar
  35. 35.
    Barker DJ, Stuckey DC (1999) A review of soluble microbial products (SMP) in wastewater treatment systems. Water Res 33:3063–3082Google Scholar
  36. 36.
    Trabelsi L, Ouada HB, Zili F, Mazhoud N, Ammar J (2013) Evaluation of Arthrospira platensis extracellular polymeric substances production in photoautotrophic, heterotrophic and mixotrophic conditions. Folia Microbiol 58:39–45Google Scholar
  37. 37.
    Trabelsi L, Ouada HB, Bacha H, Ghoul M (2009) Combined effect of temperature and light intensity on growth and extracellular polymeric substances production by the cyanobacterium Arthrospira platensis. J Appl Phycol 21:405–412Google Scholar
  38. 38.
    Guillaume-Cogne JB, Gros-Dussap CG (2003) Identification of a metabolic network structure representative of Arthrospira (spirulina) platensis metabolism. Biotechnol Bioeng 84:667–676Google Scholar
  39. 39.
    Markou G, Angelidaki I, Georgakakis D (2012) Microalgal carbohydrates: an overview of the factors influencing carbohydrates production, and of main bioconversion technologies for production of biofuels. Appl Microbiol Biotechnol 96:631–645Google Scholar
  40. 40.
    Dang NM, Lee K (2018) Decolorization of organic fertilizer using advanced oxidation process and its application for microalgae cultivation. J Ind Eng Chem 59:297–303Google Scholar
  41. 41.
    Mandal S, Mallick N (2009) Microalga Scenedesmus obliquus as a potential source for biodiesel production. Appl microbiol biotechnol 84:281–291Google Scholar
  42. 42.
    Ratha SK, Babu S, Renuka N, Prasanna R, Prasad RB, Saxena AK (2013) Exploring nutritional modes of cultivation for enhancing lipid accumulation in microalgae. J Basic Microbiol 53:440–450Google Scholar
  43. 43.
    Mendoza H, Jiménez del Río M, García-Reina G, Ramazanov Z (1996) Low temperature induced β-carotene and fatty acid synthesis, and ultrastructural reorganization of the chloroplast in Dunaliella salina (Chlorophyta). Eur J Phycol 31:329–331Google Scholar
  44. 44.
    Mendoza H, Martel A, Jiménez del Río M, García-Reina G (1999) Oleic acid is the main fatty acid related with carotenogenesis in Dunaliella salina. J Appl Phycol 11:15–19Google Scholar
  45. 45.
    Wang H, Xiong H, Hui Z, Zeng X (2012) Mixotrophic cultivation of Chlorella pyrenoidosa with diluted primary piggery wastewater to produce lipids. Bioresour Technol 104:215–220Google Scholar
  46. 46.
    Bamgboye AI, Hansen AC (2008) Prediction of cetane number of biodiesel fuel from the fatty acid methyl ester composition. Int Agrophys 22:21–29Google Scholar
  47. 47.
    Ramos MJ, Fernandez CM, Casas A, Rodriguez L, Perez A (2009) Influence of fatty acid composition of raw materials on biodiesel properties. Bioresour Technol 100:261–268Google Scholar
  48. 48.
    UNE-EN 14214 (2003) Automotive fuels – fatty acid methyl esters (FAME) for diesel engines — requirement methods, European Committee for Standardization (CEN), Brussels, BelgiumGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Center for Water Resource Cycle ResearchKorea Institute of Science and TechnologySeoulSouth Korea
  2. 2.Department of Civil and Environmental EngineeringYonsei UniversitySeoulSouth Korea
  3. 3.Department of Civil Engineering and Landscape ArchitectureSuncheon Jeil CollegeSuncheonSouth Korea

Personalised recommendations