Skip to main content
SpringerLink
Log in

Effects of Nitrogen and Phosphorus on Biochemical Composition of Microalgae Polyculture Harvested from Phycoremediation of Piggery Wastewater Digestate

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

The effects of nitrogen (N) and/or phosphorus (P) starvation on the biochemical composition of native microalgae Chlorella spp. polyculture obtained from the phycoremediation of swine wastewaters were investigated. Microalgae-specific growth rate of 1.2 day−1 was achieved (30.3 mg L−1 day−1). PO4 −2 and NH3 were completely removed from swine digestate effluent after 3 and 11 days, respectively. Microalgae harvested immediately after nutrient removal showed high protein (56–59 %) and carbohydrate (25–34 %) but low lipid (1.8–3 %) contents. Depletion of N or P alone stimulated carbohydrate production at the expenses of proteins. Significant lipid accumulation from 3 % ± 0.5 to 16.3 % ± 0.8 was reached only after 25 days following N and P starvation as demonstrated by Nile red-stained cells. Regarding to the effects of harvesting methods on cellular biochemical composition, circumstantial evidences indicate that coagulation–flocculation with tannin may lead to lower protein and lipid amounts but increased carbohydrate content (p < 0.01) as compared to centrifugation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Mezzari, M. P., Da Silva, M. L. B., Nicoloso, R. S., Ibelli, A. M. G., Bortoli, M., Viancelli, A., & Soares, H. M. (2013). Assessment of N2O emission from a photobioreactor treating ammonia-rich swine wastewater digestate. Bioresource Technology, 149, 327–332. doi:10.1016/j.biortech.2013.09.065.

    Article  CAS  Google Scholar 

  2. Wang, H., Xiong, H., Hui, Z., & Zeng, X. (2012). Mixotrophic cultivation of Chlorella pyrenoidosa with diluted primary piggery wastewater to produce lipids. Bioresource Technology, 104, 215–220. doi:10.1016/j.biortech.2011.11.020.

    Article  CAS  Google Scholar 

  3. Prajapati, S. K., Kaushik, P., Malik, A., & Vijay, V. K. (2013). Phycoremediation and biogas potential of native algal isolates from soil and wastewater. Bioresource Technology, 135, 232–238.

    Article  CAS  Google Scholar 

  4. Choi, H. J., Seung, C., & Lee, M. (2014). Effect of the N/P ratio on biomass productivity and nutrient removal from municipal wastewater. Bioprocess and Biosystems Engineering, 38, 761–766. doi:10.1007/s00449-014-1317-z.

    Article  Google Scholar 

  5. Choi, H. J. (2014). Effect of optical panel distance in a photobioreactor for nutrient removal and cultivation of microalgae. World Journal of Microbiology and Biotechnology, 30, 2015–2023. doi:10.1007/s11274-014-1626-z.

    Article  CAS  Google Scholar 

  6. Gupta, P. L., & Choi, S. L. H. (2015). A mini review: photobioreactors for large scale algal cultivation. World Journal of Microbiology and Biotechnology, 31(9), 1409–1417. doi:10.1007/s11274-015-1892-4.

    Article  CAS  Google Scholar 

  7. Rasala, B. A., & Mayfield, S. P. (2014). Photosynthetic biomanufacturing in green algae: production of recombinant proteins for industrial, nutritional, and medical uses. Photosynthesis Research, 123(3), 227–239.

    Article  Google Scholar 

  8. Zhu, S., Huang, W., Xu, J., Wang, Z., Xu, J., & Yuan, Z. (2014). Metabolic changes of starch and lipid triggered by nitrogen starvation in the microalga Chlorella zofingiensis. Bioresource Technology, 152, 292–298. doi:10.1016/j.biortech.2013.10.092.

    Article  CAS  Google Scholar 

  9. Ho, S. H., Huang, S. W., Chen, C. Y., Hasunuma, T., Kondo, A., & Chang, J. S. (2013). Characterization and optimization of carbohydrate production from an indigenous microalga Chlorella vulgaris FSP-E. Bioresource Technology, 135, 157–165.

    Article  CAS  Google Scholar 

  10. Ho, S. H., Chen, C. Y., & Chang, J. S. (2012). Effect of light intensity and nitrogen starvation on CO2 fixation and lipid/carbohydrate production of an indigenous microalga Scenedesmus obliquus CNW-N. Bioresource Technology, 113, 244–252.

    Article  CAS  Google Scholar 

  11. Khalil, Z. I., Asker, M. M. S., El-Sayed, S., & Kobbia, I. A. (2010). Effect of pH on growth and biochemical responses of Dunaliella bardawil and Chlorella ellipsoidea. World Journal of Microbiology and Biotechnology, 26(7), 1225–1231.

    Article  CAS  Google Scholar 

  12. Roleda, M. Y., Slocombe, S. P., Leakey, R. J. G., Day, J. G., Bell, E. M., & Stanley, M. S. (2013). Effects of temperature and nutrient regimes on biomass and lipid production by six oleaginous microalgae in batch culture employing a two-phase cultivation strategy. Bioresource Technology, 129, 439–449.

    Article  CAS  Google Scholar 

  13. Ruangsomboon, S., Ganmanee, M., & Choochote, S. (2013). Effects of different nitrogen, phosphorus, and iron concentrations and salinity on lipid production in newly isolated strain of the tropical green microalga, Scenedesmus dimorphus KMITL. Journal of Applied Phycology, 25(3), 867–874.

    Article  CAS  Google Scholar 

  14. Sun, X., Cao, Y., Xu, H., Liu, Y., Sun, J., Qiao, D., & Cao, Y. (2014). Effect of nitrogen-starvation, light intensity and iron on triacylglyceride/carbohydrate production and fatty acid profile of Neochloris oleoabundans HK-129 by a two-stage process. Bioresource Technology, 155, 204–212. doi:10.1016/j.biortech.2013.12.109.

    Article  CAS  Google Scholar 

  15. Lowrey, J., Brooks, M. S., & McGinn, P. J. (2014). Heterotrophic and mixotrophic cultivation of microalgae for biodiesel production in agricultural wastewaters and associated challenges. Journal of Applied Phycology, 27(4), 1–14. doi:10.1007/s10811-014-0459-3.

    Google Scholar 

  16. Lee, S. J., Kim, S. B., Kim, J. E., Kwon, G. S., Yoon, B. D., & Oh, H. M. (1998). Effects of harvesting method and growth stage on the flocculation of the green alga Botryococcus braunii. Letters in Applied Microbiology, 27(1), 14–18.

    Article  Google Scholar 

  17. Liang, K., Zhang, Q., Gu, M., & Cong, W. (2013). Effect of phosphorus on lipid accumulation in freshwater microalga Chlorella sp. Journal of Applied Phycology, 25(1), 311–318.

    Article  CAS  Google Scholar 

  18. Li, Y., Han, F., Xu, H., Mu, J., Chen, D., Feng, B., & Zeng, H. (2014). Potential lipid accumulation and growth characteristic of the green alga Chlorella with combination cultivation mode of nitrogen (N) and phosphorus (P). Bioresource Technology, 174, 24–32. doi:10.1016/j.biortech.2014.09.142.

    Article  CAS  Google Scholar 

  19. Coward, T., Lee, J. G. M., & Caldwell, G. S. (2014). Harvesting microalgae by CTAB-aided foam flotation increases lipid recovery and improves fatty acid methyl ester characteristics. Biomass and Bioenergy, 67, 354–362.

    Article  CAS  Google Scholar 

  20. Borges, L., Morón-Villarreyes, J. A., D’Oca, M. G. M., & Abreu, P. C. (2011). Effects of flocculants on lipid extraction and fatty acid composition of the microalgae Nannochloropsis oculata and Thalassiosira weissflogii. Biomass and Bioenergy, 35(10), 4449–4454.

    Article  CAS  Google Scholar 

  21. Bonato, A.L.V., do Valle, C.B., Jank, L., Resende, R.M.S., & Leguizamon, G.O.C. (2002). Extração de DNA genômico de Brachiaria e Panicum maximum. Campo Grande, MS, BRAZIL. doi: ISSN 1516–9308.

  22. White, T. J., Bruns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications, 18, 315–322.

    Google Scholar 

  23. Koetschan, C., Hackl, T., Müller, T., Wolf, M., Förster, F., & Schultz, J. (2012). ITS2 database IV: interactive taxon sampling for internal transcribed spacer 2 based phylogenies. Molecular Phylogenetics and Evolution, 63(3), 585–588. doi:10.1016/j.ympev.2012.01.026.

    Article  CAS  Google Scholar 

  24. Hall, J. D., Fucikova, K., Lo, C., Lewis, L. A., & Karol, K. G. (2010). An assessment of proposed DNA barcodes in freshwater green algae. Cryptogamie Algologie, 31(4), 529–555.

    Google Scholar 

  25. APHA. (2012). Standard methods for the examination for water and wastewater. (D. American Water Works Association, Washington, Ed.) (22nd Ed.).

  26. Porra, R. J., Thompson, W. a., & Kriedemann, P. E. (1989). Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochimica et Biophysica Acta (BBA) - Bioenergetics, 975(3), 384–394. doi:10.1016/S0005-2728(89)80347-0.

    Article  CAS  Google Scholar 

  27. Held, P., & Raymond, K. (2011). Determination of algal cell lipids using Nile red—using microplates to monitor neutral lipids in Chlorella vulgaris. Biofuel Research, 8, 1–5.

    Google Scholar 

  28. Mezzari, M. P., da Silva, M. L. B., Pirolli, M., Perazzoli, S., Steinmetz, R. L. R., Nunes, E. O., & Soares, H. M. (2014). Assessment of a tannin-based organic polymer to harvest Chlorella vulgaris biomass from swine wastewater digestate phycoremediation. Water Science and Technology, 70(5), 888–894.

    Article  CAS  Google Scholar 

  29. Chu, F. F., Chu, P. N., Cai, P. J., Li, W. W., Lam, P. K. S., & Zeng, R. J. (2013). Phosphorus plays an important role in enhancing biodiesel productivity of Chlorella vulgaris under nitrogen deficiency. Bioresource Technology, 134, 341–346.

    Article  CAS  Google Scholar 

  30. Chu, F. F., Chu, P. N., Shen, X. F., Lam, P. K. S., & Zeng, R. J. (2014). Effect of phosphorus on biodiesel production from Scenedesmus obliquus under nitrogen-deficiency stress. Bioresource Technology, 152, 241–246.

    Article  CAS  Google Scholar 

  31. Ho, S. H., Nakanishi, A., Ye, X., Chang, J. S., Hara, K., Hasunuma, T., & Kondo, A. (2014). Optimizing biodiesel production in marine Chlamydomonas sp. JSC4 through metabolic profiling and an innovative salinity-gradient strategy. Biotechnology for Biofuels, 7(1), 97. doi:10.1186/1754-6834-7-97.

    Article  Google Scholar 

  32. Roopnarain, A., Gray, V. M., & Sym, S. D. (2014). Phosphorus limitation and starvation effects on cell growth and lipid accumulation in Isochrysis galbana U4 for biodiesel production. Bioresource Technology, 156, 408–411.

    Article  CAS  Google Scholar 

  33. AOCS. (2013). Official methods and recommended practices of the AOCS. (D. Firestone, Ed.) (6th ed.). Champaign: American Oil Chemists’ Society.

    Google Scholar 

  34. AOAC. (1990). Official methods of analysis of AOAC International. (Kenneth Helrich, Ed.) (15th ed.). Washington: Association of Official Analytical Chemists.

    Google Scholar 

  35. BCAA. (2009). Ash or mineral matter. In Brazilian Compendium of Animal Nutrition (p. 204). São José do Rio Preto, SP, Brazil.

  36. Bi, Z., & He, B. B. (2013). Characterization of microalgae for the purpose of biofuel production. Transactions of the ASABE, 56(4), 1529–1539.

    CAS  Google Scholar 

  37. Pancha, I., Chokshi, K., George, B., Ghosh, T., Paliwal, C., Maurya, R., & Mishra, S. (2014). Nitrogen stress triggered biochemical and morphological changes in the microalgae Scenedesmus sp. CCNM 1077. Bioresource Technology, 156, 146–154. doi:10.1016/j.biortech.2014.01.025.

    Article  CAS  Google Scholar 

  38. Ikaran, Z., Suárez-Alvarez, S., Urreta, I., & Castañón, S. (2015). The effect of nitrogen limitation on the physiology and metabolism of Chlorella vulgaris var L3. Algal Research, 10, 134–144. doi:10.1016/j.algal.2015.04.023.

    Article  Google Scholar 

  39. Bondioli, P., Della Bella, L., Rivolta, G., Chini Zittelli, G., Bassi, N., Rodolfi, L., Casini, D., Prussi, M., Chiaramonti, D., & Tredici, M. R. (2012). Oil production by the marine microalgae Nannochloropsis sp. F&M-M24 and Tetraselmis suecica F&M-M33. Bioresource Technology, 114, 567–572.

    Article  CAS  Google Scholar 

  40. Peccia, J., Haznedaroglu, B., Gutierrez, J., & Zimmerman, J. B. (2013). Nitrogen supply is an important driver of sustainable microalgae biofuel production. Trends in Biotechnology, 31(3), 134–138.

    Article  CAS  Google Scholar 

  41. Bellou, S., Baeshen, M. N., Elazzazy, A. M., Aggeli, D., Sayegh, F., & Aggelis, G. (2014). Microalgal lipids biochemistry and biotechnological perspectives. Biotechnology Advances, 32(6), 1476–1493. doi:10.1016/j.biotechadv.2014.10.003.

    Article  CAS  Google Scholar 

  42. Zhao, Y., Yu, Z., Song, X., & Cao, X. (2009). Biochemical compositions of two dominant bloom-forming species isolated from the Yangtze River Estuary in response to different nutrient conditions. Journal of Experimental Marine Biology and Ecology, 368(1), 30–36.

  43. Said, H. A. (2009). Changes in levels of cellular constituents of Dunaliella parva associated with inorganic phosphate depletion. Middle-East Journal of Scientific Research, 4(2), 44–49.

    Google Scholar 

Download references

Acknowledgments

This research was supported by CAPES-EMBRAPA (#001/2011) and EMBRAPA (#02.12.08.004.00.05). We thank Dr. Mariana Groke for the photomicrographs and Dr. Bruno dos Santos A. F. Brazil for microalgae identification.

Author information

Authors and Affiliations

  1. Department of Chemical Engineering, Federal University of Santa Catarina, Florianópolis, SC, 88040-900, Brazil

    William Michelon, Mateus Pirolli, Jean Michel Prandini & Hugo Moreira Soares

  2. EMBRAPA Swine and Poultry, P.O. Box 321, Concórdia, SC, 89700-000, Brazil

    Marcio Luis Busi Da Silva

  3. Biotechnology and Sciences Program, West University of Santa Catarina, Videira, SC, 89560-000, Brazil

    Melissa Paola Mezzari

Authors
  1. William Michelon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marcio Luis Busi Da Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Melissa Paola Mezzari
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mateus Pirolli
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jean Michel Prandini
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hugo Moreira Soares
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Marcio Luis Busi Da Silva.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Michelon, W., Da Silva, M.L.B., Mezzari, M.P. et al. Effects of Nitrogen and Phosphorus on Biochemical Composition of Microalgae Polyculture Harvested from Phycoremediation of Piggery Wastewater Digestate. Appl Biochem Biotechnol 178, 1407–1419 (2016). https://doi.org/10.1007/s12010-015-1955-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-015-1955-x

Keywords

Search

Navigation