Skip to main content

Influence of Exogenous CO2 on Biomass and Lipid Accumulation of Microalgae Auxenochlorella protothecoides Cultivated in Concentrated Municipal Wastewater

Applied Biochemistry and Biotechnology volume 166, pages 1661–1673 (2012)Cite this article

Abstract

The effects of exogenous CO2 on the growth and lipid accumulation of a local screened facultative heterotrophic microalgae strain Auxenochlorella protothecoides (UMN280) as well as nutrient removal from concentrated municipal wastewater stream (centrate) were examined in this study. A 12-day batch experiment was conducted with CO2 aeration at three levels, namely, 0%, 1%, and 5% (v/v) CO2 mixed with air, under light intensity of 60 μmol/(m2 @@s). A two-stage growth pattern was observed. The first stage (first–fifth day) was dominated by heterotrophic growth in which organic carbon was the main carbon source. The second stage (6th–12th day) was dominated by autotrophic growth in which exogenous CO2 had a positive effect on algal biomass accumulation. The addition of 5% CO2 was better than that of 1% CO2 on the biomass and lipid production. The uptakes of nutrients were similar between injection and no injection of CO2, except on phosphorus removal which was affected by the acidification of CO2.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Lau, P. S., Tam, N. F. Y., & Wong, Y. S. (1996). Wastewater nutrients removal by Chlorella vulgaris: optimization through acclimation. Environmental Technology, 17(2), 183–189.

    Article  CAS  Google Scholar 

  2. Hernandez, J. P., de-Bashan, L. E., & Bashan, Y. (2005). Starvation enhances phosphorus removal from wastewater by the microalga Chlorella spp. Co-immobilized with Azospirillum brasilense. Enzyme and Microbial Technology, 38(2006), 190–198.

    Google Scholar 

  3. Li, Y., Chen, Y. F., Chen, P., Min, M., Zhou, W., Martinez, B., et al. (2011). Characterization of a microalgae Chlorella sp. well adapted to highly concentrated municipal wastewater in nutrient removal and biodiesel production. Bioresource Technology, 102(8), 5138–5144.

    Article  CAS  Google Scholar 

  4. Wang, L., Wang, Y., Chen, P., & Ruan, R. (2010). Semi-continuous cultivation of Chlorella vulgaris for treating undigested and digested dairy manure. Applied Biochemistry and Biotechnology, 162(8), 2324–2332.

    Article  CAS  Google Scholar 

  5. Mulbry, W. W., & Wilkie, A. C. (2001). Growth of benthic freshwater algae on dairy manure. Journal of Applied Phycology, 13, 301–306.

    Article  Google Scholar 

  6. Spatharis, S., Danielidis, D. B., & Tsirtsis, G. T. (2007). Recurrent Pseudo-nitzschia calliantha (Bacillariophyceae) and Alexandrium insuetum (Dinophyceae) winter blooms induced by agricultural runoff. Harmful Algae, 6(6), 811–822.

    Article  Google Scholar 

  7. Kumar, A., Ergas, S., Yuan, X., Sahu, A., Zhang, Q., Dewulf, J., et al. (2010). Enhanced CO2 fixation and biofuel production via microalgae: recent developments and future directions. Trends in Biotechnology, 28, 371–380.

    Article  CAS  Google Scholar 

  8. Andrade, M. R., & Costa, J. A. V. (2007). Mixotrophic cultivation of microalga Spirulina platensis using molasses as organic substrate. Aquaculture, 264(1–4), 130–134.

    Article  Google Scholar 

  9. Lee, H. Y., Lee, S. Y., & Park, B. K. (1989). The estimation of algal yield parameters associated with mixotrophic and photoheterotrophic growth under batch cultivation. Biomass, 18(2), 153–160.

    Article  Google Scholar 

  10. Xu, F., Hu, H., Cong, W., Cai, Z., & Ouyang, F. (2004). Growth characteristics and eicosapentaenoic acid production by Nannochloropsis sp. in mixotrophic conditions. Biotechnology Letters, 26(1), 51–53.

    Article  CAS  Google Scholar 

  11. Martínez, F., & OrÚs, M. I. (1991). Interactions between glucose and inorganic carbon metabolism in Chlorella vulgaris strain UAM101. Plant Physiology, 95, 1150–1155.

    Article  Google Scholar 

  12. Kobayashi, M., Kakizono, T., Yamagichi, K., Nishio, N., & Nagai, S. (1992). Growth and astaxanthin formation of Haematococcus pluvialls in heterotrophic and mixotrophic conditions. Journal of Fermentation and Bioengineering, 74(1), 17–20.

    Article  CAS  Google Scholar 

  13. Chiu, S., Kao, C., Chen, C., Kuan, T., Ong, S., & Lin, C. (2008). Reduction of CO2 by a high-density culture of Chlorella sp. in semicontinuous photobioreactor. Bioresource Technology, 99(9), 3389–3396.

    Article  CAS  Google Scholar 

  14. Yang, Y., & Gao, K. (2003). Effects of CO2 concentrations on the freshwater microalgae Chlamydomonas reinhardtii, Chlorella pyrenoidosa and Scenedesmus obliquus (Chlorophyta). Journal of Applied Phycology, 15, 379–389.

    Article  CAS  Google Scholar 

  15. Kodama, M., Ikemoto, H., & Miyachi, S. (1993). A new species of highly CO2-tolerant fast-growing marine microalga suitable for high-density culture. Journal of Marine Biotechnology, 1(1), 21–25.

    Google Scholar 

  16. Ingram, L. O., Calder, J. A., Van Baalen, C., Plucker, F. E., & Parker, P. L. (1973). Role of reduced exogenous organic compounds in the physiology of the blue-green bacteria (algae): photoheterotrophic growth of a “heterotrophic” blue-green bacterium. Journal of Bacteriology, 114(2), 695–700.

    CAS  Google Scholar 

  17. Baalan, C. V., Pulich, W. M., & Brandeis, M. G. (1973). Heterotrophic growth of the microalgae. CRC Critical Reviews in Microbiology, 2(2), 229–254.

    Article  Google Scholar 

  18. Darley, W. M., Wimpee, B. B., & Ohlman, C. T. (1981). Heterotrophic and photoheterotrophic utilization of lactate by the diatom, Cylindrotheca fusiformis. British Phycological Journal, 16, 423–428.

    Article  Google Scholar 

  19. Zhou, W., Li, Y., Min, M., Hu, B., Chen, P., & Ruan, R. (2011). Local bioprospecting for high-lipid producing microalgal strains to be grown on concentrated municipal wastewater for biofuel production. Bioresource Technology. doi:10.1016/j.biortech.2011.04.038.

  20. Huss, V. A., Ciniglia, C., Cennamo, P., Cozzolino, S., Pinto, G., & Pollio, A. (2002). Phylogenetic relationships and taxonomic position of Chlorella-like isolates from low pH environments (pH < 3.0). BMC Evolutionary Biology, 2, 13.

    Article  Google Scholar 

  21. Rippka, R., Deruelles, J., Waterbury, J., Herdman, M., & Stanier, R. (1979). Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Journal of General Microbiology, 111, 1–61.

    Google Scholar 

  22. APHA, AWWA, WEF. (1995). Standard methods for the examination of water and wastewater (19th ed.). Washington: American Public Health Association.

    Google Scholar 

  23. Hach. Procedure manual. (2008) Hach, Loveland, CO.

  24. Folch, J., Lees, M., & Sloane Stanley, G. H. (1956) A simple method for the isolation and purification of total lipides from animal tissues. Journal of Biological Chemistry. 497–509.

  25. Tredici, M. R., Papuzzo, T., & Tomasell, L. (1986). Outdoor mass culture of Spirulina platensis in sea-water. Applied Microbiology and Biotechnology, 24(1), 47–50.

    Article  Google Scholar 

  26. Chen, F., & Zhang, Y. (1997). High cell density mixotrophic culture of Spirulina platensis on glucose for phycocyanin production using a fed-batch system. Enzyme and Microbial Technology, 20(3), 21–224.

    Google Scholar 

  27. Agusti, S., Duarte, C. M., & Kalff, J. (1987). Algal cell size and the maximum density and biomass of phytoplankton. Limnology and Oceanography, 32(4), 983–986.

    Article  Google Scholar 

  28. James, C. M., Al-Khars, A. M., & Chorbani, P. (1988). pH dependent growth of Chlorella in a continuous culture system. Journal of the World Aquaculture Society, 19, 27–35.

    Article  Google Scholar 

  29. Pitter, P. (1976). Determination of biological degradability of organic substrances. Water Research, 10(3), 231–235.

    Article  CAS  Google Scholar 

  30. Amblard, C., Couture, P., & Bourdier, G. (1990). Effects of a pulp and paper mill effluent on the structure and metabolism of perphytic algae in experimental streams. Aquatic Toxicology, 18(3), 137–161.

    Article  CAS  Google Scholar 

  31. Morris, I., Yentsch, C. M., & Yentsch, C. S. (1971). Relationship between light carbon dioxide fixation and dark carbon dioxide fixation by marine algae. Limnology and Oceanography, 16(6), 854–858.

    Article  Google Scholar 

  32. OECD: Eutrophication of waters. (1982). Monitoring, assessment and control. Pairs: OECD Publications.

    Google Scholar 

  33. González, L. E., Cañizares, R. O., & Baena, S. (1997). Efficiency of ammonia and phosphorus removal from a colombian agroindustrial wastewater by the microalgae Chlorella vulgaris and Scenedesmus dimorphus. Bioresource Technology, 60, 259–262.

    Article  Google Scholar 

  34. Zhou, A., Tang, H., & Wang, D. (2005). Phosphorus adsorption on natural sediments: modeling and effects of pH and sediment composition. Water Research, 39(7), 1245–1254.

    Article  CAS  Google Scholar 

  35. Boyd, C. E. (1982). Water quality management for pond fish culture. Amsterdam: Elsevier Scientific.

    Google Scholar 

  36. Ferrara, R. A., & Avci, C. B. (1982). Nitrogen dynamics in waste stabilization ponds. Journal of the Water Pollution Control Federation, 54(4), 361–369.

    CAS  Google Scholar 

  37. Srinath, E. G., & Loehr, R. C. (1974). Ammonia desorption by diffusion aeration. Journal of the Water Pollution Control Federation, 46(8), 1939–1957.

    CAS  Google Scholar 

  38. Meron, A. (1971). Kinetics of algal systems in waste treatment field studies. University of California, Berkeley, FWQA, USDOI, PB 206812.

  39. Huppe, H. C., & Turpin, D. H. (1994). Integration of carbon and nitrogen metabolism in plant and algal cells. Annual Review of Plant Physiology and Plant Molecular Biology, 45, 577–607.

    Article  CAS  Google Scholar 

  40. Amory, A. M., Vanlerberghe, G. C., & Turpin, D. H. (1991). Demonstration of both a photosynthetic and nonphotosynthetic CO2 requirement for NH +4 assimilation in the green alga Selenastrum minutum. Plant Physiology, 95, 192–96.

    Article  CAS  Google Scholar 

  41. Thacker, A., & Syrctt, P. J. (1972). The assimilation of nitrate and ammonium by Chlamydomonas reinhardtii. The New Phytologist, 71, 423–433.

    Article  CAS  Google Scholar 

  42. Christie, W. (2003). Lipid analysis, isolation, separation, identification and structural analysis of lipids (3rd ed.). Dundee: MRS Lipids Analysis Unit, Scottish Crop Research Institute, Invergowrie.

    Google Scholar 

  43. Ogbonna, J. C., Masui, H., & Tanaka, H. (1997). Sequentical heterotrophic/autotrophic cultivation—an efficient method of producing Chlorella biomass for health food and animal feed. Journal of Applied Physics, 9(4), 359–366.

    Google Scholar 

Download references

Acknowledgement

The study was supported by grants from the University of Minnesota Initiative for Renewable Energy and the Environment (IREE) and Metropolitan Council Environmental Services (MCES), as well as the Legislative-Citizen Commission on Minnesota Resource (LCCMR). The authors are also grateful to Robert C. Polta and Adam Sealock of Saint Paul MCES Wastewater Treatment Plant for helping with the sample collection.

Author information

Affiliations

  1. Center for Biorefining and Department of Bioproducts and Biosystems Engineering, University of Minnesota, 1390 Eckles Ave, St. Paul, MN, 55108, USA

    Bing Hu, Min Min, Wenguang Zhou, Yecong Li, Michael Mohr, Yanling Cheng, Yuhuan Liu, Xiangyang Lin, Paul Chen & Roger Ruan

  2. Biological Systems Engineering, Washington State University, Richland, WA, 99354, USA

    Hanwu Lei

  3. Biomass Energy Center and State Key Laboratory of Food Science, Nanchang University, Jiangxi, 330047, China

    Yuhuan Liu & Roger Ruan

  4. College of Bioscience and Biotechnology, Fuzhou University, Fujian, 350108, China

    Xiangyang Lin

Authors
  1. Bing Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Min Min
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wenguang Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yecong Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michael Mohr
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yanling Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hanwu Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yuhuan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Xiangyang Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Paul Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Roger Ruan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Roger Ruan.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Hu, B., Min, M., Zhou, W. et al. Influence of Exogenous CO2 on Biomass and Lipid Accumulation of Microalgae Auxenochlorella protothecoides Cultivated in Concentrated Municipal Wastewater. Appl Biochem Biotechnol 166, 1661–1673 (2012). https://doi.org/10.1007/s12010-012-9566-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-012-9566-2

Keywords

  • Microalgae
  • Mixotrophic growth
  • Concentrated municipal wastewater
  • Nutrient removal
  • CO2