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Journal of Applied Phycology

, Volume 31, Issue 1, pp 249–254 | Cite as

Interference of starch accumulation in microalgal cell growth measurement

  • Marcella Fernandes de SouzaEmail author
  • Marcoaurélio Almenara Rodrigues
  • Elba Pinto da Silva Bon
  • Suely Pereira Freitas
Article

Abstract

Starch accumulation in microalgae is a subject of growing interest in the energy sector since this polysaccharide can be used as feedstock to produce ethanol and biogas. Microalgal cell growth dynamics are commonly monitored by measuring the optical density (OD) of the culture, which then is converted to either biomass dry weight or cell concentration with the aid of standard curves. However, starch accumulation causes changes in the cell morphology, which may impair the use of OD for the measurement of cell growth. Therefore, this study investigated the correlation of OD to cell concentration and dry weight during the cultivation of Chlorella sorokiniana, a starch-accumulating green microalga. A linear correlation between OD and cell concentration was observed only during the exponential and the decelerating growth phases. The OD increase during the stationary phase was related to starch accumulation rather than to cell growth. Similarly, a lack of correlation between OD and biomass dry weight occurred during the stationary growth phase, as the increase of dry weight, due to the pronounced starch accumulation, was greater than the OD changes. Thus, this study shows that the use of OD to measure microalgal cell growth in starch-accumulating cultures can be misleading. The direct measurement of cell concentration and dry weight is advisable for monitoring the full cell growth cycle and accurately calculating biomass productivity.

Keywords

Chlorella sorokiniana Cell growth measurement Carbohydrate accumulation Optical density Dry weight Cell counting 

Notes

Funding information

This work was financed by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) project “Microalgae: production, characterization, and fractioning to obtain biofuels and bioproducts with bioactive potential” [Grant Number 4074812013].

References

  1. Bhatnagar A, Bhatnagar M, Chinnasamy S, Das KC (2010) Chlorella minutissima—a promising fuel alga for cultivation in municipal wastewaters. Appl Biochem Biotechnol 161:523–536CrossRefGoogle Scholar
  2. Borowitzka MA (2013) High-value products from microalgae—their development and commercialisation. J Appl Phycol 25:743–756CrossRefGoogle Scholar
  3. Chen C-Y, Zhao X-Q, Yen H-W, Ho S-H, Cheng C-L, Lee D-J, Bai F-W, Chang J-S (2013) Microalgae-based carbohydrates for biofuel production. Biochem Eng J 78:1–10CrossRefGoogle Scholar
  4. Chioccioli M, Hankamer B, Ross IL (2014) Flow cytometry pulse width data enables rapid and sensitive estimation of biomass dry weight in the microalgae Chlamydomonas reinhardtii and Chlorella vulgaris. PLoS One 9:e97269CrossRefGoogle Scholar
  5. Chojnacka K, Noworyta A (2004) Evaluation of Spirulina sp. growth in photoautotrophic, heterotrophic and mixotrophic cultures. Enzym Microb Technol 34:461–465CrossRefGoogle Scholar
  6. Dragone G, Fernandes BD, Abreu AP, Vicente AA, Teixeira JA (2011) Nutrient limitation as a strategy for increasing starch accumulation in microalgae. Appl Energy 88:3331–3335CrossRefGoogle Scholar
  7. Feng Y, Li C, Zhang D (2011) Lipid production of Chlorella vulgaris cultured in artificial wastewater medium. Bioresour Technol 102:101–105CrossRefGoogle Scholar
  8. Gifuni I, Olivieri G, Pollio A, Franco TT, Marzocchella A (2017) Autotrophic starch production by Chlamydomonas species. J Appl Phycol 29:105–114CrossRefGoogle Scholar
  9. Griffiths MJ, Garcin C, van Hille RP, Harrison STL (2011) Interference by pigment in the estimation of microalgal biomass concentration by optical density. J Microbiol Methods 85:119–123CrossRefGoogle Scholar
  10. Ho SH, Huang SW, Chen CY, Hasunuma T, Kondo A, Chang JS (2013a) Bioethanol production using carbohydrate-rich microalgae biomass as feedstock. Bioresour Technol 135:191–198CrossRefGoogle Scholar
  11. Ho SH, Li PJ, Liu CC, Chang JS (2013b) Bioprocess development on microalgae-based CO2 fixation and bioethanol production using Scenedesmus obliquus CNW-N. Bioresour Technol 145:142–149CrossRefGoogle Scholar
  12. John RP, Anisha GS, Nampoothiri KM, Pandey A (2011) Micro and macroalgal biomass: a renewable source for bioethanol. Bioresour Technol 102:186–193CrossRefGoogle Scholar
  13. Li T, Zheng Y, Yu L, Chen S (2014) Mixotrophic cultivation of a Chlorella sorokiniana strain for enhanced biomass and lipid production. Biomass Bioenergy 66:204–213CrossRefGoogle Scholar
  14. Liang Y, Sarkany N, Cui Y (2009) Biomass and lipid productivities of Chlorella vulgaris under autotrophic, heterotrophic and mixotrophic growth conditions. Biotechnol Lett 31:1043–1049CrossRefGoogle Scholar
  15. Lichtenthaler HK, Buschmann C (2001) Chlorophylls and carotenoids: measurement and characterization by UV-VIS spectroscopy. In: Wrolstad RE, Acree,TE, An H, Decker EA, Penner MH, Reid DS, Schwartz SJ, Shoemaker CF, Sporns P (eds) Current Protocols in Food Analytical Chemistry (CPFA). John Wiley and Sons, New York, F4.3.1-F4.3.8Google Scholar
  16. Lichtenthaler H, Wellburn A (1983) Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592CrossRefGoogle Scholar
  17. Mata TM, Martins AA, Caetano NS (2010) Microalgae for biodiesel production and other applications: a review. Renew Sust Energ Rev 14:217–232CrossRefGoogle Scholar
  18. Mayers JJ, Vaiciulyte S, Malmhäll-Bah E et al (2018) Identifying a marine microalgae with high carbohydrate productivities under stress and potential for efficient flocculation. Algal Res 31:430–442Google Scholar
  19. Megazyme (2017) Total starch assay procedure (amyloglucosidase/alpha-amylase method). http://secure.megazyme.com/files/BOOKLET/K-TSTA_1107_ DATA.pdf 2011. Accessed 1 Jan 2017
  20. Moheimani NR, Borowitzka MA, Isdepsky A, Fon Sing S (2013) Standard methods for measuring growth of algae and their composition. In: Borowitzka MA, Moheimani NR (eds) Algae for biofuels and energy. Springer, Dordrecht, pp 265–284CrossRefGoogle Scholar
  21. Mondal M, Ghosh A, Oinam G, Tiwari ON, Gayen K, Halder GN (2017) Biochemical responses to bicarbonate supplementation on biomass and lipid productivity of Chlorella sp. BTA9031 isolated from coalmine area. Environ Prog Sustain Energy 36:1498–1506Google Scholar
  22. Nichols HW, Bold HC (1965) Trichosarcina polymorpha Gen. et Sp. Nov. J Phycol 1:34–38CrossRefGoogle Scholar
  23. Podevin M, Fotidis IA, De Francisci D, Møller P, Angelidaki I (2017) Detailing the start-up and microalgal growth performance of a full-scale photobioreactor operated with bioindustrial wastewater. Algal Res 25:101–108CrossRefGoogle Scholar
  24. Richmond A, Hu Q (eds) (2013) Handbook of microalgal culture: applied phycology and biotechnology. Wiley, ChichesterGoogle Scholar
  25. Rodríguez-López M (1966) Utilization of sugars by Chlorella under various conditions. J Gen Microbiol 43:139–143CrossRefGoogle Scholar
  26. Samiee-Zafarghandi R, Karimi-Sabet J, Abdoli MA, Karbassi A (2018) Increasing microalgal carbohydrate content for hydrothermal gasification purposes. Renew Energy 116:710–719.dCrossRefGoogle Scholar
  27. Sayre R (2010) Microalgae: the potential for carbon capture. Bioscience 60:722–727CrossRefGoogle Scholar
  28. Sheehan J, Dunahay T, Benemann J, Roessler P (1998) A look back at the U.S. Department of Energy's Aquatic Species Program - Biodiesel from Algae. National Renewable Energy Laboratory,, Golden, Colorado. Report NREL/TP-580-24190, pp 1–328Google Scholar
  29. Spolaore P, Joannis-Cassan C, Duran E, Isambert A (2006) Commercial applications of microalgae. J Biosci Bioeng 101:87–96CrossRefGoogle Scholar
  30. Vieira Salla AC, Margarites AC, Seibel FI, Holz LC, Brião VB, Bertolin TE, Colla LM, Costa JAV (2016) Increase in the carbohydrate content of the microalgae Spirulina in culture by nutrient starvation and the addition of residues of whey protein concentrate. Bioresour Technol 209:133–141CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Marcella Fernandes de Souza
    • 1
    Email author
  • Marcoaurélio Almenara Rodrigues
    • 1
    • 2
  • Elba Pinto da Silva Bon
    • 1
  • Suely Pereira Freitas
    • 3
  1. 1.Bioethanol Laboratory, Chemistry InstituteFederal University of Rio de JaneiroRio de JaneiroBrazil
  2. 2.Laboratory of Plant Biotechnology and PhotosynthesisFederal University of Rio de JaneiroRio de JaneiroBrazil
  3. 3.Laboratory for the Processing of Plant FeedstocksFederal University of Rio de JaneiroRio de JaneiroBrazil

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