Skip to main content
Log in

Isolation of high-lipid content strains of the marine microalga Tetraselmis suecica for biodiesel production by flow cytometry and single-cell sorting

  • Published:
Journal of Applied Phycology Aims and scope Submit manuscript

Abstract

Biodiesel from algae is considered an alternative for a third generation of biofuels. However, most microalgae are not lipogenic during fast growth periods, but high-lipid content occurs at resting stages. Microalgae biomass production for biodiesel needs continuous high volumetric and aerial yields and large amount of neutral lipid in the biomass. These requirements are similar to demanding a marathon runner to be obese. We show that by using cell sorting capabilities of flow cytometers, in combination with the lipid-soluble fluorescent dye Nile Red, we can isolate and select cells with a high and stable lipid content. In our study, we were able to select the equivalent of a stable “fat marathon runner” through three sorting events obtained from wild populations of Tetraselmis suecica.

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

Access this article

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

  • Chen W, Zhang C, Song L, Sommerfeld M, Hu Q (2009) A high throughput Nile Red method for quantitative measurement of neutral lipids in microalgae. J Microbiol Meth 77:41–47

    Article  CAS  Google Scholar 

  • Chisti Y (2007) Biodiesel from microalgae. Biotechnol Adv 25:294–306

    Article  PubMed  CAS  Google Scholar 

  • Chini Zitelli G, Rodolfi L, Biondi N, Tredici MR (2006) Productivity and photosynthetic efficiency of outdoor cultures of Tetraselmis suecica in annular columns. Aquaculture 261:932–943

    Article  Google Scholar 

  • Coder DM (1997) Assessment of cell viability. In: Robinson PJ, Darzynkiewics Z, Dobrucki J et al (eds) Current protocols in cytometry. Wiley, Somerset, pp 9.2.1–9.2.14

    Google Scholar 

  • Comas J, Vives-Rego J (1997) Assesment the effects of gramicidin, formaldehyde and surfactants on Escherichia coli by flow cytometry using nucleic acid and membrane potential dyes. Cytometry 29:58–64

    Article  PubMed  CAS  Google Scholar 

  • Cooksey KE, Guckert JB, Williams SA, Callis PR (1987) Fluorometric determination of the neutral lipid-content of microalgal cells using Nile Red. J Microbiol Meth 6:333–345

    Article  CAS  Google Scholar 

  • de la Jara A, Mendoza H, Martel A, Molina C, Nordstro L, de la Rosa V, Díaz R (2003) Flow cytometric determination of lipid content in a marine dinoflagellate Cryptecodinium cohnii. J Appl Phycol 15:433–438

    Article  Google Scholar 

  • Dismukes C, Carrieri D, Bennette N, Ananyev GM, Posewitz MC (2008) Aquatic phototrophs: efficient alternatives to land-based crops for biofuels. Curr Opin Biotechnol 19:235–240

    Article  PubMed  CAS  Google Scholar 

  • Elsey D, Jameson D, Raleigh B, Cooney MJ (2007) Fluorescent measurement of microalgal neutral lipids. J Microbiol Meth 68:639–642

    Article  CAS  Google Scholar 

  • Griffiths MJ, Harrison STL (2009) Lipid productivity as a key characteristic for choosing algal species for biodiesel production. J Appl Phycol 21:493–507

    Article  CAS  Google Scholar 

  • Hu Q, Sommerfeld M, Jarvis E, Ghirardi M, Posewitz M, Seibert M, Darzins A (2008) Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant J 54:621–639

    Article  PubMed  CAS  Google Scholar 

  • Lopes da Silva T, Amarelo Santos C, Reis A (2009) Multi-parameter flow cytometry as tool to monitor heterotrophic microalgal batch fermentations for oil production towards biodiesel. Biotechnol Bioprocess Eng 14:330–337

    Article  Google Scholar 

  • McGinnis KM, Dempster TA, Sommerfeld MR (1997) Characterization of the growth and lipid content of the diatom Chaetoceros muelleri. J Appl Phycol 9:19–24

    Article  CAS  Google Scholar 

  • Mendoza-Guzmán H, de la Jara A, Carmona-Duarte L, Freijanes-Presmanes K (2009) Estimate by means of flow cytometry of variation in composition of fatty acids from Tetraselmis suecica in response to culture conditions. Aquacult Int 18:189–199

    Article  Google Scholar 

  • Pultz O (2001) Photobioreactors: production systems for phototrophic microorganisms. Appl Microbiol Biotechnol 57:287–297

    Article  Google Scholar 

  • Resina-Pelfort O, Comas-Riu J, Vives-Rego J (2001) Effect of deflected droplet electrostatic cell sorting on the viability and exoproteolytic activity of bacterial cultures and marine bacterioplankton. Syst Appl Microbiol 24:31–36

    Article  PubMed  CAS  Google Scholar 

  • Rodolfi L, Zittelli GC, Bassi N, Padovani G, Biondi N, Bonini G, Tredici MR (2008) Microalgae for oil: strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor. Biotechnol Bioeng 102:100–112

    Article  Google Scholar 

  • Sensen CW, Heimann K, Melkonian M (1993) The production of clonal and axenic cultures of microalgae using fluorescence-activated cell sorting. Europ J Phycol 28:93–97

    Article  Google Scholar 

  • Williams PJLEB (2007) Biofuel: microalgae cut the social and ecological costs. Nature 450:478

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work has been supported by the “Consorcio Estratégico Nacional de Investigación Técnica” (CENIT)—“Proyecto de Investigación para el Impulso del Biodiesel en España” (PIIBE), of the Spanish Government (coordinated by Repsol-YPF). We would like to thank Dr J.L. Gómez-Pinchetti for his thoughtful comments and advice during the development of this study.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to María F. Montero.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Montero, M.F., Aristizábal, M. & García Reina, G. Isolation of high-lipid content strains of the marine microalga Tetraselmis suecica for biodiesel production by flow cytometry and single-cell sorting. J Appl Phycol 23, 1053–1057 (2011). https://doi.org/10.1007/s10811-010-9623-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10811-010-9623-6

Keywords

Navigation