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Applied Microbiology and Biotechnology

, Volume 61, Issue 5–6, pp 545–551 | Cite as

Interactions between irradiance and nutrient availability during astaxanthin accumulation and degradation in Haematococcus pluvialis

  • J. Fábregas
  • A. Domínguez
  • A. Maseda
  • A. Otero
Original Paper

Abstract

Fully synchronised germination of Haematococcus pluvialis astaxanthin-replete aplanospores was induced by transfer to nitrogen-sufficient conditions under either high or low light intensities, and growth, pigment content and nitrogen consumption were monitored during the cell cycle. No germination of the aplanospores was achieved in the absence of nitrate, even when cells were transferred at low light intensities. On the other hand, cell density and chlorophyll concentration increased dramatically and astaxanthin concentration decreased in N-sufficient cultures due to the germination of 100% of the aplanospores, as demonstrated by flow cytometry. No significant effect of light intensity was observed on the degradation of astaxanthin during germination. In germinated cultures, nitrogen was depleted more rapidly under high light conditions, which resulted in earlier entry into the aplanospore stage and accumulation of astaxanthin. Germination of aplanospores accompanied by astaxanthin degradation could also be obtained in the dark in nutrient-sufficient conditions although at a much lower efficiency. The results demonstrate that nutrient availability is the main factor controlling the transition between red and green stages of H. pluvialis, with astaxanthin being accumulated only when cell division has ceased. High light levels accelerate the process by increasing the rate of nutrient depletion and providing more energy for astaxanthin synthesis.

Keywords

Carotenoid Astaxanthin High Light Intensity High Light Condition Photoprotective Role 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work was supported by the EC project FAIR 97–1518: "Development of Microalgal Pigments for Aquaculture". We are grateful to F. Hoffman-La Roche Ltd. for kindly providing Astaxanthin and Zeaxanthin standards and to Dr. A. Cid and C. Rioboo from the University of A Coruña for flow cytometry analysis.

References

  1. Borowitzka MA, Huisman JM, Osborn A (1991) Culture of astaxanthin-producing green alga Haematococcus pluvialis. 1. Effects of nutrients on growth and cell type. J Appl Phycol 3:295–304Google Scholar
  2. Boussiba S (2000) Carotenogenesis in the green alga Haematococcus pluvialis: cellular physiology and stress response. Physiol Plant 108:111–117CrossRefGoogle Scholar
  3. Boussiba S, Vonshak A (1991) Astaxanthin accumulation in the green alga Haematococcus pluvialis. Plant Cell Physiol 32:1077–1082Google Scholar
  4. Chaumont D, Thépenier C (1995) Carotenoid content in growing cells of Haematococcus pluvialis during a sunlight cycle. J Appl Phycol 7:529–537Google Scholar
  5. Clesceri LS, Greenberg AE, Trussell RR (eds) (1989) Standard methods for the examination of water and wastewater, 17th edn. American Public Health Association, Washington, D.C.Google Scholar
  6. Droop MR (1954) Conditions governing haematochrome formation and loss in the alga Haematococcus pluvialis Flotow. Arch Mikrobiol 20:391–397Google Scholar
  7. Droop MR (1955) Carotenogenesis in Haematococcus pluvialis. Nature 175:42Google Scholar
  8. Elliot AM (1934) Morphology and life history of Haematococcus pluvialis. Arch Protistenkd 82:250–271Google Scholar
  9. Fábregas J, Domínguez A, García-Álvarez D, Lamela T, Otero A (1998) Induction of astaxanthin accumulation by nitrogen and magnesium deficiencies in Haematococcus pluvialis. Biotechnol Lett 20:623–626CrossRefGoogle Scholar
  10. Fábregas J, Domínguez A, Regueiro M, Maseda A, Otero A (2000) Optimization of culture medium for the continuous cultivation of the microalga Haematococcus pluvialis. Appl Microbiol Biotechnol 53:530–535CrossRefPubMedGoogle Scholar
  11. Fábregas J, Otero A, Maseda A, Domínguez A (2001) Two-stage cultures for the production of astaxanthin from Haematococcus pluvialis. J Biotechnol 89:65–71CrossRefPubMedGoogle Scholar
  12. Fan L, Vonshak A, Boussiba S (1994) Effect of temperature and irradiance on growth of Haematococcus pluvialis (Chlorophyceae). J Phycol 30:829–833Google Scholar
  13. Goodwin TW, Jamikorn M (1954) Studies in carotenogenesis. 11. Carotenoid synthesis in the alga Haematococcus pluvialis. Biochem J 57:376–381Google Scholar
  14. Grünewald K, Hagen C, Braune W (1997). Secondary carotenoid accumulation in flagellates of the green alga Haematococcus lacustris. Eur J Phycol 32:387–392CrossRefGoogle Scholar
  15. Grung M, D'Souza FML, Borowitzka M, Liaaen-Jensen S (1992) Algal carotenoids 51. Secondary carotenoids 2. Haematococcus pluvialis aplanospores as a source of (3S, 3′S)-astaxanthin esters. J Appl Phycol 4:165–171Google Scholar
  16. Hagen C, Braune W, Björn LO (1994) Functional aspects of secondary carotenoids in Haematococcus lacustris (Volvocales). III Action as a "sunshade". J Phycol 30:241–248Google Scholar
  17. Hagen C, Grünewald K, Xyländer M, Rothe E (2001) Effect of cultivation parameters on growth and pigment biosynthesis in flagellated cells of Haematococcus pluvialis. J Appl Phycol 13:79–87CrossRefGoogle Scholar
  18. Harker M, Tsavalos AJ, Young AJ (1996) Factors responsible for astaxanthin formation in the chlorophyte Haematococcus pluvialis. Bioresour Technol 55:207–214CrossRefGoogle Scholar
  19. Johnson EA, An GH (1991) Astaxanthin from microbial sources. Crit Rev Biotechnol 11:297–326Google Scholar
  20. Jyonouchi H, Hill RJ, Tomita Y, Good RA (1991) Studies of immunomodulating actions of carotenoids. I. Effects of β-carotene and astaxanthin on murine lymphocyte functions and cell surface marker expression in vitro culture system. Nutr Cancer 16:93–105PubMedGoogle Scholar
  21. Kobayashi M (2000) In vivo antioxidant role of astaxanthin under oxidative stress in the green alga Haematococcus pluvialis. Appl Microbiol Biotechnol 54:550–555CrossRefPubMedGoogle Scholar
  22. Kobayashi M, Kakizono T, Nishio N, Nagai S (1992) Effects of light intensity, light quality, and illumination cycle on astaxanthin formation in a green alga, Haematococcus pluvialis. J Ferment Bioeng 74:61–63Google Scholar
  23. Kobayashi M, Kakizono T, Nagai S (1993) Enhanced carotenoid biosynthesis by oxidative stress in acetate-induced cysts cells of a green unicellular alga, Haematococcus pluvialis. Appl Environ Microbiol 59:867–873Google Scholar
  24. Kobayashi M, Kurimura Y, Tsuji Y (1997a) Light-independent, astaxanthin production by the green microalga Haematococcus pluvialis under salt stress. Biotechnol Lett 19:507–509Google Scholar
  25. Kobayashi M, Kurimura Y, Kakizono T, Nishio N, Tsuji Y (1997b) Morphological changes in the life cycle of the green alga Haematococcus pluvialis. J Ferment Bioeng 84:94–97CrossRefGoogle Scholar
  26. Kraay GW, Zapata M, Veldhuis MJW (1992) Separation of chlorophylls c1, c2, and c3 of marine phytoplankton by reversed-phase-C18-high-performance liquid chromatography. J Phycol 28:708–712Google Scholar
  27. Pringsheim EG (1966) Nutritional requirements of Haematococcus pluvialis and related species. J Phycol 2:1–7Google Scholar
  28. Renstrøm B, Borch G, Skulberg OM, Liaaen-Jensen S (1981) Optical purity of (3S,3′S)-astaxanthin from Haematococcus pluvialis. Phytochemistry 20:2561–2564CrossRefGoogle Scholar
  29. Santos MF, Mesquita JF (1984) Ultrastructural study of Haematococcus lacustris (Girod.) Rostafinski (Volvocales) I. Some aspects of carotenogenesis. Cytologia 49:215–228Google Scholar
  30. Tanaka T, Makita H, Ohnishi M, Mori H, Satoh K, Hara A (1995) Chemoprevention of rat oral carcinogenesis by naturally occurring xanthophylls, astaxanthin and canthaxanthin. Cancer Res 55:4059–4064PubMedGoogle Scholar
  31. Yong YYR, Lee YK (1991) Do carotenoids play a photoprotective role in the cytoplasm of Haematococcus lacustris (Chlorophyta)? Phycologia 30:257–261Google Scholar
  32. Zlotnik (Shmerler) I, Sukenik A, Dubinsky Z (1993) Physiological and photosynthetic changes during the formation of red aplanospores in the Chlorophyte Haematococcus pluvialis. J Phycol 29:463–469Google Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • J. Fábregas
    • 1
  • A. Domínguez
    • 1
  • A. Maseda
    • 1
  • A. Otero
    • 1
  1. 1.Laboratorio de Microbiología, Facultad de FarmaciaUniversidad de SantiagoSantiagoSpain

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