Journal of Applied Phycology

, Volume 18, Issue 6, pp 769–781 | Cite as

Purification of the blue-green pigment “marennine” from the marine tychopelagic diatom Haslea ostrearia (Gaillon/Bory) Simonsen

  • Jean-Bernard Pouvreau
  • Michèle Morançais
  • Guillaume Massé
  • Philippe Rosa
  • Jean-Michel Robert
  • Joël Fleurence
  • Pierre PondavenEmail author
Original Article


The diatom Haslea ostrearia that lives in oyster ponds has the distinctive feature of synthesizing “marennine”, a blue-green pigment of which the chemical nature still remains unknown. This pigment is responsible for the greening of oyster gills. Here, we report a new method for extraction and purification of intracellular (accumulated in the apex of the cell) and extracellular (released into the external medium) forms of the pigment. Intracellular marennine is obtained by extraction from blue algal pellets with a carbonate buffer. The extract is then centrifuged and filtered. Extracellular marennine is obtained by clarification of blue-coloured culture medium. Both extracts are then purified by a semi-preparative process, using ultrafiltration through membranes and anion-exchange chromatography. This procedure allows us to produce native pigment displaying the degree of purity required to enter upon the molecular characterisation of marennine. By this process, about 35% of the initial amount of pigment can be recovered. If necessary, this method could be easily scaled up to a larger production system to accommodate potential industrial applications.


Diatom Haslea ostrearia Microalgae Marennine Pigment Ultrafiltration 



Ethylen Diamine Tetraacetic Acid


Ethylen Glycol-bis (2-aminoethylether)-N,N,N′,N′-Tetraacetic Acid


Phenyl Methane Sulfonyl Fluoride


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bachrach E (1935) Le bleuissement des diatomées et le verdissement des huîtres. Rev Trav Inst Pêches Mar 8: 112–123Google Scholar
  2. Bergé JP, Bourgougnon N, Alban S, Pojer F, Billaudel S, Chermann JC, Robert JM Franz G (1999) Antiviral and anticoagulant activities of a water-soluble fraction of the marine diatom Haslea ostrearia. Planta Medica 65: 604–609PubMedCrossRefGoogle Scholar
  3. Carbonnelle D, Pondaven P, Morançais M, Massé G, Bosch S, Jacquot C, Briand G, Robert JM, Roussakis C (1999) Antitumor and antiproliferative effects of an aqueous extract from the marine diatom Haslea ostrearia (Gaillon) Simonsen against solid tumors: lung carcinoma (NSCLC-N6), kidney carcinoma (E39) and melanoma (M96) cell lines. Anticancer Res 19: 621–624PubMedGoogle Scholar
  4. Lankester ER (1886) On green oysters. Quart J Microsc Sci 26: 71–94Google Scholar
  5. Méléder V, Barillé L, Launeau P, Carrère V, Rincé Y (2003) Spectrometric constraint in analysis of benthic diatom biomass using monospecific cultures. Remote Sens Environ 88: 386–400CrossRefGoogle Scholar
  6. Moreau J (1970) Contribution aux recherches écologiques sur les claires à huîtres du bassin de Marennes-Oléron. Rev Trav Inst Pêches Mar 34: 381–462Google Scholar
  7. Mouget JL, Tremblin G, Morant-Manceau A, Morançais M, Robert JM (1999) Long-term photoacclimation of Haslea ostrearia (Bacillariophyta): effect of irradiance on growth rate, pigment content and photosynthesis. Eur J Phycol 34: 109–115CrossRefGoogle Scholar
  8. Mouget JL, Rosa P, Vachoux C, Tremblin G (2005) Enhancement of marennine production by blue light in the diatom Haslea ostrearia. J Appl Phycol 17: 437–445CrossRefGoogle Scholar
  9. Nassiri Y, Robert JM, Rincé Y, Ginsburger-Vogel T (1998) The cytoplasmic fine structure of the diatom Haslea ostrearia (Bacillariophycae) in relation to marennine production. Phycologia 37(2): 84–91CrossRefGoogle Scholar
  10. Neuville D, Daste PH (1970) Observations concernant la production du pigment bleu par la diatomée Navicula ostrearia (Gaillon) Bory maintenue en culture unialgale. C R Acad Sci Paris 271(Série D): 2389–2391Google Scholar
  11. Neuville D, Daste PH (1972) Production du pigment bleu par la diatomée Navicula ostrearia (Gaillon) Bory, maintenue en culture uni-algale sur un milieu synthétique carencé en azote nitrique. C R Acad Sci Paris 274: 2030–2033.Google Scholar
  12. Neuville D, Daste PH (1973) Sur la singularité de la production d'un pigment bleu-vert par la diatomée Navicula ostrearia (Gaillon) Bory. C R Acad Sci Paris 276(Série D): 3469–3472Google Scholar
  13. Neuville D, Daste PH (1978) Recherches sur le déterminisme de la production de marennine par la diatomée marine Navicula ostrearia (Gaillon) Bory en culture in vitro. Rev Gen Bot 85: 255–303Google Scholar
  14. Puységur H (1880) Notice sur la cause du verdissement des huîtres. Rev Mar Colon 64: 248–256Google Scholar
  15. Ranson G (1927) L'absorption de matières organiques dissoutes par la surface extérieure du corps chez les animaux aquatiques. Ann Inst Ocean IV: 49–174Google Scholar
  16. Robert JM, Hallet JN (1981) Absorption spectrum in vivo of the blue pigment marennine of the pennate diatom Navicula ostrearia (Bory). J Exp Bot 32: 341–345Google Scholar
  17. Robert JM (1983) Fertilité des claires ostréicoles et verdissement: utilisation de l'azote par les diatomées dominantes. PhD Thesis. University of Nantes, Nantes, France.Google Scholar
  18. Robert JM, Rouillard I (1991) Production en masse de l'algue unicellulaire Haslea ostrearia Simonsen sur eau souterraine salée: application au verdissement des huîtres de claires. In: Compte-rendu de fin d'étude au profit de SODEXAL (Société d'Expérimentation des Algues) La Guérinière. pp 43–46.Google Scholar
  19. Robert JM, Morançais M, Pradier E, Mouget JL, Tremblin G (2002) Extraction and quantitative analysis of the blue-green pigment “marennine” synthesized by the diatom Haslea ostrearia. J Appl Phycol 14: 299–305CrossRefGoogle Scholar
  20. Schubert H, Tremblin G, Robert JM, Sagert S, Rincé Y (1995) In vivo fluorescence measurement of photosynthesis of Haslea ostrearia Simonsen in relation to marennine content. Diatom Res 10: 341–349Google Scholar
  21. Simonsen R (1974) The diatom plankton of the Indian Ocean expedition of R/V Meteor 1964–1965. Forsch. Ergebnisse Reihe D 19: 1–107Google Scholar
  22. Tremblin G, Robert JM (1996) Comportement photosynthétique de Haslea ostrearia en relation avec sa pigmentation bleue. C R Acad Sci Paris 319: 933–944Google Scholar
  23. Tremblin G, Cannuel R, Mouget JL, Rech M, Robert JM (2000) Change in light quality due to a blue-green pigment, marennine, released in oyster-ponds: effect on growth and photosynthesis in two diatoms, Haslea ostrearia and Skeletonema costatum. J Appl Phycol 12: 557–566CrossRefGoogle Scholar
  24. Turpin V, Robert JM, Goulletquer P (1999) Limiting nutrients of oyster pond seawaters in the Marennes-Oléron region for Haslea ostrearia: applications to the mass production of the diatom in mesocosm experiments. Aquat Living Resour 12: 335–342CrossRefGoogle Scholar
  25. Vandanjon L, Jaouen P, Rossignol N, Quéméneur F, Robert JM (1999) Concentration and desalting by membrane processes of a natural pigment produced by the marine diatom Haslea ostrearia Simonsen. J Biotechnol 70: 393–402CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Jean-Bernard Pouvreau
    • 1
  • Michèle Morançais
    • 1
  • Guillaume Massé
    • 2
  • Philippe Rosa
    • 1
  • Jean-Michel Robert
    • 1
  • Joël Fleurence
    • 1
  • Pierre Pondaven
    • 1
    Email author
  1. 1.EMI, EA2663, ISOMer-UFR SciencesUniversité de Nantes Nantes Atlantique UniversitésBrittanyFrance
  2. 2.Petroleum and Environmental Geochemistry Group, School of Environmental SciencesUniversity of PlymouthPlymouthGreat Britain

Personalised recommendations