A culture method for microalgal forms using two-tier vessel providing carbon-dioxide environment: studies on growth and carotenoid production

  • Usha Tripathi
  • R. Sarada
  • G.A. Ravishankar


A culture method was developed for photoautotrophic culture of Haematococcus pluvialis, Chlorella vulgaris, Scenedesmus obliquus, Spirulina platensis, Nostoc and Stigonema in a two-tier flask consisting of nutrient media in the upper chamber and CO2 generating buffer mixture (KHCO3/K2CO3) in the lower chamber. The concentration of buffer mixture was varied to obtain desired levels of CO2. CO2 at 2.0% (v/v) level enhanced growth and chlorophyll content over control cultures (without CO2 supplementation) in all microalgal species. Haematococcus pluvialis culture in BBM and KM1 media showed 6.71- and 2.07-fold increase in biomass yields with astaxanthin productivity at 7.26 and 7.48 mg l−1 level respectively. CO2 supplementation to C. vulgaris and S. obliquus cultures resulted in 5.97- and 7.30-folds increase in biomass with 2–3 fold increase in chlorophyll and carotenoid contents over their respective controls. Similarly 2–3 fold increase in chlorophyll and carotenoid contents were observed in Sp. platensis, Nostoc and Stigonema spp. This culture methodology will provide information on CO2 requirement for growth of algae and metabolite production and also facilitates studies on the influence of light and temperature conditions.

Astaxanthin autotrophic carotenoid Chlorella vulgaris Haematococcus pluvialis heterotrophic mixotrophic Nostoc Scenedesmus obliquus Spirulina platensis Stigonema two-tier vessel 


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  1. Akimoto, M., Yamada, H., Ohtaguchi, K. & Koide, K. 1997 Photoautotrophic cultivation of the green alga Chlamydomonas reinharditii as a method for carbon dioxide fixation and α-linolenic acid production. J. Assoc. Oil Chem. Soc. 74, 181–183.Google Scholar
  2. Benemann, J.R. 1990 Microalgae products and production — An overview. In Developments in Industrial Microbiology, Vol. 31, ed. Pierce, G.E. pp. 247–256. Amsterdam: Elsevier.Google Scholar
  3. Benemann, J.R. 1992 Microalgae aquaculture feed. Journal of Applied Phycology 4, 233–245.Google Scholar
  4. Ding, S.Y., Chee, W.S., Yong, Y.Y., Bee, P.L. & Lee, Y.K. 1994 Production strategies of astaxanthin by growing of Haematococus lacustris cells. In Algal Biotechnology in the Asia Pacific Region, ed. Phang, S.M. Lee, Y.K. Borowitzka, M.A. & Whitton, B.A. pp. 28–32. KaulaLumpur: University of Malaya. ISBN No.–983–808–026–8.Google Scholar
  5. Herdman, M. 1988 Cellular differentiation: akinetes. In Methods in Enzymology, Vol. 167 ed. Packer, L. & Glazer, A.N. London: Academic press, pp. 222–232.Google Scholar
  6. Hüsemann, W. & Barz, W. 1977 Photoautotrophic growth and photosynthesis in cell suspension cultures of Chenopodium rubrum. Physiologia Plantarum 40, 77–81.Google Scholar
  7. Jensen, A. 1978 Chlorophylls and carotenoids. In Handbook of Phycological Methods, ed. Hellebust, J.A. & Craigc, J.S. pp. 59–70. Cambridge: Cambridge University Press.Google Scholar
  8. Kanz, T. & Bold, H.C. 1969 Physiological studies: 9 Morphological and Taxonomic Investigations of Nostoc and Anabaena in Culture. University of Texas publ. No.6924, University of Texas, Austin Texas.Google Scholar
  9. Kobayashi, M., Kakizono, T. & Nagai, S. 1991 Astaxanthin production by a green alga, Haematococcus pluvialis accompanied with morphological changes in acetate media. Journal of Fermentation Bioengineering 71, 335–339.Google Scholar
  10. Lichtenthaler, H.K. 1987 Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. In Methods in Enzymology, Vol. 148, ed. Packer, L. & Douce, R. pp. 350–382. New York: Academic Press.Google Scholar
  11. Rippka, R., Deruelles, J., Warterbury, J.B., Herdman, M. & Stanier, R.Y. 1979 Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Journal of General Microbiology 111, 1–61.Google Scholar
  12. Usha Tripathi, Sarada, R., Ramachandra Rao, S. & Ravishankar, G.A. 1999 Production of astaxanthin in Haematococcus pluvialis cultured in various media. Bioresource Technology 68, 197–199.Google Scholar
  13. Venkataraman, L.V. & Becker, E.W. 1985. Biotechnology and utilization of algae: the Indian experience. DST, New Delhi.Google Scholar
  14. Zarrouk, C. 1966 Contribution a 1 etude dune cyanophyceae. Influence de divers facteurs physiques et chimiques sur la croissance et la photosynthese de Spirulina maxima (Setch et Gardner) Geitler, These Sciences, Paris.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Usha Tripathi
    • 1
  • R. Sarada
    • 1
  • G.A. Ravishankar
    • 1
  1. 1.Department of Plant Cell BiotechnologyCentral Food Technological Research InstituteMysoreIndia

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