World Journal of Microbiology and Biotechnology

, Volume 23, Issue 9, pp 1209–1215 | Cite as

Study of docosahexaenoic acid production by the heterotrophic microalga Crypthecodinium cohnii CCMP 316 using carob pulp as a promising carbon source

  • Ana Mendes
  • Pedro Guerra
  • Vânia Madeira
  • Francisco Ruano
  • Teresa Lopes da Silva
  • Alberto Reis
Original Paper

Abstract

In this work, carob pulp syrup was used as carbon source in C. cohnii fermentations for docosahexaenoic acid production. In preliminary experiments different carob pulp dilutions supplemented with sea salt were tested. The highest biomass productivity (4 mg/lh) and specific growth rate (0.04/h) were observed at the highest carob pulp dilution (1:10.5 (v/v), corresponding to 8.8 g/l glucose). Ammonium chloride and yeast extract were tested as nitrogen sources using different carob pulp syrup dilutions, supplemented with sea salt as growth medium. The best results were observed for yeast extract as nitrogen source. A C. cohnii fed-batch fermentation was carried out using diluted carob pulp syrup (1:10.5 v/v) supplemented with yeast extract and sea salt. The biomass productivity was 420 mg/lh, and the specific growth rate 0.05/h. Under these conditions the DHA concentration and DHA production volumetric rate attained 1.9 g/l and 18.5 mg/lh respectively after 100.4 h. The easy, clean and safe handling of carob pulp syrup makes this feedstock a promising carbon source for large-scale DHA production from C. cohnii. In this way, this carob industry by-product could be usefully disposed of through microbial production of a high value fermentation product.

Keywords

Carob pulp syrup DHA Microalga Crypthecodinium cohnii Omega-3 polyunsaturated fatty acid 

References

  1. Albergaria H, Roseiro J, Collaço MT (1999) Technological aspects and kinetic analysis of microbial gum production in carob. Agro Food Industry Hi-Tech 10:24–26Google Scholar
  2. Borowitzka MA (1992) Algal biotechnology products and processes: matching science and economics. J Appl Phycol 4:267–279CrossRefGoogle Scholar
  3. De Swaaf ME, Rijk TC, Eggink G et al. (1999) Optimisation of docosahexaenoic acid production in batch cultivations by Crypthecodinium cohnii. J Biotechnol 70:185–192CrossRefGoogle Scholar
  4. De Swaaf ME, Sijtsma L, Pronk JT (2003a) High-cell density fed-batch cultivation of the docosahexaenoic acid producing marine alga Crypthecodinium cohnii. Biotechnol Bioeng 81: 666–672CrossRefGoogle Scholar
  5. De Swaaf M, Pronk JT, Sijtsma L (2003b) Fed-batch cultivation of docosahexaenoic-acid-producing marine alga Crypthecodinium cohnii on ethanol. Appl Microbiol Biotechnol 61:40–43Google Scholar
  6. De Swaaf ME, Sijtsma L (2004) Biotechnological production and applications of the ω−3 polyunsaturated fatty acid docosahexaenoic acid. Appl Microbiol Biotechnol 64:146–153CrossRefGoogle Scholar
  7. Guillard RL (1960) A mutant of Chlamydomonas moewusii lacking contractile vacuoles. J Protozool 7:262–269Google Scholar
  8. Guillard RL (1975) Culture of phytoplankton for feeding marine invertebrates. In: Smith WL, Chanley MH (eds) Culture of marine invertebrate animals, Plenum Press, New York, p 26Google Scholar
  9. Guillard RL, Ryther JH (1962) Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt and Detonula confervacea Cleve. Can J Microbiol 8:229–239CrossRefGoogle Scholar
  10. Jiang Y, Chen F, Liang S-Z (1999) Production potential of docosahexaenoic acid by the heterotrophic marine dinoflagellate Crypthecodinium cohnii. Process Biochem 34:633–637CrossRefGoogle Scholar
  11. Jiang Y, Chen F (2000a) Effects of temperature and temperature shift on docosahexaenoic acid production by the marine microalga Crypthecodinium cohnii. J Am Oil Chem Soc 77:613–617CrossRefGoogle Scholar
  12. Jiang Y, Chen F (2000b) Effects of medium glucose concentration and pH on docosahexaenoic acid content of heterotrophic Crypthecodinium cohnii. Proc Biochem 35:1205–1209CrossRefGoogle Scholar
  13. Lepage G, Roy CC (1986) Direct transesterification of all classes of lipids in a one-step reaction. J Lipid Res 27:114–119Google Scholar
  14. Molina E, Fernández J, Acién FG et al. (2001) Tubular photobioreactor design for algal cultures. J Biotechnol 92:113–131CrossRefGoogle Scholar
  15. Petit MD, Pinilla JH (1995) Production and purification of a sugar syrup from carob pods: Lebensm-Wiss u-Technol 28:145–152Google Scholar
  16. Ratledge C (2004) Fatty acid biosynthesis in microorganisms being used for single cell oil production. Biochemie 86:807–815CrossRefGoogle Scholar
  17. Roseiro JC, Gírio FM, Collaço MT (1991) Yield improvements in Carob sugar extraction. Process Biochem 26:179–182CrossRefGoogle Scholar
  18. Sijtsma L, Anderson A, Ratledge C (2005) Alternative carbon sources for heterotrophic production of docosahexaenoic acid by the marine alga Crypthecodinium cohnii. In: Cohen Z, Ratledge C (eds) Single cell oils, AOCS PRESS, Champaign, p 107Google Scholar
  19. Tuttle RC, Loeblich AR (1975) An optimal growth medium for the dinoflagellate Crypthecodimium cohnii. Phycologia 14:1–8Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Ana Mendes
    • 1
  • Pedro Guerra
    • 1
  • Vânia Madeira
    • 1
  • Francisco Ruano
    • 1
  • Teresa Lopes da Silva
    • 1
  • Alberto Reis
    • 1
  1. 1.Instituto Nacional de Engenharia, Tecnologia e Inovação (INETI), Departamento de BiotecnologiaUnidade de Bioengenharia e BioprocessosLisboaPortugal

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