Journal of Applied Phycology

, Volume 6, Issue 1, pp 13–20 | Cite as

Halotolerance studies onChlamydomonas reinhardtii: glycerol excretion by free and immobilized cells

  • R. León
  • F. Galván


The freshwater green algaChlamydomonas reinhardtii can tolerate a maximum saline concentration of 200 mM NaCl. In response to this osmotic shock, the cells accumulated during the first 24 h 15% of the total glycerol synthesized as osmoregulatory metabolite, to provide the corresponding osmotic balance. After this period all glycerol synthesized was excreted to the medium, 4 g L-1 at 120 h in optimal conditions, before cell degradation occurred. This excretion was about 2-fold higher in Ca-alginate entrapped cells in the presence of 250 mM NaCl. It was concluded that immobilized cells may be of biotechnological interest for continuous glycerol photoproduction in air-lift bioreactors.

Key words

Alginate Chlamydomonas glycerol halotolerance immobilization 


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  1. Arnon DI (1949) Copper enzymes in isolated choloroplasts: polyphenol oxidase inBeta vulgaris. Plant Physiol. 24: 1–15.Google Scholar
  2. Bailliez C, Largeau C, Casadevall E (1985) Growth and hydrocarbon production ofBotryococcus braunii immobilized in calcium alginate gel. Appl. Microbiol. Biotechnol. 23: 99–105.Google Scholar
  3. Ben-Amotz A, Avron M (1981) Glycerol and β-carotene metabolism in the halotolerant algaDunaliella: a model system for biosolar energy consersion. Trends Biochem. Sci. 6: 297–299.Google Scholar
  4. Ben-Amotz A, Avron M (1983) Accumulation of metabolites by halotolerant algae and its industrial potential. Ann. Rev. Microbiol. 37: 95–119.Google Scholar
  5. Bispin B, Baumann U, Hellfors H, Rehm HJ (1990) Formation of polyols by immobilized fungi. In de Bont JAM, Visser J, Mattiasson B, Tramper J (eds), Physiology of Immobilized Cells. Elsevier, Amsterdam: 387–392.Google Scholar
  6. Borowitzka MA, Borowitzka LJ (1988)Dunaliella. In Borowitzka MA, Borowitzka LJ (eds), Micro-algal Biotechnology. Cambridge U.P., Cambridge: 27–58.Google Scholar
  7. Brouers M, Hall DO (1986) Ammonia and hydrogen production by immobilized cyanobacteria. J. Biotechnol. 3: 307–321.Google Scholar
  8. Chen BU, Chi CH (1981) Process development and evaluation for algal glycerol production. Biotechnol. Bioengng. 23: 1267–1287.Google Scholar
  9. Chitlary E, Pick U (1991) Regulation of glycerol synthesis in response to osmotic changes inDunaliella. Plant Physiol. 96: 50–60.Google Scholar
  10. Gamboa A, Alfaro S, Reynoso T (1985) Taurine induction of cation tolerance inChlamydomonas reinhardtii. Comp. Biochem. Physiol. 81A: 491–493.Google Scholar
  11. Greenway H, Munns R (1980) Mechanisms of salt tolerance in nonhalophytes. Ann. Rev. Microbiol. 31: 149–190.Google Scholar
  12. Grizeau D, Navarro JM (1986) Glycerol production byDunaliella tertiolecta immobilized within Ca-alginate beads. Biotechnol. Lett. 8: 261–264.Google Scholar
  13. Hellebust JA (1976) Osmoregulation. Ann. Rev. Plant Physiol. 27: 485–505.Google Scholar
  14. Husic HD, Tolbert NE (1986) Effect of osmotic stress on carbon metabolism inChlamydomonas reinhardtii. Plant Physiol. 82: 594–596.Google Scholar
  15. Marker (1972) The use of acetone and methanol in the estimation of chlorophyll in the presence of phaeophytin. Freshwater Biol. 2: 361–385.Google Scholar
  16. Merchuk JC (1990) Why use air-lift bioreactors? Trends Biotechnol. 8: 66–71.Google Scholar
  17. Neale PJ, Melis A (1989) Salinity-stress enhances photoinhibition of photosynthesis inChlamydomonas reinhardtii. J. Plant Physiol. 134: 619–622.Google Scholar
  18. Rao KK, Hall DO (1992) Immobilized photosynthetic systems. Applications in Biotechnology. In: Barber J, Guerrero MG, Medrano H (eds), Trend in Photosynthesis Research. Intercept, Andover: 135–147.Google Scholar
  19. Reynoso T, Gamboa A (1982) Salt tolerance in the freshwater algaChlamydomonas reinhardtii: effect of proline and taurine. Comp. Biochem. Physiol. 73A: 95–99.Google Scholar
  20. Santos-Rosa F, Galván F (1989) Ammonium photoproduction by free and immobilized cells ofChlamydomonas reinhardtii. Appl. Microbiol. Biotechnol. 31: 55–58.Google Scholar
  21. Santos-Rosa F, Galván F, Vega JM (1989) Biological viability ofChlamydomonas reinhardtii cells entrapped in alginate beads for ammonium photoproduction. J. Biotechnol. 9: 209–220.Google Scholar
  22. Smidsrod O, Skjak-Braek G (1990) Alginate as immobilization matrix for cells. Trends Biotechnol. 8: 71–78.Google Scholar
  23. Sueoka N, Chiang KS, Kates JR (1967) Deoxyribonucleic acid replication in meiosis ofChlamydomonas reinhardtii. Isotopic transfer experiments with a strain producing eight zoospores. J. mol. Biol. 25: 47–66.Google Scholar
  24. Tramper J (1990) Conversions by immobilized cells versus traditional fermentations. In de Bont JAM, Visser J, Mattiasson B, Tramper J (eds), Physiology of Immobilized Cells. Elsevier, Amsterdam: 1–14.Google Scholar
  25. Trevan MD, Mak AL (1988) Immobilized algae and their potential for use as biocatalysts. Trends Biotechnol. 6: 68–72.Google Scholar
  26. Vijalkishore P, Karanth NG (1987) Glycerol production by fermentation a fed-batch approach. Biotechnol. Bioengng: 30: 325–328.Google Scholar
  27. Vilchez C, Galván F, Vega JM (1991) Glycolate photoproduction by free and alginate-entrapped cells ofChlamydomonas reinhardtii. Appl. Microbiol. Biotechnol. 35: 716–719.Google Scholar
  28. Wegmann K, Ben-Amotz A, Avron M (1980) Effect of temperature on glycerol retention in the halotolerant algaeDunaliella andAsteromonas. Plant Physiol. 66, 1196–1197.Google Scholar
  29. Wieland OH (1984) Glycerol: UV-method. In Bergmeyer J, Braβ1 M (eds), Methods of Enzymatic Analysis. Metabolites 1: Carbohydrates (Vol. VI). Verlag-Chemie, Weinheim: 504–510.Google Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • R. León
    • 1
  • F. Galván
    • 1
  1. 1.Departamento de Bioquímica Vegetal y Biología Molecular, Facultad de Química, Universidad de SevillaSevillaSpain

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