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Downstream processing and purification of eicosapentaenoic (20:5n-3) and arachidonic acids (20:4n-6) from the microalga Porphyridium cruentum

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Bioseparation

Abstract

Eicosapentaenoic acid (FPA, 20:5n-3) and arachidonic acid (AA, 20:4n-3)were obtained from the microalga Porphyridium cruentum by a three-stepprocess: fatty acid extraction by direct saponification of biomass,polyunsaturated fatty acid (PUFA) concentration by urea inclusion complexingand EPA isolation by high-performance liquid chromatography (HPLC). Twosolvents were tested for direct saponification of lipids in biomass. Themost efficient solvent, ethanol (96% v/v), extracted 75% ofthe fatty acids. PUFAs concentration by urea inclusion employed a urea/fattyacid ratio of 4:1 wt/wt at the crystallization temperatures of 4°C and28°C. Concentration factors were similar at both temperatures, but theEPA and AA recoveries were higher at 28°C (67.7% and 61.8%for the two acids, respectively). EPA and AA were purified from this PUFAconcentrate using analytical scale HPLC and the best results of thisseparation were scaled up to preparative level (4.7 i. d. × 30 cmcompression radial cartridge). A 94.3% pure EPA fraction and a81.4% pure AA fraction were obtained. Suitability of severalmicroalgae (Porphyridium cruentum, Phaeodactylum tricornutum and Isochrysisgalbana) and cod liver oil as sources of highly pure PUFAs, mainly EPA, wascompared.

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References

  • Ahern TJ, Katoh S and Sada E (1983) Arachidonic acid production by the red alga Porphyridium cruentum. Biotechnol. Bioeng. 25: 1057–1070.

    Google Scholar 

  • Belter PA, Cussler EL and Hu WS (1988) Elution chromatography. In Belter PA, Cussler EL and Hu WS (eds) Bioseparation, Downstream Processing for Biotechnology. (pp. 209–213). Wiley-Interscience Publication, John Wiley & Sons, New York.

    Google Scholar 

  • Braden LM and Carroll KK (1986) Dietary polyunsaturated fat in relation to mammary carcinogenesis in rats. Lipids 21: 285-288.

    Google Scholar 

  • Cartens M, Molina Grima E, Robles Medina A, Giménez Giménez A and Ibáñez González MJ (1996) Eicosapentaenoic acid (20:5n-3) from the marine microalga Phaeodactylum tricornutum. J. Am. Oil Chem. Soc. 72: 1–7.

    Google Scholar 

  • Chrétiennot-Dinet MJ (1990) Atlas du Phytoplancton Marin, Vol. 3, (pp. 85–86). Centre National de la Recherche Scientifique (ed.), Paris.

    Google Scholar 

  • Cohen Z and Cohen S (1991) Preparation of eicosapentaenoic acid (EPA) concentrate from Porphyridium cruentum. J. Am. Oil Chem. Soc. 68: 16–19.

    Google Scholar 

  • Dyerberg J (1986) Linoleate-derived polyunsaturated fatty acids and prevention of atherosclerosis. Nutr. Rev. 44: 125–134.

    Google Scholar 

  • García Sánchez JL, Molina Grima E, García Camacho F, Sánchez Pérez JA and Giménez Giménez A (1993) Cuantificación de ácidos grasos a partir de biomasa microalgal. Grasas y Aceites} 44}: 348–3

    Google Scholar 

  • Innis SM (1991) Essential fatty acids in growth and development. Prog. Lip. Res. 30: 39–103.

    Google Scholar 

  • Kates M (1988) Separation of lipid mixtures. In: Techniques of Lipidology: Isolation, Analysis and Identification of Lipids, RH Burdon and PH Van Knippenberg (eds) 2nd edition, (pp. 186–278). Elsevier, Amsterdam.

    Google Scholar 

  • Koletzko B, Decsi T and Demmelmair H (1996) Arachidonic acid supply and metabolism in human infants born at full term. Lipids 31: 79–83.

    Google Scholar 

  • Koletzko B, Schmidt E, Bremer HJ, Haung M and Harzer G (1989) Effects of dietary long-chain polyunsaturated fatty acids on the essential fatty acid status of premature infants. Eur. J. Pediatr. 148: 669–675.

    Google Scholar 

  • Lepage G and Roy CC (1984) Improved recovery of fatty acid through direct transesterification without prior extraction or purification. J. Lipid Res. 25: 1391–1396.

    Google Scholar 

  • Molina Grima E, Robles Medina A, Giménez Giménez A, Sánchez Pérez JA, García Camacho F and García Sánchez JL (1994) Comparison between extraction of lipids and fatty acids from microalgal biomass. J. Am. Oil Chem. Soc. 71: 955–959.

  • Molina Grima E, Robles Medina A, Giménez Giménez A and Ibáñez González MJ (1996) Gram-scale purification of eicosapentaenoic acid (EPA, 20:5n-3) from wet Phaeodactylum tricornutumUTEX 640 biomass. J. Appl. Phycol. 8: 359–367.

    Google Scholar 

  • Molina Grima E, Robles Medina A, Giménez Giménez A and Ibáñez González MJ (1997) Obtención de ácido eicosapentaenoico de alta pureza a partir de la microalga Phaeodactylum tricornutummediante un proceso de tres etapas. Spanish patent no }P-960209

  • Ramus J (1972) The production of extracellular polysaccharides by the unicellular red alga Porphyridium aerugineum. J. Phycol. 8: 97–111.

    Google Scholar 

  • Nichols BW and Appleby RS (1969) The distribution and biosynthesis of arachidonic acid in algae. Phytochemistry 8: 1907–1915.

    Google Scholar 

  • Proceeding of International Conference on Highly Unsaturated Fatty Acids in Nutrition and Disease Prevention (1996) Barcelona, Spain, 4-6 November.

  • Rambjor GS, Walen AI, Windsor SL and Harris WS (1996) Eicosapentaenoic acid is primarily responsible for hypotriglyceridemic effect of fish oil in humans. Lipids 31 (suppl S): 45–49.

    Google Scholar 

  • Richardson B, Orcutt D, Schwerner H, Martínez C and Wickline H (1969) Effect of nitrogen limitation on the growth and composition of unicellular algae in continous culture. Appl. Microb. 18: 245–250.

    Google Scholar 

  • Robles Medina A, Giménez Giménez A, García Camacho F, Sánchez Pérez JA, Molina Grima E and Contreras Gómez A (1995a) Concentration and purification of stearidonic, eicosapentaenoic, and docosahexaenoic acids from cod liver oil and the marine microalga Isochrysis galbana. J. Am. Oil Chem. Soc. 72: 575–583.

    Google Scholar 

  • Robles Medina A, Giménez Giménez A, Molina Grima E and García Sánchez JL (1995b) Obtención de concentrados de ácidos grasos poliinsaturados por el método de los compuestos de inclusión de urea. Grasas y Aceites} 42}: 174–1

    Google Scholar 

  • Shimizu S, Shinmen Y, Kawashima H, Akimoto K and Yamada H (1987) Production of C-20 polyunsaturated fatty acids by fungi. Proceeding of World Conference on Biotechnology for the Fats and Oils Industry, (pp. 1000–1006). AOCS, Champaign, IL.

  • Shinmen Y, Shimizu S, Akimoto D, Kawashima H and Yamada H (1989) Production of arachidonic acid by Mortierellafungi. Selection of a potent producer and optimization of culture conditions for large-scale production. Appl. Microbiol. Biotechnol. 31: 11–16.

    Google Scholar 

  • Simopoulos AP (1991) Omega-3 fatty acids in health and disease and in growth and development. Am. J. Clin. Nutr. 54: 438–463.

    Google Scholar 

  • Singh MD and Chandra MD (1988) Biochemical and cellular effects of fish and fish oils. Prog. Food Nutr. Sci. 12: 371-419.

  • WHO/FAO (l977) Dietary fats and oils in human nutrition. Report of an expert consultation. FAO, Rome.

  • Yongmanitchai W and Ward OP (1991) Screening of algae for potential alternative sources of eicosapentaenoic acid. Phytochemistry 30: 2963-2967.

    Google Scholar 

  • Ziboh VA (1991) ω3 polyunsaturated fatty acid constituents of fish oil and the management of skin inflammatory and scaly disorders. In: AP Simopoulos, RR Kifer, RE Martin and SM Barlow (eds) Health Effects of ω3 Polyunsaturated Fatty Acid in Seafoods. Vol. 66 (pp. 425-435). Word Rev. Nutr. Diet.

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Authors and Affiliations

  1. Departamento de Ingeniería Química, Universidad de Almería, 04071, Almería, Spain

    A. Giménez Giménez, M.J. Ibáñez González, A. Robles Medina, E. Molina Grima, S. García Salas & L. Esteban Cerdán

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  1. A. Giménez Giménez
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  2. M.J. Ibáñez González
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  3. A. Robles Medina
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  4. E. Molina Grima
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  5. S. García Salas
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  6. L. Esteban Cerdán
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Giménez Giménez, A., Ibáñez González, M., Robles Medina, A. et al. Downstream processing and purification of eicosapentaenoic (20:5n-3) and arachidonic acids (20:4n-6) from the microalga Porphyridium cruentum. Bioseparation 7, 89–99 (1998). https://doi.org/10.1023/A:1008021330785

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