Purification of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) From Fish Oil Using HPLC Method and Investigation of Their Antibacterial Effects on Some Pathogenic Bacteria

Purification of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) From Fish Oil Using HPLC Method and Investigation of Their Antibacterial Effects on Some Pathogenic Bacteria

The aim of this study was to purified eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) essential oils from trout oil using high performance liquid chromatography (HPLC) method, and bioconverted EPA and DHA into bioconverted EPA (bEPA) and bioconverted DHA (bDHA) extracts by P. aeruginosa PR3. Moreover, in vitro antibacterial activity of bEPA and bDHA was investigated using disc diffusion methods and minimum inhibitory concentration (MIC). EPA and DHA concentration of 11.1% and 15.9% in trout oil increased in 58.64% and 40.33% after HPLC optimisation, respectively. In this study, EPA and DHA enriched products were obtained which are to be used as valuable supplements for food and pharmaceutical purposes. The bioconverted EPA and DHA exhibited antibacterial activities against two Gram-positive bacteria (Listeria monocytogenes ATCC 7677 and Staphylococcus aureus ATCC 29213) and six Gram-negative bacteria (Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC700603, Enterococcus faecalis ATCC 29212, Aeromonas hydrophila NCIMB 1135 and Salmonella paratyphi A NCTC 13). Inhibition zones and MIC value of bEPA and bDHA against bacterial strains ranged from 7 to 12 mm and from 350 to 2350 μg/mL, respectively. Our results suggested that the crude extracts of bioconversion of EPA and DHA by P. aeruginosa PR3 can be considered as promising antimicrobials in improving food safety by controlling foodborne pathogens.

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  • BAJPAIA, V.K., SHINA, S.Y., KIMB, H.R., KANGA, S.C., 2008. Anti-fungal action of bioconverted eicosapentaenoic acid (bEPA) against plant pathogens. Industrial Crops and Products 27, 136-141.
  • CASTRO, I.A., TIRAPEGUIA, J., SILVA, R.S.S.F., CUTRIM, J.S., 2004. Sensory evaluation of a milk formulation supplemented with ω-3 polyunsaturated fatty acids and soluble fibres. Food Chem, 85 (4): 503 - 512.
  • ECKEL, R.H., KRAUSS, R.M., 1998. American Heart Association Call to Action: Obesity as a Major Risk Factor for Coronary Heart Disease. Journal of The American Heart Association. 97:2099-2100.
  • GIBNEY, M.J., 1997. Incorporation of ω-3 polyunsaturated fatty acids to processed food. British Journal of Nutrition, 78: 193 -195.
  • GORDON, D.T., RATLIFF, V., 1992. The implications of omega-3 fatty acids in human healty, Advances in Seafood Biochemistry Composition and Quality, Ed. By George L. Flick, 406 pp. 69-98.
  • HOSOKAWA, M., HOU, C.T., WEISLEDER, D., 2003. Bioconversion of ω-3 and ω-6 PUFA by Clavibacter sp. ALA2. Journal of the American Oil Chemists' Society 80, 1085–1091.
  • HOU, C.T., 1995. Microbial oxidation of unsaturated fatty acids. In: Laskin, A.I. (Ed.), Advances in Applied Microbiology, vol. 41. Academic Press, Orlando, pp. 1–23.
  • HOU, C.T., BAGBY, M.O., 1991. Production of a new compound, 7,10-dihydroxy- 8(E)-octadecenoic acid from oleic acid by Pseudomonas sp. PR3. Journal of Industrial Microbiology 7, 123–130.
  • ICHIBARA, K., SHIBAHARA, A., YAMAMOTO, K. ve NAKAYAMA, T. 1996. An Improved Method for Rapid Analysis of The Fatty Acids of Glycerolipids, Lipids, 31: 535-539.
  • KIM, H., GARDNER, H.W., HOU, C.T., 2000. Production of isomeric 9,10,13 (9,12,13)-trihydroxy-11E (10E)-octadecenoic acid from linoleic acid by Pseudomonas aeruginosa PR3. Journal of Industrial Microbiology and Biotechnology 25, 109–115.
  • KOLANOWSKI, W., LAUFENBERG, G., 2006. Enrichment of food products with polyunsaturated fatty acids by fish oil addition. Europea Food Research Technology, 222: 472 - 477.
  • KOLANOWSKI, W., SWIDERSKI, F., BERGER, S., 1999. Possibility of fish oil application for food products enrichment with omega-3 PUFA. International Journal of Food Science and Nutritions, 50: 39 - 49.
  • KUO, T.M., KIM, H., HOU, C.T., 2001. Production of a novel compound, 7,10,12- trihydroxy-8(E)-octadecenoic acid from ricinoleic acid by Pseudomonas aeruginosa PR3. Current Microbiology 43, 198–203.
  • KUO, T.M., MANTHEY, L.K., HOU, C.T., 1998. Fatty acid bioconversions by Pseudomonas aeruginosa PR3. Journal of the American Oil Chemists' Society 75, 875–879.LEAF, A., WEBER, P. C., 1988. Cardiovaskular effekts of n-3 fatty acids, N. Engl. J. Med., 318, 549-557.
  • LOVEGROVE JA, BROOKS CN, MURPHY MC, GOULD BJ, WILLIAMS CM., 1997. Use of manufactured foods enriched with fish oils as a means of increasing long-chain n-3 polyunsaturated fatty acid intake. British Journal of Nutrition, 78: 223 - 236.
  • MURRAY, P.R., BARON, E.J., PFALLER, M.A., TENOVER, F.C., YOLKE, R.H., 1995. Manual of Clinical Microbiology, 6th ed. ASM, Washington, DC.
  • NIETO, S., CÓRDOBA, A.M., SANHUEZA, J., VALENZUELA, A., 1997. Obtention of highly purified fractions of eicosapentaenoic acid and docosahexaenoic acid from sardine oil by silver-resin chromatography: A semi-preparative procedure. Grasas y Aceites, 48, 197-199.
  • PIGOTT, G.M., B.W. TUCKER., 1990. Seafood effects of technology on nutrition. Marcel Dekker, Inc. New York.
  • SHIN, S.Y., BAJPAI, V.K., KIM, H.R., KANG, S.C., 2006. Antibacterial activity of bioconverted eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) against food borne pathogenic bacteria. International Journal of Food Microbiology, 113, 233–236.
  • TAUTWEIN, E.A., 2001. Omega-3 fatty acids-physiological and technical aspects for their use in food. European Journal of Lipid Science and Technology, 103:45-52.