Enrichment of Eicosapentaenoic Acid from Sardine Oil with ∆5-Olefinic Bond Specific Lipase from Bacillus licheniformis MTCC 6824

Chakraborty, Kajal and Paulraj, R (2008) Enrichment of Eicosapentaenoic Acid from Sardine Oil with ∆5-Olefinic Bond Specific Lipase from Bacillus licheniformis MTCC 6824. Journal of Agricultural and Food Chemistry, 55. pp. 1428-1433.

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    Abstract

    Lipase derived from Bacillus licheniformis MTCC 6824 was purified to homogeneity by anion exchange chromatography on Amberlite IRA 410 (Cl-) and gel filtration using Sephadex G-100 as judged by denaturing polyacrylamide gel electrophoresis. The purified lipase was used for hydrolysis of triacylglycerol in sardine oil to enrich ∆5-polyunsaturated fatty acids (∆5-PUFAs) namely, arachidonic acid (5,8,11,14-eicosatetraenoic acid, ARA, 20:4n-6) and eicosapentaenoic acid (5,8,11,14,17- eicosapentaenoic acid, EPA, 20:5n-3). The individual fatty acids were determined as fatty acid methyl esters (FAMEs) by gas–liquid chromatography and gas chromatography–mass spectroscopy as FAMEs and N-acyl pyrrolidides. The enzyme exhibited hydrolytic resistance toward ester bonds of ∆5-PUFAs as compared to those of other fatty acids and was proved to be effective for increasing the concentration of EPA and ARA from sardine oil. Utilizing this fatty acid specificity, EPA and ARA from sardine oil were enriched by lipase-mediated hydrolysis followed by urea fractionation at 4 °C. The purified lipase produced the highest degree of hydrolysis for SFAs and MUFAs (81.5 and 72.3%, respectively, from their initial content in sardine oil) after 9 h. The profile of conversion by lipase catalysis showed a steady increase up to 6 h and thereafter plateaued down. Lipase-catalyzed hydrolysis of sardine oil followed by urea adduction with methanol provided free fatty acids containing 55.4% EPA and 5.8% ARA, respectively, after complexation of saturated and less unsaturated fatty acids. The combination of enzymatic hydrolysis and urea complexation proved to be a promising method to obtain highly concentrated EPA and ARA from sardine oil.

    Item Type: Article
    Uncontrolled Keywords: ∆5-Olefinic bond specific lipase; sardine oil; PUFAs; EPA; ARA
    Subjects: Fish Biotechnology > Bioactive compound
    Divisions: CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division
    Subject Area > CMFRI > CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division
    CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division
    Subject Area > CMFRI-Kochi > Marine Biotechnology, Fish Nutrition and Health Division
    Depositing User: Arun Surendran
    Date Deposited: 26 Feb 2014 10:13
    Last Modified: 09 Sep 2015 15:58
    URI: http://eprints.cmfri.org.in/id/eprint/9952

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