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Preparation of Triacylglycerols Rich in Omega-3 Fatty Acids from Sardine Oil Using a Rhizomucor miehei Lipase: Focus in the EPA/DHA Ratio

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Abstract

The increasing evidence on the differential biochemical effects of eicosapentaenoic acid (EPA)/docosahexaenoic acid (DHA) raises the need of n-3 highly unsaturated fatty acid concentrates with different amounts of these fatty acids. In the present work, physicochemical and enzymatic techniques were combined to obtain acylglycerols, mainly triacylglycerols (TAG), rich in n-3 fatty acids. Sardine oil was obtained by washing sardine (Sardina pilchardus) mince with a NaHCO3 solution, hydrolyzed in a KOH–ethanol solution, and concentrated with urea. The esterification reaction was performed in the stoichiometric proportion of substrates for re-esterification to TAG, with 10 % level of Rhizomucor miehei lipase based on the weight of substrates, without any solvent, during 48 h. This procedure led to approximately 88 % of acylglycerols, where more than 66 % were TAG and the concentration of n-3 fatty acids was higher than 60 %, the EPA and DHA ratio (EPA/DHA) was 4:1. The content of DHA in the unesterifed fraction (free fatty acids) increased from 20 to 54 %, while the EPA level in the same fraction decreased from 33 to 12.5 % (EPA/DHA ratio ≈1:4). Computational methods (density functional theory calculations) have been carried out at the B3LYP/6-31G(d,p) level to explain some of the experimental results.

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Authors and Affiliations

  1. Department of Biochemistry and CEDOC, Faculty of Medical Sciences, Universidade Nova de Lisboa (UNL), Campo Mártires da Pátria, 130, 1169-056, Lisbon, Portugal

    Paulo Bispo

  2. DivAv, IPMA, Av. Brasília, 1449-006, Lisbon, Portugal

    Irineu Batista & Narcisa Maria Bandarra

  3. GIRM, Laboratory of Separation and Reaction Engineering, ESTM, Instituto Politécnico de Leiria, Santuário N.ª Sra. dos Remédios, Apartado 126, 2524-909, Peniche, Portugal

    Raul J. Bernardino

Authors
  1. Paulo Bispo
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  2. Irineu Batista
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  3. Raul J. Bernardino
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  4. Narcisa Maria Bandarra
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Correspondence to Paulo Bispo.

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Bispo, P., Batista, I., Bernardino, R.J. et al. Preparation of Triacylglycerols Rich in Omega-3 Fatty Acids from Sardine Oil Using a Rhizomucor miehei Lipase: Focus in the EPA/DHA Ratio. Appl Biochem Biotechnol 172, 1866–1881 (2014). https://doi.org/10.1007/s12010-013-0616-1

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