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Purification of Free DHA by Selective Esterification of Fatty Acids from Tuna Oil Catalyzed by Rhizopus oryzae Lipase

  • Original Paper
  • Published:
Journal of the American Oil Chemists' Society

Abstract

Tuna fish oil contains 25–30 % docosahexaenoic acid (DHA) and is one of the richest sources of DHA. The present paper investigates the enrichment of DHA by selective esterification of fatty acids obtained from hydrolysis of tuna fish oil catalyzed by Rhizopus oryzae lipase (ROL). The fatty acid mixture obtained after hydrolysis of tuna fish oil, referred to as tuna-FFA contained 26 % DHA. For purification/concentration of DHA in free fatty acids, selective esterification of the fatty acid mixtures with butanol was carried out using ROL in a water-organic solvent system. The best reaction parameters found in this study were pH 7, temperature 35 °C, agitation speed 800 rpm and a fatty acid to solvent (iso-octane) ratio of 1:1.32 (w/v). Also, the effects of other parameters such as type of alcohol, type of enzyme, alcohol to fatty acid ratio, enzyme to fatty acid ratio were studied to determine the most suitable reaction conditions. Exactly 76.2 % of tuna-FFA was esterified in 24 h, under the most suitable reaction conditions and the DHA content in the fatty acid fraction rose from 26 to 86.9 % with 80 % recovery of DHA, after selective esterification. The DHA content of fatty acids in butyl esters was found to be 13.6 %.

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Abbreviations

DHA:

Docosahexaenoic acid

EPA:

Eicosapentaenoic acid

FAME:

Fatty acid methyl ester(s)

FFA:

Free fatty acid(s)

GC:

Gas chromatography

GLA:

Gamma-linolenic acid

PUFA:

Poly unsaturated fatty acid(s)

ROL:

Rhizopus oryzae lipase

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Acknowledgments

One of the authors, Ms. Kriti Bhandari would like to acknowledge the Council of Scientific and Industrial Research (CSIR), New Delhi for providing her with a senior Research Fellowship (SRF). The authors also would like to acknowledge the financial support through Canada Research Chair Program and facilities provided by the Chemical and Biological Engineering Department, University of Saskatchewan, Canada for carrying out the experimental work.

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Authors and Affiliations

  1. Department of Chemical Engineering, Malaviya National Institute of Technology, Jaipur, 302017, India

    Kriti Bhandari, S. P. Chaurasia & Alok Gupta

  2. Catalysis and Chemical Engineering Laboratories, Department of Chemical Engineering, University of Saskatchewan, Saskatoon, SK, S7N 5A9, Canada

    Kriti Bhandari & A. K. Dalai

Authors
  1. Kriti Bhandari
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  2. S. P. Chaurasia
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  3. A. K. Dalai
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  4. Alok Gupta
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Corresponding author

Correspondence to S. P. Chaurasia.

Electronic supplementary material

Below is the link to the electronic supplementary material.

11746_2013_2322_MOESM1_ESM.tif

Supplementary Fig. 1 Effect of pH on the esterification of fatty acids with butanol using ROL. Experimental conditions: 0.85 g tuna-FFA, 0.44 g butanol, 1.12 ml iso-octane, temperature 35 °C, agitation speed 800 rpm, reaction time 1 h, ROL 40 mg dissolved in 0.5 ml buffer. (tif 39.9 kb)

11746_2013_2322_MOESM2_ESM.tif

Supplementary Fig. 2 Effect of the fatty acid to buffer ratio on esterification with butanol using ROL. Experimental conditions: 0.85 g tuna-FFA, 0.44 g butanol, temperature 35 °C, agitation speed 800 rpm, reaction time 1 h, ROL 40 mg dissolved in buffer, pH 7 and iso-octane 1.12 ml. (tif 48.2 kb)

11746_2013_2322_MOESM3_ESM.tif

Supplementary Fig. 3 Effect of temperature on the esterification of fatty acids with butanol using ROL. Experimental conditions: 0.85 g tuna-FFA, 0.44 g butanol, 1.12 ml iso-octane, pH 7, agitation speed 800 rpm, reaction time 1 h, ROL 40 mg dissolved in 0.5 ml buffer. (tif 38.0 kb)

11746_2013_2322_MOESM4_ESM.tif

Supplementary Fig. 4 Effect of speed of agitation on the esterification of fatty acids with butanol using ROL. Experimental conditions: 0.85 g tuna-FFA, 0.44 g butanol, 1.12 ml iso-octane, pH 7, temperature 35 °C, reaction time 1 h, ROL 40 mg dissolved in 0.5 ml buffer. (tif 38.3 kb)

11746_2013_2322_MOESM5_ESM.tif

Supplementary Fig. 5 Effect of solvent concentration on the esterification of fatty acids with butanol using ROL. Experimental conditions: 0.85 g tuna-FFA, 0.44 g butanol, 1.12 ml iso-octane, pH 7, temperature 35 °C, agitation speed 800 rpm, reaction time 1 h, ROL 40 mg dissolved in 0.5 ml buffer. (tif 71.5 kb)

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Bhandari, K., Chaurasia, S.P., Dalai, A.K. et al. Purification of Free DHA by Selective Esterification of Fatty Acids from Tuna Oil Catalyzed by Rhizopus oryzae Lipase. J Am Oil Chem Soc 90, 1637–1644 (2013). https://doi.org/10.1007/s11746-013-2322-0

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  • DOI: https://doi.org/10.1007/s11746-013-2322-0

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