Abstract
The transesterification of soybean lecithin with ethyl esters of polyunsaturated fatty acids (eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)) using immobilized lipase from Rhizomucor miehei was tested in the presence or absence of organic solvent (hexane) and additives (urea with Ca2+ or Mg2+). The reaction was carried out at a water concentration of 4 % and ethyl ester to phospholipid mass ratio of 3:1. After 24 h of reaction without solvent or additive, fatty acid incorporation reached 29.1 % and thereafter increased only slightly. After 48 h, incorporation was highest in the presence of Mg2+, urea, and solvent. After 72 h, it was highest with Mg2+ and urea in the presence or absence of solvent (56.8 and 45.7 %, respectively). Incorporation of EPA and DHA was thus initially fast without solvent and additive, but was increased after prolonged reaction in the presence of Mg2+ and urea with or without solvent. These results are innovative and promising since they show that immobilized Rhizomucor miehei lipase has potential as a biocatalyst for interesterification reactions without solvent.
Similar content being viewed by others
References
Chojnacka, A., Gładkowski, W., Kiełbowicz, G., & Wawrzenczyk, C. (2009). Enzymatic enrichment of egg-yolk phosphatidylcholine with alpha-linolenic acid. Biotechnology Letters, 31, 705–709.
Schuchardt, J. P., Schneider, I., Meyer, H., Neubronner, J., von Schacky, C., & Hahn, A. (2011). Incorporation of EPA and DHA into plasma phospholipids in response to different omega-3 fatty acid formulations a comparative bioavailability study of fish oil vs. krill oil. Lipids in Health and Disease, 10, 145.
Guo, Z., Vikbjerg, A. F., & Xu, X. (2005). Enzymatic modification of phospholipids for functional applications and human nutrition. Biotechnology Advances, 23, 203–259.
Peng, L., Xu, X., Mu, H., Høy, C. E., & Adler-Nissen, J. (2002). Production of structured phospholipids by lipase-catalyzed acidolysis: optimization using response surface methodology. Enzyme and Microbial Technology, 31, 523–532.
Vikbjerg, A. F., Mu, H., & Xu, X. (2005). Lipase-catalyzed acyl exchange of soybean phosphatidylcholine in n-hexane: a critical evaluation of both acyl incorporation and product recovery. Biotechnology Progress, 21, 397–404.
Hita, E., Robles, A., Camacho, B., González, P. A., Esteban, L., Jiménez, M. J., Munío, M. M., & Molina, E. (2009). Production of structured triacylglycerols by acidolysis catalyzed by lipases immobilized in a packed bed reactor. Biochemical Engineering Journal, 46, 257–264.
Totani, Y., & Hara, S. (1991). Preparation of polyunsaturated phospholipids by lipase catalyzed transesterification. Journal of the American Oil Chemists’ Society, 68, 848–851.
Ghazali, H. M., Hamidah, S., & Che Man, Y. B. (1995). Enzymatic transesterification of palm olein with nonspecific and 1,3-specific lipases. Journal of the American Oil Chemists’ Society, 72, 633–639.
Rønne, T. H., Pedersen, L. S., & Xu, X. (2005). Triglyceride selectivity of immobilized Thermomyces lanuginosa lipase in interesterification. Journal of the American Oil Chemists’ Society, 82, 737–743.
Zainal, Z., & Yusoff, M. S. A. (1999). Enzymatic interesterification of palm stearin and palm kernel olein. Journal of the American Oil Chemists’ Society, 76, 1003–1008.
Otero, C., López-Hernandez, A., Garcia, H. S., Hernández-Martín, E., & Hill, C. G., Jr. (2006). Continuous enzymatic transesterification of sesame oil and a fully hydrogenated fat: effects of reaction conditions on product characteristics. Biochemical Engineering Journal, 94, 877–887.
Vikbjerg, A. F., Mu, H., & Xu, X. (2005). Parameters affecting incorporation and byproduct formation during the production of structured phospholipids by lipase-catalyzed acidolysis in solvent-free system. Journal of Molecular Catalysis B: Enzymatic, 36, 14–21.
Otero, C., Arcos, J. A., Berrendero, M. A., & Torres, C. (2001). Emulsifiers from solid and liquid polyols: different strategies for obtaining optimum conversions and selectivities. Journal of Molecular Catalysis B: Enzymatic, 11, 883–892.
Haraldsson, G. G., & Thorarensen, A. (1999). Preparation of phospholipids highly enriched with n-3 polyunsaturated fatty acids by lipase. Journal of the American Oil Chemists’ Society, 76, 1143–1149.
Snijder, H. J., & Dijkstra, B. W. (2000). Bacterial phospholipase A: structure and function of an integral membrane phospholipase. Biochimica et Biophysica Acta, 1488, 91–101.
Arpigny, J. L., & Jaeger, K. E. (1999). Bacterial lipolytic enzymes: classification and properties. Biochemical Journal, 343, 177–183.
Tanaka, A., Sugimoto, H., Muta, Y., Mizuno, T., Senoo, K., & Obata, H. (2003). Differential scanning calorimetry of the effects of Ca2+ on the thermal unfolding of Pseudomonas cepacia lipase. Bioscience Biotechnology and Biochemistry, 67, 207–210.
Kim, H., Choi, H., Kim, M., Sohn, C., & Oh, T. (2002). Expression and characterization of Ca2+ independent lipase from Bacillus pumilus B26. Biochimica et Biophysica Acta, 1583, 205–212.
Hernández, J. A., Aguilar, A. B., Portillo, B., López-Gómez, E., Beneyto, J. M., & Gárcia-Legaz, M. F. (2003). The effect of calcium on the antioxidant enzymes from salt-treated loquat and anger plants. Functional Plant Biology, 30, 1127–1137.
Rodrigues, R. C., & Fernandez-Lafuente, R. (2010). Lipase from Rhizomucor miehei as a biocatalyst in fats and oils modification. Journal of Molecular Catalysis B: Enzymatic, 66, 15–32.
Marsaoui, N., Laplante, S., Raies, A., & Naghmouchi, K. (2013). Incorporation of omega-3 polyunsaturated fatty acids into soybean lecithin: effect of amines and divalent cations on transesterification by lipases. World Journal of Microbiology and Biotechnology, 29, 2233–2338.
Christie, W.W. (2003). Analysis of conjugated linoleic acid—an overview. In: Advances in Conjugated Linoleic Acid Research. 21–12.
Yahya, A. R. M., Anderson, W. A., & Moo-Young, M. (1998). Ester synthesis in lipase catalyzed reactions. Enzyme and Microbial Technology, 23, 438–450.
Pepin, P., & Lortie, R. (1991). Influence of water activity on the enantioselective esterification of (R, S)-ibuprofen by Candida antarctica lipase B in solventless media. Biochemical Engineering Journal, 63, 502–505.
Bornscheuer, U., Herar, A., Kreye, L., Wendel, V., Capewell, A., & Meyer, H. H. (1993). Factors affecting the lipase catalyzed transesterification reactions of 3-hydroxy esters in organic solvents. Tetrahedron: Asymmetry, 4, 1007–1016.
Kim, I. H., Garcia, H. S., & Hill, C. G., Jr. (2010). Synthesis of structured phosphatidylcholine containing n-3 PUFA residues via acidolysis mediated by immobilized phospholipase A1. Journal of the American Oil Chemists’ Society, 87, 1293–1299.
Garcia, H. S., Kim, I., Lopez-Hernandez, A., & Hill, C. G., Jr. (2008). Enrichment of lecithin with n-3 fatty acids by acidolysis using immobilized phospholipase A1. Grasas y Aceites, 59, 368–374.
Adlercreutz, D., Budde, H., & Wehtje, E. (2002). Synthesis of phosphatidylcholine with defined fatty acid in the sn-1 position by lipasecatalyzed esterification and transesterification reaction. Biochemical Engineering Journal, 78, 403–411.
Egger, D., Wehtje, E., & Adlercreutz, P. (1997). Characterization and optimization of phospholipase A2 catalyzed synthesis of phosphatidylcholine. Biochimica et Biophysica Acta, 1343, 76–84.
Dong, H., Gao, S., Han, S., & Cao, S. (1999). Purification and characterization of a Pseudomonas sp. lipase and its properties in non-aqueous media. Biotechnology and Applied Biochemistry, 30, 251–256.
Surinenaite, B., Bendikiene, V., Juodka, B., Bachmatova, I., & Marcinkevichiene, L. (2002). Characterization and physicochemical properties of a lipase from Pseudomonas mendocina 3121–1. Biotechnology and Applied Biochemistry, 36, 47–55.
Pogori, N., Cheikyoussef, A., & Wang, D. (2008). Production and biochemical characterization of an extra cellular lipase from Rhizopus chinensis CCTCC M201021. Journal of Biotechnology, 7, 710–717.
Dave, R., & Madamwar, D. (2006). Esterification in organic solvents by lipase immobilized in polymer of PVA-alginate-boric acid. Process Biochemistry, 41, 951–955.
Silva, J. E. S., & Jesus, P. C. (2003). Evaluation of the catalytic activity of lipases immobilized on chrysotile for esterification. Anais da Academia Brasileira de Ciências, 75, 157–162.
Foresti, M. L., Alimenti, G. A., & Ferreira, M. L. (2005). Interfacial activation and bioimprinting of Candida rugosa lipase immobilized on polypropylene: effect on the enzymatic activity in solvent-free ethyl oleate synthesis. Enzyme and Microbial Technology, 36, 338–349.
Ye, R., Pyo, S. H., & Hayes, D. G. (2010). Lipase-catalyzed synthesis of saccharide-fatty acid esters using suspensions of saccharide crystals in solvent-free media. Journal of the American Oil Chemists’ Society, 87, 281–293.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Marsaoui, N., Naghmouchi, K., Baah, J. et al. Incorportation of Ethyl Esters of EPA and DHA in Soybean Lecithin Using Rhizomucor miehei Lipase: Effect of Additives and Solvent-Free Conditions. Appl Biochem Biotechnol 176, 938–946 (2015). https://doi.org/10.1007/s12010-015-1621-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-015-1621-3