Abstract
Oils with a high DAG (1,3-DAG) content have attracted considerable attention as a healthful food oil component. In this study, we report on the synthesis of 1,3-DAG from a mixture of FA, constituted largely of oleic and linoleic acids, using an immobilized 1,3-regioselective lipase from Rhizomucor miehei in a solvent-free system. The kinetics of 1,3-DAG production from FA and glycerol were investigated on the basis of a simplified model, taking into consideration the acyl migration reaction, the removal of water, and glycerol dissolution in the oil phase in addition to the esterification reactions. Both the yield of 1,3-DAG and the purity of DAG were evaluated under a variety of experimental conditions, including reaction temperature, pressure, and amount of enzyme present. When either the reaction temperature or the amount of enzyme used was increased, the 1,3-DAG production rate increased, but yield remained relatively constant. The 1,3-DAG yield as well as the purity of DAG gradually decreased because of the enhancement of acyl migration at later stages of the reaction after the 1,3-DAG concentration reached a maximum. Vacuum was important for attaining high yields of 1,3-DAG. Under conditions of a high vacuum (1 mm Hg) at 50°C, 1.09 M 1,3-DAG was produced from 1.29 M glycerol and 2.59 MFA in an 84% yield and in 90% purity.
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References
D'Alonzo, R.P., W.J. Kozarek, and R.L. Wade, Glyceride Composition of Processed Fats and Oils As Determined by Glass Capillary Gas Chromatography, J. Am. Oil Chem. Soc. 59: 292–295 (1982).
Wähnelt, S., D. Meusel, and M. Tülsner, Influence of Diglyceride on the Phase-Behavior of Edible Fats, Fat. sci. Technol. 93:117–121 (1991).
Berger, M., and M.P. Schneider, Regioisomerically Pure Mono-and Diacylglycerols as Synthetic Building Blocks, 95: 169–175 (1993).
Haftendorn, R., and R. Ulbrich-Hofmann, Synthesis of 2-Modified 1,3-Diacylglycerols, Tetrahedron 51:1177–1186 (1995).
Hara, K., K. Onizawa, H. Honda, T. Ide, K. Otsuji, and M. Murata, Dietary Diacylglycerol-Dependent Reduction in Serum Triacylglycerol Concentration in Rats, Ann. Nutr. Metab. 37:185–191 (1993).
Nagao, T., H. Watanabe, N. Goto, K. Onizawa, H. Taguchi, N. Matsuo, T. Yasukawa, R. Tsusima, H. Shimasaki, and H. Itakura, Dietary Diacylglycerol Suppresses Accumulation of Body Fat Compared to Triacylglycerol in Men in a Double-Blind Controlled Trial, J. Nutr. 130:792–797 (2000).
Watanabe, H., K. Onizawa, H. Taguchi, M. Kobori, H. Chiba, S. Naito, N. Matuo, T. Yasukawa, M. Hattori, and H. Shimasaki, Nutritional Characterization of Diacylglycerols in Rats, J. Jpn. Oil Chem. Soc. 46:301–307 (1997).
Murata, M., K. Hara, and T. Ide, Alteration by Diacylglycerols of the Transport and Fatty Acid Composition of Lymph Chylomicrons in Rats, Biosci. Biotechnol. Biochem. 58:1416–1419 (1994).
Sontang, N.O.V., New Developments in the Fatty Acid Industry in America, J. Am. Oil Chem. Soc. 61:229–232 (1984).
Plou, J.F., M. Barandiárn, V.M. Calvo, A. Ballesteros, and E. Paster, High-Yield Production of Mono-and Di-oleylglycerol by Lipase-Catalyzed Hydrolysis of Triolein, Enzyme Microb. Technol. 18:66–71 (1996).
Yamane, T., S.T. Kang, K. Kawahara, and Y. Koizumi, High-Yield Diacylglycerol Formation by Solid-Phase Enzymatic Glycerolysis of Hydrogenated Beef Tallow, J. Am. Oil Chem. Soc. 71: 339–342 (1994).
Berger, M., K. Laumen, and M.P. Schneider, Enzymatic Esterification of Glycerol I. Lipase-Catalyzed Synthesis of Regioisomerically Pure 1,3-sn-Diacylglycerols, 69:955–960 (1992).
Rosu, R., M. Yasui, Y. Iwasaki, and T. Yamane, Enzymatic Synthesis of Symmetrical 1,3-Diacylglycerols by Direct Esterification of Glycerol in Solvent-Free System, 76:839–843 (1999).
Official Methods and Recommended Practices of the American Oil Chemists' Society, 5th edn., AOCS Press, Champaign, IL, 1997, Method Ca 5a-40.
Ibid., Official Methods and Recommended Practices of the American Oil Chemists' Society, 5th edn., AOCS Press, Champaign, IL, 1997, Method Ca 11b-91.
Lortie, R., M. Trani, and F. Ergan, Kinetic Study of the Lipase-Catalyzed Synthesis of Triolein, Biotechnol. Bioeng. 41:1021–1026 (1993).
Hirota, Y., J. Kohori, and Y. Kawahara, Preparation of Diglycerides, European Patent 0307154 B2 (1988).
Dudal, Y., and R. Lortie, Influence of Water Activity on the Synthesis of Triolein Catalyzed by Immobilized Mucor meihei Lipase, Biotechnol. Bioeng. 45:129–134 (1995).
Zaks, A., and A.M. Klibanov, The Effect of Water on Enzyme Action in Organic Media, J. Biol. Chem. 263:8017–8021 (1988).
Marty, A., V. Dossat, and J.-S. Condoret, Continuous Operation of Lipase-Catalyzed Reactions in Nonaqueous Solvents: Influence of the Production of Hydrophilic Compounds, Biotechnol. Bioeng. 56:232–237 (1997).
Xu, X., A.R.H. Skands, C.-E. Høy, H. Mu, S. Balchen, and J. Adler-Nissen Production of Specific-Structured Lipids by Enzymatic Interesterification: Elucidation of Acyl Migration by Response Surface Design, J. Am. Oil Chem. Soc. 75:1179–1186 (1998).
Xu, X., H. Mu, A.R.H. Skand, C.-E. Høy, and J. Adler-Nissen, Parameters Affecting Diacylglycerol Formation During the Production of Specific-Structured Lipid by Lipase-Catalyzed Interesterification, 76:175–181 (1999).
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Watanabe, T., Shimizu, M., Sugiura, M. et al. Optimization of reaction conditions for the production of DAG using immobilized 1,3-regiospecific lipase lipozyme RM IM. J Amer Oil Chem Soc 80, 1201–1207 (2003). https://doi.org/10.1007/s11746-003-0843-5
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DOI: https://doi.org/10.1007/s11746-003-0843-5