• A. Huesca-Toral
  • A. López-Hernández
  • J. O. Angulo-Guerrero
  • C. G. Hill, Jr.
  • H. S. García
Keywords: conjugated linoleic acd, triacylglycerides, enzymatic polyesterification, temperature, molar ratio


Conjugated linoleic acid (CLA) is a term used to designate a mixture of positional and geometrical isomers of linoleic acid. Several of these isomers have been associated with important biological roles, including anti carcinogenic activity. Basically, the natural sources of CLA are dairy and meat products. However, its presence in such foods is in very small quantities. The present work was undertaken to evaluate the effect of temperature (40, 50, 60 and 70°C), molar ratio of substrates (3:1 and 4:1, free fatty acid: glycerol), and two commercially available biocatalysts [Candida antarctica fraction B (Chyrazyme L-2), and Rhizomucor miehei (IM-60)], on the rate of production of triacylglycerols (TAG) from glycerol and CLA via enzyme-mediated polyesterification reactions. The formation of acylglycerols and the consumption of the free fatty acids were monitored by periodic withdrawal of samples for analysis by HPLC. Best treatments were selected by the Tukey’s multiple comparison test (p<0.05). The conditions which produced maximum synthesis of TAG (76% ±2.7 moles of the original fatty acid were esterified to form TAG), were those in which free fatty acid was used in molar excess (4:1 mole ratio of CLA to glycerol) and high temperatures (60 and 70°C) were employed. For these conditions, there were no significant differences between the results obtained with either commercial enzyme. For both enzymes, a linear relationship was observed between the production of the TAG and the reaction temperature


Akoh, C.C. y Min, P.B. (1998). Food Lipids. Chemistry, Nutrition and Biotechnology. Marcel Dekker, Inc. EUA.

Arcos, J., Otero, C. y Hill Jr., C.G. (1998). Rapid enzymatic production of acylglycerols from conjugated linoleic acid and glycerol in a solvent-free system. Biotechnology Letters 6, 617-621.

Arcos, J.A., García, H.S. y Hill Jr., C.G. (2000). Continuous enzymatic esterification of glycerol with (poly) unsaturated fatty acids in a packed-bed reactor. Biotechnology and Bioengineering 5, 563-570.

Cerdán, L.E., Medina, R.A., Jiménez, G.A., Ibáñez, G.M.J. y Molina, G.E. (1998). Síntesis of polyunsaturated fatty acidenriched triglycerides by lipase-catalysed esterification. Jounal of the American Oil Chemists’ Society 10, 1329-1337.

Ergan, F. Trani, M. y André, G. (1990). Production of glycerydes from glycerol and fatty acid by immobilized lipases in non-aqueous media. Biotechnology and Bioengineering 35, 195-200.

García, H.S., Storkson, J.M., Pariza, M.W. y Hill Jr., C.G. (1998). Enrichment of butteroil with conjugated linoleic acid via enzymatic interesterification (acidolysis) reactions. Biotechnology Letters 4, 393-395.

Haraldsson, G., Gudmundsson, G. y Almarsson, O. (1995). The synthesis of homogeneous triglycerides of eicosapentaenoic acid and docosahexaenoic acid by lipase. Tetrahedron Letter. 36, 941-952.

Kosugi, Y. y Azuma, N. (1994). Synthesis of triacylglycerol from polyunsaturated fatty acid by immobilized lipase. Journal of the American Oil Chemists’ Society 12, 1397-1403.Li, Z. y Ward, P. (1993). Enzyme catalyzed production of vegetable oils containing omega-3 polyunsaturated fatty acid. Biotechnology Letters 15, 185-188.

Liu, J., Lee, T., Bobik, Jr E., Guzman-Harty, M.,y Hastilow, C. (1993). Quantitative determination of monoglycerides and diglycerides by high-performance liquid chromatography and evaporative lightscattering detection. Journal of the American Oil Chemists’ Society 4, 343- 347.

Lortie, R., Trani, M. y Ergan, F. (1992). Kinetic study of the lipase-catalyzed synthesis of triolein. Biotechnology and Bioenginering 41, 1021-1026.

Malcata, F., Reyes, H., Garcia, H.S., Amundson C.H. y Hill Jr., C.G. (1992). Kinetics and mechanisms of reactions catalysed by immobilized lipases. Enzyme and Microbial Technology 14, 426-446.

Marangoni, A.G. y Rousseau, D. (1995). Engineering triacylglycerols: the role of interesterification. Trends in Food Science & Technology 6, 329-335.

Martínez, C. E. (2001). Empleo de lipasas en solventes orgánicos para preparar glicéridos enriquecidos con ácido linoleico conjugado (CLA). Tesis de Doctorado Instituto Tecnológico de Veracruz. México.

Noriega, J. A.. 2002. Cinética de esterificación enzimática del ácido eicosapentaenoico y ácido docosahexaenoico del aceite de sardina (Sardinops sagax caeruleus). Tesis de Maestría. Universidad de Sonora. México.

Otero, C., Arcos, J.A., García, H.S. y Hill, C.G. Jr. (1999). Enzymatic synthesis and hydrolysis reactions of acylglycerols in solvent-free systems. En: Methods in Biotechnology, Enzymes in Nonaqueous Solvents: Methods and Protocols. (E. N. Vulfson, P.J. Halling, y H. L. Holland, eds.), Pp. 479-492. Humana Press Inc., Totowa, NJ.

Robles-Medina, A., Esteban-Cerdán, L., Jiménez-Giménez, A., Camacho-Paez, B., Ibáñez- González, M. J. y Molina-Grima, E. (1999). Lipase-catalyzed esterification of glycerol and polyunsaturated fatty acids from fish and microalgae oil. Journal of Biotechnology 70, 379-391.

Selmi, B., Gontier, E., Ergan, F. y Thomas, D. (1998). Effects of fatty acid chain length and unsaturation number on triglyceride synthesis catalyzed by immobilized lipase in solvent-free medium. Enzyme and Microbial Technology 23, 182-186.

Segel, I.H. (1975). Effects of pH and temperature. Capitulo 11, en: Enzyme kinetics: behavior and analysis of rapid equilibrium and steady-state enzyme systems. Pp 926-929. John Wiley & Sons. Nueva York.

Shuler, M.L. y Kargi, F. (1992). Bioprocess Engineering. 1a. ed. Prentice Hall, Englewood Cliffs, N.J. Pp 58-67.
How to Cite
Huesca-Toral, A., López-Hernández, A., Angulo-Guerrero, J. O., Hill, Jr., C. G., & García, H. S. (2020). SYNTHESIS OF CLA-ENRICHED TRIACYLGLYCEROLS BY ENZYMATIC POLYESTERIFICATION IN A SOLVENT-FREE MEDIUM. Revista Mexicana De Ingeniería Química, 4(1), 75-87. Retrieved from