Grasas y Aceites, Vol 66, No 4 (2015)

Glycerolysis of sardine oil catalyzed by a water dependent lipase in different tert-alcohols as reaction medium

Á. G. Solaesa
Department of Biotechnology and Food Science (Chemical Engineering Section), University of Burgos, Spain

M. T. Sanz
Department of Biotechnology and Food Science (Chemical Engineering Section), University of Burgos, Spain

R. Melgosa
Department of Biotechnology and Food Science (Chemical Engineering Section), University of Burgos, Spain

S. L. Bucio
Department of Biotechnology and Food Science (Chemical Engineering Section), University of Burgos, Spain

S. Beltrán
Department of Biotechnology and Food Science (Chemical Engineering Section), University of Burgos, Spain


The production of monoacylglycerol rich in polyunsaturated fatty acids (PUFA) via enzymatic glycerolysis of sardine oil in a homogeneous system was evaluated. Reactions were conducted in two different tert-alcohols. Based on the phase equilibrium data, the amount of solvent added to create a homogeneous system has been calculated and optimized. The immobilized lipase used in this work was Lipozyme RM IM from Rhizomucor miehei, a water dependent lipase. The amount of water added as well as other reaction parameters were studied to evaluate the optimum conditions for monoacylglycerol obtencion. An initial reactant mole ratio glycerol to sardine oil 3:1, 12 wt% of water based on glycerol content and 10 wt% of lipase loading (based on weight of reactants), achieved a MAG yield of around 70%, with nearly 28 wt% PUFA, with low free fatty acid content (lower than 18 wt%).


Fish oil; Glycerolysis; Lipase; Monoacylglycerol; Rhizomucor miehei; Tert-alcohols

Full Text:



AOAC Official Method 991.39. Fatty Acids in Encapsulated Fish Oils and Fish Oil Methyl and Ethyl Esters. 1995.

Blanco M, Sotelo CG, Chapela MJ, Pérez-Martín RI. 2007. Towards sustainable and efficient use of fishery resources: present and future trends. Trends Food Sci. Tech. 18, 29–36.

Bornscheuer UT. 1995. Lipase-catalyzed syntheses of monoacylglycerols. Enzyme Microb. Technol. 17, 578–586.

Cheirsilp B, Kaewthong W, H-Kittikun A. 2007. Kinetic study of glycerolysis of palm olein for monoacylglycerol production by immobilized lipase. Biochem. Eng. J. 35, 71–80.

Damstrup ML, Jensen T, Sparsø FV, Kiil SZ, Jensen AD, Xu X. 2005. Solvent optimization for efficient enzymatic monoacylglycerol production based on a glycerolysis reaction. J. Am. Oil Chem. Soc. 82, 559–564.

Feltes MMC, de Oliveira D, Block JM, Ninow JL. 2013. The Production, Benefits, and Applications of Monoacylglycerols and Diacylglycerols of Nutritional Interest. Food Bioprocess Tech. 6, 17–35.

Fregolente PBL,Fregolente LV, Pinto GMF, Batistella BC, Wolf-Maciel MR, Filho RM. 2008. Monoglycerides and diglycerides synthesis in a solvent-free system by lipase-catalyzed glycerolysis. Appl. Biochem. Biotechnol. 146, 165–172. PMid:18421596

Fureby AM, Virto C, Adlercreutz P, Mattiasson B. 1996. Acyl group migrations in 2-monoolein. Biocatal. Biotransform. 14, 89–111.

Hernandez EM. 2014. Issues in fortification and analysis of omega-3 fatty acids in foods. Lipid Technol. 26, 103–106.

Jin J, Li D, Zhu XM, Adhikari P, Lee K-T, Lee J-H. 2011. Production of diacylglycerols from glycerol monooleate and ethyl oleate through free and immobilized lipase-catalyzed consecutive reactions. New Biotechnol. 28, 190–195. PMid:20951847

Kim SK, Mendis E. 2006. Bioactive compounds from marine processing byproducts - A review. Food Res. Int. 39, 383–393.

Kristensen JB, Xu X, Mu H. 2005. Diacylglycerol synthesis by enzymatic glycerolysis: Screening of commercially available lipases. J. Am. Oil Chem. Soc. 82, 329–334.

Krüger RL, Valério A, Balen M, Ninow JL, Oliveira JV, de Oliveira D, Corazza ML. 2010. Improvement of mono and diacylglycerol production via enzymatic glycerolysis in tert-butanol system. Eur. J. Lipid Sci. Technol. 112, 921–927.

Majid N, Cheirsilp B. 2012. Optimal conditions for the production of monoacylglycerol from crude palm oil by an enzymatic glycerolysis reaction and recovery of carotenoids from the reaction product. Int. J. Food Sci. Technol. 47, 793–800.

Nichols PD, McManus A, Krail K, Sinclair AJ, Miller M. 2014. Recent advances in omega-3: Health benefits, Sources, Products and bioavailability. Nutrients. 6, 3727–3733. PMid:25255830 PMCid:PMC4179185

Pawongrat R, Xu X, H-Kittikun A. 2008. Physico-enzymatic production of monoacylglycerols enriched with very-long-chain polyunsaturated fatty acids. J. Sci. Food Agric. 88, 256–262.

Rendón X, López-Munguía A, Castillo E. 2001. Solvent engineering applied to lipase-catalyzed glycerolysis of triolein. J. Am. Oil Chem. Soc. 78, 1061–1066.

Riddick JA, Bunger WB, Sakano TK. 1986. Organic Solvents, Physical Properties and Methods of Purification. Wiley. New York.

Sidhu KS. 2003. Health benefits and potential risks related to consumption of fish or fish oil. Regul. Toxicol. Pharm. 38, 336–344. PMid:14623484

Singh AK, Mukhopadhyay M. 2012. Olive oil glycerolysis with the immobilized lipase Candida antarctica in a solvent free system. Grasas Aceites. 63, 202–208.

Solaesa ÁG, Bucio SL, Sanz MT, Beltrán S, Rebolleda S. 2013. Liquid-liquid equilibria for systems glycerol + sardine oil + tert-alcohols. Fluid Phase Equilib. 356, 284–290.

Solaesa ÁG, Bucio SL, Sanz MT, Beltrán S, Rebolleda S. 2014. Characterization of Triacylglycerol Composition of Fish Oils by Using Chromatographic Techniques. J. Oleo Sci. 63, 449–460. PMid:24770476

Torres C, Lin B, Hill Jr CG. 2002. Lipase-catalyzed glycerolysis of an oil rich in eicosapentaenoic acid residues. Biotechnol. Lett. 24, 667–673.

Voll F, Kru.ger RL, de Castilhos F, Filho LC, Cabral V, Ninow J, Corazza ML. 2011. Kinetic modeling of lipase-catalyzed glycerolysis of olive oil. Biochem. Eng. J. 56, 107–115.

Weber N, Mukherjee KD. 2004. Solvent-free lipase-catalyzed preparation of diacylglycerols. J. Agric. Food Chem. 52, 5347–5353. PMid:15315368

Wongsakul S, Prasertsan P, Bornscheuer UT, H-Kittikun A. 2003. Synthesis of 2-monoglycerides by alcoholysis of palm oil and tuna oil using immobilized lipases. Eur. J. Lipid Sci. Technol. 105, 68–73.

Xu X. 2000. Production of specific-structured triacylglycerols by lipase-catalyzed reactions: a review. Eur. J. Lipid Sci. Technol. 102, 287–303.<287::AID-EJLT287>3.0.CO;2-Q

Xu X. 2003. Engineering of enzymatic reactions and reactors for lipid modification and synthesis. Eur. J. Lipid Sci. Technol. 105, 289–304.

Yang T, Rebsdorf M, Engelrud U, Xu X. 2005a. Enzymatic production of monoacylglycerols containing polyunsaturated fatty acids through an efficient glycerolysis system. J. Agric. Food Chem. 53, 1475–1481. PMid:15740027

Yang T, Rebsdorf M, Engelrud U, Xu X. 2005b. Monoacylglycerol synthesis via enzymatic glycerolysis using a simple and efficient reaction system. J. Food Lipids. 12, 299–312.

Yeoh CM, Choong TSY, Abdullah LC, Yunus R, Siew WL. 2009. Influence of silica gel in production of diacylglycerol via enzymatic glycerolysis of palm olein. Eur. J. Lipid Sci. Technol. 111, 599–606.

Zhong N, Li L, Xu X, Cheong L, Li B, Hu S, Zhao X. 2009. An Efficient Binary Solvent Mixture for Monoacylglycerol Synthesis by Enzymatic Glycerolysis. J. Am. Oil Chem. Soc. 86, 783–789.

Copyright (c) 2015 Consejo Superior de Investigaciones Científicas (CSIC)

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Contact us

Technical support