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Enzymatic Synthesis of Medium- and Long-Chain Triacylglycerols (MLCT): Optimization of Process Parameters Using Response Surface Methodology

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Abstract

The main objective of this study was to understand the effects and relationship amongst four factors, which are reaction temperature, reaction time, enzyme load, and substrate mole ratio with the purpose of producing healthy functional cooking oil for long-term dietary treatment. Lipozyme RM IM lipase-catalyzed esterification of medium- and long-chain triacylglycerols (MLCT) from glycerol and mixtures of capric and oleic acid was optimized using response surface methodology (RSM) with a five-level, four-factorial design. Reaction temperature, reaction time, and substrate mole ratio strongly affected MLCT synthesis (P < 0.01). However, enzyme load did not have a significant (P > 0.01) effect on MLCT yield. Comparison between predicted and experimental value from central composite rotatable design optimization procedures revealed good correlation, implying that the reduced cubic polynomial model with backward elimination statistically expressed the percent MLCT yield obtained. The optimum MLCT yield was 59.76% by using 10 wt% enzyme load, reaction temperature of 70°C, reaction time of 14 h, and substrate mole ratio of 3.5:1. Experiments to confirm the predicted results using the optimal parameters showed an MLCT yield of 56.35% (n = 2). The choice on the types of fatty acids used in MLCT optimization work greatly influenced the physical and chemical properties of MLCT oil produced. The refined MLCT oil characteristics study showed this oil is suitable to be used for cooking/frying purposes as a high-value added product.

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References

  • AOCS (1997). Official methods and recommended practices of the American oil chemist’s society (5th ed.). Champaign, USA: American Oil Chemist’s Society Press.

    Google Scholar 

  • Aoyama, T. (2003). Fats and oils composition for reducing lipids in blood. U. S. Patent No 0170368 A1.

  • Bendixen, H., Flint, H. A., Raben, A., Høy, C. E., Mu, H., Xu, X., et al. (2002). Effect of 3 modified fats and a conventional fat on appetite, energy intake, energy expenditure and substrate oxidation in healthy men. American Journal of Clinical Nutrition, 75, 47–56.

    CAS  Google Scholar 

  • Deniz, B., & İsmail, H. B. (2007). Modelling and optimization I: usability of response surface methodology. Journal of Food Engineering, 78, 836–845.

    Article  Google Scholar 

  • Fomuso, L. B., & Akoh, C. C. (1997). Enzymatic modification of triolein: incorporation of caproic and butyric acids to produce reduced-calorie structured lipids. Journal of the American Oil Chemists’ Society, 74, 269–272.

    Article  CAS  Google Scholar 

  • Fu, X., & Parkin, K. L. (2004). Selectivity of fatty acid incorporation into acylglycerols in esterification reactions using Rhizomucor miehei and Burkholderia cepacia lipases. Food Research International, 37, 651–657.

    Article  CAS  Google Scholar 

  • Garnacho-Montero, J., Ortiz-Leyba, C., Jiménez-Jiménez, F. J., Garcia-Garmendia, J. L., Jiménez-Jiménez, L. M., Garnacho-Montero, M. C., et al. (2002). Clinical and metabolic effects of two lipid emulsions on the parenteral nutrition of septic patients. Nutrition, 18, 134–138.

    Article  CAS  Google Scholar 

  • Hassel, C. (1993). Nutritional implication on fat substitutes. Cereal Foods World, 38, 142–144.

    CAS  Google Scholar 

  • Jennings, B. H., & Akoh, C. C. (2001). Lipase catalyzed modification of fish oil to incorporate capric acid. Food Chemistry, 72, 273–278.

    Article  CAS  Google Scholar 

  • Kasai, M., Nosaka, N., Maki, H., Negishi, S., Aoyama, T., Nakamura, M., et al. (2003). Effect of dietary medium- and long-chain triacylglycerols (MLCT) on accumulation of body fat in healthy humans. Asia Pacific Journal of Clinical Nutrition, 12, 151–160.

    CAS  Google Scholar 

  • Kawashima, A., Nagao, T., Watanabe, Y., Kobayashi, T., Ikeda, I., Tominaga, Y., et al. (2004). Preparation of regioisomers of structured TAG consisting of one mole of CLA and two moles of caprylic acid. Journal of the American Oil Chemists’ Society, 81, 1013–1020.

    Article  CAS  Google Scholar 

  • Kawashima, A., Shimada, Y., Yamamoto, M., Sugihara, A., Nagao, T., Komemushi, S., et al. (2001). Enzymatic synthesis of high-purity structured lipids with caprylic acid at 1,3-positions and polyunsaturated fatty acid at 2-position. Journal of the American Oil Chemists’ Society, 78, 611–616.

    Article  CAS  Google Scholar 

  • Kim, B. H., & Akoh, C. C. (2005). Modelling of lipase-catalyzed acidolysis of sesame oil and caprylic acid by response surface methodology: optimization of reaction conditions by considering both acyl incorporation and migration. Journal of Agriculture and Food Chemistry, 53, 8033–8037.

    Article  CAS  Google Scholar 

  • Kristensen, J. B., Xu, X., & Mu, H. (2005). Process optimization using response surface design and pilot plant production of dietary diacylglycerols by lipase-catalyzed glycerolysis. Journal of Agriculture and Food Chemistry, 53, 7059–7066.

    Article  CAS  Google Scholar 

  • Lai, H. S., & Chen, W. J. (2000). Effects of medium-chain and long-chain triacylglycerols in pediatric surgical patients. Nutrition, 16, 401–406.

    Article  CAS  Google Scholar 

  • Lin, M. T., Yeh, S. L., Kuo, M. L., Liaw, K. Y., Lee, P. H., Chang, K. J., et al. (2002). Effects of medium-chain triglyceride in parenteral nutrition on rats undergoing gastrectomy. Clinical Nutrition, 21, 39–43.

    Article  CAS  Google Scholar 

  • Lo, S. K., Baharin, B. S., Tan, C. P., & Lai, O. M. (2004). Analysis of 1,2(2,3)- and 1,3-positional isomers of diacylglycerols from vegetable oils by reversed-phase high-performance liquid chromatography. Journal of Chromatographic Science, 42, 145–154.

    CAS  Google Scholar 

  • Manohar, B., & Divakar, S. (2004). Applications of surface plots and statistical design to selected lipase-catalyzed esterification reactions. Process Biochemistry, 39, 847–853.

    Article  CAS  Google Scholar 

  • Matsuo, T., Matsuo, M., Taguchi, N., & Takeuchi, H. (2001). The thermic effect is greater for structured medium- and long-chain triacylglycerols versus long-chain triacylglycerols in healthy young women. Metabolisme, 50, 125–130.

    CAS  Google Scholar 

  • Matsuo, T., & Takeuchi, H. (2004). Effects of structured medium- and long-chain triacylglycerols in diets with various levels of fat on body fat accumulation in rats. British Journal of Nutrition, 91, 219–225.

    Article  CAS  Google Scholar 

  • Matulka, R. A., Noguchi, O., & Nosaka, N. (2006). Safety evaluation of a medium- and long-chain triacylglycerol oil produced from medium-chain triacylglycerols and edible vegetable oil. Food and Chemical Toxicology, 44, 1530–1538.

    Article  CAS  Google Scholar 

  • Mu, H., Kurvinen, J. P., Kallio, H., Xu, X., & Høy, C. E. (2001). Quantitation of acyl migration during lipase-catalyzed acidolysis, and of the regioisomers of structured triacylglycerols formed. Journal of the American Oil Chemists’ Society, 78, 959–964.

    Article  CAS  Google Scholar 

  • Mu, H., & Porsgaard, T. (2005). The metabolism of structured triacylglycerols. Progress in Lipid Research, 44, 430–448.

    Article  CAS  Google Scholar 

  • Myers, R. H., & Montgomery, D. C. (1995). Response surface methodology: Process and product optimization using design experiments. New York, USA: John Wiley & Sons.

    Google Scholar 

  • Negishi, S., Itakura, M., Arimoto, S., Nagasawa, T., & Tsuchiya, K. (2003). Measurement of foaming of frying oil and effect of the composition of TG on foaming. Journal of the American Oil Chemists’ Society, 80, 471–474.

    Article  CAS  Google Scholar 

  • Podlaha, O., & Töregård, B. (1989). Some new observations on the equivalent carbon numbers of triglycerides and relationship between changes in equivalent carbon number and molecular structure. Journal of Chromatography, 482, 215–226.

    Article  CAS  Google Scholar 

  • PORIM Test Methods (1995). Palm Oil Research Institute of Malaysia, Ministry of Primary Industries, Malaysia.

  • Rubin, M., Moser, A., Vaserberg, N., Greig, F., Levy, Y., Spivak, H., et al. (2000). Structured triacylglycerol emulsion, containing both medium- and long-chain fatty acids, in long-term home parenteral nutrition: a double-blind randomized cross-over study. Nutrition, 16, 95–100.

    Article  CAS  Google Scholar 

  • Seaton, T. B., Welle, S. T., Warenko, M., & Campbell, R. G. (1986). Thermic effect of medium-chain and long-chain triglycerides in man. American Journal of Clinical Nutrition, 44, 630–634.

    CAS  Google Scholar 

  • Shinohara, H., Ogawa, A., Kasai, M., & Aoyama, T. (2005). Effect of randomly interesterified triacylglycerols containing medium- and long-chain fatty acids on energy expenditure and hepatic fatty acid metabolism in rats. Bioscience, Biotechnology and Biochemistry, 69, 1811–1818.

    Article  CAS  Google Scholar 

  • Shinohara, H., Shimada, H., Noguchi, O., Kubota, F., & Aoyama, T. (2002). Effect of medium-chain fatty acids-containing dietary oil on hepatic fatty acid oxidation enzyme activity in rats. Journal of Oleo Science, 51, 621–626.

    CAS  Google Scholar 

  • Socha, P., Koletzko, B., Demmelmair, H., Jankowska, I., Stajniak, A., Bednarska-Makaruk, M., et al. (2007). Short-term effects of parenteral nutrition of cholestatic infants with lipid emulsions based on medium-chain and long-chain triacylglycerols. Nutrition, 23, 121–126.

    Article  CAS  Google Scholar 

  • Swift, L. L., Hill, J. O., Peters, J. C., & Grene, H. L. (1992). Plasma lipids and lipoproteins during 6 d of maintenance feeding with long-chain, medium-chain and mixed-chain triglycerides. American Journal of Clinical Nutrition, 56, 881–886.

    CAS  Google Scholar 

  • Takeuchi, H., Itakura, M., Kubota, F., & Taguchi, N. (2004). Oil or fat composition, U. S. Patent No 0191391 A1.

  • Warner, K., Orr, P., & Glynn, M. (1997). Effect of the fatty acid composition of oils and flavour. Journal of the American Oil Chemists’ Society, 74, 347–356.

    Article  CAS  Google Scholar 

  • Wolff, J. P., Mordret, F. X., & Dieffenbacher, A. (1991). Determination of triglycerides in vegetable oils in terms of their partition numbers by high performance liquid chromatography. Pure and Applied Chemistry, 63, 1173–1182.

    Article  CAS  Google Scholar 

  • Xu, X. (2000a). Production od specific-structured triacylglycerols by lipase-catalyzed reactions: a review. European Journal of Lipid Science and Technology, 102, 287–303.

    Article  CAS  Google Scholar 

  • Xu, X., Balchen, S., Høy, C. E., & Adler-Nissen, J. (1998). Pilot batch production of specific-structured lipids by lipase-catalyzed interesterification: Preliminary study on incorporation and acyl migration. Journal of the American Oil Chemists’ Society, 75, 301–308.

    Article  CAS  Google Scholar 

  • Xu, X., Fomuso, L. B., & Akoh, C. C. (2000b). Synthesis of structured triacylglycerols by lipase-catalyzed acidolysis in a packed bed bioreactor. Journal of Agriculture and Food Chemistry, 48, 3–10.

    Article  CAS  Google Scholar 

  • Zhao, H., Lu, Z., Bie, X., Lu, F., & Liu, Z. (2007). Lipase catalyzed acidolysis of lard with capric acid in organic solvent. Journal of Food Engineering, 78, 41–46.

    Article  CAS  Google Scholar 

Download references

Acknowledgement

The authors gratefully acknowledge Mr Azmi and Ms Mary Goh from MARDI, Serdang and Mr Govalan, Mr Radha Krishnan and Mr Vijay Krishnan from Golden Jomalina, Banting for their kind technical assistance. We thank Golden Hope Sdn. Bhd. for the financial support of this work.

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Correspondence to C. P. Tan.

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Koh, S.P., Tan, C.P., Lai, O.M. et al. Enzymatic Synthesis of Medium- and Long-Chain Triacylglycerols (MLCT): Optimization of Process Parameters Using Response Surface Methodology. Food Bioprocess Technol 3, 288–299 (2010). https://doi.org/10.1007/s11947-008-0073-y

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  • DOI: https://doi.org/10.1007/s11947-008-0073-y

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