The Enzyme Kinetic of Lipase Catalysed Acidolysis of Used Cooking Palm Oil


  • Shakirah S. Sulaiman School of Chemical Engineering, Faculty of Engineering, Universiti Teknologi Malaysia, Johor, Malaysia
  • Nor Athirah Zaharudin School of Chemical Engineering, Faculty of Engineering, Universiti Teknologi Malaysia, Johor, Malaysia
  • Roslina Rashid School of Chemical Engineering, Faculty of Engineering, Universiti Teknologi Malaysia, Johor, Malaysia
  • Siti Marsilawati Mohamed Esivan School of Chemical Engineering, Faculty of Engineering, Universiti Teknologi Malaysia, Johor, Malaysia



Used cooking palm oil, Enzymatic acidolysis, Kinetic


Enzymatic acidolysis of various fats and oils has successfully shown a promising ability to alter specific positions of lipids and incorporate desirable fatty acids at specific positions. However, used cooking oil utilization for enzymatic acidolysis is still lacking scientific investigation. Hence, this study aims to utilize used cooking palm oil (UCPO) by enhancing the oleic acid content via enzymatic acidolysis using immobilized C. rugosa lipase. The optimum substrate molar ratio (mol/mol) was identified based on the analysis of the peroxide, iodine, and acid values. Then, kinetic parameters, Km and Vmax for the enzymatic acidolysis of UCPO were calculated. Substrate molar ratio of 1:1, 1:2, 1:3, 1:4, and 1:5 mol/mol (oil: acid) was varied to evaluate the peroxide, acid, and iodine values. The reaction conditions such as reaction temperature (50 °C), reaction time (24 hours), enzyme concentration (0.05 g), agitation speed (250 rpm), and pH (7) were fixed throughout the experiment. The Michaelis-Menten kinetic model was selected to describe the kinetic of enzymatic acidolysis of UCPO. The result showed that incorporation of oleic acid in UCPO has been successfully achieved with the increased in IV value from 39.47 I2/g to 110.53 I2/g. The optimum substrate molar ratio obtained was 1:3 mol/mol with the highest iodine value of 110.53 I2/g. The best linear regression approach is Lineweaver-Burk plot with the values of Vmax and Km were 34.5658 μmol/ml.min and 0.06 mol/L, respectively. The enzyme activity for C. rugosa lipase was obtained at 0.5804 μmol/


Abed, S. M., Wei, W., Ali, A. H., Korma, S. A., Mousa, A. H., Hassan, H. M., Jin, Q., Wang, X. 2018. Synthesis of Structured Lipids Enriched with Medium-Chain Fatty Acids Via Solvent-Free Acidolysis of Microbial Oil Catalyzed by Rhizomucor miehei Lipase. LWT, 93, 306–315.

Badoei-dalfard, A., Karami, Z., Malekabadi, S. 2019. Construction of CLEAs-lipase on magnetic graphene oxide nanocomposite: An efficient nanobiocatalyst for biodiesel production. Bioresource Technology. 278, 473-476.

Bronsky, J., Campoy, C., Embleton, N., Fewtrell, M., Mis, N. F., Gerasimidis, K., Hojsak, I., Hulst, J., Indrio, F., Lapillonne, A., Molgaard, C., Moltu, S. J., Verduci, E., Vora, R, Domellöf, M. 2019. Palm Oil and Betapalmitate in Infant Formula: A Position Paper by the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) Committee on Nutrition. Journal of Pediatric Gastroenterology and Nutrition. 68(5): 742-760.

Chhetri, A., Watts, K., Islam, M. 2008. Waste cooking oil as an alternate feedstock for biodiesel production. Energies, 117-128.

De Santi, A., Monti, S., Barcaro, G., Zhang, Z., Barta, K., Deuss, P. J. 2021. New Mechanistic Insights into the Lignin β-O-4 Linkage Acidolysis with Ethylene Glycol Stabilization. ACS Sustainable Chemical Engineering. 9(5): 2388–2399.

Devi, N. A., Radhika, G. B., Bhargavi, R. J. 2017. Lipase Catalyzed Transesterification of Ethyl Butyrate in nhexane— A Kinetic Study. Journal of Food Science and Technology. 54(9): 2871–2877.

Elias, N., Abdul Wahab, R., Chandren, S., Lau, W. J. 2021. Performance of Candida rugosa Lipase Supported on Nanocellulose-Silica Reinforced Polyethersulfone Membrane for the Synthesis of Pentyl Valerate: Kinetic, Thermodynamic, and Regenerability Studies. Molecular Catalysis. 514: 111852.

Hamam, F., Shahidi, F. 2004. Lipase-catalyzed Acidolysis of Algal Oils with Capric Acid: Optimization of Reaction Conditions Using Response Surface Methodology, Journal of Food Lipids. 11(2): 147–163.

Heater, B. S., Chan, W. S., Lee, M. M., Chan, M. K. 2019. Directed Evolution of a Genetically Encoded Immobilized Lipase for The Efficient Production of Biodiesel from Waste Cooking Oil. Biotechnology for Biofuels. 12(1): 1–14.

Ifeduba, E. A., & Akoh, C. C. 2014. Modification of Stearidonic Acid Soybean Oil by Immobilized Rhizomucor miehei Lipase to Incorporate Caprylic Acid. Journal of the American Oil Chemists’ Society. 91(6): 953–965.

Jurid, L. S., Zubairi, S. I., Mohd Kasim, Z., Ab. Kadir, I. A. 2020. The Effect of Repetitive Frying on Physicochemical Properties of Refined, Bleached and Deodorized Malaysian Tenera Palm Olein During Deep-Fat Frying. Arabian Journal of Chemistry. 13: 6149-6160.

Kuo, C. H., Huang, C. Y., Lee, C. L., Kuo, W. C., Hsieh, S. L., Shieh, C. J. 2020. Synthesis of DHA/EPA Ethyl Esters Via Lipase-Catalyzed Acidolysis Using Novozym® 435: A Kinetic Study. Catalysts, 10(5), 565.

Liu, N., Ren, G., Faiza, M., Li, D., Cui, J., Zhang, K., Yao, X., Zhao, M. 2022. Comparison of conventional and green extraction methods on oil yield, physicochemical properties, and lipid compositions of pomegranate seed oil. Journal of Food Composition and Analysis. 114, 104747.

Manzoor, S., Masoodi, F. A., Rashid, R., Dar, M.M. 2022. Effect of Apple Pomace-Based Antioxidants on the Stability of Mustard Oil During Deep Frying of French Fries. LWT. 163: 113576.

Mariana, R. R., Susanti, E., Hidayati, L., Abdul Wahab, R. 2020. Analysis of Peroxide Value, Free Fatty Acid, and Water Content Changes in Used Cooking Oil from Street Vendors in Malang. International Conference on Life Sciences and Technology, 22 April 2020.

Moazeni, F., Chen, Y. C., Zhang, G. 2019. Enzymatic transesterification for biodiesel production from used cooking oil, a review. Journal of Cleaner Production, 216, 117–128.

Mohd Fadzel, F., Salimon, J., Derawi, D. 2021. Low-Energy Separation Technique on Purification of Unsaturated Fatty Acids of Palm Stearin using Methanol Crystallization. Sains Malaysiana. 50(1): 151-160.

Peng, L., Tan, W., Lu, Y., Yao, A., Zheng, D., Li L., Xiao, J., Li, L., Li, Q., Zhou, S. F., Zhan, G. 2022. Convenient Immobilization of α-L Rhamnosidase on Ceriumbased Metal-Organic Frameworks Nanoparticles for Enhanced Enzymatic Activity and Recyclability. The European Society Journal for Catalysis. 14(3): e202101489.

Ren, T., Qi, W., Su, R., He, Z. 2019. Promising Techniques for Depolymerization of Lignin into Value-added Chemicals. The European Society Journal for Catalysis. 11(2): 639-654.

Tiefenbacher, K. F. 2017. Chapter Three - Technology of Main Ingredients—Sweeteners and Lipids. Wafer and Waffle: Processing and Manufacturing. Pages 123-225.

Wang, X., Zou, S., Miu, Z., Jin, Q., Wang, X. 2019. Enzymatic Preparation of Structured Triacylglycerols with Arachidonic and Palmitic Acids at the sn-2 Position for Infant Formula Use. Food Chemistry. 283, 331-337.

Wanyonyi, W. C., Onyari, J. M., Shiundu, P. M., Mulaa, F. J. 2017. Biodegradation And Detoxification of Malachite Green Dye Using Novel Enzymes from Bacillus Cereus Strain KM201428: Kinetic and Metabolite Analysis. Energy Procedia. 119: 38–51.

Yusuff, A.S., Bhonsle, A. K., Bangwal, D. P., Atray, N. 2021. Development of a Barium-Modified Zeolite Catalyst for Biodiesel Production from Waste Frying Oil: Process Optimization by Design of Experiment. Renewable Energy, 177, 1253-1264.

Zaharudin, N. A., Remzi, N. S., Rashid, R., Mohamed Esivan, S. M., Idris, A., Othman, N. 2018. Oleic Acid Enhancement in Used Frying Palm Oil Via Enzymatic Acidolysis. Malaysian Journal of Analytical Science, 22(4), 633-641.

Zakaria, Z. A. (Ed.). (2018). Sustainable Technologies for The Management of Agricultural Wastes. Applied Environmental Science and Engineering for a Sustainable Future. Published.