• Vol 9, No 2 (2018)
  • Chemical Engineering

The Effect of Papain Enzyme Dosage on the Modification of Egg-yolk Lecithin Emulsifier Product through Enzymatic Hydrolysis Reaction

Setiadi Setiadi, Nurul Hidayah

Cite this article as:
Setiadi, S., Hidayah, N., 2018. The Effect of Papain Enzyme Dosage on the Modification of Egg-yolk Lecithin Emulsifier Product through Enzymatic Hydrolysis Reaction. International Journal of Technology. Volume 9(2), pp. 380-389
Setiadi Setiadi Teknik Kimia, Universitas Indonesia
Nurul Hidayah Teknik Kimia, Universitas Indonesia
Email to Corresponding Author


Lecithin is needed as a bioemulsifier product in stabilizing agents for the food, pharmaceutical and cosmetic industries due to its renewability and as it is environmentally friendly. In the food industry, most of the emulsifiers used are the oil-in-water (O/W) type. Lecithin can be seen as a promising emulsifier product because it is extracted from egg yolk and modified by enzymatic hydrolysis reaction using the papain enzyme. This modification will change the molecular structure of the compound, which makes lecithin more stable in the oil-in-water type of emulsion. This study aims to determine the optimum amount of papain enzyme used in the hydrolysis reaction to achieve the most stable O/W lecithin emulsion type. The results show that the breaking of a single fatty acid chain from the structure of lecithin can be demonstrated by FTIR instrumentation. The fatty acids detected from the lecithin structure are shown at wavenumber 1699.45 cm-1 (C=O), 1231.44 cm-1 (C-O), 1422.45 cm-1 (C-O-H), 1092.85 cm-1 (C-C), 665.89 cm-1 (CH2), and 3400.57 (-OH in carboxylate). Determination of the modified lecithin yield was made by several tests, namely a stability test, and tests for acid value, surface tension and zeta potential. From the results of tests, the emulsion stability for the O/W type was achieved in modified-lecithin using a 4% papain enzyme dosage, with a stability duration of up to 31 hours. The lowest acid number was achieved in modified-lecithin using a 2% papain enzyme dosage with value of 10.40. The lowest surface tension was obtained in modified-lecithin using a 2% papain enzyme dosage with a surface tension value of 48.68 dyne/cm. The zeta potential of the modified-lecithin using a 2% papain enzyme had a value of -94.8 mV. These results show that the enzymatic hydrolysis of lecithin using a papain enzyme is clearly able to enhance the emulsifier properties of the lecithin produced.

Emulsifier; Enzymatic hydrolysis; Extraction; Lecithin; Papain enzyme


The papain enzyme from crude papaya latex is an effective enzyme for lecithin modification in order to improve its availability and affordability. In addition, the reaction of enzymatic hydrolysis on egg yolk lecithin can be confirmed based on the FTIR spectrum. The purified egg yolk lecithin resulted in a yield of 8.24%. The optimum amount of papain enzyme dosage to establish the best characteristic of modified lecithin was 2%, based on the weight of egg yolk lecithin. The hydrolysis reaction of egg yolk lecithin using the papain enzyme increased the stability of lecithin in an O/W emulsion to up to 31 hours, with a zeta potential value of -94.8 mV. It also decreased the surface tension of lecithin to 48.68 dyne/cm as more polar lecithin was produced after the hydrolysis reaction. The acid value of modified lecithin decreased to 10.40 due to the decreasing fatty acid content. In further research, it is advised to use a larger amount of water during the lecithin modification process.


The authors would like to acknowledge the grant “Publikasi Internasional Terindeks untuk Tugas Akhir Mahasiswa” (PITTA) Universitas Indonesia with Grant number 832/UN2.R3.1/HKP.05.00/ 2017 in the year 2017, for its financial support during the research.


Al-Abayaji, M. A., 2015. Extraction and Determination of Iraqi Boiled Egg Yolk Constituents, and Characterization of Lecithin. Department of Chemistry, Ibn Al Haitham College of Education.

Asomaning, J., Curtis, J.M., 2017. Enzymatic Modification of Egg Lecithin to Improve Properties, Food Chemistry, Volume 220, pp. 385–392

Banat, I.M., Makkar, R.S., Cameotra, S.S., 2000. Potential Commercial Applications of Microbial Surfactants. Applied Microbiology and Biotechnology, Volume 53(5), pp. 495–508.

Biotechnology Industry Organization, 2011. Biotechnology Solutions for Renewable Specialty Chemicals & Food Ingredients. Washington, DC: Biotechnology Industry Organization.

Cabezas, D., Madoery, R.M., Tomás, B.D.M.C., 2012. Emulsifying Properties of Different Modified Sunflower Lecithins. Journal of the American Oil Chemists' Society, Volume 89(2), pp. 335–361

Dastghieb, S.M.M., Amoozegar, M.A., Elahi, E., Asad, S., Banat, I.M., 2008. Bioemulsifier Production by a Halothermophilic Bacillus Strain with Potential Applications in Microbially Enhanced Oil Recovery. Biotechnol Lett, Volume 30(2), pp. 263–270

Estiasih, T., Kgs, A., Ginting, E., Priyanto, A.D., 2013. Modification of Soy Crude Lecithin by Partial Enzymatic Hydrolysis using Phosholipase A1. International Food Research Journal, Volume 20(2), pp. 843–849

Giordani, R., Moulin, A., Verger, R., 1991. Tributyroylglycerol Hydrolase Activity in Carica Papaya and Other Lattices. Phytochemistry, Volume 30(4), pp. 1069–1072

Jiang, Y., Noh, S.K., Koo, S., 2001. Egg Phosphatidylcholine Decreases the Lymphatic Absorption of Cholesterol in Rats. The Journal of Nutrition, Volume 131(9), pp. 2358–2363

Joshi, A., Paratkar, S.G., Thorat, B., 2006. Modification of Lecithin by Physical, Chemical and Enzymatic Methods. European Journal of Lipid Science and Technology, Volume 108(4), pp. 363–373

Lamour, G., Hamraoui, A., Buvailo, A., Xing, Y., Keuleyan, S., Prakash, V., Bafrooei, A.E., Borguet E., 2010. Contact Angle Measurements using a Simplified Experimental Setup. Journal of Chemical Education, Volume 87(12), pp. 1403–1407

Lopes, E.M., Castellane, T.C.L., Moretto, C., Lemos, E.G.D.M., Souza, J.A.M.D., 2014. Emulsification Properties of Bioemulsifiers Produced by Wild-type and Mutant Bradyrhizobium elkanii Strains. Journal of Bioremediation & Biodegradation, Volume 5(6), pp. 1–6

Nasir, M.I., Bernards, M.A., Charpentier, P.A., 2007. Acetylation of Soybean Lecithin and Identification of Components for Solubility in Supercritical Carbon Dioxide. Journal of Agricultural and Food Chemistry, Volume 55(5), pp. 1961–1969

Nieuwenhuyzen, W.V., Tomas, M.C., 2008. Update on Vegetable Lecithin and Phospoholipid Technologies. European Journal of Lipid Science and Technology, Volume 110(5), pp. 472–486

Nimalaratne, C., Wu, J., 2015. Hen Egg as an Antioxidant Food Commodity: A Review. Nutrients, Volume 7(10), pp. 8274–8293

Nitschke, M., Costa, Siddhartha G.V.A.O., 2007. Biosurfactants in Food Industry. Trends in Food Science & Technology, Volume 18(5), pp. 252–259

Palacios, L.E., Wang, T., 2005. Egg Yolk Lecithin Fractionation and Characterization. J.American Oil Chemists' Society, Volume 82(8), pp. 571–578

Reckziegel, Y., 2014. Characterization and Comparison of the Functionality of Fractionated Lecithin from Different Sources. Master Thesis of Science in Food Technology. Faculty of Bioscience Engineering, Universiteit Gent.

Schneider, M., 2001. Phospholipids for Functional Food. European Journal of Lipid Science and Technology, Volume 103(2), pp. 98–101

Shah, A.K.M.A., Nagao, T., Kurihara, H., Takahashi, K., 2017. Production of a Health-beneficial Food Emulsifier by Enzymatic Partial Hydrolysis of Phospholipids Obtained from the Head of Autumn Chum Salmon. Journal of Oleo Science, Volume 66(2), pp. 147–155

Sharma, S., Kanwar, S.S., 2014. Organic Solvent Tolerant Lipases and Applications. The Scientific World Journal. Volume 2014, pp. 115

Song, J.K., Han, J.J., Rhee, J.S., 2005. Phospholipases: Occurrence and Production in Microorganisms, Assay for High-throughput Screening, and Gene Discovery from Natural and Man-made Diversity. Journal of the American Oil Chemists’ Society, Volume 82(10), pp. 691–705

Vikbjerg, A.F., 2006. Enzyme Catalyzed Production of Phospholipids with Modified Fatty Acid Profile, Denmark: BioCentrum-DTU Technical University of Denmark

Vikbjerg, A.F, Rusig, J.Y., Jonsson, G., Mu, H., Xu, X., 2006. Comparative Evaluation of the Emulsifying Properties of Phosphatidylcholine after Enzymatic Acyl Modification. J. Agric. Food Chem, Volume 54(9), pp. 3310–3316