Immobilization of lipase onto <i>Cyperus Papyrus</i> L. for biodiesel production by transesterification and hydrolysis-esterification


  • Sireerat Charuchinda Faculty of Science, Chulalongkorn University, Biofuels by Biocatalysts Research Unit, Chulalongkorn University and Center for Petroleum, Petrochemicals, and Advanced Materials, Chulalongkorn University
  • Piyanan Suthianthong Faculty of Science, Chulalongkorn University
  • Warawut Chulalaksananukul Faculty of Science, Chulalongkorn University and iofuels by Biocatalysts Research Unit, Chulalongkorn University


Cyperus papyrus L., Candida rugosa lipase, Immobilization, Biodiesel, Transesterification, Hydrolysis-esterification


Lipase of Candida rugosa was immobilized on biomass support, plant fiber from Cyperus papyrus L., for biodiesel synthesis. The two immobilization techniques investigated in this study were physical adsorption and covalent binding with 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC). Results showed that the prepared immobilized lipase by physical adsorption with adding heptane presented higher protein loading, lipase activity and degree of immobilization than that by physical adsorption in phosphate buffer solution and by covalent binding. This immobilized lipase was further applied as biocatalyst for biodiesel synthesis by transesterification and hydrolysis-esterification. The results showed that this immobilized lipase preferred to hydrolyze triglyceride in palm oil to be fatty acid in water as medium. Then, the obtained fatty acid could be a good substrate to react with alcohol for biodiesel synthesis by esterification route. So, the biodiesel synthesis yield by hydrolysis-esterification was higher than transesterification. Nevertheless, the results revealed that bioethanol was found to be a better substrate than methanol for biodiesel synthesis via enzymatic hydrolysis-esterification.


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Gerpen, J. V. (2005). Biodiesel processing and production. Fuel Process. Technol. 86(10): 1097-1107.

Sugunan, S. & Gopinath, S. (2007). Enzymes immobilized on montmorillonite K 10: Effect of adsorption and grafting on the surface properties and the enzyme activity. Appl. Clay Sci. 35(1-2): 67-75.

Lee, D.H., Park, C.H., Yeo, J.M. & Kim, S.W. (2006). Lipase immobilization on silica gel using a cross-linking method. J. Ind. Eng. Chem. 12(5): 777-782.

Minovska, V., Winkelhausen, E. & Kuzmanova, S. (2005). Lipase immobilized by different techniques on various support materials applied in oil hydrolysis. J. Serb. Chem. Soc. 70(4): 609-624.

Varavinit, S., Chaokasem, N. & Shobsngob, S. (2001). Covalent immobilization of a glucoamylase to bagasse dialdehyde cellulose. World J. Microb. Biot. 17(7):721-725.

Iqbal, M. & Saeed, A. (2005). Novel method for cell immobilization and its application for production of organic acid. Lett. Appl. Microbiol. 40(3): 178- 182.

Shimada, Y., Watanabe, Y., Sugihara, A., and Tominaga, Y. (2002). Enzymatic alcoholysis for biodiesel fuel production and application of the reaction to oil processing. J. Mol. Catal. B-Enzym. 17(3-5): 133-142.

Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.

Gao, Y., Tan, T.W., Nie, K.L. & Wang, F. (2002). Immobilization of lipase on macroporous resin and its application in synthesis of biodiesel in low aqueous media. Chin. J. Biotech. 22(1): 114-118.

Bastida, A., Sabuquillo, P., Armiscn, P. Fernandez-Lafuente, R., Huguet, J. & Guisan, J.M. (1998). A single step purification, immobilization, and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports. Biotechnol. Bioeng. 58(5): 486-493.

Iso, M., Chen. B., Eguchi. M., Kudo, T. & Shrestha, S. (2001). Production of biodiesel fuel from triglycerides and alcohol using immobilized lipase. J. Mol. Catal. B-Enzym. 16(1): 53-58.

Palomo, J.M., Munoz, G., Fernandez-Lorente, G., Mateo, C., Fernandez-Lafuente, R. & Guisan, J.M. (2002). Interfacial adsorption of lipases on very hydrophobic support (octadecylSepabeads): immobilization, hyperactivation and stabilization of the open form of lipases. J. Mol. Catal. BEnzym. 19-20(Special Issue): 279-286.

Hung, T.C., Giridhar R., Chiou, S.H., and Wu, W.T. (2003). Binary immobilization of Candida rugosa lipase on chitosan. J. Mol. Catal. B-Enzym. 26(1-2): 69 -78.

Chui, W.K., and Wan, L.S. (1997). Prolonged retention of cross-linked trypsin in calcium alginate microspheres. J. Microencapsul. 14(1):51-61.

Piamtongkam, R., Duquesne, S., Barbe, S., Bordes, F., André, I., Marty, A. & Chulalaksananukul, W. (2011). Enantioselectivity of Candida rugosa lipases (Lip1, Lip3 and Lip4) towards 2- bromo phenylacetic acid octyl esters controlled by a single amino acid. Biotechnol. Bioeng. 108(8): 1749-1756.




How to Cite

S. Charuchinda, P. Suthianthong, and W. Chulalaksananukul, “Immobilization of lipase onto <i>Cyperus Papyrus</i> L. for biodiesel production by transesterification and hydrolysis-esterification”, J Met Mater Miner, vol. 21, no. 1, Aug. 2022.



Original Research Articles