ISOLATION AND PURIFICATION OF LIPASE FROM COCOA BEANS (Theobroma cacao. L.) OF CLONE PBC 159

https://doi.org/10.22146/ijc.21654

Ratna Agung Samsumaharto(1*)

(1) Faculty of Biology, Setia Budi University, Jl. Let. Jend. Sutoyo Mojosongo – Surakarta 57127
(*) Corresponding Author

Abstract


Lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) was extracted and purified from acetone dry powder of cocoa (Theobroma cacao. L.) of clone PBC 159 extract. The Lipase  from AcDP of cocoa beans was used for purification using 40-60 and 60-80% ammonium sulphate precipitation. The resulted indicated 44.73 and 60.51-fold purification with 26.74 and 33.31% recovery lipase activity (yield), respectively. The crude lipase enzyme from both precipitation were eluted, producing a single peak after applying through Sephacryl S-200 chromatography. The purified enzyme had a uniform specific activity throughout the final chromatography peak. The results from SDS-PAGE analysis showed that the molecular weight of the lipase enzyme was in between 45-66 kDa.


Keywords


isolation; purification; lipase; cocoa beans

Full Text:

Full Text Pdf


References

[1] Bloch, A.K., 1960, Lipid Metabolism, John Wiley and Sons Inc., New York.

[2] Scheupp, C., Kermasha, S., Michalski, M.C. and Morin, A., 1997, Process Biochem., 32, 3, 2225-2232.

[3] Antonian, E., 1988, Lipids, 23, 12, 1-5.

[4] Mozaffar, Y. and Weete, J.D., 1993, Lipid, 28, 5, 377-381.

[5] Kirchhoff, P.M., Biehl, B., and Crone, G., 1989, Food Chem., 31, 295-311.

[6] Seow, T.K., Jinap S., Lee, K.H. and Lee, P.M., 1994, J. Food Sci. Technol., 31, 6, 511-513.

[7] Green, A.A. and Hughes, 1955, Protein Fractionation on The Basis of Solubility In Aqueous of Salts and Organic Solvent. In: Collowick, S.P. and Kaplan, N.O. (eds) Methods of Enzymology 1. Preparation and Assay of Enzyme. Academic Press, New York. 67-90.

[8] Lowry, O.H., Rossbrough, N.J., Farr, A.L. and Randall, R.J., 1951, J. Biol. Chem. 76, 460-463.

[9] Kwon, D.Y. and Rhee, J.S., 1986, American Oil Chem. And Sci. 63(1), 89-92.

[10] Boyer, R.F., 1986, Modern Experimental Biochemistry. Addison-Wesley Publishing Company. 362-363.

[11] Laemmli, U.K., 1970, Nature, 227, 680.

[12] Gill, J. and Pharish, J.H., 1997, Biochem. Edu., 25, 1, 1-5.



DOI: https://doi.org/10.22146/ijc.21654

Article Metrics

Abstract views : 1705 | views : 1272


Copyright (c) 2010 Indonesian Journal of Chemistry

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

 

Indonesian Journal of Chemistry (ISSN 1411-9420 /e-ISSN 2460-1578) - Chemistry Department, Universitas Gadjah Mada, Indonesia.

Web Analytics View The Statistics of Indones. J. Chem.