PARTIAL CHARACTERIZATION of lipase from COCOA BEANS (Theobroma cacao. L.) of clone PBC 159

https://doi.org/10.22146/ijc.21604

Ratna Agung Samsumaharto(1*)

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

Abstract


A study was carried out to characterize the cocoa lipase from cocoa beans (Theobroma cacao, L.) of clone PBC 159. The optimum temperature of cocoa lipase was 30-40 °C and the pH optimum was 7.0-8.0. The moleculer weight of the lipase enzyme was in between 45-66 kDa. The results indicate that Km value for cocoa bean lipase was 2.63 mM, when trimyristin was used as a substrate. The incubation of cocoa bean lipase with triolein and tributyrin (as substrate) yielded Km of 11.24 and 35.71 mM, respectively. The Vmax value obtained from the incubation of the lipase with a wide range of substrates, including tributyrin, trimyristin and triolein, are expressed as µmole acid/min/mg protein for cocoa lipase. Vmax values decreased with the increase in the triacylglycerol chain-length, with Vmax values of 27.78, 13.16 and 11.63 µmole acid/min/mg protein when incubated with tributyrin, trimyristin and triolein, respectively. Inhibition of lipase occurred in the presence of diisopropyl flourophosphate, N-bromosuccinimide and 5,5-dithiobis-(-2-nitrobenzoic acid).


Keywords


characterization; lipase; cocoa beans

Full Text:

Full Text Pdf


References

[1] Scheupp, C., Kermasha, S., Michalski, M.C., and A. Morin, 1997, Process Biochemistry. 32 (3), 2225-2232.

[2] Jensen, R.G., Dejong, F.A., and Clark, R.M., 1983, Lipids, 18 (3), 239.

[3] Ziegleder, V.G., and Biehl, B., 1988, Analysis of Cocoa Flavour Components And Flavour Precursors. In: Lickens, H.J., Jackson, J.F. (eds) Modern Methods Of Plants Analysis Of Non-alcoholic Beverages. Springer-Verlag, Berlin. 321-390.

[4] Antonian, E., 1988, Lipids, 23 (12), 1-5

[5] Jensen, R.G., 1983, Lipids, 18 (9), 650-657.

[6] Shabtai, Y., and Mishne, N.D., 1992, J. Appl. Environ. Microbiol., 58 910, 174-180.

[7] Abigor, D.R., Opute, F.I., Opuku, A.R., and Osagie, A.U., 1985, J. Sci. Food Agric., 36, 599-606.

[8] Hassan, N.H., 1995, Penulenan Separa dan Pencirian Ensim Lipase dari Mesokarp Buah Kelapa Sawit (Elaeis quineensis, Jacq.). Thesis Project. Universiti Putra Malaysia, Serdang.

[9] Muto, S., and Beevers, H., 1974, Plant Physiol. 54: 23-28.

[10] Ory, R.L., 1968, Lipids. 26 (92), 370-386.

[11] Ory, R.L., Yatsu, L.Y., and Kircher, H.W., 1968, Arc. Biochem. Biophys., 264, 255-264.

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

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

[14] Green, A.A., and Hughes, H., 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.

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

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

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

[18] Patkar, S., and Bjorkling, F., 1994, Lipase Inhibitors. In: Lipase; Their Structure, Biochemistry and Application, eds P. Woolley and S.B. Petersen. Cambridge University Press, Cambridge, UK. 207-224.

[19] Liu, W.H., Beppu, T., and Arima, K., 1977, J. Agric. Biol. Chem., 41, 131-135.



DOI: https://doi.org/10.22146/ijc.21604

Article Metrics

Abstract views : 1545 | views : 1975


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.