Partial Purification and Some Biochemical Properties of Lipase from Germinating Oil Palm Seeds
Abstract
Lipases hydrolyze lipids to yield fatty acids and glycerol which are used as raw materials in food, cosmetics and pharmaceutical industries. This study was conducted to characterize lipase from germinating oil palm seeds. The lipase was extracted, purified and the biochemical properties were studied. Lipase assay was carried out using p- ntrophenyl palmitate as the substrate. The lipase was subjected to 80% ammonium sulphate precipitation, purified on ion exchange chromatography column using DEAE Sephadex A50 resin and gel filteration on Sephadex G-100. Relative molecular weight of the enzyme was determined on a calibrated gel filtration column using Sephadex G-100 while the subunit molecular weight was estimated on 10% Sodium Dodecyl Sulphate Polycarylamide gel electrophoresis. Kinetic parameters (Km and Vmax), effect of temperature, pH, and sensitivity to metal ions as well as substrate specificity of the lipase were studied. The lipase had a specific activity of 27.78 μmol/min/mg, a purification fold of 30.32%. The relative molecular weight of the enzyme was estimated to be 40kDa, while the Km and Vmax values were 2.5mM and 32.6 μmol/min/ml respectively. ZnCl2, KCl, EDTA and MnCl2, inhibited the enzyme activity, the highest inhibition of 8.53% was observed with EDTA while its activity was enhanced by CaCl2. Optimal pH was 7.0 and the temperature for maximal lipase activity was 45oC. The enzyme hydrolyzed coconut oil at about twice the rate of palm kernel oil and palm oil, and at a higher rate than olive oil and raphia oil.
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Abigor, R.D., Uadia, P.O., Foglia, T.A., Haas, M.J., Scott K. and Savary B.J.(2003). Partial purification and properties of lipase from germinating seeds of Jatropha cucas. Journal of the American Oil Chemist’s Society, 79 (11): 1123.
Boland W, Frobl, C. and Lorenz, M. (1991). Estrolytic and lipolytic enzymes in organic synthesis. Synthesis, 12: 1049 – 1072.
Brabcová, J., Zarevúcka, M. and Macková, M. (2010). Differences in hydrolytic abilities of two crude lipases from Geotrichum candidum 4013. Yeast. 27: 1029–1038.
Brockerhoff, H. and Jensen, R.G. (1974). Lipolytic Enzymes, 1st Edn., Academic Press, New York, pp. 330.
Doonan, S. (1996) Protein purification protocols. Methods in Molecular Biology 59:135-140.
Ejedegba, B. O., Onyeneke, E. C. and Oviasogie, P. O. (2007). Characteristics of lipase isolated from coconut (Cocos nucifera Iinn) seed under different nutrient treatments, African Journal of Biotechnology 6:723.
Enujiugha,V. N., Thani, F. A., Sanni, J. M. and Abigor, R. D. (2004). Lipase activity in dormant seeds of African oil bean (Pentaclethra macrophylla Benth). Food Chemistry 88(3): 405-408.
Forgia, T. A, Conkerton, E. J. and Philip, E. (1995). Regio selective analysis of Tricicylglycerols by lipase enzymes with attractive applications for stepwise hydrolysis of triolein using Candida rugosa lipase in biphasic oil-water system. Angewandte Chemie International Edition 37:1608-1633.
Freire, G. D. M., and Castilho, F. L. (2008). Lipases em Biocatalise. In: Bon et al. (org). Enyimas em biotechnologia: Producao Applicacao e Mercado, Rio d Janeiro, Interciencia.
Ibrahim, C. O. (1996). Lipolytic enzyme technology for the palm oil industry proceedings of 1996 PORIM International Oil Congress-Chemistry, Technology, Soap and Detergent Industry Conference, 23-28 Sept., 1996 at hotel Isana, Kuala Lumpur, Malaysia, pp.133-149.
Kambourova M, Kirilova N, Mandeva R, and Derekova A.(2003). Purification and properties of thermostable lipase from a thermophilic Bacillus stearothermophilus MC 7. Journal of Molecular Catalysis B: Enzyme. 22(5): 307-313.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680-685.
Lin, Y. H., and Huang, A. H. C. (1983). Lipase in the lipid bodies of the cotyledons of Rape and Mustard seedings. Archives of Biochemistry and Biophysics 225:300-369.
Lotrakul, P. and Dharmsthiti, S., (1997). Lipase production by Aeromonassobria LP004 in a medium containing whey and soybean meal. World Journal of Microbiology and Biotechnology 13: 163–166.
Lowry, O. H., Rosebrough, N. J. and Farr, A. L. (1951). Folin- phenol reagent. Journal of Biological Chemistry 193: 265-275.
Monnet, T. Y., Pamphile, K. B. K., Soro, R.Y., Due, E. A. and Kouame, L. P. (2012). Biochemical characterization of lipase activities in dormant seeds of T. catappa. Journal of Applied Bioscience 33: 71-81.
Ohnishi, K., Yoshida, Y., Toita, J and Sekiguchi, J. (1994). Purification and characterization of a novel lipolytic enzyme from Aspergillus oryzae. Journal of Fermentation and Bioengineering 78(6): 413–419.
Pancreac’h, G. and Baratti, J. C. (1996). Hydrolysis of p-nitrophenol pamitate in n-heptane by Pseudomonas cepacia lipase: a simple test for the determination of lipase activity in organic media. Enzyme and Microbial Technology 18: 412–422.
Rahman, R. N. Z. R. A., Baharum, S. N., Basri, M. and Salleh, A. B.(2005). High-yield purification of an organic solvent-tolerant lipase from Pseudomonas sp. strain S5. Analytical Biochemistry. 341(2): 267-274.
Ratna A. S. (2008). Partial characterization of lipase from Cocoa Beans (Theobroma cacao. l.) of Clone PBC 159. Indonesia Journal of Chemistry 8(3): 448-453.
Sadeghipour, H. R. and Bhatla, S. C. (2003). Light-enhanced Oil Body Mobilization in Sunflower Seedlings Accompanies Faster Protease Action on Oleosins. Plant Physiology and Biochemistry. 41: (4) 309 .
Sagiroglu, A. and Arabaci, N. (2005). Purification and characterization of lipase from sunflower seed. Preparative Biochemistry and Biotechnology. 35: (1) 37.
Sharma, R., Soni, S. K., Vohra, R. M., Gupta, L. K. and Gupta, J. K. (2002). Purification and characterisation of a thermostable alkaline lipase from a new thermophilic Bacillus sp. RSJ-1. Process Biochemistry 37(10): 1075-1084.
Sharon, C., Nakazato, M., Ogawa, H. I. and Kato, Y. (1998). Lipase-induced hydrolysis of castor oil: effect of various metals, Journal of Industrial Microbiology and Biotechnology 21(6): 292-295.
Sugihara, A., Shimada, Y., Tominaga, Y. (1988). Purification and characterization of Aspergillus niger lipase. Agriculture and Biological Chemistry 52: 1591-1592.
Svendson. A. (2000). Lipase protein Engineering. Biochemica et Biophysica Acta 1543: 223-238.
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