Biochemical Studies of Lipase from Germinating Oil Seeds (Glycine max)
P. P. Gadge, S. D. Madhikar, J. N. Yewle, U. U. Jadhav, A. D. Chougale, V. P. Zambare and M. V. Padul
DOI : 10.3844/ajbbsp.2011.141.145
American Journal of Biochemistry and Biotechnology
Volume 7, Issue 3
Problem statement: Lipase is one of the important enzymes in food, pharmaceutical, detergent and biofuels industries. Search for the lipase with distinct features, possibly from germinating seeds, is of interest for industrial applications. Approach: The lipase produced by soybean oil seeds was partially purified and characterized in terms of the optimal pH and temperature for activity as well as substrate specificity. Results: The lipase was extracted and partially purified from germinating soybean seeds using chilled acetone and ammonium sulfate precipitation. Partially purified and dialyzed enzyme profile was observed on native-Polyacrylamide Gel Electrophoresis (PAGE). The lipase was optimally active at pH 8 and temperature of 24°C. In the presence of Ca2+ and Mg2+ enhance the activity at low concentration, while the Hg2+ and Ethylene Diaminotetracetic Acid (EDTA) showed inhibitory effect. The enzyme was found to be metalloenzyme. Enzyme kinetics with olive oil emulsion substrate showed km and vmax of 7.67 mg and 0.0125 µm mL min-1, respectively. Conclusion: The mettaloenzyme enzyme was able to attack specifically on oil in seeds to generate free fatty acids as the major end product. This understanding may help in devising efficient methods to overcome the problem of soybean seed oil in stability.
© 2011 P. P. Gadge, S. D. Madhikar, J. N. Yewle, U. U. Jadhav, A. D. Chougale, V. P. Zambare and M. V. Padul. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.