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Identification, Purification and Characterization of Lipase from Germinating Oil Seeds (Brassica napus L.)

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Author(s): N.K. Sana | I. Hossin | E.M. Haque | Ranajit Kumar Shaha

Journal: Pakistan Journal of Biological Sciences
ISSN 1028-8880

Volume: 7;
Issue: 2;
Start page: 246;
Date: 2004;
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Keywords: Germinating | brassica napus L | lipase | hydrolyzing lipid | km value

ABSTRACT
Ionic effect on the degradation of oil seeds (Brassica napus L) storage lipid was investigated during germination. Lipid degradating enzyme lipase was identified in germinating Brassica napus L and the enzyme was purified to homogeneity by extraction, Sephadex G-50, DEAE and CM-cellulose column chromatography. The enzyme was purified to 67.59 fold with the final specific activity of 39.10 mmol min-1 mg-1 at 37�C using triolein as a substrate. The purified enzyme showed a single band when it was subjected to SDS / PAGE electrophoresis. The SDS/PAGE electrophoresis indicated a molecular mass of 34 kDa for this lipase which corresponding to the high-resolution gel filtration. The optimum pH and temperature for hydrolysis of Oleic acid were 7.0 and 37�C, respectively. In the presence of Ca 2+ and Bi 3+, the lipase activity was dramatically enhanced by 165 and 124%, respectively. Fe3+, Fe2+, Zn2+, Hg2+ and Cu2+ could inhibit this lipase but Al 3+, Pb 2+ showed no influence on hydrolysis activity. Km of this enzyme was 0.23 mM.
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