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Purification and characterization of acid phosphatase from a germinating black gram (Vigna mungo L.) seedling

Author(s): Asaduzzaman A.K.M. | Rahman Habibur M. | Yeasmin Tanzima

Journal: Archives of Biological Sciences
ISSN 0354-4664

Volume: 63;
Issue: 3;
Start page: 747;
Date: 2011;
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Keywords: Germination | seedling | Vigna mungo L | acid phosphatase | black gram | purification | characterization and Km Value

An acid phosphatase has been isolated and purified from an extract of a germinating black gram seedling. The method was accomplished by gel filtration of a germinating black gram seedling crude extract on sephadex G-75 followed by ion exchange chromatography on DEAE cellulose. The acid phosphatase gave a single band on SDS-polyacrylamide slab gel electrophoresis. The molecular weight of the acid phosphatase determined by SDS-polyacrylamide slab gel electrophoresis was estimated to be 25 kDa. The purified enzyme showed maximum activity at pH 5 and at temperature of 55°C. Mg2+, Zn2+ and EDTA had an inhibitory effect on the activity of the acid phosphatase. Black gram seedling acid phosphatase was activated by K+, Cu2+ and Ba2+. The Km value of the enzyme was found to be 0.49 mM for pNPP as substrate.

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