Bommisetty Padmakar
Sri Krishnadevaraya University, India
Pagadala Madhavilatha
Sri Krishnadevaraya University, India
Vadde Ramakrishna
Sri Krishnadevaraya University, India
ABSTRACT
The present study reported the purification, characterization of alkaline protease and its possible involvement in moblization of storage proteins. The mobilization of seed storage proteins represents one of the most important post-germinative events in the growth and development of seedling. The proteolytic enzymes play a central role in the biochemical mechanism of germination. The alkaline protease from the cotyledons of 4-day old germinating Indian bean seedlings was purified to 198 folds by a four step procedure comprising-crude extract, (NH4)2SO4 fractionation, DEAE-cellulose and finally casein-alginate affinity chromatography. The alkaline protease was shown to be homogeneity, as attested by a single protein band on both native PAGE and SDS-PAGE. It is a monomeric enzyme with molecular mass of 40 kDa and exhibited sharp pH optima at 8.8 with casein. It has been characterized by seeing the effect of various inhibitors and metal cofactors on the alkaline protease activity. The enzyme activity was markedly increased with Zn and significantly inhibited by the metal chelating agents-EDTA and 1, 10-phenanthroline. These results suggest that the alkaline protease is a metalloenzyme.
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Bommisetty Padmakar, Pagadala Madhavilatha and Vadde Ramakrishna, 2005. Purification of Alkaline Protease from the Cotyledons of Germinating Indian Bean (Dolichos lablab L. var lignosus) Seeds. Biotechnology, 4: 365-369.
DOI: 10.3923/biotech.2005.365.369
URL: https://scialert.net/abstract/?doi=biotech.2005.365.369
DOI: 10.3923/biotech.2005.365.369
URL: https://scialert.net/abstract/?doi=biotech.2005.365.369
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