Abstract
The positive correlation between the developments of acid, neutral and alkaline proteases (azocaseinolytic) with protein depletion suggest the involvement of these proteases in the degradation of proteins in germinating Indian bean. These proteases increased in the early stages of germination and decreased later. However, the activity of acid proteases was higher throughout the germination period compared with the activities of neutral and alkaline proteases. The acid protease from the cotyledons of 4-day old germinating Indian bean seedlings was purified to 152 folds by a five step procedure comprising - crude extract from cotyledons, (NH4)2SO4 fractionation, DEAE-cellulose, CM-cellulose and finally casein-agarose affinity chromatography. The molecular mass of acidic protease is 32 kDa. Key words: Dolichos lablab, proteolytic enzymes, acidic protease, purification, protein mobilization, germination.
Highlights
The plant seed is an organ of propagation and dispersal and the major plant tissue harvested by humankind
The positive correlation between the developments of acid, neutral and alkaline proteases with protein depletion suggest the involvement of these proteases in the degradation of proteins in germinating Indian bean
The mobilization of storage proteins in germinating Indian bean seeds, as well in other plant sources, is initiated by endoproteases which convert the water insoluble storage proteins into soluble peptides that can be further hydrolyzed to amino acids by exopeptidases (Callis, 1995; Mikola, 1983; Shutov and Vaintraub, 1987)
Summary
The plant seed is an organ of propagation and dispersal and the major plant tissue harvested by humankind. The mobilization of seed storage proteins represents one of the most important post-germinative events in the growth and development of seedling. To study the mechanism of protein mobilization process, many have undertaken the task of purifying and characterizing a variety of proteases and peptidases, some of which occur only transiently in germinating seeds (Ashton, 1976; Davy et al, 1998; Shutov and Vaintraub, 1987). It is important to establish its role in protein degradation and the natural substrates need to be study in vitro. The realization of such approaches obviously requires purification of seed proteases, or at least their separation from each other.
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