Characterization of soybean endopeptidase activity using exogenous and endogenous substrates.

Endopeptidase activity in mature soybean seeds (Glycine max), has been measured using an exogenous substrate, [(125)I]iodoinsulin B chain. On the basis of pH optimum and the use of specific proteinase inhibitors, two distinct endopeptidase activities can be identified in both the embryonic axis and the cotyledons. One activity is characteristic of a neutral/alkaline metalloendopeptidase(s) and the other of an acidic carboxylendopeptidase(s). Neither activity is membrane-bound. The metalloendopeptidase(s), most probably working with neutral expopeptidases also present in the tissues, is capable of degrading certain subunits of the storage proteins. The beta subunit of conglycinin and additional seed polypeptides remain resistant to degradation. The carboxylpeptidase activity displayed a different specificity towards endogenous substrates; in particular, an acid-soluble polypeptide of apparent molecular weight 30,000 appeared to be the principal substrate for limited proteolytic degradation by the proteinase(s). Soybean agglutinin remained resistant to degradation by either class of endopeptidases.