Some properties of the allantoate amidohydrolase from French bean seedlings.

Allantoate degradation was demonstrated in the extracts of ungerminated seeds and roots, stems and leaves in germinated seedlings of French bean (Phaseolus vulgaris L.). Activity of allantoate-degrading enzyme could only be measured when phenylhydrazine was included in the assay mixture. Partial purification of allantoate-degrading enzyme from seedlings was performed and two fractions with allantoate-degrading enzyme activity were obtained. The molecular mass of the first fraction was over 200 kD and that of the second one was 13.5 kD. The allantoate-degrading enzyme with small molecular weight contained no activity of either ureidoglycolate-degrading enzyme or urease. From the stoichiometry of the reaction catalyzed by the allantoate-degrading enzyme with small molecular weight it followed that the enzyme was allantoate amidohydrolase (EC 3.5.3.9). The optimal pH for the allantoate amidohydrolase was 8.5. Mn(2+) ions were essential for enzymatic activity. Glyoxylate and glycolate strongly inhibited the enzyme activity. The lysine and tryptophan residues were essential to the enzymatic catalysis; thiol group and tyrosyl residues were not involved in the enzyme catalysis.