Ureidoglycolate amidohydrolase from developing French bean fruits (Phaseolus vulgaris [L.].).
Ureidoglycolate is an intermediate of allantoin catabolism in ureide-transporting legumes. This report describes the first purification of ureidoglycolate degrading activity (UGDA) from plant tissue in which the enzyme has been separated from urease. The enzyme from developing fruits of Phaseolus vulgaris has been purified 48-fold to give a preparation free of allantoinase and urease activity. UGDA was inhibited by EDTA while the Vmax was increased in the presence of Mn2+. The Km values for ureidoglycolate in the presence and the absence of Mn2+ were 2.0 and 5.4 mM, respectively. In the absence of Mn2+ UGDA was heat labile at 40 degrees C, but in the presence of Mn2+ the activity was stable up to temperatures of 60 degrees C. The Mr of UGDA was determined to be 300,000 by gel filtration chromatography and the pH optimum ranged from pH 7.0 to 8.5. Ammonia was determined to be the nitrogen-containing product of UGDA by a microdiffusion assay. This enzyme should therefore be described as ureidoglycolate amidohydrolase. The activity was shown to be associated with peroxisomes by fractionation of a crude extract on a sucrose density gradient. The products of ureidoglycolate degradation are glyoxylate, ammonia, and presumably carbon dioxide, which can be readily utilized by pathways of metabolism that are known to be present in this organelle.
Wells
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Department of Agricultural Chemistry, University of Sydney, New South Wales, Australia.
Lees
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D019760:Amidine-Lyases; D002850:Chromatography, Gel; D004492:Edetic Acid / Q000494:pharmacology; D007887:Fabaceae / Q000201:enzymology* / Q000254:growth & development; D006863:Hydrogen-Ion Concentration; D008190:Lyases / Q000302:isolation & purification / Q000378:metabolism*; D008345:Manganese / Q000502:physiology; D008670:Metals / Q000494:pharmacology; D009584:Nitrogen / Q000378:metabolism; D009928:Organ Specificity; D010946:Plants, Medicinal
D008670:Metals; D008345:Manganese; D004492:Edetic Acid; D008190:Lyases; D019760:Amidine-Lyases; C033757:ureidoglycollate lyase; D009584:Nitrogen