Abstract
A ureidoglycolate-degrading activity was analysed in different tissues of French bean (Phaseolus vulgaris L.) plants during development. Activity was detected in all the tissues analysed, although values were very low in seeds before germination and in cotyledons. After radicle emergence, the activity increased due to high activity present in the axes. The highest levels of specific activity were found in developing fruits, from which the enzyme was purified and characterised. This is the first ureidoglycolate-degrading activity that has been purified to homogeneity from a ureide legume. The enzyme was purified 280 fold, and the specific activity for the pure enzyme was 4.4 units mg−1, which corresponds to a turnover number of 1,055 min−1. The native enzyme has a molecular mass of 240 kDa and consists of six identical or similar-sized subunits each of 38 kDa. The activity of the purified enzyme was completely dependent on manganese and asparagine. The enzyme exhibited hyperbolic, Michaelian kinetics for ureidoglycolate with a K m value of 3.9 mM. This enzyme has been characterised as a ureidoglycolate urea-lyase (EC 4.3.2.3).
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Abbreviations
- DAI:
-
Days after imbibition
- DAG:
-
Days after germination
- Tea:
-
Triethanolamine
- Tes:
-
N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid
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Acknowledgements
This work was supported by Ministerio de Ciencia y Tecnología (BOS2003-01595) and Plan Andaluz de Investigación (CVI-115). We thank the Ministerio de Educación, Cultura y Deporte for the award of predoctoral fellowships to A. Muñoz and M.J. Raso and the Ministerio de Ciencia y Tecnología for the award of a “Ramón y Cajal” contract to P. Piedras. We thank Anne Edwards the critical reading of the manuscript.
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Muñoz, A., Raso, M.J., Pineda, M. et al. Degradation of ureidoglycolate in French bean (Phaseolus vulgaris) is catalysed by a ubiquitous ureidoglycolate urea-lyase. Planta 224, 175–184 (2006). https://doi.org/10.1007/s00425-005-0186-8
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DOI: https://doi.org/10.1007/s00425-005-0186-8