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Catalytic analysis of a recombinant d-hydantoinase from Agrobacterium tumefaciens

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Abstract

The d-hydantoinase gene of a wild strain of Agrobacterium tumefaciens BQL9 had 99.78% nucleotide sequence identity with other available Agrobacterium genes. The resulting amino acid sequence showed two important substitutions affecting two α-helixes in the secondary structure of the protein. The union of Mn2+ to the protein was essential for activating the enzyme and was independent of the temperature. d-Hydantoinase only was inactivated in the presence of 70 mM EDTA and at over 40 °C. The enzyme showed both hydantoinase and pyrimidinase activities, but only with the d-enantiomers of the substrates. Activity was greater for substrates with apolar groups in the number 5 carbon atom of the hydantoin. The native structure of the N-terminal end of this d-hydantoinase proved to be indispensable to its enzymatic activity.

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Authors and Affiliations

  1. Departamento de Química-Física, Bioquímica y Química Inorgánica Edificio C.I.T.E. I, Universidad de Almería, Almería, 04120, Spain

    Josefa María Clemente-Jiménez, Sergio Martínez-Rogríguez, Lydia Mingorance-Cazorla, Francisco Javier Las Heras-Vázquez & Felipe Rodríguez-Vico

  2. Bioferma Murcia S.A., Polígono Industrial `Las Salinas', Avda. Europa s/n, Alhama de Murcia, Murcia, 30840, Spain

    Santiago De La Escalera-Hueso

Authors
  1. Josefa María Clemente-Jiménez
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  2. Sergio Martínez-Rogríguez
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  3. Lydia Mingorance-Cazorla
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  4. Santiago De La Escalera-Hueso
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  5. Francisco Javier Las Heras-Vázquez
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  6. Felipe Rodríguez-Vico
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Correspondence to Felipe Rodríguez-Vico.

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Clemente-Jiménez, J.M., Martínez-Rogríguez, S., Mingorance-Cazorla, L. et al. Catalytic analysis of a recombinant d-hydantoinase from Agrobacterium tumefaciens . Biotechnology Letters 25, 1067–1073 (2003). https://doi.org/10.1023/A:1024115220304

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