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Engineering Cyclic Amidases for Non-natural Amino Acid Synthesis

  • Francisco Javier Las Heras-VázquezEmail author
  • Josefa María Clemente-Jiménez
  • Sergio Martínez-Rodríguez
  • Felipe Rodríguez-Vico
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 794)

Abstract

Hydantoinases/dihydropyrimidinases are important biotechnological enzymes involved in the production of α- and β-amino acids. Their isolation from new sources with different substrate specificities, improved activity, enantioselectivity, or higher stability continues to be of great industrial interest. Here, we provide a detailed description of how to produce high quantities of the recombinant hydantoinase/dihydropyrimidinase enzyme from Sinorhizobium meliloti CECT4114 (SmeDhp). Several techniques are combined to obtain this goal, from cloning to activity measurement by HPLC.

Key words

α- and β-Amino acids Hydantoinases/dihydropyrimidinases Cloning Overexpression Metal-affinity HPLC 

Notes

Acknowledgements

This work was supported by the Spanish Ministry of Education and Science and European Regional Development Fund (ERDF) through the project BIO2007-67009, the Andalusian Regional Council of Innovation, Science and Technology through the projects CV7-02651 and TEP4691 and the European Science Foundation COST Action CM0701.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Francisco Javier Las Heras-Vázquez
    • 1
    Email author
  • Josefa María Clemente-Jiménez
    • 1
  • Sergio Martínez-Rodríguez
    • 1
  • Felipe Rodríguez-Vico
    • 1
  1. 1.Departamento de Química-Física, Bioquímica y Química InorgánicaUniversidad de AlmeríaAlmeríaSpain

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