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Efficient Enzymatic Production of the Bacterial Second Messenger c-di-GMP by the Diguanylate Cyclase YdeH from E. coli

Zähringer, Franziska ; Massa, Claudia ; Schirmer, Tilman

In: Applied Biochemistry and Biotechnology, 2011, vol. 163, no. 1, p. 71-79

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    Titre
    • Efficient Enzymatic Production of the Bacterial Second Messenger c-di-GMP by the Diguanylate Cyclase YdeH from E. coli
    Auteur
    • Zähringer, Franziska. Core Program Structural Biology and Biophysics, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland
    • Massa, Claudia. Core Program Structural Biology and Biophysics, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland
    • Schirmer, Tilman. Core Program Structural Biology and Biophysics, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Applied Biochemistry and Biotechnology, 2011, vol. 163, no. 1, p. 71-79. Humana Press Inc
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:317045
    Summary
    Cyclic di-GMP (c-di-GMP) is an almost universal bacterial second messenger involved in the regulation of cell surface-associated traits and the persistence of infections. GGDEF and EAL domain-containing proteins catalyse c-di-GMP synthesis and degradation, respectively. We report the enzymatic large-scale synthesis of c-di-GMP, making use of the GGDEF domain-containing protein YdeH from Escherichia coli. Overexpression and purification of YdeH have been established, and the conditions for c-di-GMP synthesis were optimised. In contrast to the chemical synthesis of c-di-GMP, enzymatic c-di-GMP production is a one-step reaction that can easily be performed with the equipment of a standard biochemical lab. The protocol allows the production of milligram amounts of c-di-GMP within 1day and paves the way for extensive biochemical and biophysical studies on c-di-GMP-mediated processes