Efficient Enzymatic Production of the Bacterial Second Messenger c-di-GMP by the Diguanylate Cyclase YdeH from E. coli
| Autor: | Claudia Massa, Franziska Zähringer, Tilman Schirmer |
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| Rok vydání: | 2010 |
| Předmět: |
Cell
Bioengineering medicine.disease_cause Second Messenger Systems Applied Microbiology and Biotechnology Biochemistry Chemical synthesis Escherichia coli medicine Cyclic GMP Molecular Biology chemistry.chemical_classification biology Escherichia coli Proteins Gene Expression Regulation Bacterial General Medicine Enzymatic synthesis GGDEF domain Protein Structure Tertiary medicine.anatomical_structure Enzyme chemistry Second messenger system biology.protein Diguanylate cyclase Phosphorus-Oxygen Lyases Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 163:71-79 |
| ISSN: | 1559-0291 0273-2289 |
| DOI: | 10.1007/s12010-010-9017-x |
| Popis: | 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 1 day and paves the way for extensive biochemical and biophysical studies on c-di-GMP-mediated processes. |
| Databáze: | OpenAIRE |
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