Efficient enzymatic production of the bacterial second messenger c-di-GMP by the diguanylate cyclase YdeH from E. coli

Appl Biochem Biotechnol. 2011 Jan;163(1):71-9. doi: 10.1007/s12010-010-9017-x. Epub 2010 Jun 29.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cyclic GMP / biosynthesis*
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / isolation & purification
  • Escherichia coli Proteins / metabolism*
  • Gene Expression Regulation, Bacterial
  • Phosphorus-Oxygen Lyases / chemistry
  • Phosphorus-Oxygen Lyases / genetics
  • Phosphorus-Oxygen Lyases / isolation & purification
  • Phosphorus-Oxygen Lyases / metabolism*
  • Protein Structure, Tertiary
  • Second Messenger Systems*


  • Escherichia coli Proteins
  • Phosphorus-Oxygen Lyases
  • diguanylate cyclase
  • Cyclic GMP