Journal
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 163, Issue 1, Pages 71-79Publisher
SPRINGER
DOI: 10.1007/s12010-010-9017-x
Keywords
c-di-GMP; Diguanylate cyclase; GGDEF domain; Enzymatic synthesis; E. coli
Funding
- Swiss National Science Foundation [3100A0-105587]
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available