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Differential activity profiles of translation inhibitors in whole‐cell and cell‐free approaches
Author(s) -
Weidlich M.,
Klammt C.,
Bernhard F.,
Karas M.,
Stein T.
Publication year - 2008
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2007.02281.x
Subject(s) - translation (biology) , cell , biology , computational biology , differential (mechanical device) , chemistry , biochemistry , physics , gene , messenger rna , thermodynamics
Abstract Aims: Evaluation of the activity profiles of standard prokaryotic translation inhibitors with different physicochemical properties under whole‐cell and cell‐free conditions. Methods and Results: The minimal inhibitory concentration values (cell‐free/whole‐cell μ g ml −1 ) for three aminoglycosides (neomycin, 0·01/6·92; paromomycin, 0·7/1·96; streptomycin 1·45/1·57), three macrolides (erythromycin, 1·53/56·9; josamycin, 1·61/87·7; oleandomycin, 5·12/565·9), chloramphenicol (11·9/3·04), and two tetracyclines (tetracycline hydrochloride, not determined/0·63; minocycline hydrochloride, 2·53/1·09), towards Escherichia coli A19 cells were determined with a microtitre plate‐based broth dilution method and compared with values determined in a coupled transcription/translation system based on a S30 extract of the same E. coli strain (cell‐free) for the production of the green fluorescent protein. Conclusions: The analysed prokaryotic translation inhibitors showed substance‐specific activity profiles under cell‐free vs whole‐cell conditions that are explainable by the physicochemical properties of the molecules. Significance and Impact of the Study: This study shows the advantages and limits of cell‐ free transcription/translation (CFTT) experiments for the discovery of novel antimicrobials. The main advantage is the direct access of the target structures (ribosomes) for the inhibitors, and our results provide an estimation of the concentration necessary to detect new agents. The main limitations are that the inhibitory properties of different agents in CFTT experiments do not necessarily reflect their growth inhibition activity in cell cultures.