Glycopeptide antibiotics: a mechanism-based screen employing a bacterial cell wall receptor mimetic.
The evolution of a highly targeted screening program for the discovery of antibiotics of the glycopeptide (vancomycin) class is described. A holistic approach was utilized which optimized not just screening techniques but also the selection of candidate producer cultures and their growth under conditions which enhanced production of target compounds. Two screen techniques were utilized; differential inhibition of a vancomycin-resistant strain and its susceptible parent, and a specific antagonism screen using the reversal of glycopeptide activity by a tripeptide analog of the glycopeptide receptor, diacetyl-L-lysyl-D-alanyl-D-alanine. The latter screen was 2- to 32-fold more sensitive to known glycopeptides than the former, and was absolutely specific, yielding no false positive responses. The use of the tripeptide antagonism assay, combined with optimized culture selection and growth conditions yielded novel glycopeptide antibiotics at a rate of 1 per 320 cultures screened. With a holistic approach to screening and properly optimized techniques, large numbers of cultures do not need to be examined in order to discover novel antibiotics.
D000192:Actinomycetales / Q000032:analysis; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002473:Cell Wall / Q000187:drug effects*; D004352:Drug Resistance, Microbial; D005189:False Positive Reactions; D006020:Glycopeptides / Q000494:pharmacology*; D008722:Methods; D008826:Microbial Sensitivity Tests; D014640:Vancomycin / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D006020:Glycopeptides; D014640:Vancomycin