| The ability of bacteria to develop or acquire resistance to multiple antibiotics plays a major role in morbidity and mortality of bacterial infections. In many cases resistance is conferred by the presence of genes coding for antibiotic modifying enzymes, often present in plasmids. It was postulated that a strategy for combating antibiotic resistance may be to inhibit antibiotic modifying enzyme activity with peptide inhibitors. The model enzyme used for this study was aminoglycoside (6')-N-acetyltransferase type Ib [AAC(6')-Ib], which catalyzes the transfer of an acetyl group from acetyl coenzyme A to the 6' amino group of several clinically relevant aminoglycosides, rendering them ineffective. Multiple phage display techniques, including subtractive biopanning and solution phase biopanning, were used to identify putative inhibitory oligopeptides. Candidate AAC(6')-Ib specific peptides, identified by alignment studies, were subjected to an ELISA binding assay to gauge their binding affinity and synthesized. An in vitro inhibition assay was performed to determine if acetylation activity was inhibited in the presence of putative peptide inhibitors. Although, no inhibitory peptides were identified, the findings from this study will contribute to the effective use of phage display and the ELISA binding assay in the identification of peptide inhibitors. |
