An in vivo study of novel bioactive peptides that inhibit the growth of Escherichia coli

We have created a system in which synthetically produced novel bioactive pe ptides can be expressed in vivo in Escherichia coli. Twenty thousand of the se peptides were screened and 21 inhibitors were found that could inhibit t he growth of E. coli on minimal media. The inhibitors could be placed into one of two groups, 1-day inhibitors, which were partially inhibitory, and 2 -day inhibitors, which were completely inhibitory. Sequence analysis showed that two of the most potent inhibitors were actually peptide-protein chime ras in which the peptides had become fused to the 63 amino acid Rop protein which was also contained in the expression vector used in this study. Give n that Rop is known to form an incredibly stable structure, it could be ser ving as a stabilizing motif for these peptides. Sequence analysis of the pr edicted coding regions from the next 10 most inhibitory peptides showed tha t four of the 10 peptides contained one or more proline residues either at or very near the C-terminal end of the peptide which could act to prevent d egradation by peptidases. Collectively, based on what we observed in our sc reen of synthetic bioactive peptides that could prevent the growth of E. co li and what has been learned from structural studies of naturally occurring bioactive peptides, the presence of a stabilizing motif seems to be import ant for small peptides, if they are to be biologically active.