Construction and Activity Testing of a Modular Fusion Peptide against Enterococcus faecalis

Abstract

The emergence of antibiotic resistance in enterococci is a great concern encountered worldwide. Almost all enterococci exhibit significant levels of resistance to penicillin, ampicillin, semi-synthetic penicillin and most cephalosporins, primarily due to the expression of low-affinity penicillin-binding proteins. The development of new and novel antibacterial agents against enterococci is a significant need of the hour. In this research, we have constructed a modular peptide against Enterococcus faecalis. The enzymatic domain of the constructed peptide BP404 is from the bacteriocin BacL1 and the cell wall binding domain from endolysin PlyV12 of phage ϕ1. The protein BP404 was found to be active against two tested strains of Enterococcus faecalis, with a reduction in cell density amounting to 85% and 65%. The cell wall binding assay confirms the binding of the protein to Enterococcus faecalis, which was not seen towards the control strain Escherichia coli, invariably pointing to the specificity of BP404. To the best of our knowledge, this is one of the first instances of the development of a chimeric peptide against Enterococcus faecalis. This study points out that novel proteins can be genetically engineered against clinically relevant enterococci.

An Enzybiotic Cocktail Effectively Disrupts Preformed Dual Biofilm of Staphylococcus aureus and Enterococcus faecalis

“…Almost 100% of the protein was bound to the cell pellet and no protein was seen in the unbound supernatant fraction (Figure 3A: Lane 2 and Lane 4). In corroboration with a previously observed result described by Manoharadas et al [34], BP404 was able to bind to the E. faecalis clinical strain at a much lower rate, with only approximately 20% of the protein bound to the cell pellet (Figure 3B: Lane 1). Similarly, BP404 was also able to bind to S. aureus Rumba, but at a very low intensity (approximately 15-20%, Figure 3B: Lane 3).…”

“…In addition, we also wanted to test if protein P16-17/100 was able to bind to S. aureus Rumba and E. faecalis clinical isolate. It was earlier shown that BP404 was able to bind to E. faecalis clinical isolate, but not very efficiently [34]. As shown in Figure 3A, protein P16-17/100 could efficiently bind to both E. faecalis clinical isolate (Figure 3A: Lane 1) and S. aureus Rumba (Figure 3A: Lane 3).…”

“…In this work, we evaluated the potential of two chimeric enzybiotics in disrupting a dual biofilm formed by S. aureus Rumba, a veterinary isolate, and E. faecalis, a clinical isolate. One of the chimeric enzybiotics used in this study, BP404, was earlier found to be active against E. faecalis ATCC 29,212 and E. faecalis clinical isolate [34], with a higher activity exhibited against E. faecalis clinical isolate. However, the study was performed against planktonic cells of E. faecalis.…”

“…BP404 is comprised of a catalytic domain at the N-terminal targeting a critical peptidoglycan bond. However, the C-terminal was composed of a cell-wall-targeting domain from the phage φ1 endolysin PlyV12 [34], expected to bind to E. faecalis strains. This observation of the activity of BP404 against S. aureus Rumba was surprising.…”

“…Our earlier studies have shown that an engineered lysin, BP404, was active against E. faecalis [34]. In this study, we evaluated the activity of BP404 combined with another engineered endolysin, P16-17/100, against a mixed biofilm formed by a veterinary isolate of S. aureus and a clinical E. faecalis isolate.…”