Expression and delivery of an endolysin to combat Clostridium perfringens

Abstract

Clostridium perfringens is a cause for increasing concern due to its responsibility for severe infections both in humans and animals, especially poultry. To find new control strategies to treat C. perfringens infection, we investigated the activity and delivery of a bacteriophage endolysin. We identified a new endolysin, designated CP25L, which shows similarity to an N-acetylmuramoyl-l-alanine amidase domain and is distinct from other C. perfringens endolysins whose activity has been demonstrated in vitro. The cp25l gene was cloned and expressed in Escherichia coli, and the gene product demonstrated lytic activity against all 25 C. perfringens strains tested. The probiotic strain Lactobacillus johnsonii FI9785 was engineered to deliver the endolysin to the gastrointestinal tract. The integration of the nisRK two-component regulatory system from the Lactococcus lactis nisin A biosynthesis operon into the chromosome of L. johnsonii allowed constitutive expression of the endolysin under the control of the nisA promoter (PnisA), while the use of a signal peptide (SLPmod) led to successful secretion of the active endolysin to the surrounding media. The high specificity and activity of the endolysin suggest that it may be developed as an effective tool to enhance the control of C. perfringens by L. johnsonii in the gastrointestinal tract.

Intravitreal Injection of the Chimeric Phage Endolysin Ply187 Protects Mice from Staphylococcus aureus Endophthalmitis

“…The endolysins are highly specific to the host bacterium (genus) and have evolved to bind to unique and essential bacterial cell wall targets. In recent years, phage endolysins have attracted considerable interest as novel antibacterial agents (18) and have been used to treat a variety of bacterial infections, as demonstrated by the increasing number of experimental animal studies (19)(20)(21)(22). Similarly, human studies have documented the use of live antistaphylococcal phages for the treatment of conjunctivitis and blepharitis in 28 patients, with no side effects (23,24).…”

Bacteriophages and bacteriophage-derived endolysins as potential therapeutics to combat Gram-positive spore forming bacteria

“…perfringens strains in vitro and a limited level of cross species activity against a strain of B. cereus and B. subtilis (Gervasi et al . ).…”

“…The expression of a Cl. perfringens endolysin (CP25L) within a probiotic Lactobacillus lactis strain presents an interesting alternative to the use of a replicating phage cocktail (Seal 2013;Gervasi et al 2014) and has demonstrated activity against multiple Cl. perfringens strains in vitro and a limited level of cross species activity against a strain of B. cereus and B. subtilis (Gervasi et al 2014).…”

Non-antibiotic strategies for the control of necrotic enteritis in poultry

“…or gut bacteria. The recombinant probiotic was used for delivering the endolysin to the gastrointestinal tract of poultry where the active endolysin was efficiently secreted, suggesting that it may be helpful for controlling enteric clostridiosis (Gervasi et al, 2013b). Recently, a newly-identified specific anticlostridial phage endolysin CP25L was derived from a C. perfringens prophage (Gervasi et al, 2013a).…”