Abstract
As the incidence of antibiotic-resistant bacteria has become increased, phage endolysins are believed as one of the promising alternatives to antibiotics. However, the discovery of potent endolysin is still challenging because it is labor intensive and difficult to obtain a soluble form with high lytic activity. In this respect, the modular structures of Gram-positive endolysins can provide an opportunity to develop novel endolysins by domain rearrangement. In this study, a random domain swapping library of four different endolysins from phages infecting Staphylococcus aureus was constructed and screened to obtain engineered endolysins. The novel chimeric endolysin, Lys109 was selected and characterized for its staphylolytic activity. Lys109 exhibited greater bacterial cell lytic activity than its parental endolysins against staphylococcal planktonic cells and biofilms, showing highly improved activity in eliminating S. aureus from milk and on the surface of stainless steel. These results demonstrate that a novel chimeric endolysin with higher activity and solubility can be developed by random domain swapping and that this chimeric endolysin has a great potential as an antimicrobial agent.
Highlights
Staphylococcus aureus is a Gram-positive bacterium that threatens human and animal health, causing staphylococcal food poisoning and a wide range of infectious diseases, including skin infections, pneumonia, meningitis, endocarditis, and osteomyelitis (Lowy, 1998; De Lencastre et al, 2007)
The clear zones were visualized on the agar plate depending on their activities, whereas clones without pBAD33_SPN1S lysRz did not show a clear zone (Figure 1C). These results suggest that SPN1S lysRz-induced lysis of E. coli allowed active chimeric endolysins to form clear zone on agar plates containing target bacteria and that this system can be used as a method for the successful screening of novel chimeric endolysins
There was a single clone containing a LysSAP4 cysteine- and histidinedependent amidohydrolase/peptidase (CHAP) domain. These results suggest that CHAP domain is necessary to degrade S. aureus cell wall peptidoglycan, and this is consistently observed in other chimeric endolysins to control S. aureus (Daniel et al, 2010; Schmelcher et al, 2012c; Yang et al, 2014)
Summary
Staphylococcus aureus is a Gram-positive bacterium that threatens human and animal health, causing staphylococcal food poisoning and a wide range of infectious diseases, including skin infections, pneumonia, meningitis, endocarditis, and osteomyelitis (Lowy, 1998; De Lencastre et al, 2007). The strong biofilm-forming ability of S. aureus has aggravated problems in the food and medical industries (Lewis, 2001; Otto, 2012). For these reasons, there is an urgent need to create new antimicrobials to combat S. aureus (Foster, 2004). Endolysins are bacteriophage-encoded peptidoglycan hydrolases produced by bacteriophages at the end of their replication cycle to breakdown peptidoglycans of the bacterial cell wall, resulting in the release of viral progeny (Schmelcher et al, 2012a).
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