Cloning and Characterization of an Endolysin LysSA120 as a Potential Staphylococcus Biofilm-Removing Agent
Natalia N Golosova1, Yana A Khlusevich1, Bogdana I Kravchuk1
1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia.
Abstract:
Staphylococcus aureus, including methicillin- and vancomycin-resistant variants (MRSA, VRSA), causes infections that are increasingly difficult to treat with conventional antibiotics. One of the approaches to developing new therapeutics to treat staphylococcal infections is the use of bacteriophages specific to these bacteria or the lytic enzymes of such bacteriophages, which are capable of hydrolyzing the cell walls of these bacteria. Phage-encoded endolysins offer an alternative promising class of antimicrobial agents. In this study, LysSA120, a 250-amino-acid endolysin encoded by the S. aureus podophage vB_SauP_120, was cloned, expressed in Escherichia coli, and characterized. The domain organization and tertiary structure of LysSA120 were predicted. Recombinant LysSA120 hydrolyzed cell walls were obtained from S. aureus, S. epidermidis, S. haemolyticus, S. warneri, S. auricularis and S. saprophyticus. It was shown that treatment of S. aureus planktonic cells with endolysin LysSA120 led to reduced viability. Furthermore, LysSA120 could hydrolyze mature biofilm formed by VRSA. The lytic spectrum and antibiofilm activity of LysSA120 warrant its further evaluation as an enzybiotic against drug-resistant staphylococcal infections.
Insights
LysSA120, a novel endolysin derived from Staphylococcus aureus bacteriophage vB_SauP_120, effectively reduced bacterial viability and degraded mature biofilms. This enzybiotic shows promise for treating drug-resistant staphylococcal infections.
Area of Science:
- Microbiology
- Biotechnology
- Antimicrobial Research
Background:
- Staphylococcus aureus infections, including methicillin-resistant (MRSA) and vancomycin-resistant (VRSA) strains, pose significant treatment challenges due to antibiotic resistance.
- Bacteriophage-derived endolysins are emerging as a promising alternative antimicrobial strategy targeting bacterial cell walls.
Purpose of the Study:
- To clone, express, and characterize LysSA120, an endolysin from the S. aureus podophage vB_SauP_120.
- To evaluate the antimicrobial and antibiofilm activity of LysSA120 against various staphylococcal species, including resistant strains.
Main Methods:
- LysSA120 was cloned and expressed in Escherichia coli.
- Recombinant LysSA120 activity was tested against cell walls of multiple staphylococcal species.
- The effect of LysSA120 on S. aureus planktonic cell viability and VRSA mature biofilm hydrolysis was assessed.
Main Results:
- Recombinant LysSA120 demonstrated hydrolytic activity against cell walls of S. aureus and several other staphylococcal species.
- Treatment with LysSA120 significantly reduced the viability of S. aureus planktonic cells.
- LysSA120 effectively hydrolyzed mature biofilms formed by vancomycin-resistant S. aureus (VRSA).
Conclusions:
- LysSA120 exhibits broad lytic activity against staphylococci and potent antibiofilm properties.
- The findings support further investigation of LysSA120 as a potential enzybiotic for combating drug-resistant staphylococcal infections.
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