Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus Bacteriophage
Antibiotic-resistant bacteria (ARB) have spread widely and rapidly, with their increased occurrence corresponding with the increased use of antibiotics. Infections caused by Staphylococcus aureus have a considerable negative impact on human and livestock health. Bacteriophages and their peptidoglyca...
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MDPI AG
2018-02-01
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| Series: | Pharmaceuticals |
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| Online Access: | http://www.mdpi.com/1424-8247/11/1/25 |
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| author | Jumpei Fujiki Tomohiro Nakamura Takaaki Furusawa Hazuki Ohno Hiromichi Takahashi Junya Kitana Masaru Usui Hidetoshi Higuchi Yasunori Tanji Yutaka Tamura Hidetomo Iwano |
| author_facet | Jumpei Fujiki Tomohiro Nakamura Takaaki Furusawa Hazuki Ohno Hiromichi Takahashi Junya Kitana Masaru Usui Hidetoshi Higuchi Yasunori Tanji Yutaka Tamura Hidetomo Iwano |
| author_sort | Jumpei Fujiki |
| collection | DOAJ |
| description | Antibiotic-resistant bacteria (ARB) have spread widely and rapidly, with their increased occurrence corresponding with the increased use of antibiotics. Infections caused by Staphylococcus aureus have a considerable negative impact on human and livestock health. Bacteriophages and their peptidoglycan hydrolytic enzymes (endolysins) have received significant attention as novel approaches against ARB, including S. aureus. In the present study, we purified an endolysin, Lys-phiSA012, which harbors a cysteine/histidine-dependent amidohydrolase/peptidase (CHAP) domain, an amidase domain, and a SH3b cell wall binding domain, derived from a polyvalent S. aureus bacteriophage which we reported previously. We demonstrate that Lys-phiSA012 exhibits high lytic activity towards staphylococcal strains, including methicillin-resistant S. aureus (MRSA). Analysis of deletion mutants showed that only mutants possessing the CHAP and SH3b domains could lyse S. aureus, indicating that lytic activity of the CHAP domain depended on the SH3b domain. The presence of at least 1 mM Ca2+ and 100 µM Zn2+ enhanced the lytic activity of Lys-phiSA012 in a turbidity reduction assay. Furthermore, a minimum inhibitory concentration (MIC) assay showed that the addition of Lys-phiSA012 decreased the MIC of oxacillin. Our results suggest that endolysins are a promising approach for replacing current antimicrobial agents and may contribute to the proper use of antibiotics, leading to the reduction of ARB. |
| first_indexed | 2024-12-18T11:09:18Z |
| format | Article |
| id | doaj.art-c6126b2a32c54ee9a50bfba857ee69c8 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-12-18T11:09:18Z |
| publishDate | 2018-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-c6126b2a32c54ee9a50bfba857ee69c82022-12-21T21:10:02ZengMDPI AGPharmaceuticals1424-82472018-02-011112510.3390/ph11010025ph11010025Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus BacteriophageJumpei Fujiki0Tomohiro Nakamura1Takaaki Furusawa2Hazuki Ohno3Hiromichi Takahashi4Junya Kitana5Masaru Usui6Hidetoshi Higuchi7Yasunori Tanji8Yutaka Tamura9Hidetomo Iwano10Laboratory of Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Food Microbiology and Food Safety, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Veterinary Hygiene, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanDepartment of Bioengineering, Tokyo Institute of Technology, Yokohama 226-8502, JapanLaboratory of Food Microbiology and Food Safety, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanLaboratory of Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu 069-8501, JapanAntibiotic-resistant bacteria (ARB) have spread widely and rapidly, with their increased occurrence corresponding with the increased use of antibiotics. Infections caused by Staphylococcus aureus have a considerable negative impact on human and livestock health. Bacteriophages and their peptidoglycan hydrolytic enzymes (endolysins) have received significant attention as novel approaches against ARB, including S. aureus. In the present study, we purified an endolysin, Lys-phiSA012, which harbors a cysteine/histidine-dependent amidohydrolase/peptidase (CHAP) domain, an amidase domain, and a SH3b cell wall binding domain, derived from a polyvalent S. aureus bacteriophage which we reported previously. We demonstrate that Lys-phiSA012 exhibits high lytic activity towards staphylococcal strains, including methicillin-resistant S. aureus (MRSA). Analysis of deletion mutants showed that only mutants possessing the CHAP and SH3b domains could lyse S. aureus, indicating that lytic activity of the CHAP domain depended on the SH3b domain. The presence of at least 1 mM Ca2+ and 100 µM Zn2+ enhanced the lytic activity of Lys-phiSA012 in a turbidity reduction assay. Furthermore, a minimum inhibitory concentration (MIC) assay showed that the addition of Lys-phiSA012 decreased the MIC of oxacillin. Our results suggest that endolysins are a promising approach for replacing current antimicrobial agents and may contribute to the proper use of antibiotics, leading to the reduction of ARB.http://www.mdpi.com/1424-8247/11/1/25antibiotic resistantmultidrug resistantantimicrobial agentphage therapybacteriophageendolysinstaphylococciStaphylococcus aureus |
| spellingShingle | Jumpei Fujiki Tomohiro Nakamura Takaaki Furusawa Hazuki Ohno Hiromichi Takahashi Junya Kitana Masaru Usui Hidetoshi Higuchi Yasunori Tanji Yutaka Tamura Hidetomo Iwano Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus Bacteriophage Pharmaceuticals antibiotic resistant multidrug resistant antimicrobial agent phage therapy bacteriophage endolysin staphylococci Staphylococcus aureus |
| title | Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus Bacteriophage |
| title_full | Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus Bacteriophage |
| title_fullStr | Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus Bacteriophage |
| title_full_unstemmed | Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus Bacteriophage |
| title_short | Characterization of the Lytic Capability of a LysK-Like Endolysin, Lys-phiSA012, Derived from a Polyvalent Staphylococcus aureus Bacteriophage |
| title_sort | characterization of the lytic capability of a lysk like endolysin lys phisa012 derived from a polyvalent staphylococcus aureus bacteriophage |
| topic | antibiotic resistant multidrug resistant antimicrobial agent phage therapy bacteriophage endolysin staphylococci Staphylococcus aureus |
| url | http://www.mdpi.com/1424-8247/11/1/25 |
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