{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T16:55:34Z","timestamp":1792601734453,"version":"4.5.0"},"reference-count":45,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,4,19]],"date-time":"2021-04-19T00:00:00Z","timestamp":1618790400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation of China","award":["31570175"],"award-info":[{"award-number":["31570175"]}]},{"name":"National Natural Science Foundation of China","award":["31770192"],"award-info":[{"award-number":["31770192"]}]},{"name":"National Natural Science Foundation of China","award":["32070187"],"award-info":[{"award-number":["32070187"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Lysins, including chimeric lysins, have recently been explored as novel promising alternatives to failing antibiotics in treating multi-drug resistant (MDR) pathogens, including methicillin-resistant Staphylococcus aureus (MRSA). Herein, by fusing the CHAP (cysteine, histidine-dependent amidohydrolase\/peptidase) catalytic domain from the Ply187 lysin with the non-SH3b cell-wall binding domain from the LysSA97 lysin, a new chimeric lysin ClyC was constructed with Ca2+-enhanced bactericidal activity against all S. aureus strains tested, including MRSA. Notably, treating S. aureus with 50 \u03bcg\/mL of ClyC in the presence of 100 \u03bcM Ca2+ lead to a reduction of 9 Log10 (CFU\/mL) in viable bacterial number, which was the first time to observe a lysin showing such a high activity. In addition, the effective concentration of ClyC could be decreased dramatically from 12 to 1 \u03bcg\/mL by combination with 0.3 \u03bcg\/mL of penicillin G. In a mouse model of S. aureus bacteremia, a single intraperitoneal administration of 0.1 mg\/mouse of ClyC significantly improved the survival rates and reduced 2 Log10 (CFU\/mL) of the bacterial burdens in the organs of the infected mice. ClyC was also found stable after lyophilization without cryoprotectants. Based on the above observations, ClyC could be a promising candidate against S. aureus infections.<\/jats:p>","DOI":"10.3390\/antibiotics10040461","type":"journal-article","created":{"date-parts":[[2021,4,19]],"date-time":"2021-04-19T11:21:38Z","timestamp":1618831298000},"page":"461","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["A Highly Active Chimeric Lysin with a Calcium-Enhanced Bactericidal Activity against Staphylococcus aureus In Vitro and In Vivo"],"prefix":"10.3390","volume":"10","author":[{"given":"Xiaohong","family":"Li","sequence":"first","affiliation":[{"name":"CAS Key Laboratory of Emerging Pathogens and Biosafety, Centre for Biosafety Mega-Sciences, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"},{"name":"College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shujuan","family":"Wang","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Emerging Pathogens and Biosafety, Centre for Biosafety Mega-Sciences, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"},{"name":"College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8467-8959","authenticated-orcid":false,"given":"Raphael","family":"Nyaruaba","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Emerging Pathogens and Biosafety, Centre for Biosafety Mega-Sciences, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"},{"name":"College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China"},{"name":"Sino-Africa Joint Research Center, Chinese Academy of Sciences, Wuhan 430074, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huan","family":"Liu","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Emerging Pathogens and Biosafety, Centre for Biosafety Mega-Sciences, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"},{"name":"College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6750-1465","authenticated-orcid":false,"given":"Hang","family":"Yang","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Emerging Pathogens and Biosafety, Centre for Biosafety Mega-Sciences, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9948-8880","authenticated-orcid":false,"given":"Hongping","family":"Wei","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Emerging Pathogens and Biosafety, Centre for Biosafety Mega-Sciences, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Favero, M.S. (2003). Outbreaks of community-associated methicillin-resistant Staphylococcus aureus skin infections. Infect. Control Hosp. Epidemiol., 24.","DOI":"10.1086\/infeconthospepid.24.10.787"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Pollitt, E.J.G., Szkuta, P.T., Burns, N., and Foster, S.J. (2018). Staphylococcus aureus infection dynamics. PLoS Pathog., 14.","DOI":"10.1371\/journal.ppat.1007112"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1373","DOI":"10.1080\/14760584.2016.1179583","article-title":"Staphylococcus aureus: The current state of disease, pathophysiology and strategies for prevention","volume":"15","author":"Dayan","year":"2016","journal-title":"Expert Rev. Vaccines"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1016\/S1471-4892(03)00109-7","article-title":"The evolution of a resistant pathogen\u2013the case of MRSA","volume":"3","author":"Enright","year":"2003","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1093\/femsre\/fux007","article-title":"Antibiotic resistance in Staphylococcus aureus. Current status and future prospects","volume":"41","author":"Foster","year":"2017","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Mulani, M.S., Kamble, E.E., Kumkar, S.N., Tawre, M.S., and Pardesi, K.R. (2019). Emerging Strategies to Combat ESKAPE Pathogens in the Era of Antimicrobial Resistance: A Review. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.00539"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Gondil, V.S., Harjai, K., and Chhibber, S. (2020). Endolysins as emerging alternative therapeutic agents to counter drug-resistant infections. Int. J. Antimicrob. Agents, 55.","DOI":"10.1016\/j.ijantimicag.2019.11.001"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1007\/s00253-012-4484-3","article-title":"Peptidoglycan hydrolases-potential weapons against Staphylococcus aureus","volume":"96","author":"Szweda","year":"2012","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e01923-17","DOI":"10.1128\/mBio.01923-17","article-title":"Are Phage Lytic Proteins the Secret Weapon To Kill Staphylococcus aureus","volume":"9","year":"2018","journal-title":"MBio"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/j.ijmm.2010.04.002","article-title":"Bacteriophage endolysins: A novel anti-infective to control Gram-positive pathogens","volume":"300","author":"Fischetti","year":"2010","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e02629-16","DOI":"10.1128\/AAC.02629-16","article-title":"Pharmacokinetics and Tolerance of the Phage Endolysin-Based Candidate Drug SAL200 after a Single Intravenous Administration among Healthy Volunteers","volume":"61","author":"Jun","year":"2017","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e02616-18","DOI":"10.1128\/AAC.02616-18","article-title":"Postantibiotic and Sub-MIC Effects of Exebacase (Lysin CF-301) Enhance Antimicrobial Activity against Staphylococcus aureus","volume":"63","author":"Oh","year":"2019","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1159\/000473872","article-title":"Successful Treatment of Chronic Staphylococcus aureus-Related Dermatoses with the Topical Endolysin Staphefekt SA.100: A Report of 3 Cases","volume":"9","author":"Pasmans","year":"2017","journal-title":"Case Rep. Dermatol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e02043-18","DOI":"10.1128\/AAC.02043-18","article-title":"ClyJ Is a Novel Pneumococcal Chimeric Lysin with a Cysteine- and Histidine-Dependent Amidohydrolase\/Peptidase Catalytic Domain","volume":"63","author":"Yang","year":"2019","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1111\/1574-6968.12104","article-title":"Chimeric Ply187 endolysin kills Staphylococcus aureus more effectively than the parental enzyme","volume":"342","author":"Mao","year":"2013","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Yang, H., Linden, S.B., Wang, J., Yu, J., Nelson, D.C., and Wei, H. (2015). A chimeolysin with extended-spectrum streptococcal host range found by an induced lysis-based rapid screening method. Sci. Rep., 5.","DOI":"10.1038\/srep17257"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3019","DOI":"10.3390\/v7062758","article-title":"A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability","volume":"7","author":"Swift","year":"2015","journal-title":"Viruses"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2743","DOI":"10.1128\/AAC.02526-12","article-title":"Novel Bacteriophage Lysin with Broad Lytic Activity Protects against Mixed Infection by Streptococcus pyogenes and Methicillin-Resistant Staphylococcus aureus","volume":"57","author":"Gilmer","year":"2013","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e01379-14","DOI":"10.1128\/mBio.01379-14","article-title":"Engineered Endolysin-Based \u201cArtilysins\u201d To Combat Multidrug-Resistant Gram-Negative Pathogens","volume":"5","author":"Briers","year":"2014","journal-title":"MBio"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1080\/1040841X.2020.1809346","article-title":"Advanced engineering of third-generation lysins and formulation strategies for clinical applications","volume":"46","author":"Paeshuyse","year":"2020","journal-title":"Crit. Rev. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e02291-18","DOI":"10.1128\/AAC.02291-18","article-title":"The Antistaphylococcal Lysin, CF-301, Activates Key Host Factors in Human Blood To Potentiate Methicillin-Resistant Staphylococcus aureus Bacteriolysis","volume":"63","author":"Indiani","year":"2019","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1545","DOI":"10.1093\/clinchem\/24.9.1545","article-title":"Measurement of free calcium ion in capillary blood and serum","volume":"24","author":"Christiansen","year":"1978","journal-title":"Clin. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.biochi.2019.04.005","article-title":"Kinetics of inactivation of staphylolytic enzymes: Qualitative and quantitative description","volume":"162","author":"Filatova","year":"2019","journal-title":"Biochimie"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Gu, J., Feng, Y., Feng, X., Sun, C., Lei, L., Ding, W., Niu, F., Jiao, L., Yang, M., and Li, Y. (2014). Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented \u201cEF-Hand-Like\u201d Calcium-Binding Phage Lysin. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004109"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4452","DOI":"10.1128\/JB.181.15.4452-4460.1999","article-title":"Evidence for a Holin-Like Protein Gene Fully Embedded Out of Frame in the Endolysin Gene of Staphylococcus aureus Bacteriophage 187","volume":"181","author":"Loessner","year":"1999","journal-title":"J. Bacteriol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1111\/1751-7915.12166","article-title":"Construction of a chimeric lysin Ply187N-V12C with extended lytic activity against staphylococci and streptococci","volume":"8","author":"Dong","year":"2015","journal-title":"Microb. Biotechnol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5225","DOI":"10.3390\/v7102870","article-title":"Isolation and Genome Characterization of the Virulent Staphylococcus aureus Bacteriophage SA97","volume":"7","author":"Chang","year":"2015","journal-title":"Viruses"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1007\/s00253-016-7747-6","article-title":"Characterization of a novel cell wall binding domain-containing Staphylococcus aureus endolysin LysSA97","volume":"101","author":"Chang","year":"2017","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"L799","DOI":"10.1152\/ajplung.2000.279.5.L799","article-title":"Synergistic and additive killing by antimicrobial factors found in human airway surface liquid","volume":"279","author":"Singh","year":"2000","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.2217\/fmb.12.97","article-title":"Bacteriophage endolysins as novel antimicrobials","volume":"7","author":"Schmelcher","year":"2012","journal-title":"Future Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Yang, H., Zhang, H., Wang, J., Yu, J., and Wei, H. (2017). A novel chimeric lysin with robust antibacterial activity against planktonic and biofilm methicillin-resistant Staphylococcus aureus. Sci. Rep., 7.","DOI":"10.1038\/srep40182"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1111\/j.1751-7915.2011.00263.x","article-title":"Domain shuffling and module engineering of Listeria phage endolysins for enhanced lytic activity and binding affinity","volume":"4","author":"Schmelcher","year":"2011","journal-title":"Microb. Biotechnol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1128\/AAC.01793-13","article-title":"Novel Chimeric Lysin with High-Level Antimicrobial Activity against Methicillin-Resistant Staphylococcus aureus In Vitro","volume":"58","author":"Yang","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4621","DOI":"10.1128\/AAC.00126-14","article-title":"Intravitreal injection of the chimeric phage endolysin Ply187 protects mice from Staphylococcus aureus endophthalmitis","volume":"58","author":"Singh","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4808","DOI":"10.1128\/JB.186.14.4808-4812.2004","article-title":"Identification of a Broadly Active Phage Lytic Enzyme with Lethal Activity against Antibiotic-Resistant Enterococcus faecalis and Enterococcus faecium","volume":"186","author":"Yoong","year":"2004","journal-title":"J. Bacteriol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.vetmic.2007.12.004","article-title":"Characterization of a bacteriophage lysin (Ply700) from Streptococcus uberis","volume":"130","author":"Celia","year":"2008","journal-title":"Vet. Microbiol."},{"key":"ref_37","first-page":"232","article-title":"Expression and Antibacterial Activity of CHAP Catalytic Domain of Staphylococcus aureus Phage Lysin Ply187","volume":"32","author":"Wu","year":"2016","journal-title":"Biotechnol. Bull."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2671","DOI":"10.1128\/AAC.02972-15","article-title":"Novel Engineered Peptides of a Phage Lysin as Effective Antimicrobials against Multidrug-Resistant Acinetobacter baumannii","volume":"60","author":"Thandar","year":"2016","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"445","DOI":"10.4161\/viru.2.5.17724","article-title":"Staphylococcus aureus biofilms: Properties, regulation, and roles in human disease","volume":"2","author":"Archer","year":"2011","journal-title":"Virulence"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1007\/s12250-019-00192-3","article-title":"Bacteriophages and Lysins in Biofilm Control","volume":"35","year":"2020","journal-title":"Virol. Sin."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.jbiotec.2008.09.003","article-title":"Antimicrobial activity of a chimeric enzybiotic towards Staphylococcus aureus","volume":"139","author":"Manoharadas","year":"2009","journal-title":"J. Biotechnol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1603","DOI":"10.1128\/AAC.01625-09","article-title":"Synergism between a Novel Chimeric Lysin and Oxacillin Protects against Infection by Methicillin-Resistant Staphylococcus aureus","volume":"54","author":"Daniel","year":"2010","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.1016\/j.ijbiomac.2019.11.042","article-title":"Recombinant engineered phage-derived enzybiotic in Pichia pastoris X-33 as whole cell biocatalyst for effective biocontrol of Vibrio parahaemolyticus in aquaculture","volume":"154","author":"Srinivasan","year":"2020","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1023\/A:1012180707283","article-title":"Rational Design of Stable Lyophilized Protein Formulations: Some Practical Advice","volume":"14","author":"Carpenter","year":"1997","journal-title":"Pharm. Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"D387","DOI":"10.1093\/nar\/gkn750","article-title":"The SWISS-MODEL Repository and associated resources","volume":"37","author":"Kiefer","year":"2009","journal-title":"Nucleic Acids Res."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/10\/4\/461\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:49:42Z","timestamp":1760161782000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/10\/4\/461"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,19]]},"references-count":45,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["antibiotics10040461"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics10040461","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,19]]}}}