{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T07:44:56Z","timestamp":1791791096927,"version":"4.3.4"},"reference-count":57,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T00:00:00Z","timestamp":1791676800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T00:00:00Z","timestamp":1791676800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100004160","name":"Mazandaran University of Medical Sciences","doi-asserted-by":"publisher","award":["14548"],"award-info":[{"award-number":["14548"]}],"id":[{"id":"10.13039\/501100004160","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["iubmb.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech and App Biochem"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    The emergence of antibiotic\u2010resistant pathogens, particularly methicillin\u2010resistant\n                    <jats:italic>Staphylococcus aureus<\/jats:italic>\n                    (MRSA), represents a major public health threat, with limited treatment options. Bacteriophage\u2010derived endolysins and their engineered chimeric derivatives have attracted attention as alternative antibacterial agents due to their rapid bacteriolytic activity, target specificity, and low resistance propensity. ASEnd was constructed by fusing the enzymatically active LYZ2 domain from\n                    <jats:italic>S. aureus<\/jats:italic>\n                    phage \u03c6MR11 with the SH3b\u2010type cell wall\u2013binding domain from phage K via a flexible (G4S)\n                    <jats:sub>3<\/jats:sub>\n                    linker. Structural integrity was assessed in silico using Ramachandran plot analysis, ERRAT, Verify3D, and ProSA\u2010web. The recombinant protein was expressed in\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    BL21 (DE3) and purified by immobilized metal affinity chromatography. Antibacterial activity was evaluated against MRSA, methicillin\u2010susceptible\n                    <jats:italic>S. aureus<\/jats:italic>\n                    (MSSA), and other bacterial strains using disk diffusion, turbidity reduction, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. pH tolerance, storage stability, and cytotoxicity toward HEK\u2010293 cells were also examined. ASEnd was obtained as a soluble 34.7\u00a0kDa protein with favorable structural metrics (&gt;99% of residues in favored Ramachandran regions). The protein exhibited potent activity against\n                    <jats:italic>S. aureus<\/jats:italic>\n                    , producing inhibition zones up to 19\u00a0mm and rapidly reducing turbidity at low concentrations. ASEnd demonstrated MIC and MBC values of 1.56 and 3.125\u00a0\u00b5g\/mL, respectively, representing a 16\u2010fold improvement over LYZ2. The enzyme retained activity between pH 6 and 8, but declined significantly below pH 5, and gradual loss was observed during storage at 4\u00b0C (21%, 42%, and 78% reductions after 1, 7, and 14\u00a0days), with complete inactivation after 3\u00a0weeks. ASEnd showed no cytotoxicity toward HEK\u2010293 cells and displayed selective activity against MRSA and MSSA, with limited cross\u2010reactivity against\n                    <jats:italic>Streptococcus pyogenes<\/jats:italic>\n                    (2.4\u2010fold lower) and no effect on\n                    <jats:italic>E. coli<\/jats:italic>\n                    . These findings identify ASEnd as a rationally designed chimeric endolysin with enhanced potency and selectivity against\n                    <jats:italic>S. aureus<\/jats:italic>\n                    and a favorable safety profile. Further stability optimization, clinical isolate testing, and in vivo validation are required to establish its therapeutic potential.\n                  <\/jats:p>","DOI":"10.1002\/bab.70212","type":"journal-article","created":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T06:48:57Z","timestamp":1791787737000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Design, Characterization, and Efficacy of ASEnd: A Novel Chimeric Endolysin for Targeting Methicillin\u2010Resistant\n                    <i>Staphylococcus aureus<\/i>"],"prefix":"10.1002","author":[{"given":"Sarvenaz","family":"Janbaz","sequence":"first","affiliation":[{"name":"Department of Medical Biotechnology, School of Advanced Technologies in Medicine Mazandaran University of Medical Sciences Sari Iran"},{"name":"Molecular and Cell Biology Research Center Faculty of Medicine Mazandaran University of Medical Sciences Sari Iran"},{"name":"Student Research Committee, Faculty of Advanced Technologies in Medicine Mazandaran University of Medical Sciences Sari Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amir","family":"Savardashtaki","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology School of Advanced Medical Sciences and Technologies Shiraz University of Medical Sciences Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marziyeh","family":"Asadi","sequence":"additional","affiliation":[{"name":"Cellular and Molecular Research Center Gerash University of Medical Sciences Gerash Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mortaza","family":"Taheri\u2010Anganeh","sequence":"additional","affiliation":[{"name":"Cellular and Molecular Research Center Cellular and Molecular Medicine Research Institute Urmia University of Medical Sciences Urmia Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehrdad","family":"Gholami","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Virology Faculty of Medicine Mazandaran University of Medical Sciences Sari Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seyed Ehsan","family":"Enderami","sequence":"additional","affiliation":[{"name":"Immunogenetics Research Center School of Medicine, Mazandaran University of Medical Sciences  Sari Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amir","family":"Maleksabet","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, School of Advanced Technologies in Medicine Mazandaran University of Medical Sciences Sari Iran"},{"name":"Molecular and Cell Biology Research Center Faculty of Medicine Mazandaran University of Medical Sciences Sari Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,11]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms24098182"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2023.127969"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.idc.2020.04.003"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.3390\/v12111340"},{"issue":"11","key":"e_1_2_9_6_1","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.3390\/antibiotics10111277","article-title":"Endolysin, a Promising Solution against Antimicrobial Resistance","volume":"10","author":"Rahman M. U.","year":"2021","journal-title":"Antibiotics (Basel)"},{"issue":"4","key":"e_1_2_9_7_1","doi-asserted-by":"crossref","first-page":"148","DOI":"10.3390\/antibiotics9040148","article-title":"Development of Staphylococcus Enzybiotics: The Ph28 Gene of Staphylococcus epidermidis Phage PH15 Is a Two\u2010Domain Endolysin","volume":"9","author":"Muharram M. M.","year":"2020","journal-title":"Antibiotics (Basel)"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2018.08.001"},{"issue":"1","key":"e_1_2_9_9_1","article-title":"Recombinant Endolysins as Potential Therapeutics Against Antibiotic\u2010Resistant Staphylococcus aureus: Current Status of Research and Novel Delivery Strategies","volume":"31","author":"Haddad Kashani H.","year":"2017","journal-title":"Clinical Microbiology Reviews"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01616\u201006"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dku552"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.2217\/fmb.12.97"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1086\/374001"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.3390\/v10040182"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s42770\u2010023\u201001219\u20104"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574\u20106968.2008.01152.x"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12866\u2010023\u201003002\u20109"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.0c00812"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2020.615887"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010019\u201042435\u2010z"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.gene.2009.04.023"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.46.6.2017\u20102020.2002"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt381"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471\u20102105\u20109\u201040"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2010.5"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv342"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt458"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1107\/S1600576725001669"},{"issue":"4","key":"e_1_2_9_29_1","first-page":"281","article-title":"Specific Targeting of Recombinant Human Pancreatic Ribonuclease 1 Using Gonadotropin\u2010Releasing Hormone Targeting Peptide Toward Gonadotropin\u2010Releasing Hormone Receptor\u2010Positive Cancer Cells","volume":"46","author":"Maleksabet A.","year":"2021","journal-title":"Iranian Journal of Medical Sciences"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10989\u2010023\u201010584\u20105"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574\u20106968.2009.01541.x"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471\u20102180\u201012\u201033"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/48.suppl_1.5"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.32.3.374"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1111\/1574\u20106968.12104"},{"key":"e_1_2_9_36_1","doi-asserted-by":"crossref","DOI":"10.3389\/fmicb.2020.574739","article-title":"Engineered Lysins With Customized Lytic Activities Against Enterococci and Staphylococci","volume":"11","author":"Binte Muhammad Jai H. S.","year":"2020","journal-title":"Frontiers in Microbiology"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics10060722"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.188.7.2463\u20102472.2006"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.3390\/ph3041139"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.5.1431\u20101437.2001"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2008.09.012"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep40182"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574\u20106968.2008.01308.x"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.20.7161\u20107164.2005"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2008.09.003"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2017.00290"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.1089\/mdr.2012.0025"},{"key":"e_1_2_9_48_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01625\u201009"},{"key":"e_1_2_9_49_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2012.00043"},{"key":"e_1_2_9_50_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.02927"},{"key":"e_1_2_9_51_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms26094376"},{"key":"e_1_2_9_52_1","doi-asserted-by":"publisher","DOI":"10.3390\/v10110638"},{"key":"e_1_2_9_53_1","doi-asserted-by":"publisher","DOI":"10.1111\/apm.12099"},{"key":"e_1_2_9_54_1","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.000032"},{"key":"e_1_2_9_55_1","doi-asserted-by":"crossref","first-page":"1632","DOI":"10.3389\/fmicb.2017.01632","article-title":"Real\u2010Time Assessment of Staphylococcus aureus Biofilm Disruption by Phage\u2010derived Proteins","volume":"8","author":"Guti\u00e9rrez D.","year":"2017","journal-title":"Frontiers in Microbiology"},{"issue":"4","key":"e_1_2_9_56_1","doi-asserted-by":"crossref","first-page":"182","DOI":"10.3390\/v10040182","article-title":"Characterization of a New Staphylococcus aureus Kayvirus Harboring a Lysin Active against Biofilms","volume":"10","author":"Melo L. D. R.","year":"2018","journal-title":"Viruses"},{"key":"e_1_2_9_57_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2026.152528"},{"key":"e_1_2_9_58_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00203\u2010026\u201004846\u20101"}],"container-title":["Biotechnology and Applied Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bab.70212","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/bab.70212","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bab.70212","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T06:48:59Z","timestamp":1791787739000},"score":1,"resource":{"primary":{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/10.1002\/bab.70212"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,11]]},"references-count":57,"alternative-id":["10.1002\/bab.70212"],"URL":"https:\/\/doi.org\/10.1002\/bab.70212","archive":["Portico"],"relation":{},"ISSN":["0885-4513","1470-8744"],"issn-type":[{"value":"0885-4513","type":"print"},{"value":"1470-8744","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,11]]},"assertion":[{"value":"2025-12-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-08","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-10-11","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"bab.70212"}}