{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T09:54:30Z","timestamp":1791194070925,"version":"4.1.0"},"reference-count":60,"publisher":"American Society for Microbiology","issue":"11","license":[{"start":{"date-parts":[[2013,11,1]],"date-time":"2013-11-01T00:00:00Z","timestamp":1383264000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2013,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Phage endolysins are murein hydrolases that break the bacterial cell wall to provoke lysis and release of phage progeny. Recently, these enzymes have also been recognized as powerful and specific antibacterial agents when added exogenously. In the pneumococcal system, most cell wall associated murein hydrolases reported so far depend on choline for activity, and Cpl-7 lysozyme constitutes a remarkable exception. Here, we report the improvement of the killing activity of the Cpl-7 endolysin by inversion of the sign of the charge of the cell wall-binding module (from \u221214.93 to +3.0 at neutral pH). The engineered variant, Cpl-7S, has 15 amino acid substitutions and an improved lytic activity against\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Streptococcus pneumoniae<\/jats:named-content>\n            (including multiresistant strains),\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Streptococcus pyogenes<\/jats:named-content>\n            , and other pathogens. Moreover, we have demonstrated that a single 25-\u03bcg dose of Cpl-7S significantly increased the survival rate of zebrafish embryos infected with\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">S. pneumoniae<\/jats:named-content>\n            or\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">S. pyogenes<\/jats:named-content>\n            , confirming the killing effect of Cpl-7S\n            <jats:italic>in vivo<\/jats:italic>\n            . Interestingly, Cpl-7S, in combination with 0.01% carvacrol (an essential oil), was also found to efficiently kill Gram-negative bacteria such as\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Escherichia coli<\/jats:named-content>\n            and\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Pseudomonas putida<\/jats:named-content>\n            , an effect not described previously. Our findings provide a strategy to improve the lytic activity of phage endolysins based on facilitating their pass through the negatively charged bacterial envelope, and thereby their interaction with the cell wall target, by modulating the net charge of the cell wall-binding modules.\n          <\/jats:p>","DOI":"10.1128\/aac.01372-13","type":"journal-article","created":{"date-parts":[[2013,8,20]],"date-time":"2013-08-20T01:58:37Z","timestamp":1376963917000},"page":"5355-5365","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":106,"title":["Improving the Lethal Effect of Cpl-7, a Pneumococcal Phage Lysozyme with Broad Bactericidal Activity, by Inverting the Net Charge of Its Cell Wall-Binding Module"],"prefix":"10.1128","volume":"57","author":[{"given":"Roberto","family":"D\u00edez-Mart\u00ednez","sequence":"first","affiliation":[{"name":"Departamento de Microbiolog\u00eda Molecular y Biolog\u00eda de las Infecciones, Centro de Investigaciones Biol\u00f3gicas, CSIC, Madrid, Spain"},{"name":"CIBER de Enfermedades Respiratorias, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H\u00e9ctor","family":"de Paz","sequence":"additional","affiliation":[{"name":"Departamento de Microbiolog\u00eda Molecular y Biolog\u00eda de las Infecciones, Centro de Investigaciones Biol\u00f3gicas, CSIC, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noem\u00ed","family":"Bustamante","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica-F\u00edsica Biol\u00f3gica, Instituto Qu\u00edmica-F\u00edsica Rocasolano, CSIC, Madrid, Spain"},{"name":"CIBER de Enfermedades Respiratorias, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ernesto","family":"Garc\u00eda","sequence":"additional","affiliation":[{"name":"Departamento de Microbiolog\u00eda Molecular y Biolog\u00eda de las Infecciones, Centro de Investigaciones Biol\u00f3gicas, CSIC, Madrid, Spain"},{"name":"CIBER de Enfermedades Respiratorias, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margarita","family":"Men\u00e9ndez","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica-F\u00edsica Biol\u00f3gica, Instituto Qu\u00edmica-F\u00edsica Rocasolano, CSIC, Madrid, Spain"},{"name":"CIBER de Enfermedades Respiratorias, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro","family":"Garc\u00eda","sequence":"additional","affiliation":[{"name":"Departamento de Microbiolog\u00eda Molecular y Biolog\u00eda de las Infecciones, Centro de Investigaciones Biol\u00f3gicas, CSIC, Madrid, Spain"},{"name":"CIBER de Enfermedades Respiratorias, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00109-009-0557-x"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.2174\/157489107782497290"},{"key":"e_1_3_3_4_2","first-page":"93","article-title":"Pneumococcal conjugate vaccine for childhood immunization\u2014WHO position paper","volume":"82","author":"WHO","year":"2007","unstructured":"WHO. 2007. Pneumococcal conjugate vaccine for childhood immunization\u2014WHO position paper. Wkly. Epidemiol. Rec. 82:93\u2013104.","journal-title":"Wkly. Epidemiol. Rec."},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.2011.00284.x"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-0691.2012.03938.x"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cis359"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.061038398"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature01026"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.2174\/138920312804871193"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1066869"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2007.08.002"},{"key":"e_1_3_3_13_2","first-page":"149","volume-title":"Enzybiotics: antibiotic enzymes as drugs and therapeutics","author":"Borysowski J","year":"2010","unstructured":"BorysowskiJG\u00f3rskiA. 2010. Anti-staphylococcal lytic enzymes, p 149\u2013172. In VillaTGVeiga-CrespoP (ed), Enzybiotics: antibiotic enzymes as drugs and therapeutics. John Wiley & Sons, Inc., Hoboken, NJ."},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.186.14.4808-4812.2004"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02526-12"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.femsre.2004.05.002"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(90)90116-9"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.154559"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0046654"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkg485"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.49.11.4789-4792.2005"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00137-07"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/0006-3002(60)90205-5"},{"key":"e_1_3_3_24_2","volume-title":"Molecular cloning: a laboratory manual","author":"Sambrook J","year":"2001","unstructured":"SambrookJRussellDW. 2001. Molecular cloning: a laboratory manual, 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.","edition":"3"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.4.1074"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.12.6832"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M407067200"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M802808200"},{"key":"e_1_3_3_29_2","first-page":"90","volume-title":"Analytical ultracentrifugation in biochemistry and polymer science","author":"Laue TM","year":"1992","unstructured":"LaueTMShahBDRidgewayTMPelletierSL. 1992. Computer-aided interpretation of analytical sedimentation data for proteins, p 90\u2013125. In HardingSERoweAJHortonJC (ed), Analytical ultracentrifugation in biochemistry and polymer science. Royal Society of Chemistry, Cambridge, United Kingdom."},{"key":"e_1_3_3_30_2","volume-title":"The PyMOL molecular graphics system","author":"DeLano WL","year":"2005","unstructured":"DeLanoWL. 2005. The PyMOL molecular graphics system. DeLano Scientific LLC, South San Francisco, CA."},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-10-168"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jasms.2008.04.016"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)63393-9"},{"key":"e_1_3_3_34_2","volume-title":"Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, document","author":"CLSI","year":"2006","unstructured":"CLSI. 2006. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, document M7-A7. Clinical and Laboratory Standards Institute, Wayne, PA."},{"key":"e_1_3_3_35_2","volume-title":"A guide for the laboratory use of zebrafish (Danio rerio)","author":"Westerfield M","year":"2007","unstructured":"WesterfieldM. 2007. The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio), 5th ed. University of Oregon Press, Eugene, OR.","edition":"5"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.244160"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0044135"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2003.03569.x"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2004.03.022"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2003.10.020"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1002\/jps.20853"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-13-53"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkr1065"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01783-07"},{"key":"e_1_3_3_45_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00284-009-9379-x"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1993.tb06078.x"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.180.2.210-217.1998"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.67.4.686-723.2003"},{"key":"e_1_3_3_49_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2011.08.005"},{"key":"e_1_3_3_50_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2672.2010.04931.x"},{"key":"e_1_3_3_51_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1203472109"},{"key":"e_1_3_3_52_2","doi-asserted-by":"publisher","DOI":"10.1177\/153537020623100402"},{"key":"e_1_3_3_53_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijmm.2010.04.002"},{"key":"e_1_3_3_54_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2008.09.012"},{"key":"e_1_3_3_55_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00890-10"},{"key":"e_1_3_3_56_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.19.5709-5717.2001"},{"key":"e_1_3_3_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01148-06"},{"key":"e_1_3_3_58_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2009.01979.x"},{"key":"e_1_3_3_59_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2010.02398.x"},{"key":"e_1_3_3_60_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.78.12.7458"},{"key":"e_1_3_3_61_2","doi-asserted-by":"publisher","DOI":"10.3109\/1040841X.2013.1763219"}],"updated-by":[{"DOI":"10.1128\/aac.02743-14","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2014,4,10]],"date-time":"2014-04-10T00:00:00Z","timestamp":1397088000000}}],"container-title":["Antimicrobial Agents and Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.01372-13","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.01372-13","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,21]],"date-time":"2022-02-21T22:35:17Z","timestamp":1645482917000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AAC.01372-13"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,11]]},"references-count":60,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2013,11]]}},"alternative-id":["10.1128\/AAC.01372-13"],"URL":"https:\/\/doi.org\/10.1128\/aac.01372-13","relation":{},"ISSN":["0066-4804","1098-6596"],"issn-type":[{"value":"0066-4804","type":"print"},{"value":"1098-6596","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,11]]},"assertion":[{"value":"2013-06-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2013-08-09","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2013-10-11","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}