{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T13:40:33Z","timestamp":1790775633230,"version":"4.1.0"},"reference-count":36,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2020,1,20]],"date-time":"2020-01-20T00:00:00Z","timestamp":1579478400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Robert Koch Institute"},{"name":"German Ministry of Health","award":["1369\u2013402"],"award-info":[{"award-number":["1369\u2013402"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,4,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Objectives<\/jats:title>\n                  <jats:p>To investigate the carbapenem resistance mechanism of a carbapenem-resistant clinical Pseudomonas aeruginosa isolate.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>A carbapenem-resistant P. aeruginosa isolate was recovered from a tracheal swab from a patient of a general ward in central Germany. Various phenotypic tests confirmed production of a carbapenemase that could not be identified further by PCR. A novel bla gene was identified by WGS and its carbapenemase activity was verified by heterologous expression in an Escherichia coli cloning strain. Kinetic parameters of the novel \u03b2-lactamase were determined by spectrophotometric measurements using purified enzyme.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>WGS confirmed the presence of a novel class A carbapenemase. The novel bla gene was named GPC-1 (GPC standing for German Pseudomonas Carbapenemase) and exhibited 77% amino acid identity to BKC-1. WGS also showed that blaGPC-1 was located on the chromosome surrounded by multiple ISs as part of a 26\u2009kb genetic island. Heterologous expression of GPC-1 in E. coli TOP10 led to increased MICs of penicillins, oxyimino-cephalosporins, aztreonam and imipenem, but not of meropenem or ertapenem. Spectrophotometric measurements supported the MIC studies, but detected a slight hydrolysis of ertapenem and meropenem when using high concentrations of purified enzyme.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>The biochemical characterization of GPC-1 emphasizes the ongoing emergence of novel carbapenemases. Strains expressing a weak carbapenemase like GPC-1 might go unrecognized by routine diagnostics due to low MICs for the bacterial strains producing such enzymes.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/jac\/dkz536","type":"journal-article","created":{"date-parts":[[2019,12,5]],"date-time":"2019-12-05T12:09:54Z","timestamp":1575547794000},"page":"911-916","source":"Crossref","is-referenced-by-count":15,"title":["GPC-1, a novel class A carbapenemase detected in a clinical Pseudomonas aeruginosa isolate"],"prefix":"10.1093","volume":"75","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5975-0429","authenticated-orcid":false,"given":"Jennifer","family":"Schauer","sequence":"first","affiliation":[{"name":"German National Reference Centre for Multidrug-resistant Gram-negative Bacteria, Department of Medical Microbiology, Ruhr-University Bochum, Universit\u00e4tsstra\u00dfe 150, 44801 Bochum, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00f6ren G","family":"Gatermann","sequence":"first","affiliation":[{"name":"German National Reference Centre for Multidrug-resistant Gram-negative Bacteria, Department of Medical Microbiology, Ruhr-University Bochum, Universit\u00e4tsstra\u00dfe 150, 44801 Bochum, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Hoffmann","sequence":"first","affiliation":[{"name":"SYNLAB MVZ Kassel GmbH, Kurt-Wolters-Stra\u00dfe 2\u20134, 34125 Kassel, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lars","family":"Hupfeld","sequence":"first","affiliation":[{"name":"SYNLAB MVZ Kassel GmbH, Kurt-Wolters-Stra\u00dfe 2\u20134, 34125 Kassel, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6403-5646","authenticated-orcid":false,"given":"Niels","family":"Pfennigwerth","sequence":"first","affiliation":[{"name":"German National Reference Centre for Multidrug-resistant Gram-negative Bacteria, Department of Medical Microbiology, Ruhr-University Bochum, Universit\u00e4tsstra\u00dfe 150, 44801 Bochum, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,1,20]]},"reference":[{"key":"2020031310474049000_dkz536-B1","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1038\/nrmicro3380","article-title":"Molecular mechanisms of antibiotic resistance","volume":"13","author":"Blair","year":"2015","journal-title":"Nat Rev Microbiol"},{"key":"2020031310474049000_dkz536-B2","doi-asserted-by":"crossref","first-page":"2624","DOI":"10.1128\/AAC.43.11.2624","article-title":"Involvement of an active efflux system in the natural resistance of Pseudomonas aeruginosa to aminoglycosides","volume":"43","author":"Aires","year":"1999","journal-title":"Antimicrob Agents Chemother"},{"key":"2020031310474049000_dkz536-B3","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1093\/jac\/47.3.247","article-title":"Of Pseudomonas, porins, pumps and carbapenems","volume":"47","author":"Livermore","year":"2001","journal-title":"J Antimicrob Chemother"},{"key":"2020031310474049000_dkz536-B4","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1128\/AAC.43.2.400","article-title":"Interplay between chromosomal \u03b2-lactamase and the MexAB-OprM efflux system in intrinsic resistance to \u03b2-lactams in Pseudomonas aeruginosa","volume":"43","author":"Masuda","year":"1999","journal-title":"Antimicrob Agents Chemother"},{"key":"2020031310474049000_dkz536-B5","doi-asserted-by":"crossref","first-page":"4062","DOI":"10.1128\/AAC.00148-07","article-title":"Resistance mechanisms of multiresistant Pseudomonas aeruginosa strains from Germany and correlation with hypermutation","volume":"51","author":"Henrichfreise","year":"2007","journal-title":"Antimicrobial Agents Chemother"},{"key":"2020031310474049000_dkz536-B6","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1128\/CMR.8.4.557","article-title":"\u03b2-Lactamases in laboratory and clinical resistance","volume":"8","author":"Livermore","year":"1995","journal-title":"Clin Microbiol Rev"},{"key":"2020031310474049000_dkz536-B7","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1098\/rstb.1980.0049","article-title":"The structure of \u03b2-lactamases","volume":"289","author":"Ambler","year":"1980","journal-title":"Philos Trans R Soc Lond B Biol Sci"},{"key":"2020031310474049000_dkz536-B8","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1517\/13543784.17.2.131","article-title":"Metallo beta lactamases in Pseudomonas aeruginosa and Acinetobacter species","volume":"17","author":"Gupta","year":"2008","journal-title":"Expert Opin Investig Drugs"},{"key":"2020031310474049000_dkz536-B9","doi-asserted-by":"crossref","first-page":"5046","DOI":"10.1128\/AAC.00774-09","article-title":"Characterization of a new metallo-\u03b2-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India","volume":"53","author":"Yong","year":"2009","journal-title":"Antimicrobial Agents Chemother"},{"key":"2020031310474049000_dkz536-B10","first-page":"263","article-title":"Bericht des Nationalen Referenzzentrums (NRZ) f\u00fcr gramnegative Krankenhauserreger","author":"Pfennigwerth","year":"2018","journal-title":"Epidemiol Bull"},{"key":"2020031310474049000_dkz536-B11","year":"2013","edition":"Twenty-Third"},{"key":"2020031310474049000_dkz536-B12","doi-asserted-by":"crossref","first-page":"5648","DOI":"10.1128\/JCM.43.11.5648-5652.2005","article-title":"Sensitive EDTA-based microbiological assays for detection of metallo-\u03b2-lactamases in nonfermentative gram-negative bacteria","volume":"43","author":"Marchiaro","year":"2005","journal-title":"J Clin Microbiol"},{"key":"2020031310474049000_dkz536-B13","doi-asserted-by":"crossref","first-page":"3129","DOI":"10.1128\/JCM.43.7.3129-3135.2005","article-title":"Detection of Pseudomonas aeruginosa producing metallo-\u03b2-lactamases in a large centralized laboratory","volume":"43","author":"Pitout","year":"2005","journal-title":"J Clin Microbiol"},{"key":"2020031310474049000_dkz536-B14","doi-asserted-by":"crossref","first-page":"3846","DOI":"10.1128\/JCM.01299-13","article-title":"A convenient method to screen for carbapenemase-producing Pseudomonas aeruginosa","volume":"51","author":"Fournier","year":"2013","journal-title":"J Clin Microbiol"},{"key":"2020031310474049000_dkz536-B15","year":"2018"},{"key":"2020031310474049000_dkz536-B16","year":"2019"},{"key":"2020031310474049000_dkz536-B17","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1089\/cmb.2012.0021","article-title":"SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing","volume":"19","author":"Bankevich","year":"2012","journal-title":"J Comput Biol"},{"key":"2020031310474049000_dkz536-B18","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1186\/1471-2164-9-75","article-title":"The RAST server: rapid annotations using subsystems technology","volume":"9","author":"Aziz","year":"2008","journal-title":"BMC Genomics"},{"key":"2020031310474049000_dkz536-B19","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/s41587-019-0036-z","article-title":"SignalP 5.0 improves signal peptide predictions using deep neural networks","volume":"37","author":"Almagro Armenteros","year":"2019","journal-title":"Nat Biotechnol"},{"key":"2020031310474049000_dkz536-B20","first-page":"406","article-title":"The neighbor-joining method: a new method for reconstructing phylogenetic trees","volume":"4","author":"Saitou","year":"1987","journal-title":"Mol Biol Evol"},{"key":"2020031310474049000_dkz536-B21","doi-asserted-by":"crossref","first-page":"1596","DOI":"10.1093\/molbev\/msm092","article-title":"MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0","volume":"24","author":"Tamura","year":"2007","journal-title":"Mol Biol Evol"},{"key":"2020031310474049000_dkz536-B22","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/0378-1119(94)90237-2","article-title":"Construction of improved Escherichia-Pseudomonas shuttle vectors derived from pUC18\/19 and sequence of the region required for their replication in Pseudomonas aeruginosa","volume":"148","author":"West","year":"1994","journal-title":"Gene"},{"key":"2020031310474049000_dkz536-B23","doi-asserted-by":"crossref","first-page":"2891","DOI":"10.1093\/jac\/dkz295","article-title":"Genetic and biochemical characterization of FRI-3, a novel variant of the Ambler class A carbapenemase FRI-1","volume":"74","author":"Schauer","year":"2019","journal-title":"J Antimicrob Chemother"},{"key":"2020031310474049000_dkz536-B24","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.marpolbul.2016.06.086","article-title":"Antibiotic and metal resistance in a ST395 Pseudomonas aeruginosa environmental isolate: a genomics approach","volume":"110","author":"Teixeira","year":"2016","journal-title":"Mar Pollut Bull"},{"key":"2020031310474049000_dkz536-B25","doi-asserted-by":"crossref","first-page":"5159","DOI":"10.1128\/AAC.00158-15","article-title":"Characterization of BKC-1 class A carbapenemase from Klebsiella pneumoniae clinical isolates in Brazil","volume":"59","author":"Nicoletti","year":"2015","journal-title":"Antimicrob Agents Chemother"},{"key":"2020031310474049000_dkz536-B26","doi-asserted-by":"crossref","first-page":"2190","DOI":"10.1101\/gr.2482804","article-title":"The Pseudomonas aeruginosa PA01 gene collection","volume":"14","author":"Labaer","year":"2004","journal-title":"Genome Res"},{"key":"2020031310474049000_dkz536-B27","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1093\/jac\/dkm226","article-title":"Class A carbapenemases","volume":"60","author":"Walther-Rasmussen","year":"2007","journal-title":"J Antimicrob Chemother"},{"key":"2020031310474049000_dkz536-B28","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1002\/(SICI)1097-0134(199701)27:1<47::AID-PROT6>3.0.CO;2-K","article-title":"A disulfide bridge near the active site of carbapenem-hydrolyzing class A \u03b2-lactamases might explain their unusual substrate profile","volume":"27","author":"Raquet","year":"1997","journal-title":"Proteins"},{"key":"2020031310474049000_dkz536-B29","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1007\/BF01690097","article-title":"Detection of genes coding for extended-spectrum SHV \u03b2-lactamases in clinical isolates by a molecular genetic method, and comparison with the E-test","volume":"15","author":"Nuesch-Inderbinen","year":"1996","journal-title":"Eur J Clin Microbiol Infect Dis"},{"key":"2020031310474049000_dkz536-B30","doi-asserted-by":"crossref","first-page":"22\u2009196","DOI":"10.1074\/jbc.M116.749648","article-title":"Role of the conserved disulfide bridge in class A carbapenemases","volume":"291","author":"Smith","year":"2016","journal-title":"J Biol Chem"},{"key":"2020031310474049000_dkz536-B31","doi-asserted-by":"crossref","first-page":"1448","DOI":"10.1099\/mic.0.033639-0","article-title":"Acquisition of multidrug resistance transposon Tn6061 and IS6100-mediated large chromosomal inversions in Pseudomonas aeruginosa clinical isolates","volume":"156","author":"Coyne","year":"2010","journal-title":"Microbiology"},{"key":"2020031310474049000_dkz536-B32","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.1093\/jac\/dkq163","article-title":"IS6100-mediated genetic rearrangement within the complex class 1 integron In104 of the Salmonella genomic island 1","volume":"65","author":"Targant","year":"2010","journal-title":"J Antimicrob Chemother"},{"key":"2020031310474049000_dkz536-B33","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1128\/MMBR.00048-05","article-title":"ISCR elements: novel gene-capturing systems of the 21st century?","volume":"70","author":"Toleman","year":"2006","journal-title":"Microbiol Mol Biol Rev"},{"key":"2020031310474049000_dkz536-B34","doi-asserted-by":"crossref","first-page":"1633","DOI":"10.1128\/AAC.50.5.1633-1641.2006","article-title":"Interplay of efflux system, ampC, and oprD expression in carbapenem resistance of Pseudomonas aeruginosa clinical isolates","volume":"50","author":"Quale","year":"2006","journal-title":"Antimicrob Agents Chemother"},{"key":"2020031310474049000_dkz536-B35","doi-asserted-by":"crossref","first-page":"2219","DOI":"10.1128\/AAC.00816-09","article-title":"Efflux pumps, OprD porin, AmpC \u03b2-lactamase, and multiresistance in Pseudomonas aeruginosa isolates from cystic fibrosis patients","volume":"54","author":"Tomas","year":"2010","journal-title":"Antimicrob Agents Chemother"},{"key":"2020031310474049000_dkz536-B36","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1042\/bj2760269","article-title":"A standard numbering scheme for the class A \u03b2-lactamases","volume":"276","author":"Ambler","year":"1991","journal-title":"Biochem J"}],"container-title":["Journal of Antimicrobial Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/jac\/article-pdf\/75\/4\/911\/32903482\/dkz536.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/jac\/article-pdf\/75\/4\/911\/32903482\/dkz536.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,3,13]],"date-time":"2020-03-13T14:48:06Z","timestamp":1584110886000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jac\/article\/75\/4\/911\/5710707"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,1,20]]},"references-count":36,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2020,1,20]]},"published-print":{"date-parts":[[2020,4,1]]}},"URL":"https:\/\/doi.org\/10.1093\/jac\/dkz536","relation":{},"ISSN":["0305-7453","1460-2091"],"issn-type":[{"value":"0305-7453","type":"print"},{"value":"1460-2091","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2020,4]]},"published":{"date-parts":[[2020,1,20]]}}}