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Cloning and expression of the \u03b2-lactamase gene content of this isolate in<jats:italic>Escherichia coli<\/jats:italic>DH10B identified a chromosome-encoded oxacillinase, OXA-40, that differed by one or two amino acid changes from OXA-24, -25, and -26 and an AmpC-type cephalosporinase. The OXA-40 \u03b2-lactamase had a mainly narrow-spectrum hydrolytic profile, but it included ceftazidime and imipenem. Its activity was resistant to inhibition by clavulanic acid, tazobactam, sulbactam, and, like most of the other carbapenem-hydrolyzing oxacillinases, NaCl. OXA-40 had an FGN triad replacing a YGN motif at class D \u03b2-lactamase (DBL) positions 144 to 146. Site-directed DNA mutagenesis leading to a Phe-to-Tyr change at DBL position 144 in OXA-40 gave a mutant enzyme with increased hydrolytic activity against most \u03b2-lactams, including imipenem. Conversely, with a gene encoding the narrow-spectrum oxacillinase OXA-1 as the template, a nucleotide substitution leading to a Tyr-to-Phe change in the YGN motif of OXA-1 gave a mutant enzyme with decreased hydrolytic activity without an increase in carbapenem-hydrolyzing activity. Thus, the Phe residue in the FGN motif was not associated with carbapenem-hydrolyzing activity by itself but instead was associated with weak overall hydrolytic activity. Finally, this Phe residue in OXA-40 explained resistance to inhibition by NaCl whereas a Tyr residue in motif YGN was related to susceptibility to NaCl.<\/jats:p>","DOI":"10.1128\/aac.47.1.268-273.2003","type":"journal-article","created":{"date-parts":[[2002,12,24]],"date-time":"2002-12-24T04:39:08Z","timestamp":1040704748000},"page":"268-273","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":121,"title":["Genetic and Functional Analysis of the Chromosome-Encoded Carbapenem-Hydrolyzing Oxacillinase OXA-40 of<i>Acinetobacter baumannii<\/i>"],"prefix":"10.1128","volume":"47","author":[{"given":"Claire","family":"He\u0301ritier","sequence":"first","affiliation":[{"name":"Service de Bacte\u0301riologie-Virologie, Ho\u0302pital de Bice\u0302tre, Assistance Publique\/Ho\u0302pitaux de Paris, Faculte\u0301 de Me\u0301decine Paris-Sud, 94275 Le Kremlin-Bice\u0302tre, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laurent","family":"Poirel","sequence":"additional","affiliation":[{"name":"Service de Bacte\u0301riologie-Virologie, Ho\u0302pital de Bice\u0302tre, Assistance Publique\/Ho\u0302pitaux de Paris, Faculte\u0301 de Me\u0301decine Paris-Sud, 94275 Le Kremlin-Bice\u0302tre, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Aubert","sequence":"additional","affiliation":[{"name":"Service de Bacte\u0301riologie-Virologie, Ho\u0302pital de Bice\u0302tre, Assistance Publique\/Ho\u0302pitaux de Paris, Faculte\u0301 de Me\u0301decine Paris-Sud, 94275 Le Kremlin-Bice\u0302tre, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrice","family":"Nordmann","sequence":"additional","affiliation":[{"name":"Service de Bacte\u0301riologie-Virologie, Ho\u0302pital de Bice\u0302tre, Assistance Publique\/Ho\u0302pitaux de Paris, Faculte\u0301 de Me\u0301decine Paris-Sud, 94275 Le Kremlin-Bice\u0302tre, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/41.5.576"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/43.1.127"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.2.583-588.2001"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.6.1615-1620.2001"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.8.1924-1930.1997"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.9.2.148"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.46.6.2004-2006.2002"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.9.3299-3305.2000"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.2.428-432.2000"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.6.1556-1561.2000"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.6.1211"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.3.710-714.2001"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/38.2.245"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1992.tb00894.x"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.1.196-199.2000"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1159\/000238887"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj2920555"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.14.6874"},{"key":"e_1_3_2_20_2","doi-asserted-by":"crossref","first-page":"865","DOI":"10.2174\/1381612805666230112185101","volume":"5","year":"1999","unstructured":"Naas, T., and P. 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