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The R214 residue, located in the \u00df5-\u00df6 loop, is crucial for the carbapenemase activity, as it stabilizes carbapenems in the active site and maintains the shape of the active site through interactions with D159. In this study, we have characterized a novel variant of OXA-48, OXA-933 with a single D159N change. To evaluate the importance of this residue, point mutations were generated (D159A, D159G, D159K, and D159W), kinetic parameters of OXA-933, OXA-48 D159G, and OXA-48 D159K were determined and compared to those of OXA-48 and OXA-244. The\n                      <jats:italic>bla<\/jats:italic>\n                      <jats:sub>OXA-933<\/jats:sub>\n                      gene was borne on Tn\n                      <jats:italic>2208<\/jats:italic>\n                      , a 2,696-bp composite transposon made of two IS\n                      <jats:italic>1<\/jats:italic>\n                      elements surrounded by 9 bp target site duplications and inserted into a non-self-transmissible plasmid pOXA-933 of 7,872 bp in size. Minimal inhibitory concentration values of\n                      <jats:italic>E. coli<\/jats:italic>\n                      expressing the\n                      <jats:italic>bla<\/jats:italic>\n                      <jats:sub>OXA-933<\/jats:sub>\n                      gene or of its point mutant derivatives were lower for carbapenems (except for D159G) as compared to those expressing the\n                      <jats:italic>bla<\/jats:italic>\n                      <jats:sub>OXA-48<\/jats:sub>\n                      gene. Steady-state kinetic parameters revealed lower catalytic efficiencies for expanded spectrum cephalosporins and carbapenems. A detailed structural analysis confirmed the crucial role of D159 in shaping the active site of OXA-48 enzymes by interacting with R214. Our work further illustrates the remarkable propensity of OXA-48-like carbapenemases to evolve through mutations at positions outside the \u03b25-\u03b26 loop, but interacting with key residues of it.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aac.00180-24","type":"journal-article","created":{"date-parts":[[2024,3,25]],"date-time":"2024-03-25T08:45:39Z","timestamp":1711356339000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":4,"title":["Role of amino acid 159 in carbapenem and temocillin hydrolysis of OXA-933, a novel OXA-48 variant"],"prefix":"10.1128","volume":"68","author":[{"given":"Mariam","family":"Rima","sequence":"first","affiliation":[{"name":"Team ReSIST, INSERM U1184, School of Medicine Universit\u00e9 Paris-Saclay, LabEx LERMIT, Le Kremlin-Bic\u00eatre, 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