{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T23:20:02Z","timestamp":1787008802391,"version":"build-2736575974"},"reference-count":38,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2025,1,1]],"date-time":"2025-01-01T00:00:00Z","timestamp":1735689600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100001407","name":"Department of Biotechnology, GOI","doi-asserted-by":"publisher","award":["BT\/PR40383\/BCE\/8\/1561\/2020"],"award-info":[{"award-number":["BT\/PR40383\/BCE\/8\/1561\/2020"]}],"id":[{"id":"10.13039\/501100001407","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Albert David Limited, India","award":["#TN: AP: 204"],"award-info":[{"award-number":["#TN: AP: 204"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,1,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Verona-integron-metallo-\u03b2-lactamase (VIM-2) is one of the most widespread class B \u03b2-lactamase responsible for \u03b2-lactam resistance. Although active-site residues help in metal binding, the residues nearing the active-site possess functional importance. Here, to decipher the role of such residues in the activity and stability of VIM-2, the residues E146, D182, N210, S207, and D213 were selected through in-silico analyses and substituted with alanine using site-directed mutagenesis. The effects of substitution mutations were assessed by comparing the changes in \u03b2-lactam susceptibility pattern of Escherichia coli host cell expressing VIM-2 and its mutated proteins. VIM-2_N210A enhanced the susceptibility of the host by \u223c4\u20138 folds against penicillins and cephalosporins, while the expression of VIM-2_D182A radically increased the susceptibility of host. However, expression of VIM-2_E146A reduced the susceptibility of host by 2-fold. Further, proteins were purified to homogeneity, and VIM_N210A and VIM_D182A displayed reduced thermal stability than VIM-2. Moreover, in vitro catalytic efficiencies of VIM-2_D182A were drastically reduced against all the \u03b2-lactams tested whereas the same were moderately reduced for VIM-2_N210A. Conversely, the catalytic efficiency was marginally altered for VIM_E146A. Overall, we infer that both N210A and D182A substitutions negatively affect the performance of VIM-2 by influencing substrate specificity and stability, respectively.<\/jats:p>","DOI":"10.1093\/femsle\/fnaf007","type":"journal-article","created":{"date-parts":[[2025,1,10]],"date-time":"2025-01-10T07:19:10Z","timestamp":1736493550000},"source":"Crossref","is-referenced-by-count":2,"title":["Conserved ancillary residues situated proximally to the VIM-2 active-site affect its metallo\n                    <i>\u03b2<\/i>\n                    -lactamase activity"],"prefix":"10.1093","volume":"372","author":[{"given":"Diamond","family":"Jain","sequence":"first","affiliation":[{"name":"Department of Bioscience and Biotechnology, Indian Institute of Technology Kharagpur , Kharagpur 721302 ,","place":["India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tejavath","family":"Ajith","sequence":"additional","affiliation":[{"name":"Department of Bioscience and Biotechnology, Indian Institute of Technology Kharagpur , Kharagpur 721302 ,","place":["India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jyoti","family":"Verma","sequence":"additional","affiliation":[{"name":"Advanced Technology Development Centre, Indian Institute of Technology Kharagpur , Kharagpur 721302 ,","place":["India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Debasmita","family":"Chatterjee","sequence":"additional","affiliation":[{"name":"Department of Bioscience and Biotechnology, Indian Institute of Technology Kharagpur , Kharagpur 721302 ,","place":["India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6403-032X","authenticated-orcid":false,"given":"Anindya S","family":"Ghosh","sequence":"additional","affiliation":[{"name":"Department of Bioscience and Biotechnology, Indian Institute of Technology Kharagpur , Kharagpur 721302 ,","place":["India"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,1,11]]},"reference":[{"key":"2025091923042293200_bib1","doi-asserted-by":"crossref","first-page":"fnz003","DOI":"10.1093\/femsle\/fnz003","article-title":"Role of non-active-site residues in maintaining New Delhi metallo-\u03b2-lactamase-1 (NDM-1) function: an approach of site-directed mutagenesis and docking","volume":"368","author":"Ali","year":"2021","journal-title":"FEMS Microbiol Lett"},{"key":"2025091923042293200_bib2","doi-asserted-by":"crossref","first-page":"7957","DOI":"10.1021\/acs.chemrev.1c00138","article-title":"Metallo-beta-lactamases in the age of multidrug resistance: from structure and mechanism to evolution, dissemination, and inhibitor design","volume":"121","author":"Bahr","year":"2021","journal-title":"Chem 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