{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T12:53:11Z","timestamp":1762865591978},"reference-count":22,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2010,4,1]],"date-time":"2010-04-01T00:00:00Z","timestamp":1270080000000},"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":[[2010,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Directed evolution by random PCR mutagenesis of the gene for the aminoglycoside 2\u2033-IIa phosphotransferase generated R92H\/D268N and N196D\/D268N mutant enzymes, resulting in elevated levels of resistance to amikacin and isepamicin but not to other aminoglycoside antibiotics. Increases in the activities of the mutant phosphotransferases for isepamicin are the result of decreases in\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            values, while improved catalytic efficiency for amikacin is the result of both a decrease in\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            values and an increase in turnover of the antibiotic. Enzymes with R92H, D268N, and D268N single amino acid substitutions did not result in elevated MICs for aminoglycosides.\n          <\/jats:p>","DOI":"10.1128\/aac.01444-09","type":"journal-article","created":{"date-parts":[[2010,2,10]],"date-time":"2010-02-10T04:54:38Z","timestamp":1265777678000},"page":"1590-1595","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":12,"title":["Mutant APH(2\u2033)-IIa Enzymes with Increased Activity against Amikacin and Isepamicin"],"prefix":"10.1128","volume":"54","author":[{"given":"Marta","family":"Toth","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hilary","family":"Frase","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph W.","family":"Chow","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases Clinical Research, Merck Research Laboratories, North Wales, Pennsylvania 19454"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Clyde","family":"Smith","sequence":"additional","affiliation":[{"name":"Stanford Synchrotron Radiation Laboratory, Stanford University, Menlo Park, California 94025"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sergei B.","family":"Vakulenko","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 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