{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T07:57:31Z","timestamp":1778054251924,"version":"3.51.4"},"reference-count":43,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2013,3,1]],"date-time":"2013-03-01T00:00:00Z","timestamp":1362096000000},"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":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2013,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A homologue of the\n            <jats:named-content content-type=\"genus-species\">Escherichia coli<\/jats:named-content>\n            penicillin acylase is encoded in the genomes of several thermophiles, including in different\n            <jats:named-content content-type=\"genus-species\">Thermus thermophilus<\/jats:named-content>\n            strains. Although the natural substrate of this enzyme is not known, this acylase shows a marked preference for penicillin K over penicillin G. Three-dimensional models were created in which the catalytic residues and the substrate binding pocket were identified. Through rational redesign, residues were replaced to mimic the aromatic binding site of the\n            <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n            penicillin G acylase. A set of enzyme variants containing between one and four amino acid replacements was generated, with altered catalytic properties in the hydrolyses of penicillins K and G. The introduction of a single phenylalanine residue in position \u03b1188, \u03b1189, or \u03b224 improved the\n            <jats:italic>\n              K\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            for penicillin G between 9- and 12-fold, and the catalytic efficiency of these variants for penicillin G was improved up to 6.6-fold. Structural models, as well as docking analyses, can predict the positioning of penicillins G and K for catalysis and can demonstrate how binding in a productive pose is compromised when more than one bulky phenylalanine residue is introduced into the active site.\n          <\/jats:p>","DOI":"10.1128\/aem.03215-12","type":"journal-article","created":{"date-parts":[[2012,12,22]],"date-time":"2012-12-22T07:03:55Z","timestamp":1356159835000},"page":"1555-1562","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":11,"title":["Engineering the Substrate Specificity of a Thermophilic Penicillin Acylase from Thermus thermophilus"],"prefix":"10.1128","volume":"79","author":[{"given":"Leticia L.","family":"Torres","sequence":"first","affiliation":[{"name":"Center for Molecular Biology Severo Ochoa, Universidad Aut\u00f3noma de Madrid, Consejo Superior de Investigaciones Cient\u00edficas (UAM-CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"\u00c1ngel","family":"Cantero","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology Severo Ochoa, Universidad Aut\u00f3noma de Madrid, Consejo Superior de Investigaciones Cient\u00edficas (UAM-CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mercedes","family":"del Valle","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology Severo Ochoa, Universidad Aut\u00f3noma de Madrid, Consejo Superior de Investigaciones Cient\u00edficas (UAM-CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anabel","family":"Marina","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology Severo Ochoa, Universidad Aut\u00f3noma de Madrid, Consejo Superior de Investigaciones Cient\u00edficas (UAM-CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fernando","family":"L\u00f3pez-Gallego","sequence":"additional","affiliation":[{"name":"Department of Biocatalysis, Institute for Catalysis and Petrochemistry, Consejo Superior de Investigaciones Cient\u00edficas (CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jos\u00e9 M.","family":"Guis\u00e1n","sequence":"additional","affiliation":[{"name":"Department of Biocatalysis, Institute for Catalysis and Petrochemistry, Consejo Superior de Investigaciones Cient\u00edficas (CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jos\u00e9","family":"Berenguer","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology Severo Ochoa, Universidad Aut\u00f3noma de Madrid, Consejo Superior de Investigaciones Cient\u00edficas (UAM-CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aurelio","family":"Hidalgo","sequence":"additional","affiliation":[{"name":"Center for Molecular Biology Severo Ochoa, Universidad Aut\u00f3noma de Madrid, Consejo Superior de Investigaciones Cient\u00edficas (UAM-CSIC), Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2003.03728.x"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-002-1113-6"},{"issue":"10","key":"e_1_3_3_4_2","first-page":"S243","article-title":"New enzymatic protecting group techniques for the construction of peptides and glycopeptides","volume":"50","author":"Waldmann H","year":"1991","unstructured":"WaldmannH BraunP KunzH. 1991. 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