{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T13:07:19Z","timestamp":1780578439947,"version":"3.54.1"},"reference-count":30,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2013,2,28]],"date-time":"2013-02-28T00:00:00Z","timestamp":1362009600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2013,4]]},"abstract":"<jats:sec><jats:label\/><jats:p>The salicylate 1,2\u2010dioxygenase (<jats:styled-content style=\"fixed-case\">SDO<\/jats:styled-content>) from the bacterium <jats:italic><jats:styled-content style=\"fixed-case\">P<\/jats:styled-content>seudaminobacter salicylatoxidans <\/jats:italic><jats:styled-content style=\"fixed-case\">BN<\/jats:styled-content>12 is a versatile gentisate 1,2\u2010dioxygenase (<jats:styled-content style=\"fixed-case\">GDO<\/jats:styled-content>) that converts both gentisate (2,5\u2010dihydroxybenzoate) and various monohydroxylated substrates. Several variants of this enzyme were rationally designed based on the previously determined enzyme structure and sequence differences between the <jats:styled-content style=\"fixed-case\">SDO<\/jats:styled-content> and the \u2018conventional\u2019 <jats:styled-content style=\"fixed-case\">GDO<\/jats:styled-content> from <jats:italic>Corynebacterium\u00a0glutamicum<\/jats:italic>. This was undertaken in order to define the structural elements that give the SDO its unique ability to dioxygenolytically cleave (substituted) salicylates. <jats:styled-content style=\"fixed-case\">SDO<\/jats:styled-content> variants <jats:styled-content style=\"fixed-case\">M<\/jats:styled-content>103<jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> G<\/jats:styled-content>106<jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> G<\/jats:styled-content>111<jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> R<\/jats:styled-content>113<jats:styled-content style=\"fixed-case\">G<\/jats:styled-content>,<jats:styled-content style=\"fixed-case\"> S<\/jats:styled-content>147<jats:styled-content style=\"fixed-case\">R<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">F<\/jats:styled-content>159<jats:styled-content style=\"fixed-case\">Y<\/jats:styled-content> were constructed and it was found that <jats:styled-content style=\"fixed-case\">G<\/jats:styled-content>106<jats:styled-content style=\"fixed-case\">A<\/jats:styled-content> oxidized only gentisate; 1\u2010hydroxy\u20102\u2010naphthoate and salicylate were not converted. This indicated that this enzyme variant behaves like previously known \u2018conventional\u2019 <jats:styled-content style=\"fixed-case\">GDO<\/jats:styled-content>s. Crystals of the <jats:styled-content style=\"fixed-case\">G<\/jats:styled-content>106<jats:styled-content style=\"fixed-case\">A SDO<\/jats:styled-content> variant and its complexes with salicylate and <jats:italic>gentisate<\/jats:italic> were obtained under anaerobic conditions, and the structures were solved and analyzed. The amino acid residue Gly106 is located inside the SDO active site cavity but does not directly interact with the substrates. Crystal structures of <jats:styled-content style=\"fixed-case\">G<\/jats:styled-content>106<jats:styled-content style=\"fixed-case\">A SDO<\/jats:styled-content> complexes with gentisate and salicylate showed a different binding mode for salicylate when compared with the wild\u2010type enzyme. Thus, salicylate coordinated in the <jats:styled-content style=\"fixed-case\">G<\/jats:styled-content>106<jats:styled-content style=\"fixed-case\">A<\/jats:styled-content> variant with the catalytically active Fe(II) ion in an unusual and unproductive manner because of the inability of salicylate to displace a hydrogen bond that was formed between <jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>rp104 and <jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>sp174 in the <jats:styled-content style=\"fixed-case\">G<\/jats:styled-content>106<jats:styled-content style=\"fixed-case\">A<\/jats:styled-content> variant. It is proposed that this type of unproductive substrate binding might generally limit the substrate spectrum of \u2018conventional\u2019 <jats:styled-content style=\"fixed-case\">GDO<\/jats:styled-content>s.<\/jats:p><\/jats:sec><jats:sec><jats:title>Database<\/jats:title><jats:p>Structural data are available in the Protein Data Bank databases under the accession numbers <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=3NST\">3NST<\/jats:ext-link>, <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=3NWA\">3NWA<\/jats:ext-link>, <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=3NVC\">3NVC<\/jats:ext-link><\/jats:p><\/jats:sec>","DOI":"10.1111\/febs.12173","type":"journal-article","created":{"date-parts":[[2013,2,6]],"date-time":"2013-02-06T09:34:26Z","timestamp":1360143266000},"page":"1643-1652","source":"Crossref","is-referenced-by-count":32,"title":["The salicylate 1,2\u2010dioxygenase as a model for a conventional gentisate 1,2\u2010dioxygenase: crystal structures of the G106A mutant and its adducts with gentisate and salicylate"],"prefix":"10.1111","volume":"280","author":[{"given":"Marta","family":"Ferraroni","sequence":"first","affiliation":[{"name":"Dipartimento di Chimica \u2018Ugo Schiff\u2019 Universit\u00e0 di Firenze Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene","family":"Matera","sequence":"additional","affiliation":[{"name":"Dipartimento di Chimica \u2018Ugo Schiff\u2019 Universit\u00e0 di Firenze Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sibylle","family":"B\u00fcrger","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Mikrobiologie Universit\u00e4t Stuttgart Stuttgart Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Reichert","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Mikrobiologie Universit\u00e4t Stuttgart Stuttgart Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lenz","family":"Steimer","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Mikrobiologie Universit\u00e4t Stuttgart Stuttgart Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Scozzafava","sequence":"additional","affiliation":[{"name":"Dipartimento di Chimica \u2018Ugo Schiff\u2019 Universit\u00e0 di Firenze Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Stolz","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Mikrobiologie Universit\u00e4t Stuttgart Stuttgart Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabrizio","family":"Briganti","sequence":"additional","affiliation":[{"name":"Dipartimento di Chimica \u2018Ugo Schiff\u2019 Universit\u00e0 di Firenze Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2013,2,28]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1021\/cr960039f"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1080\/10409230600817422"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.181.3.757-763.1999"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M313500200"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02816-09"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01188-10"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2011.11.026"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.phytochem.2003.08.016"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.23.6936-6942.2001"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2008.05.041"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1107\/S1744309106016435"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2006.05334.x"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1110\/ps.035881.108"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.60444"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10529-007-9421-7"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.14.8332"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0607048103"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-010-3033-1"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.71.7.3442-3452.2005"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1119(03)00619-X"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.07137-11"},{"key":"e_1_2_7_23_1","first-page":"203","article-title":"The generation of a 1\u2010hydroxy\u20102\u2010naphthoate 1,2\u2010dioxygenase by single point mutations of salicylate 1,2\u2010dioxygenase \u2013 rational design of mutants and the crystal structures of the A85H and W104Y variants","volume":"111","author":"Ferraroni M","year":"2012","journal-title":"J Struct Biol"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)45688-8"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja00383a012"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1995.0492"},{"key":"e_1_2_7_27_1","doi-asserted-by":"crossref","unstructured":"LeslieAGW&PowellHR(2007)Processing diffraction data with mosflm. InEvolving Methods for Macromolecular Crystallography(ReadRJ&SussmanJL) pp.41\u201351 Springer The Netherlands.","DOI":"10.1007\/978-1-4020-6316-9_4"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444994003112"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2008.91"},{"key":"e_1_2_7_31_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.10286"}],"container-title":["The FEBS Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ffebs.12173","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/febs.12173","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,30]],"date-time":"2023-10-30T18:28:15Z","timestamp":1698690495000},"score":1,"resource":{"primary":{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/10.1111\/febs.12173"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,2,28]]},"references-count":30,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2013,4]]}},"alternative-id":["10.1111\/febs.12173"],"URL":"https:\/\/doi.org\/10.1111\/febs.12173","archive":["Portico"],"relation":{},"ISSN":["1742-464X","1742-4658"],"issn-type":[{"value":"1742-464X","type":"print"},{"value":"1742-4658","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,2,28]]}}}