{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T13:01:30Z","timestamp":1762866090755,"version":"build-2065373602"},"reference-count":27,"publisher":"Wiley","issue":"16","license":[{"start":{"date-parts":[[2012,11,4]],"date-time":"2012-11-04T00:00:00Z","timestamp":1351987200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Adv Synth Catal"],"published-print":{"date-parts":[[2012,11,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The <jats:italic>Escherichia coli<\/jats:italic> esterase YbfF displays high activity towards 1,2\u2010<jats:italic>O<\/jats:italic>\u2010isopropylideneglycerol (IPG) butyrate and IPG caprylate, and prefers the <jats:italic>R<\/jats:italic>\u2010enantiomer of these substrates, producing the <jats:italic>S<\/jats:italic>\u2010enantiomer of the IPG product in excess. To improve the potential of the enzyme for the kinetic resolution of racemic esters of IPG, an enhancement of the activity and enantioselectivity would be highly desirable. Molecular docking of the <jats:italic>R<\/jats:italic>\u2010enantiomer of both IPG esters into the active site of YbfF allowed the identification of proximal YbfF active site residues. Four residues (25, 124, 185 and 235) were selected as targets for mutagenesis, in order to enhance YbfF activity and enantioselectivity towards IPG esters. Random mutagenesis at positions 25, 124, 185 and 235 yielded several best YbfF variants with enhanced activity and enantioselectivity towards IPG esters. The best YbfF mutant, W235I, exhibited a 2\u2010fold higher enantioselectivity than wild\u2010type YbfF, with an <jats:italic>E<\/jats:italic>=38 for IPG butyrate and an <jats:italic>E<\/jats:italic>=77 for IPG caprylate. Molecular docking experiments further support the enhanced enantioselectivity shown experimentally and the structural effects of this amino acid substitution on the active site of YbfF are provided. The engineered W235I mutant is an attractive catalyst for practical applications in the kinetic resolution of IPG esters.<\/jats:p>","DOI":"10.1002\/adsc.201200211","type":"journal-article","created":{"date-parts":[[2012,11,4]],"date-time":"2012-11-04T19:13:12Z","timestamp":1352056392000},"page":"3009-3015","source":"Crossref","is-referenced-by-count":15,"title":["An Esterase with Superior Activity and Enantioselectivity towards 1,2\u2010<i>O<\/i>\u2010Isopropylideneglycerol Esters Obtained by Protein Design"],"prefix":"10.1002","volume":"354","author":[{"given":"Luis F.","family":"Godinho","sequence":"first","affiliation":[]},{"given":"Carlos R.","family":"Reis","sequence":"additional","affiliation":[]},{"given":"Ronald","family":"van Merkerk","sequence":"additional","affiliation":[]},{"given":"Gerrit J.","family":"Poelarends","sequence":"additional","affiliation":[]},{"given":"Wim J.","family":"Quax","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2012,11,4]]},"reference":[{"key":"e_1_2_6_1_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.2002.tb00599.x"},{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2010.01.010"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0040-4020(01)87445-7"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01024395"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.05122-11"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2007.12.062"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.17.9813"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/35051731"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200700754"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2007.05782.x"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2009.10.001"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1039\/b610461d"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1521-3765(19981102)4:11<2324::AID-CHEM2324>3.0.CO;2-I"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200800298"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0141-0229(96)00066-X"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tetasy.2004.12.021"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.72"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201000826"},{"key":"e_1_2_6_18_3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201000826"},{"volume-title":"Molecular cloning: a laboratory manual","year":"2001","author":"Sambrook J.","key":"e_1_2_6_19_2"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.2144\/02335st03"},{"key":"e_1_2_6_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2005.01.016"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.200301509"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja00389a064"},{"key":"e_1_2_6_24_2","doi-asserted-by":"publisher","DOI":"10.1021\/jm030209y"},{"key":"e_1_2_6_25_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.10306"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioeng.2005.02.002"}],"container-title":["Advanced Synthesis &amp; Catalysis"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fadsc.201200211","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/advanced.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/adsc.201200211","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,6]],"date-time":"2025-10-06T13:45:05Z","timestamp":1759758305000},"score":1,"resource":{"primary":{"URL":"https:\/\/advanced.onlinelibrary.wiley.com\/doi\/10.1002\/adsc.201200211"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,11,4]]},"references-count":27,"journal-issue":{"issue":"16","published-print":{"date-parts":[[2012,11,12]]}},"alternative-id":["10.1002\/adsc.201200211"],"URL":"https:\/\/doi.org\/10.1002\/adsc.201200211","archive":["Portico"],"relation":{},"ISSN":["1615-4150","1615-4169"],"issn-type":[{"type":"print","value":"1615-4150"},{"type":"electronic","value":"1615-4169"}],"subject":[],"published":{"date-parts":[[2012,11,4]]}}}