{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T02:04:28Z","timestamp":1775873068808,"version":"3.50.1"},"reference-count":57,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2014,2,19]],"date-time":"2014-02-19T00:00:00Z","timestamp":1392768000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Angew Chem Int Ed"],"published-print":{"date-parts":[[2014,3,17]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Terminal\u2010selective cytochrome P450pyr has been successfully engineered through directed evolution for the subterminal hydroxylation of alkanes with excellent regio\u2010 and enantioselectivity. A sensitive colorimetric high\u2010throughput screening (HTS) assay was developed for the measurement of both the regioselectivity and the enantioselectivity of a hydroxylation reaction. By using the HTS assay and iterative saturation mutagenesis, sextuple\u2010mutant P450pyrSM1 was created for the hydroxylation of <jats:italic>n<\/jats:italic>\u2010octane (<jats:bold>1<\/jats:bold>) to give (<jats:italic>S<\/jats:italic>)\u20102\u2010octanol (<jats:bold>2<\/jats:bold>) with 98\u2009%\u2005<jats:italic>ee<\/jats:italic> and &gt;99\u2009% subterminal selectivity. The engineered P450 is the first enzyme for this type of highly selective alkane hydroxylation, being useful for the C\uf8ffH activation and functionalization of alkanes and the preparation of enantiopure alcohols. Molecular modeling provided structure\u2010based understanding of the fully altered regioselectivity and the excellent enantioselectivity. Another sextuple\u2010mutant P450pyrSM2 catalyzed the hydroxylation of propylbenzene (<jats:bold>3<\/jats:bold>) to afford (<jats:italic>S<\/jats:italic>)\u20101\u2010phenyl\u20102\u2010propanol (<jats:bold>4<\/jats:bold>) with 95\u2009%\u2005<jats:italic>ee<\/jats:italic> and 98\u2009% subterminal selectivity.<\/jats:p>","DOI":"10.1002\/anie.201311091","type":"journal-article","created":{"date-parts":[[2014,2,20]],"date-time":"2014-02-20T03:12:05Z","timestamp":1392865925000},"page":"3120-3124","source":"Crossref","is-referenced-by-count":85,"title":["Engineering of P450pyr Hydroxylase for the Highly Regio\u2010 and Enantioselective Subterminal Hydroxylation of Alkanes"],"prefix":"10.1002","volume":"53","author":[{"given":"Yi","family":"Yang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ji","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhi","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2014,2,19]]},"reference":[{"key":"e_1_2_2_1_2","unstructured":"\u00a0"},{"key":"e_1_2_2_2_2","doi-asserted-by":"publisher","DOI":"10.1039\/C0CS00067A"},{"key":"e_1_2_2_3_2","doi-asserted-by":"publisher","DOI":"10.2174\/1385272043369584"},{"key":"e_1_2_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2005.02.007"},{"key":"e_1_2_2_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.648"},{"key":"e_1_2_2_6_2","unstructured":"\u00a0"},{"key":"e_1_2_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1367-5931(02)00296-X"},{"key":"e_1_2_2_8_2","doi-asserted-by":"publisher","DOI":"10.1039\/C1CS15192D"},{"key":"e_1_2_2_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/1521-3757(20010803)113:15<2860::AID-ANGE2860>3.0.CO;2-2"},{"key":"e_1_2_2_9_3","doi-asserted-by":"publisher","DOI":"10.1002\/1521-3773(20010803)40:15<2782::AID-ANIE2782>3.0.CO;2-P"},{"key":"e_1_2_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01758-08"},{"key":"e_1_2_2_11_2","unstructured":"\u00a0"},{"key":"e_1_2_2_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/jo901046k"},{"key":"e_1_2_2_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature11117"},{"key":"e_1_2_2_14_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201000826"},{"key":"e_1_2_2_14_3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201000826"},{"key":"e_1_2_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1367-5931(02)00305-8"},{"key":"e_1_2_2_16_2","unstructured":"\u00a0"},{"key":"e_1_2_2_17_2","doi-asserted-by":"publisher","DOI":"10.1039\/b718124h"},{"key":"e_1_2_2_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201303986"},{"key":"e_1_2_2_18_3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201303986"},{"key":"e_1_2_2_19_2","unstructured":"\u00a0"},{"key":"e_1_2_2_20_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200500266"},{"key":"e_1_2_2_21_2","doi-asserted-by":"publisher","DOI":"10.1002\/adsc.201200067"},{"key":"e_1_2_2_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.1113"},{"key":"e_1_2_2_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja3073462"},{"key":"e_1_2_2_24_2","unstructured":"\u00a0"},{"key":"e_1_2_2_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/ol006735q"},{"key":"e_1_2_2_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/jo015826d"},{"key":"e_1_2_2_27_2","doi-asserted-by":"publisher","DOI":"10.1021\/ol025829s"},{"key":"e_1_2_2_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0957-4166(02)00534-7"},{"key":"e_1_2_2_29_2","doi-asserted-by":"publisher","DOI":"10.1002\/adsc.201000266"},{"key":"e_1_2_2_30_2","unstructured":"\u00a0"},{"key":"e_1_2_2_31_2","doi-asserted-by":"publisher","DOI":"10.1039\/c0cc00735h"},{"key":"e_1_2_2_32_2","doi-asserted-by":"publisher","DOI":"10.1039\/c2cc30779k"},{"key":"e_1_2_2_33_2","unstructured":"\u00a0"},{"key":"e_1_2_2_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2010.01.010"},{"key":"e_1_2_2_35_2","doi-asserted-by":"publisher","DOI":"10.1039\/c1ob05565h"},{"key":"e_1_2_2_36_2","doi-asserted-by":"publisher","DOI":"10.1002\/1615-4169(200108)343:6\/7<601::AID-ADSC601>3.0.CO;2-9"},{"key":"e_1_2_2_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00280-06"},{"key":"e_1_2_2_38_2","unstructured":"\u00a0"},{"key":"e_1_2_2_39_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200500261"},{"key":"e_1_2_2_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt744"},{"key":"e_1_2_2_41_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja0303790"},{"key":"e_1_2_2_42_2","doi-asserted-by":"publisher","DOI":"10.1039\/b110957j"},{"key":"e_1_2_2_43_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1381-1177(97)00025-8"},{"key":"e_1_2_2_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2012.11.048"},{"key":"e_1_2_2_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0957-4166(03)00438-5"},{"key":"e_1_2_2_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tet.2007.07.012"},{"key":"e_1_2_2_47_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2005.05.091"},{"key":"e_1_2_2_48_2","unstructured":"\u00a0"},{"key":"e_1_2_2_49_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.200353055"},{"key":"e_1_2_2_49_3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200353055"},{"key":"e_1_2_2_50_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201001772"},{"key":"e_1_2_2_50_3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201001772"},{"key":"e_1_2_2_51_2","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.63.1051"},{"key":"e_1_2_2_52_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-012-4643-6"}],"container-title":["Angewandte Chemie International Edition"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fanie.201311091","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/anie.201311091","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,16]],"date-time":"2023-10-16T10:01:19Z","timestamp":1697450479000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/anie.201311091"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,2,19]]},"references-count":57,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2014,3,17]]}},"alternative-id":["10.1002\/anie.201311091"],"URL":"https:\/\/doi.org\/10.1002\/anie.201311091","archive":["Portico"],"relation":{},"ISSN":["1433-7851","1521-3773"],"issn-type":[{"value":"1433-7851","type":"print"},{"value":"1521-3773","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,2,19]]}}}