{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T12:51:58Z","timestamp":1761310318865,"version":"build-2065373602"},"reference-count":37,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2006,2,7]],"date-time":"2006-02-07T00:00:00Z","timestamp":1139270400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2006,3,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The activity of the aminopeptidase P from <jats:italic>Escherichia coli<\/jats:italic> in hydrolyzing a series of organophosphonate sarin analogues (<jats:bold>1<\/jats:bold>\u2013<jats:bold>6<\/jats:bold>) was evaluated. The enzymatic rates of hydrolysis for methylphosphonate <jats:bold>1<\/jats:bold> with a methoxy group attached to the phosphorus center were 7\u2010 to 15\u2010fold higher than those for the corresponding analogues <jats:bold>2<\/jats:bold>\u2013<jats:bold>6<\/jats:bold>. Double mutant R153W\/R370L was able to hydrolyze the <jats:italic>S<\/jats:italic><jats:sub>p<\/jats:sub> enantiomer of racemic <jats:bold>1<\/jats:bold> at a considerable rate. This mutant allowed the preparation of the <jats:italic>R<\/jats:italic><jats:sub>p<\/jats:sub> isomer of the sarin analogue <jats:bold>1<\/jats:bold>. All the mutants, R370L, R153A, W88L, R153L\/R370L, and R153W\/R370L, preferred the formation of (<jats:italic>S<\/jats:italic><jats:sub>p<\/jats:sub>)\u2010<jats:bold>8<\/jats:bold> to that of the corresponding (<jats:italic>R<\/jats:italic><jats:sub>p<\/jats:sub>)\u2010<jats:bold>8<\/jats:bold> enantiomer and displayed a better enantiomeric excess of products, by 1.4\u2010 to 2\u2010fold as compared to the wild\u2010type enzyme. Enzymatic hydrolysis of <jats:italic>O,O<\/jats:italic>\u2010diisopropyl\u2010<jats:italic>p<\/jats:italic>\u2010nitrophenyl phosphate (<jats:bold>9<\/jats:bold>) in H<jats:sub>2<\/jats:sub><jats:sup>18<\/jats:sup>O led to the formation of the <jats:sup>18<\/jats:sup>O\u2010labeled <jats:italic>O,O<\/jats:italic>\u2010diisopropyl phosphate product and confirmed that the catalytic reaction starts with cleavage of the P\uf8ffO bond. From chemical and kinetic studies, the utilization of an optically pure <jats:italic>S<\/jats:italic><jats:sub>p<\/jats:sub> enantiomer of <jats:italic>O<\/jats:italic>\u2010methyl\u2010p\u2010nitrophenyl methylphosphonothioate ((<jats:italic>S<\/jats:italic><jats:sub>p<\/jats:sub>)\u2010MNMPT, <jats:bold>7<\/jats:bold>) has demonstrated that the enzymatic reaction proceeds through a displacement mechanism and generates a chiral product in situ with an inversion of stereochemical configuration at the phosphorus atom. The results also lead to the conclusion that alteration of the active site through site\u2010directed mutagenesis can result in a preference for (<jats:italic>S<\/jats:italic><jats:sub>p<\/jats:sub>)\u2010MNMPT (<jats:bold>7<\/jats:bold>) rather than the <jats:italic>R<\/jats:italic><jats:sub>p<\/jats:sub> isomer.<\/jats:p>","DOI":"10.1002\/cbic.200500412","type":"journal-article","created":{"date-parts":[[2006,2,9]],"date-time":"2006-02-09T17:56:59Z","timestamp":1139507819000},"page":"506-514","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Aminopeptidase P Mediated Detoxification of Organophosphonate Analogues of Sarin: Mechanistic and Stereochemical Study at the Phosphorus Atom of the Substrate"],"prefix":"10.1002","volume":"7","author":[{"given":"Li\u2010Fang","family":"Huang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benson","family":"Su","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shu\u2010Chuan","family":"Jao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kwang\u2010Ting","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wen\u2010Shan","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2006,3]]},"reference":[{"key":"e_1_2_6_1_2","first-page":"655","volume-title":"Casarett and Doull's Toxicology: The Basic Science of Poisons","author":"Ecobichon D. 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