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dc.contributor.authorChang, Cheng-Hsiangen_US
dc.contributor.authorWen, Hao-Yuen_US
dc.contributor.authorShie, Wen-Shiangen_US
dc.contributor.authorLu, Ching-Tingen_US
dc.contributor.authorLi, Meng-Erhen_US
dc.contributor.authorLiu, Yuan-Tingen_US
dc.contributor.authorLi, Wen-Hsuanen_US
dc.contributor.authorWu, Tung-Kungen_US
dc.date.accessioned2014-12-08T15:31:44Z-
dc.date.available2014-12-08T15:31:44Z-
dc.date.issued2013en_US
dc.identifier.issn1477-0520en_US
dc.identifier.urihttp://hdl.handle.net/11536/22457-
dc.identifier.urihttp://dx.doi.org/10.1039/c3ob40493een_US
dc.description.abstractA computational modeling/protein engineering approach was applied to probe H234, C457, T509, Y510, and W587 within Saccharomyces cerevisiae oxidosqualene-lanosterol cyclase (ERG7), which spatially affects the C-10 cation of lanosterol formation. Substitution of Trp587 to aromatic residues supported the "aromatic hypothesis" that the pi-electron-rich pocket is important for the stabilization of electron-deficient cationic intermediates. The Cys457 to Gly and Thr509 to Gly mutations disrupted the pre-existing H-bond to the protonating Asp456 and the intrinsic His234 : Tyr510 H-bond network, respectively, and generated achilleol A as the major product. An H234W/Y510W double mutation altered the ERG7 function to achilleol A synthase activity and generated achilleol A as the sole product. These results support the concept that a few-ring triterpene synthase can be derived from polycyclic cyclases by reverse evolution, and exemplify the power of computational modeling coupled with protein engineering both to study the enzyme's structure-function-mechanism relationships and to evolve new enzymatic activity.en_US
dc.language.isoen_USen_US
dc.titleProtein engineering of oxidosqualene-lanosterol cyclase into triterpene monocyclaseen_US
dc.typeArticleen_US
dc.identifier.doi10.1039/c3ob40493een_US
dc.identifier.journalORGANIC & BIOMOLECULAR CHEMISTRYen_US
dc.citation.volume11en_US
dc.citation.issue25en_US
dc.citation.spage4214en_US
dc.citation.epage4219en_US
dc.contributor.department生物科技學系zh_TW
dc.contributor.departmentDepartment of Biological Science and Technologyen_US
dc.identifier.wosnumberWOS:000319877900012-
dc.citation.woscount3-
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