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dc.contributor.authorBracco, Paula
dc.contributor.authorWijma, Hein J
dc.contributor.authorNicolai, Bastian
dc.contributor.authorRodriguez Buitrago, Jhon Alexander
dc.contributor.authorKlünemann, Thomas
dc.contributor.authorVila, Agustina
dc.contributor.authorSchrepfer, Patrick
dc.contributor.authorBlankenfeldt, Wulf
dc.contributor.authorJanssen, Dick B
dc.contributor.authorSchallmey, Anett
dc.date.accessioned2020-11-26T09:36:36Z
dc.date.available2020-11-26T09:36:36Z
dc.date.issued2020-11-04
dc.identifier.citationChembiochem. 2020 Nov 4. doi: 10.1002/cbic.202000735. Epub ahead of print.en_US
dc.identifier.pmid33145893
dc.identifier.doi10.1002/cbic.202000735
dc.identifier.urihttp://hdl.handle.net/10033/622614
dc.description.abstractCYP154C5 from Nocardia farcinica is a P450 monooxygenase able to hydroxylate a range of steroids with high regio- and stereoselectivity at the 16a-position. Using protein engineering and substrate modifications based on the crystal structure of CYP154C5, an altered regioselectivity of the enzyme in steroid hydroxylation had been achieved. Thus, conversion of progesterone by mutant CYP154C5 F92A resulted in formation of the corresponding 21-hydroxylated product 11-deoxycorticosterone in addition to 16α-hydroxylation. Using MD simulation, this altered regioselectivity appeared to result from an alternate binding mode of the steroid in the active site of mutant F92A. MD simulation further suggested that water entrance to the active site caused higher uncoupling in this mutant. Moreover, exclusive 15α-hydroxylation was observed for wild-type CYP154C5 in the conversion of 5a-androstan-3-one, lacking an oxy-functional group at C17. Overall, our data give valuable insight into the structure-function relationship of this cytochrome P450 monooxygenase for steroid hydroxylation.en_US
dc.language.isoenen_US
dc.publisherWileyen_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectbiocatalysisen_US
dc.subjectcytochrome P450 monooxygenaseen_US
dc.subjectprotein engineeringen_US
dc.subjectregioselectivityen_US
dc.subjectsteroid hydroxylationen_US
dc.titleCYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate Modifications and Protein Engineering.en_US
dc.typeArticleen_US
dc.identifier.eissn1439-7633
dc.contributor.departmentHZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.en_US
dc.identifier.journalChembiochem : a European journal of chemical biologyen_US
refterms.dateFOA2020-11-26T09:36:37Z
dc.source.journaltitleChembiochem : a European journal of chemical biology
dc.source.countryGermany


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Attribution 4.0 International
Except where otherwise noted, this item's license is described as Attribution 4.0 International