CYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate Modifications and Protein Engineering.
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Authors
Bracco, PaulaWijma, Hein J
Nicolai, Bastian
Rodriguez Buitrago, Jhon Alexander
Klünemann, Thomas
Vila, Agustina
Schrepfer, Patrick
Blankenfeldt, Wulf

Janssen, Dick B
Schallmey, Anett
Issue Date
2020-11-04
Metadata
Show full item recordAbstract
CYP154C5 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.Citation
Chembiochem. 2020 Nov 4. doi: 10.1002/cbic.202000735. Epub ahead of print.Affiliation
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.Publisher
WileyPubMed ID
33145893Type
ArticleLanguage
enEISSN
1439-7633ae974a485f413a2113503eed53cd6c53
10.1002/cbic.202000735
Scopus Count
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- Creative Commons