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Enantioselective Benzylic Hydroxylation Catalysed by P450 Monooxygenases: Characterisation of a P450cam Mutant Library and Molecular Modelling
Author(s) -
Eichler Anja,
Gricman Łukasz,
Herter Susanne,
Kelly Paul P.,
Turner Nicholas J.,
Pleiss Jürgen,
Flitsch Sabine L.
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500536
Subject(s) - monooxygenase , hydroxylation , chemistry , stereoselectivity , enantioselective synthesis , cytochrome p450 , stereochemistry , biocatalysis , directed evolution , active site , molecular dynamics , substrate (aquarium) , molecular model , cytochrome , mutant , enzyme , combinatorial chemistry , computational chemistry , catalysis , reaction mechanism , biochemistry , biology , ecology , gene
Cytochrome P450 monooxygenases can catalyse the stereoselective C−H activation of a very broad range of substrates. Prediction and control of enantioselectivity of this enzyme class is of great interest for the synthesis of high‐value chiral molecules. Here we have used a combination of molecular dynamics simulations and experimental screening to study the enantioselectivity of a library of active‐site mutants of chimeric P450cam‐RhFRed towards the benzylic hydroxylation of structurally related regioisomers of ethylmethylbenzene. Small variations either in substrate structure or in enzyme active site architecture were shown to lead to dramatic changes in enantioselectivity; this was broadly in agreement with computational predictions. In addition to validating computational approaches, these studies have provided us with a deeper understanding of effects that might control stereoselectivity in these biooxidation reactions.