Enantioselective Benzylic Hydroxylation Catalysed by P450 Monooxygenases: Characterisation of a P450cam Mutant Library and Molecular Modelling. (16th February 2016)

• Record Type: Journal Article
• Authors: Eichler, Anja; Gricman, Łukasz; Herter, Susanne; Kelly, Paul P.; Turner, Nicholas J.; Pleiss, Jürgen; Flitsch, Sabine L.
• Is Part Of: Chembiochem. Volume 17:Number 5(2016)
• Page Start: 426
• Page End: 432
• Publication Date: 2016-02-16
• Subjects: benzylic oxidation -- C−H activation -- chiral alcohols -- directed evolution -- enantioselectivity -- P450 enzymes; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201500536
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: 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. Abstract : Finding the best cast : A combination of experimental screening and molecular dynamics simulation was used to study the enantioselectivity of a library of active‐site mutants of chimeric P450cam‐RhFRed in benzylic hydroxylation of structurally related regioisomers of ethylmethylbenzene, resulting in the identification of three new P450cam mutants with excellent and opposite enantioselectivities.