Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.823147
Title: Development of cytochrome P450 monooxygenased for applications in biocatalysis
Author: Manning, Jack
ISNI:       0000 0005 0290 0456
Awarding Body: University of Manchester
Current Institution: University of Manchester
Date of Award: 2019
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Abstract:
Cytochrome P450 monooxygenases are multicomponent heme enzymes found in the majority of life on Earth. These enzymes are widespread due to their roles as detoxification catalysts, a job they are ideally suited for due to their impressive promiscuity. In addition to the metabolism of exogenous compounds, many P450s are involved in the biosynthesis of complex natural products such as steroids, lipids and antibiotics. Despite these diverse natural applications, the number of Cytochrome P450 monooxygenases used industrially remains startlingly low. This work aims to address some of the current issues with the use of P450s by introducing new enzymes with novel activities to the 'biocatalytic toolbox' as well as applying these enzymes for the formation of industrially relevant products. Here we present the cloning, expression and characterisation of three new self sufficient Cytochrome P450 monooxygenases (CYP102A25, CYP102A26 and CYP116B62) in addition to the characterisation of novel activity of the recently discovered P450 CYP116B46. We also report the successful application of this novel activity into a 2-step chemo-enzymatic cascade for the regio- and enantioselective synthesis of the widely used flavouring compound δ-decalactone. Finally, a study on controlling the regioselectivity of Cytochrome P450 monooxygenases by the rational design of substrates containing a 'functional handle' is detailed with promising results. The work presented in this thesis provides synthetic biologists with additional tools for monooxidation reactions as well as furthering the use of Cytochrome P450 monooxygenases within biocatalysis.
Supervisor: Flitsch, Sabine ; Turner, Nicholas Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.823147  DOI: Not available
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