Centre for Novel Agricultural Products

Clean Technology


Enzyme discovery - Developing a new platform technology for oxidative biocatalysis
Funded by CoEBio3
Professor Neil Bruce (CNAP), Dr Gideon Grogan (YSBL, Department of Chemistry) and Professor Sabine Flitsch (CoEBio3)

This project aims to establish a suite of technologies for the cloning, functional expression and screening of prokaryotic and eukaryotic cytochromes P450 in E. coli. Cytochromes P450 represent a largely untapped resource in the context of industrial hydroxylations. This is essentially due to previous difficulties with the heterologous expression and screening of these P450s in E. coli, and the requirement for auxiliary redox proteins for electron transfer to the important heme domain, which is responsible for substrate specificity.  We have recently developed a cloning system – LIC-RED - based on the unusual and naturally fused P450Rhf from Rhodococcus NCIMB 9784, which permits drop-in of heme domains from a variety of sources to generate libraries of redox-self sufficient CYPs, which catalyse substrate oxidation merely with the addition of NADPH. 

Enzyme Discovery

 

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Contacts

CNAP Director, Professor Simon McQueen-Mason

CNAP Manager, Dr Caroline Calvert

CNAP, Department of Biology, University of York, Wentworth Way, York YO10 5DD, UK