Award details

Using forced evolution,molecular modelling and bioinformatics to understand and exploit plant CYP74s

Reference	BBS/E/J/0000A151
Principal Investigator / Supervisor	Professor Rod Casey
Co-Investigators / Co-Supervisors	Professor Stephen Bornemann, Dr Shirley Ann Fairhurst
Institution	John Innes Centre
Department	John Innes Centre Department
Funding type	Research
Value (£)	47,406
Status	Completed
Type	Institute Project
Start date	01/07/2003
End date	30/06/2006
Duration	36 months

Abstract

This project is to increase our understanding of lipoxygenase (LOX) and a number of specific cytochrome P450 (CYP74) activities through a combination of forced evolution molecular modelling and bioinformatics. It will: 1. force the evolution of new LOX activities that efficiently use `unusual¿ PUFAs such as 16:3, ?-18:3, 18:4, 20:4, 22:5, through DNA shuffling, to provide substrates for the forced evolution of CYP74s 2. force the evolution of entirely new CYP74 activities, either on the novel hydroperoxide substrates produced by the above new LOXs or on `standard¿ fatty acid hydroperoxide substrates 3. gain understanding of the molecular basis of the differences in activity between the different CYP74s by modelling their actives sites against known P450 structures and producing mutants based on the modelling 4. increase our understanding of CYP74 activity by forcing the evolution of one CYP74 activity into another and analysing the consequent sequence differences by homology modelling Such advances will produce new understanding on the mechanisms and specificities of CYP74s, new biocatalysts and new oxylipins with potential as `green¿ plant protectants, flavours, fragrances and chemical feedstocks.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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