Details of Grant 

EPSRC Reference:	GR/R41781/01
Title:	Probing stability of structure and intermediates in a biotechnologically important mono-oxygenase.
Principal Investigator:	Munro, Professor AW
Other Investigators:	Smith, Professor WE
Researcher Co-Investigators:
Project Partners:
Department:	Biochemistry
Organisation:	University of Leicester
Scheme:	Fast Stream
Starts:	01 October 2001	Ends:	30 September 2004	Value (£):	65,201
EPSRC Research Topic Classifications:	Chemical Biology Chemical Structure
EPSRC Industrial Sector Classifications:	Chemicals Pharmaceuticals and Biotechnology No relevance to Underpinning Sectors
Related Grants:
Panel History:
Summary on Grant Application Form
The cytochromes P450 ar diversw biological catalysts with the ability to catalyse reductive scission of molecular oxygen and insertion of an atom of oxygen into an organic substrate. The reaction is acheived usually with exquisite stereo-and regio- selectivity. The fact that these enzymes can perform several reactions difficult or impossible to acheive with any efficiency by conventional organic synthesis routes means that they have great potential for industrial synthesis of fine chemicals. A drawback to their use for such processes is the fact that they can lose catalytic activity through structural denaturation. Inactivation events occur often througha a route involving non-productive activation of molecular oxygen, leading to release of damaging oxygen radicals. In this proposal, a thorough characterisation of the structural stability of a biotechnologically important P450 catalyst (P450 BM3) will be performed, examining factors underlying the loss of catalytic activity using spectroscopic probes. In particular, the P450's are known to form an inactive species (cytochrome P420) in which the catalytically fundamental ligation of the heme iron to a cysteine sulfhydryl is lost. The precise chemical nature of the P420 form remains enigmatic , and we intend to determine it's specific properties using spectroscopic methods, In addition, we will use P450 mutants which stabilise the ferrous-oxy catalyti intermediate in order to probe properties of this form, it's reactivity and the relevant importance of the non-productive collapse of the ferrous-oxy form ( compared to wild-type P450 BM3 ) in denaturation of P450 through Oxygen radical production.
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Organisation Website:	http://www.le.ac.uk