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Towards a complete structure-function description of an aromatic degradative pathway

Reference	BB/C004450/1
Principal Investigator / Supervisor	Professor David Ian Roper
Co-Investigators / Co-Supervisors	Professor Timothy Bugg, Professor Vilmos Fulop
Institution	University of Warwick
Department	Biological Sciences
Funding type	Research
Value (£)	235,653
Status	Completed
Type	Research Grant
Start date	01/04/2005
End date	31/03/2008
Duration	36 months

Abstract

Microbial enzymes represent the most diverse biocatalytic potential on Earth. The capacity of bacteria to degrade aromatic chemicals in particular provides a comprehensive library of chemical strategies, diverse enzymatic mechanisms and novel structures, which have been little studied at a molecular level. This proposal focuses on two enzymes from the 3,4-dihydroxyphenylacetate degradative pathway of E. coli, which have particular mechanistic interest and potential for biocatalysis. The first of these is a novel bifunctional decarboxylase isomerase for which we have already obtained an X-ray structure, revealing a two domain arrangement containing distinct but related catalytic sites in each domain. A structure-function study of these sites in combination with the determination of enzyme ligand complexes, is now required to define the catalytic mechanisms and strategies for substrate selection. Similarities between the two active sites, which catalyse distinct chemical transformations, may provide a model for the development of novel activities and substrate selection. The second enzyme to be studied, along with a sequence related homologue, are members of a family of stable metal ion dependent C-C bond type II aldolases which are distinct from those previously studied. We have obtained crystals and X-ray data for both and will have structures prior to the start of this project where we aim to investigate the mechanism of this class of enzymes and use the information obtained in the design of novel C-C bond forming activities. Type II aldolases are generally stable structures and have the potential as stable platforms for industrial biocatalysis applications and as part of our study we will address the use of such enzymes in the formation of novel compounds. Mechanistically there are issues relating to the involvement of the metal 2+ ion in type II enzyme catalysis that this project will also address. In collaboration with other researchers we hope to define a complete structural description of the degradative pathway for 3,4-dihydroxyphenylacetate and determine the catalytic strategies employed. This will be the first description of its type and represents a significant step forward in our understanding of natural biotransformation strategies and their potential for biotechnology applications.

Summary

unavailable

Committee	Closed Committee - Biomolecular Sciences (BMS)
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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