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Enzymes for the synthesis of chiral effects chemicals
Reference
E03668
Principal Investigator / Supervisor
Professor John Holbrook
Co-Investigators /
Co-Supervisors
Institution
University of Bristol
Department
Biochemistry
Funding type
Research
Value (£)
171,399
Status
Completed
Type
Research Grant
Start date
01/12/1994
End date
01/04/1998
Duration
40 months
Abstract
(i) Overexpress the gene and overproduce Candida methylica NAD: formate oxidoreductase (formate dehydrogenase). Determine the X-ray structure of the apoenzyme and of complexes with NAD and with NAD+ azide. Determine the mechanism and then use protein engineering techniques to enhance the catalytic rate and the solvent and thermal stability of the enzyme for use in chemoenzymic syntheses. (ii) Determine the X-ray structure of existing crystals of Lactobacillus bulgaricus D-lactate dehydrogenase and large D-2-hydroxyacid dehydrogenase - as apo enzyme and a complex with NADH and a suitable oxamate. Use protein engineering to establish the mechanism and also to enhance stability. (iii) The formate and D-2-hydroxyacid enzymes are each homodimers and the sequences suggest they will have very similar folds and intersubunit contacts. Three protein engineering strategies will be used to try to make a heterodimer with one FDH and one D-2-hydroxyacid subunit. The multienzyme complex will academically be sued to investigate the advantage or otherwise of short cofactor diffusion distances for catalysis and for efficient coenzyme utilisation in chemoenzymic synthesis of chirons.
Summary
unavailable
Committee
Closed Committee - Engineering & Biological Systems (EBS)
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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