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Molecular biology enzymology and structural biology of the biosynthesis of aminoglycoside antibiotics

Reference	BB/C505667/1
Principal Investigator / Supervisor	Dr Jonathan Spencer
Co-Investigators / Co-Supervisors	Professor Sir Tom Blundell, Dr Fanglu Huang, Dr Bojana Popovic
Institution	University of Cambridge
Department	Chemistry
Funding type	Research
Value (£)	429,225
Status	Completed
Type	Research Grant
Start date	01/10/2004
End date	29/02/2008
Duration	41 months

Abstract

We have recently sequenced part of the butirosin gene cluster for Bacillus circulans by gene walking. Twelve of the orfs were sub-cloned and soluble proteins expressed. BtrR has been characterised as a transaminase which catalyses two of the steps on the pathway to 2-deoxystreptamine. This protein has been crystallised and the structure solved to a resolution of 1.8 Angstrom. We have also recently found and sequenced the gene cluster for neomycin from Streptomyces fradiae. The availability of both sets of genes gives a good opportunity to elucidate in detail the biosynthesis of this important class of antibiotics. Of particular interest is the unique 2-hydroxy-4-amino-butyric acid moiety of butirosin, which prevents modification of the molecule by certain enzymes that confer resistance in clinical isolates. We propose that this amino acid is biosynthesised from glutamate by five enzymes found in the butirosin cluster whilst attached to a peptyl carrier protein (PCP). It is proposed to crystallise and solve the structures of these enzymes so that information can be gained on how the PCP docks with the other enzymes. Amakicin is an important semi-synthetic aminoglycoside made by modifying kanamycin with 4-amino-2-hydroxybutyrate. It would be attractive to if amikacin could be produced directly by fermentation. To achieve this we propose to move the genes responsible for 4-amino-2-hydroxybutyrate biosynthesis in butirosin into the kanamycin producing strain.

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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