Modified deacetoxycephalosporin c synthase (daocs) and x-ray...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/52 (2006.01) C12N 9/00 (2006.01) C12N 9/02 (2006.01) C12P 35/00 (2006.01)

Patent

CA 2316401

Three-dimensional crystal structure(s) of deacetoxycephalosporin C synthase (DAOCS) are described. The X-ray co-ordinates provide precise 3-dimensional information of amino acids within the structure of DAOCS. Some of these are in complexes with iron and/or substrates. Information from the structures is used to modify enzymes of the cephalosporin biosynthesis pathway including DAOCS, deacetylcephalosporin C synthase DAOC/DACS, such that they accept unnatural substrates (e.g. penicillins G, V) in order to improve the production of beta- lactam antibiotics. The structures may be used to predict the structures of other 2-oxoglutarate dependent enzymes, thereby allowing the design of inhibitors, and new catalysts for the production of e.g. oxidised amino acids/peptides. Specific modifications of amino acid residues are proposed and exemplified.

L'invention concerne la ou les structures tridimensionnelles de cristaux de la désacétoxycéphalosporine C synthase (DAOCS). Les coordonnées obtenues par rayons X fournissent des informations tridimensionnelles précises des acides aminés se trouvant dans la structure de DAOCS. Certains de ceux-ci se présentent en complexes avec du fer et/ou des substrats. Les informations tirées des structures sont utilisées pour modifier les enzymes de la voie de biosynthèse de la céphalosporine comprenant DAOCS, la désacétylcéphalosporine C synthase DAOCS/DACS, de manière qu'elles acceptent des substrats synthétiques (par exemple des pénicillines G, V) afin d'améliorer la production de .beta.-lactamines. Les structures peuvent être utilisées pour prédire les structures d'autres enzymes dépendantes de 2-oxoglutarate, permettant ainsi la conception d'inhibiteurs, et de nouveaux catalyseurs pour la production, par exemple, d'acides aminés/peptides oxydés. Des modifications spécifiques de restes d'acides aminés sont proposées et exemplifiées.

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