Description du projet
Les cyclopropane synthases comme nouveaux biocatalyseurs
Les cycles cyclopropane sont présents dans de nombreux composés pharmaceutiques et servent de blocs de construction réactifs dans la synthèse de produits chimiques. Le projet CycloproBio, financé par l’UE, explorera les enzymes cyclopropane synthases (CS) comme une nouvelle méthode sûre pour la cyclopropanation biocatalytique. L’étude caractérisera les propriétés catalytiques de la synthase des acides gras de cyclopropane (CFAS pour «cyclopropane fatty acid synthase») précédemment clonée à partir de Escherichia coli, et développera un test d’activité approprié pour étudier la spécificité de son substrat et ses caractéristiques physiochimiques. Par la suite, de nouvelles enzymes CS seront générées par mutagenèse des résidus putatifs de liaison au substrat. L’objectif du projet est d’utiliser les nouvelles données pour développer des biocatalyseurs de cyclopropanation pour la synthèse durable de produits chimiques et pharmaceutiques.
Objectif
The cyclopropane ring is an important structural motif present in many pharmaceutical compounds and widely used as reactive building blocks in the synthesis of chemicals. Many synthetic strategies to access cyclopropanes have been developed to date, most of which typically require the use of potentially explosive diazo-carbene compounds as substrates. Even recently proposed enzymatic transformations, exploiting cytochromes P450 and myoglobin, require the use of stoichiometric carbene co-substrates, thus narrowing their industrial applications. The CycloproBio project aims to explore cyclopropane synthase (CS) enzymes as a potentially mild method for biocatalytic cyclopropanation. Cyclopropane synthases are SAM dependent transferase enzymes able to catalyse cyclopropanation of unsaturated phospholipids in bacterial membranes. Even if CS are common to most bacteria, many questions on factors affecting their substrate selectivity remain unanswered. Moreover, despite their potential, CS enzymes have not been explored yet as potential biocatalysts. This project will initially characterize the cyclopropane fatty acid synthase (CFAS) from E. coli which has been cloned, expressed and purified in preliminary work. The catalytic properties of the E. coli CFAS will be studied through the development of an appropriate activity assay, the investigation of its substrate specificity and the study of its physio-chemical properties. To broaden the study, genome mining for other CS enzymes from different bacterial species will be carried out to gain insight into the factors which affect the selectivity of these enzymes. Finally, new CS enzyme variants will be generated through rational mutagenesis of putative substrate binding residues. The data generated will provide fundamental information on the CS family. The primary aim of this project is to use this data to develop cyclopropanation biocatalysts for the sustainable synthesis of chemicals and pharmaceuticals.
Champ scientifique
- natural sciencesbiological sciencesmicrobiologybacteriology
- medical and health sciencesbasic medicinepharmacology and pharmacypharmaceutical drugs
- natural sciencesbiological sciencesbiochemistrybiomoleculeslipids
- natural scienceschemical sciencescatalysisbiocatalysis
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes
Programme(s)
Régime de financement
MSCA-IF-EF-ST - Standard EFCoordinateur
WC1E 6BT London
Royaume-Uni