Method for producing dna encoding polypeptides that are...

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 15/10 (2006.01) C12N 1/21 (2006.01) C12N 15/52 (2006.01) C12N 15/62 (2006.01) C12N 15/64 (2006.01) C12N 15/66 (2006.01)

Patent

CA 2417011

The invention relates to a method for producing novel modular enzyme systems by genetic engineering, which is characterized by a cyclic in vitro gene synthesis and which allows a specific recombination of the individual, gene- encoded modular components to novel enzyme systems. Examples of such modular enzyme systems are the non-ribosomal peptide synthetases (NRPS) or polyketide synthases (PKS) of type 1, that is the amino acid sequence of such an enzyme is characterized by being composed of a repetitive sequence of identical sequence sections or sequence sections that are very similar to one another. Every single repetitive sequence is referred to as a module and every module allows the enzymatic incorporation of a specific substrate into the substance synthesized by the enzyme. The products synthesized by the NRPS and PKS enzymes are often highly valuable as pharmaceuticals, such as the penicillins, vancomycins or erythromycins. The inventive method allows for an effective production of novel genes for modular enzymes, and the gene expression of said novel genes allows for the production of novel substances.

La présente invention concerne un procédé servant à produire de nouveaux systèmes enzymatiques de structure modulaire, par la voie du génie génétique, ledit procédé se caractérisant par une synthèse de gènes in vitro se déroulant de manière cyclique, et ledit procédé permettant une nouvelle recombinaison ciblée des composants modulaires codés individuels pour donner de nouveaux systèmes enzymatiques. Des systèmes enzymatiques de structure modulaire de ce type correspondent par exemple aux synthases peptidiques non ribosomales (NRPS) ou aux synthases polycétidiques (PKS) de type 1. Cela signifie que la séquence d'acides aminés d'une enzyme de ce type se caractérise en ce qu'elle est composée d'une suite de segments de séquences identiques ou très similaires qui se répètent. Chacune de ces séquences est qualifiée de module et chaque module permet la construction enzymatique d'un substrat spécifique dans la substance synthétisée par l'enzyme. Les produits synthétisés par les enzymes NRPS et PKS ont fréquemment une grande valeur pharmaceutique, c'est le cas de la Pénicilline, de la Vancomycine ou de l'Erythromycine. Le procédé de l'invention permet de produire efficacement de nouveaux gènes d'enzymes modulaires, l'expression de ces nouveaux gènes permettant la production de nouvelles substances.

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