Process for the enzymatic production of cyclic diguanosine...

Details Process for the enzymatic production of cyclic diguanosine... Process for the enzymatic production of cyclic diguanosine...

C - Chemistry, Metallurgy

12

N

C12N 15/54 (2006.01) C07H 19/207 (2006.01) C12N 9/12 (2006.01) C12P 19/36 (2006.01)

Patent

CA 2747127

A process is disclosed for the production of cyclic di-guanosine monophosphate (c-di-GMP) without the use of protecting groups by means of an enzymatic synthesis. The process comprises the coupling of two guanosine triphosphate (GTP) molecules so as to form a c-di-GMP molecule. This is done under the influence of a mutant diguanylate cyclase (DGC) compris-ing the amino acid sequence V153M154G155G156. It has been found that the DGC is obtainable from inclusion bodies, and therewith can be made available in amounts sufficient to improve the c-di-GMP synthesis. Particularly, the latter synthesis can be conducted in a one-pot method starting from commercially available bulk chemicals and allows upscaling to a commercial produc-tion scale.

La présente invention concerne un procédé pour la production de diguanosine monophosphate cyclique (di-GMPc) sans utilisation de groupes protecteurs au moyen d'une synthèse enzymatique. Le procédé comprend le couplage de deux molécules de guanosine triphosphate (GTP) de manière à former une molécules de di-GMPc. Cela est effectué sous l'influence d'une diguanylate cyclase (DGC) mutante comprenant la séquence d'acides aminés V153M154G155G156. Il a été observé que DGC peut être obtenu à partir de corps d'inclusion, et avec ceux-ci peut être rendu disponible en quantités suffisantes pour améliorer la synthèse de di-GMPc. En particulier, cette dernière synthèse peut être conduite dans un procédé "one-pot" à partir de substances chimiques en vrac commercialisées et permet l'augmentation d'échelle à une échelle de production commerciale.

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