C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 9/10 (2006.01) C12N 15/54 (2006.01)
Patent
CA 2356597
Transglutaminase having an enzymatic activity is produced by subjecting transglutaminase which is in a denatured state to a process involving at least the following steps (a) and (b): (a) the step of forming an intermediate structure, which shows the enzymatic activity in an aqueous medium under acidic conditions, of the denatured enzyme in the course of forming the structure; and (b) the step of forming a higher-order structure, which shows the enzyme activity in an aqueous medium at a pH value within the neutral region, of the enzyme having the intermediate structure in the course of forming the structure as described above. Thus, transglutaminase obtained in a denatured state (for example, one produced by a gene recombinant microorganism) can be efficiently refolded and, therefore, highly pure transglutaminase having an enzymatic activity substantially comparable to the activity of authentic transglutaminase (i.e., transglutaminase activity) can be industrially produced. The intermediate having the intermediate structure in the course of forming the structure as described above.
Selon l'invention, on produit une transglutaminase possédant une activité enzymatique en soumettant une transglutaminase à l'état dénaturé à un procédé comprenant au moins les étapes suivantes (a) et (b) consistant: (a) à former une structure intermédiaire - possédant l'activité enzymatique dans un milieu aqueux, dans des conditions acides - de l'enzyme dénaturée, pendant la formation de la structure, et (b) à former une structure d'ordre supérieur - possédant l'activité enzymatique dans un milieu aqueux à une valeur de pH se situant dans la région neuronale - de l'enzyme possédant la structure intermédiaire, pendant la formation de la structure, comme décrit ci-dessus. Ainsi, la transglutaminase obtenue à l'état dénaturé (par exemple, une transglutaminase produite par un micro-organisme de recombinaison de gène) peut subir une renaturation efficace, ce qui permet donc de produire industriellement une transglutaminase extrêmement pure, possédant une activité enzymatique sensiblement comparable à l'activité de la transglutaminase authentique (c'est-à-dire l'activité de transglutaminase). L'intermédiaire possède la structure intermédiaire pendant la formation de la structure, comme décrit ci-dessus.
Ejima Daisuke
Ono Kunio
Yokoyama Keiichi
Ajinomoto Co. Inc.
Ogilvy Renault Llp/s.e.n.c.r.l.,s.r.l.
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