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1. WO2000040706 - PROCESS FOR PRODUCING TRANSGLUTAMINASE

Publication Number WO/2000/040706
Publication Date 13.07.2000
International Application No. PCT/JP1999/007250
International Filing Date 24.12.1999
Chapter 2 Demand Filed 28.04.2000
IPC
C12N 9/10 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
10Transferases (2.)
CPC
C12N 9/1044
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
10Transferases (2.)
1025Acyltransferases (2.3)
104Aminoacyltransferases (2.3.2)
1044Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII
C12Y 203/02013
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
YENZYMES
203Acyltransferases (2.3)
02Aminoacyltransferases (2.3.2)
02013Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII
Applicants
  • AJINOMOTO CO., INC. [JP/JP]; No. 15-1, Kyobashi 1-chome Chuo-ku Tokyo 104-8315, JP (AllExceptUS)
  • YOKOYAMA, Keiichi [JP/JP]; JP (UsOnly)
  • ONO, Kunio [JP/JP]; JP (UsOnly)
  • EJIMA, Daisuke [JP/JP]; JP (UsOnly)
Inventors
  • YOKOYAMA, Keiichi; JP
  • ONO, Kunio; JP
  • EJIMA, Daisuke; JP
Agents
  • ISHIDA, Yasumasa ; A. Kato & Associates Bohsei Building, 7th floor 20-12, Shin-Yokohama 3-chome Kohoku-ku, Yokohama-shi Kanagawa 222-0033, JP
Priority Data
10/37313128.12.1998JP
Publication Language Japanese (JA)
Filing Language Japanese (JA)
Designated States
Title
(EN) PROCESS FOR PRODUCING TRANSGLUTAMINASE
(FR) PROCEDE DE PRODUCTION DE TRANSGLUTAMINASE
Abstract
(EN)
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.
(FR)
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.
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