Process for producing transglutaminase

C - Chemistry – Metallurgy – 12 – P

Patent

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Details

C12P 21/02 (2006.01) C12N 9/10 (2006.01) C12N 9/48 (2006.01) C12N 9/52 (2006.01) C12N 15/62 (2006.01) C12N 15/63 (2006.01)

Patent

CA 2391961

The present invention relates to a process for secretory production of a foreign protein, in particular, transglutaminase by a coryneform bacterium. According to the present invention, a process is provided for the secretory production of a foreign protein, in particular, transglutaminase, by making a coryneform bacterium to produce an industrially useful foreign protein, in particular, transglutaminase and efficiently release the product extracellularly (i.e., secretory production). An intended foreign protein, in particular, transglutaminase, is produced by using an expression construct wherein the gene sequence of the intended foreign protein containing the pro-structure part, in particular, pro-transglutaminase gene sequence, is ligated to the downstream of a sequence encoding the signal peptide region from a coryneform bacterium, introducing this expressional genetic construct into a coryneform bacterium, culturing the thus transformed coryneform bacterium, and treating the extracellularly released protein with a protease, etc. to cleave and eliminate the pro-part.

Cette invention a trait à un procédé de production sécrétoire d'une protéine exogène (une transglutaminase notamment) à l'aide d'une bactérie de type corynéen. Elle porte, plus précisément, sur un procédé de production de protéine exogène (une transglutaminase notamment), lequel procédé consiste à produire une bactérie de type corynéen afin de produire une protéine exogène (une transglutaminase notamment), utile sur le plan industriel, et à expulser de manière satisfaisant le produit hors des cellules (c'est à dire une production sécrétoire). On produit une protéine exogène cible (une transglutaminase notamment) à l'aide d'un produit de recombinaison d'expression. La séquence génique de la protéine cible contenant la partie pro-structurelle (notamment une séquence génique pro-transgutaminase) est ligaturée à la partie aval d'une séquence codant le domaine peptide-signal issu de la bactérie de type corynéen. On transfère ensuite ce produit génique de recombinaison type d'expression dans la bactérie de type corynéen. On cultive alors cette bactérie de type corynéen ainsi transformée et on traite la protéine sortie des cellules à l'aide d'une protéase, etc., afin de cliver et d'éliminer la partie pro-structurelle.

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