Methods for secretory production of proteins

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/09 (2006.01) C07K 5/107 (2006.01) C07K 7/06 (2006.01) C07K 7/08 (2006.01) C07K 14/195 (2006.01) C07K 14/34 (2006.01) C07K 14/36 (2006.01) C07K 19/00 (2006.01) C12N 1/21 (2006.01) C12N 15/77 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2442679

The object of the present invention is to provide a method of producing a heterologous protein by making a coryneform bacterium to produce and efficiently extracellularly secrete (secreto-production) an industrially useful heterologous protein. According to the present invention, a genetic construct is used where a gene sequence encoding an intended protein which is ligated to the downstream of a sequence encoding the signal peptide derived from a coryneform bacterium, the gene construct is introduced into a mutant coryneform bacterium which has a capacity of secreting the heterologous protein at least 2-fold higher than the wild type Corynebacterium glutamicum ATCC13869, the mutant coryneform bacterium is cultured and the extracellularly released heterologous protein is recovered.

L'invention permet de produire une protéine exogène utile sur le plan industriel par création d'une bactérie de type corynéen afin de sécréter et de produire de manière efficace la protéine exogène. Un gène d'expression chimère, qui possède une séquence génique d'une protéine exogène cible liée à la partie avale de la séquence codant le domaine peptide-signal issue de la bactérie de type corynéen, est transféré dans le variant de la bactérie de type corynéen capable de sécréter et de produire au moins deux fois la quantité de protéine exogène par rapport au Corynebacterium glutamicum ATCC13869 de type sauvage. La bactérie de type corynéen ainsi transformée est cultivée et la protéine exogène libérée et collectée.

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