Modified e. coli enterotoxin ii signal peptide and a...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/31 (2006.01) C07K 14/245 (2006.01) C07K 14/61 (2006.01) C12N 1/21 (2006.01) C12N 15/18 (2006.01) C12N 15/62 (2006.01) C12N 15/70 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2342537

A heterologous protein is produced by: (i) culturing a microorganism transformed with an expression vector comprising a gene encoding a modified E. coli enterotoxin II signal peptide fused with the heterologous protein to produce and secrete the heterologous protein to periplasm, said modified E. coli enterotoxin II signal peptide being obtained by replacing at least one of the 2nd, 4th, 5th, 12th, 20th and 22nd amino acids of E. coli enterotoxin II signal peptide of the following amino acid sequence (SEQ ID NO: 1) with another amino acid, with the proviso that at least one of the 2nd and 4th amino acid of the modified peptide is lysine; and (ii) recovering the heterologous protein from the periplasm.

On a produit une protéine hétérologue par: (i) culture d'un micro-organisme transformé avec un vecteur d'expression comprenant un gène codant un signal peptide d'entérotoxine (II) de E. coli modifié, fusionné à la protéine hétérologue pour produire et sécréter la protéine hétérologue dans le périplasme, ledit peptide signal d'entérotoxine (II) de E. coli modifié étant obtenu par remplacement d'au moins un des seconds, quatrièmes, cinquièmes, douzièmes, vingtièmes et vingt-deuxièmes acides aminés du peptide signal d'entérotoxine (II) de E. coli de la séquence d'acides aminés (SEQ ID NO: 1) suivante Met Lys Lys Asn Ile Ala Phe Leu Leu Ala Ser Met Phe Val Phe Ser Ile Ala Thr Asn Ala Tyr Ala par un autre acide aminé, à la condition qu'au moins un des seconds et quatrièmes acides aminés du peptide modifié soit la lysine; et (ii) récupération de la protéine hétérologue du périplasme.

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