Methods for increasing the production of a recombinant...

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/57 (2006.01) C07K 14/58 (2006.01) C07K 14/595 (2006.01) C07K 14/605 (2006.01) C12N 1/19 (2006.01) C12N 9/60 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2499467

The present invention provides a host cell comprising a nucleic acid sequence encoding a recombinant polypeptide in which the production of a naturally occurring metalloprotease comprising the sequenc HXXEH has been reduced or inhibited by genetic manipulation. The present invention also relates to methods for enhancing the production of a polypeptide from a cell by disrupting the synthesis or activity of the metalloprotease. In particular, the present invention relates to methods for enhancing the secretion of recombinant polypeptides from host cells such as, but not limited to, yeast and bacterial cells. The metalloprotease is a member of the pitrilysin subfamily of proteases, characterized by comprising the sequence HXXEH, where X is any amino acid.

L'invention concerne une cellule hôte comportant une séquence d'acide nucléique codant pour un polypeptide recombinant dans lequel la production d'une métalloprotéase d'occurrence naturelle comportant une séquence de SEQ ID NO:1 a été réduite ou inhibée par manipulation génétique. La présente invention porte également sur des procédés pour augmenter la production d'un polypeptide à partir d'une cellule par rupture de la synthèse ou de l'activité de la métalloprotéase. Cette invention concerne en particulier des procédés pour accroître la sécrétion de polypeptides recombinants à partir de cellules hôtes telles que, mais sans se limiter à, des cellules de levure et des cellules bactériennes. La métalloprotéase est un membre de la sous-famille pitrilysin des protéases, caractérisée en ce qu'elle comprend la séquence HXXEH (SEQ ID NO:1), où X est un acide aminé.

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