Multiply-substituted protease variants with altered net...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/57 (2006.01) A23K 1/165 (2006.01) A61K 8/66 (2006.01) A61Q 11/00 (2006.01) C11D 3/386 (2006.01) C12N 9/54 (2006.01) C12N 9/58 (2006.01)

Patent

CA 2308113

Novel protease variants derived from the DNA sequences of naturally-occurring or recombinant non-human proteases are disclosed. The variant proteases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant protease to generate the substitution of a plurality of amino acid residues in the amino acid sequence of a precursor protease. Protease variants are provided that contain substitutions of the amino acids at one or more residue positions so that the substitution alters the charge at that position to make the charge more negative or less positive compared to a precursor protease and thus the protease variant is more effective in a low detergent concentration system than a precursor protease. Also provided are protease variants containing substitutions of the amino acids at one or more residue positions so that the substitution alters the charge at that position to make the charge more positive or less negative compared to a precursor protease and thus the protease variant is more effective in a high detergent concentration system than a precursor protease.

L'invention concerne de nouvelles variantes de protéases obtenues à partir de séquences d'ADN de protéases naturelles ou de protéases non humaines recombinées. Les protéases variantes sont généralement obtenues par modification in vitro d'une séquence d'ADN précurseur codant pour la protéase naturelle ou recombinée en vue de produire la substitution de plusieurs résidus d'acides aminés dans la séquence d'acides aminés d'une protéase précurseur. L'invention concerne des variantes de protéase qui contiennent des substitutions d'acides aminés à une ou plusieurs positions de résidus, de sorte que la substitution modifie la charge à cette position pour la rendre plus négative ou moins positive par rapport à une protéase précurseur, et par conséquent la variante de protéase est plus efficace dans un système à faible concentration de détergent qu'une protéase précurseur.

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