Stabilized enzymes and detergent compositions
AssigneeNovo Nordisk A/S
InventorsClaus Von Der Osten, Sven Branner, Allan Svendsen, Lisbeth Hedeg.ang.rd, Nina Eriksen, Maarten Robert Egmond, Eric Casteleijn
Life of this patent
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Abstract
The invention relates to stabilized subtilisin proteases wherein one or more prolines is substituted for a native amino acid at stabilizing positions defined both by the range of the dihedral angles in the primary structure present at the substitution site and by selected characteristics of the protease secondary structure in the vicinity of the substitution site, to nucleotide sequences that encode the stabilized proteases, and to host organisms that contain the nucleotide sequences encoding the stabilized proteases.
Claims19
A substantially pure stabilized subtilisin, in which the amino acid proline is substituted for a naturally occurring amino acid and which position(s) is/are not located in regions, in which the protease is characterized by possessing .alpha.-helical or .beta.-sheet structure.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN K27R+*36D+G97N+A98R+A194P+K235R+K237R+K251E+Y263F.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN K27R+*36D+G97N+A194P+K235R+K237R+K25 E+Y263F.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN K27R+*36D+G97N+A194P+K235R+K237R+Y263F.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN *36D+G97N+V104Y+H120D+A194P+G195E.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN *36D+G97N+v104Y+H120D+A194P+G195E+K235L.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN *36D+G97N+H120D+A194P+G195E.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN *36D+G97N+H120D+Al94P+G195E+K235L.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN *36D+V104Y+H120D+A194P+G195E.
A substantially pure, stabilized, subtilisin 309 having the following amino acid substitutions at positions numbered according to their alignment with the analogous amino acid position in the amino acid sequence of the mature subtilisin BPN *36D+V104Y+H120D+A194P+G195E+K235L.
Dependencies are inferred from the claim text.