Subtilisin variants and methods of use
AssigneeDanisco US Inc.
InventorsViktor Yuryevich Alekseyev, Lilia Babe, Lydia Dankmeyer, Frits Goedegebuur, Roopa Ghirnikar, Thijs Kaper, Harm Mulder, Nils Henning Redestig, Sander Van Stigt Thans
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Abstract
Disclosed herein is one or more subtilisin variant, nucleic acid encoding same, and compositions and methods related to the production and use thereof, including one or more subtilisin variant that has improved stability and/or soil removal compared to one or more reference subtilisin.
Claims18
A subtilisin variant comprising one, two, three, four, or five amino acid substitutions selected from the group consisting of X039E, X099R, X126A, X127E, and X128G and further comprises one or more additional substitutions selected from the group consisting of X024R, X037V, and X144G, wherein the variant has at least 80% identity to the amino acid sequence of SEQ ID NO:
- 1 or 8, and the amino acid positions are numbered according to SEQ ID NO:
- 1.
A subtilisin variant comprising
- (i) one, two, three, four, or five amino acid substitutions selected from the group consisting of X039E, X099R, X126A, X127E, and X128G,
- (ii) one or more additional substitutions selected from the group consisting of X024R, X037V, and X144G, and
- (iii) one or more amino acid substitutions selected from the group consisting of X074D, X114L X198A, X198G, X207Q, X211Q, X211E, X212Q, X242D, X256E;
- and
- wherein the variant has at least 80% identity to the amino acid sequence of SEQ ID NO:
- 1 or 8, and the amino acid positions are numbered according to SEQ ID NO;
- and the amino acid positions are numbered according to SEQ ID NO:
- 1.
The subtilisin variant of any preceding claim, wherein the variant comprises the substitutions:
- A037V-S039E-N074D-S099R-S126A-D127E-F128G-N198G-M211Q-N212Q;
- A037V-S039E-N074D-S099R-F128G-N198A;
- A037V-S039E-S099R-F128G;
- A037V-S039E-S099R-F128G-D144G-M211Q;
- A037V-S039E-S099R-F128G-D144G-R207Q-M211Q;
- A037V-S039E-S099R-F128G-N198A-M211Q-Q256E;
- A037V-S039E-S099R-F128G-N198G-M211E;
- A037V-S039E-S099R-F128G-N198G-Q256E;
- A037V-S039E-S099R-S126A-D127E;
- A037V-S039E-S099R-S126A-D127E-F128G-M211Q-N242D;
- A037V-S039E-S099R-S126A-D127E-F128G-N198G;
- A037V-S039E-S099R-S126A-D127E-R207Q-N212Q;
- A037V-S039E-S099R-S126A-F128G;
- A037V-S039E-S099R-S126A-F128G-D144G;
- A037V-S039E-S099R-S126A-F128G-N198G-R207Q;
- A037V-S039E-S099R-S126A-F128G-N212Q-Q256E;
- A037V-S039E-S099R-T114L-F128G-D144G;
- A037V-S039E-S099R-T114L-F128G-D144G-N242D;
- A037V-S039E-S099R-T114L-F128G-N198A-M211Q;
- A037V-S039E-S099R-T114L-F128G-R207Q-N212Q;
- A037V-S039E-S099R-T114L-S126A-F128G-Q256E;
- A037V-S099R-S126A-D127E-M211E;
- S024R-A037V-S039E-N074D-S099R-S126A-F128G;
- S024R-A037V-S039E-N074D-S099R-T114L-S126A-D127E-F128G-M211Q-N242D;
- S024R-A037V-S039E-N074D-S099R-T114L-S126A-D127E-F128G-N198G-M211Q;
- S024R-A037V-S039E-S099R-F128G;
- S024R-A037V-S039E-S099R-F128G-D144G;
- S024R-A037V-S039E-S099R-F128G-M211E;
- S024R-A037V-S039E-S099R-F128G-M211E-N242D;
- S024R-A037V-S039E-S099R-F128G-N198A-M211E;
- S024R-A037V-S039E-S099R-S126A-D127E-F128G-N198G-R207Q-M211Q-N242D;
- S024R-A037V-S039E-S099R-S126A-F128G;
- S024R-A037V-S039E-S099R-T114L-S126A-D127E-F128G-M211Q-N212Q;
- S024R-A037V-S039E-S099R-T114L-S126A-D127E-F128G-N198G-M211Q;
- S024R-A037V-S039E-S099R-T114L-S126A-D127E-F128G-R207Q-M211Q;
- S024R-S039E-N074D-S099R-F128G-M211Q;
- S024R-S039E-N074D-S099R-S126A-D127E-F128G-D144G-N198A-M211Q-N212Q;
- S024R-S039E-N074D-S099R-S126A-D127E-F128G-D144G-N198A-M211Q-N212Q-N242D;
- S024R-S039E-N074D-S099R-S126A-D127E-F128G-M211Q;
- S024R-S039E-N074D-S099R-S126A-D127E-F128G-N198A-M211Q-N212Q;
- S024R-S039E-S099R-D127E-Q256E;
- S024R-S039E-S099R-F128G;
- S024R-S039E-S099R-F128G-D144G-M211Q;
- S024R-S039E-S099R-F128G-D144G-N198A-Q256E;
- S024R-S039E-S099R-F128G-D144G-N198G-R207Q;
- S024R-S039E-S099R-F128G-D144G-N212Q-Q256E;
- S024R-S039E-S099R-F128G-M211E-N242D;
- S024R-S039E-S099R-F128G-N198G-M211Q;
- S024R-S039E-S099R-F128G-R207Q-M211Q;
- S024R-S039E-S099R-F128G-R207Q-N242D;
- S024R-S039E-S099R-S126A-D127E-F128G-M211Q-Q256E;
- S024R-S039E-S099R-S126A-F128G;
- S024R-S039E-S099R-T114L-D127E;
- S024R-S039E-S099R-T114L-S126A-D127E-F128G-N198G-M211Q;
- S024R-S039E-S099R-T114L-S126A-F128G;
- S024R-S099R-S126A-D127E-D144G;
- S024R-S099R-S126A-D127E-F128G-N198A-M211Q-N212Q;
- S024R-S099R-S126A-D127E-N198G-M211E;
- S039E-N074D-S099R-F128G-D144G-R207Q-M211Q;
- S039E-N074D-S099R-T114L-F128G-D144G-N198A;
- S039E-N074D-S099R-T114L-F128G-D144G-Q256E;
- S039E-S099R-D127E-D144G-M211Q-N212Q-N242D;
- S039E-S099R-D127E-D144G-N198G-R207Q-N212Q;
- S039E-S099R-F128G-D144G;
- S039E-S099R-F128G-D144G-M211Q-Q256E;
- S039E-S099R-F128G-D144G-N198A-R207Q;
- S039E-S099R-F128G-D144G-N198A-R207Q-N212Q;
- S039E-S099R-F128G-D144G-N198G-M211Q;
- S039E-S099R-S126A-F128G-D144G;
- S039E-S099R-T114L-D127E-D144G-M211E-N212Q;
- S039E-S099R-T 114L-F128G-D144G-N242D;
- and S099R-S126A-D127E-F128G-N198A-M211E-N212Q wherein the variant has at least 80% identity to amino acid sequence of SEQ ID NO:
- 1 or 8.
The subtilisin variant according to any of the preceding claims, wherein said variant
- (i) is derived from a Bacillus gibsonii (Bgi02446/BG46, DSM14391) subtilisin;
- (ii) has proteolytic activity;
- or
- (iii) comprises a combination of (i) and (ii).
The subtilisin variant according to any preceding claim, wherein said variant is derived from a parent or reference polypeptide with 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to SEQ ID NO:
- 1 or 8.
The subtilisin variant of any preceding claim, wherein said variant comprises an amino acid sequence with 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or less than 100% amino acid sequence identity to SEQ ID NO:
- 1 or 8.
The subtilisin variant of any preceding claim, wherein the parent subtilisin comprises a polypeptide having the amino acid sequence of SEQ ID NO:
- 1 or 8.
The subtilisin variant of any preceding claim, wherein said variant has one or more improved property when compared to a parent or reference subtilisin;
- wherein the improved property is selected from improved cleaning performance in detergent, improved stability;
- and combinations thereof.
An enzyme composition comprising one or more subtilisin variant according to any preceding claim.
Dependencies are inferred from the claim text.