US2023183669A1PendingUtilityA1

Protease variants and polynucleotides encoding same

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Assignee: NOVOZYMES ASPriority: Dec 19, 2014Filed: Nov 3, 2022Published: Jun 15, 2023
Est. expiryDec 19, 2034(~8.4 yrs left)· nominal 20-yr term from priority
Inventors:Jens Erik NielsenPernille HedeEsben Peter Friis
C11D 3/386C12N 9/54C12Y 304/21062C12Y 304/21C12N 9/6424
67
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Claims

Abstract

The present invention relates to protease variants and methods for obtaining protease variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Claims

exact text as granted — not AI-modified
1 . A protease variant comprising one or more alterations selected from the group consisting of V2H, S4K, S4H, T5K, Q6K, N16K, N16H, D17N, Q18K, Q18H, S19K, T24K, T24H, T24R, S27K, Y39R, S41K, L43H, G47K, E50Q, E50T, E50L, Q57K, S58K, N59K, D63N, G64H, S65K, T67K, Q70H, G85R, G85H, S86K, N87K, Y92R, Q97K, D108N, D108S, D108A, N109K, S111K, S111R, G112R, S114K, D116S, D116A, A118R, A118K, D126G, D126W, D126K, E127Q, E127K, E127L, E127S, S129K, R130K, T131K, T131H, G132H, G142K, G142R, S143K, S143H, S144K, D147N, D147S, D147*, D147Q, D147H, D147W, S148K, L149K, D155H, Y156R, Y156H, G159H, G174R, G174H, T177K, L184H, V185K, N186K, N186R, E194*, E194S, E194K, E194Q, N195H, N195K, V196H, Q197K, Q197H, Q197R, Q198K, N199K, G200H, D206N, D206H, F207H, N212K, A214R, A214K, T215K, D218N, D218H, D218R, Y219R, Y219H, I220K, I220H, Q222K, D225N, A233R, S234K, E236C, T241K, N245K, T246K, I247K, S248K, I264R, A267H, N268K, N268H, T269K, T269H, S270K, S272K, S274K, T278K, T278H, E279Q, E279H, V286H, V286K, Y287R, Y287H, D288N, D288K, I293R, I293K, T297K, D299R and K311R, wherein each position corresponds to SEQ ID NO: 3, and wherein the protease variant has at least 60% sequence identity to SEQ ID NO: 3. 
     
     
         2 . A protease variant of  claim 1 , which has improved wash performance compared to the parent or compared to the protease with SEQ ID NO: 3. 
     
     
         3 . A protease variant of  claim 1 , wherein the variant is selected from the group consisting of:
 a) a polypeptide having at least 60% sequence identity to the mature polypeptide of SEQ ID NO: 2;   b) a polypeptide encoded by a polynucleotide that hybridizes under medium, or high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) a sequence encoding the mature polypeptide of SEQ ID NO: 2, or (iii) the full-length complement of (i) or (ii);   c) a polypeptide encoded by a polynucleotide having at least 75% identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or a sequence encoding the mature polypeptide of SEQ ID NO: 2;   d) a fragment of the mature polypeptide of SEQ ID NO: 2, which has protease activity; and   e) a protease variant having at least 60% identity to SEQ ID NO 3, and comprising an alteration selected from the list consisting of:
 V2H, S4K, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, S173P, S175P; 
 S4K, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, S27K, I137E, S173P, S175P, F180Y, T297P; 
 A1*, V2*, P3*, S4K, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, S27K, N87K, E127Q, G132H, S173P, S175P, F180Y, Q198E, T297P; 
 S4H, S173P, S175P; 
 T5K, S173P, S175P; 
 Q6K, S173P, S175P; 
 N16K, S173P, S175P; 
 N16K, D17N, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S27K, I137E, S173P, S175P, F180Y, T297P; 
 D17N, S173P, S175P; 
 D17N, I137E, S173P, S175P, F180Y, T297P; 
 D17N, S173P, S175P, Q198E; 
 D17N, S173P, S175P, F180Y, Q198E, T297P; 
 D17N, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 D17N, S27K, I137E, S173P, S175P, F180Y, T297P; 
 Q18K, S173P, S175P; 
 Q18K, S27K, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 S19K, S173P, S175P; 
 T24K, S173P, S175P; 
 T24H, S173P, S175P; 
 S27K, S173P, S175P; 
 S27K, I137E, S173P, S175P, F180Y, T297P; 
 S27K, S173P, S175P, Q198E; 
 S27K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, S144R, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, D147N, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E50Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N87K, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, S173P, S175P, F180Y, Q198E, S270K, T297P; 
 S27K, Q97K, S173P, S175P, F180Y, Q198E, T297P; 
 D17N, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 N16H, S27K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, I137E, S144R, S173P, S175P, F180Y, T297P; 
 S27K, E127Q, I137E, S173P, S175P, F180Y, T297P; 
 S27K, I137E, S173P, G174R, S175P, F180Y, T297P; 
 S27K, Q97K, I137E, S173P, S175P, F180Y, T297P; 
 D17N, S27K, I137E, S173P, S175P, F180Y, T297P; 
 S27K, D126G, S173P, S175P, F180Y, T297P; 
 Y39R, S173P, S175P; 
 S27K, Y39R, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, Y39R, I137E, S173P, S175P, F180Y, T297P; 
 S27K, Y39R, N87K, I137E, S173P, S175P, F180Y, T297P; 
 S27K, Y39R, I137E, S144R, S173P, S175P, F180Y, T297P; 
 S27K, Y39R, I121V, S171N, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S41K, S173P, S175P; 
 S27K, S41K, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 L43H, S173P, S175P; 
 G47K, R130K, G132R, S173P, S175P; 
 E50Q, S173P, S175P; 
 E50Q, S173P, S175P, F180Y; 
 E50Q, S173P, S175P, Q198E; 
 E50Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E50Q, S173P, S175P, F180Y, Q198E, T297P; 
 E50T, S173P, S175P, F180Y; 
 E50L, S173P, S175P, F180Y; 
 G28N, Q57K, S86K, N87R, Q89R, S173P, S175P; 
 S58K, S173P, S175P; 
 S58K, S173P, S175P, F180Y, Q198E, T297P; 
 N59K, S173P, S175P; 
 D63N, S173P, S175P; 
 D63E, R69D, S173P, S175P; 
 G64H, S173P, S175P; 
 S65K, S173P, S175P; 
 S65K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, S65K, S173P, S175P, F180Y, T297P; 
 S27K, S65K, S173P, S175P, F180Y, T297P; 
 T67K, S173P, S175P; 
 Q70H, S173P, S175P, F180Y; 
 G85R, S173P, S175P, F180Y; 
 G85H, S173P, S175P, F180Y; 
 S86K, S173P, S175P; 
 N87K, S173P, S175P; 
 N87K, I137E, S173P, S175P, F180Y, T297P; 
 N87K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N87K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N87K, I137E, S173P, S175P, F180Y, T297P; 
 S27K, N87K, N109S, S111R, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N59L, T67V, N87K, G107N, I121V, S171N, S173P, G174R, S175P, F180Y, Q198E, T297P; 
   
       S27K, N59L, T67V, N87K, G107N, I121V, E127Q, S171N, S173P, G174R, S175P, F180Y, Q198E, T297P; 
       S27K, Q70N, N87K, I121V, E127Q, S171N, S173P, G174R, S175P, F180Y, Q198E, N199R, T297P;
 Y92R, S173P, S175P, F180Y; 
 Q97K, S173P, S175P; 
 Q97K, I137E, S173P, S175P, F180Y, T297P; 
 Q97K, S173P, S175P, Q198E; 
 Q97K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, Q97K, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, Q97K, I137E, S173P, S175P, F180Y, T297P; 
 S27K, N59L, T67V, Q97K, G107N, I121V, S171N, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 D108N, S173P, S175P; 
 D108S, S173P, S175P; 
 D108A, S173P, S175P; 
 S27K, N109K, S173P, S175P, F180Y, Q198E, T297P; 
 S111K, S173P, S175P; 
 S27K, N109S, S111R, S173P, S175P, F180Y, Q198E, T297P; 
 S114K, S173P, S175P, F180Y; 
 D116S, M139L, S173P, S175P; 
 A118R, S173P, S175P, F180Y; 
 S27K, S111R, A118R, S173P, S175P, F180Y, Q198E, T297P; 
 A118K, S173P, S175P, F180Y; 
 D126G, S173P, S175P, F180Y; 
 S27K, D126G, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, D126G, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 D126W, S173P, S175P, F180Y; 
 D126K, S173P, S175P, F180Y; 
 Y113T, E127Q, S173P, G174K, S175P, F180Y; 
 *109aS, E127Q, S173P, S175P, F180Y, T297P; 
 E127Q, S173P, S175P, Q198E; 
 E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, D126G, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, I137Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, S144R, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, V162S, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, V162E, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, V162Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, V162G, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, V162F, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, V162T, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, E127Q, V162Y, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N109S, S111R, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N109S, S111R, E127Q, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N59L, T67V, N87K, G107N, I121V, E127Q, S171N, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, N87K, I121V, E127Q, S171N, S173P, S175P, F180Y, T297P; 
 E127K, S173P, S175P, F180Y; 
 E127L, S173P, S175P, F180Y; 
 S129K, S173P, S175P; 
 S41K, L43K, G47K, R130K, G132R, S173P, S175P; 
 S41R, L43R, G47K, R130K, G132R, S173P, S175P; 
 T131K, S173P, S175P; 
 G132H, S173P, S175P; 
 G132H, S173P, S175P, F180Y, Q198E, T297P; 
 S143K, S173P, S175P; 
 S144K, S173P, S175P; 
 D147N, S173P, S175P; 
 D147N, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, D147N, S173P, S175P, F180Y, Q198E, T297P; 
 S27K, N109S, S111R, D147N, S173P, S175P, F180Y, Q198E, T297P; 
 D147S, S148A, L149 
 A145S, K146P, D147S, S173P, S175P; 
 A145S, K146P, D147S, S148A, S173P, S175P; 
 S144P, A145S, K146P, D147S, S148A, L149 
 D147S, L149T, S173P, S175P; 
 K146P, D147S, S148A, L149 
 A145S, K146P, D147S, S148A, L149T, S173P, S175P; 
 S27K, K146E, D147S, S171N, S173P, G174R, S175P, F180Y, G182A, Q198E, N199K, E223K, T2 97P; 
 K146*, D147*, S173P, S175P; 
 D147Q, S173P, S175P; 
 D147Q, S173P, S175P, F180Y, Q198E, T297P; 
 D147H, S173P, S175P; 
 S148K, S173P, S175P; 
 L149K, S173P, S175P; 
 D155H, S173P, S175P; 
 Y156R, S173P, S175P; 
 Y156R, S173P, S175P; 
 Y156R, S173P, S175P; 
 Y156R, S173P, S175P, Q198E; 
 Y156R, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, D17N, S27K, N59L, N87K, Y156R, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, D17N, S27K, N59L, N87K, Y156R, S173P, S175P, F180Y, Q198E, T297P; 
 Y156H, S173P, S175P; 
 G159H, S173P, S175P; 
 S27K, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, I137E, S173P, G174R, S175P, F180Y, T297P; 
 S27K, N59L, T67V, N87K, S114V, I121V, I150L, S171N, S173P, G174R, S175P, F180Y, T297P; 
 S173P, S175P, T177K; 
 S173P, S175P, F180Y, L184H; 
 I137E, S173P, S175P, F180Y, V185K, T297P; 
 I137E, S173P, S175P, F180Y, V185K, N186P, T297P; 
 I137E, S173P, S175P, F180Y, V185K, N186D, T297P; 
 I137E, S173P, S175P, F180Y, N186K, T297P; 
 S173P, S175P, N186K; 
 I137E, S173P, S175P, F180Y, N186R, T297P; 
 S173P, S175P, E194*; 
 S173P, S175P, E194*, Q198E; 
 S173P, S175P, F180Y, E194*, Q198E, T297P; 
 S173P, S175P, F180Y, E194S; 
 S27K, E127Q, S173P, S175P, F180Y, E194S, Q198E, T297P; 
 S173P, S175P, F180Y, E194K; 
 S27K, E127Q, S173P, S175P, F180Y, E194K, Q198E, T297P; 
 S27K, E127Q, S173P, S175P, F180Y, E194Q, Q198E, T297P; 
 S173P, S175P, N195H; 
 S27K, E127Q, S173P, S175P, F180Y, N195H, V196N, Q197*, Q198*, N199*, G200*, T201*, T297 P; 
 S173P, S175P, N195K; 
 S173P, S175P, V196H; 
 S173P, S175P, Q197K; 
 S27K, S173P, S175P, F180Y, Q197K, T297P; 
 S27K, S173P, S175P, F180Y, E194R, Q197K, Q198E, T297P; 
 S27K, S173P, S175P, F180Y, Q197K, Q198E, T297P; 
 S27K, E127Q, S173P, S175P, F180Y, Q197K, Q198E, T297P; 
 S27K, S171N, S173P, G174R, S175P, F180Y, Q197K, Q198E, T297P; 
 S27K, N109S, S111R, S173P, S175P, F180Y, Q197K, Q198E, T297P; 
 S27K, S173P, G174K, S175P, F180Y, L184F, Q197K, Q198E, T297P; 
 I121V, S173P, S175P, F180Y, Q197K, Q198E, T297P; 
 S173P, S175P, Q197H; 
 S173P, S175P, F180Y, Q197R; 
 S27K, S171N, S173P, G174R, S175P, F180Y, Q197R, Q198E, T297P; 
 S173P, S175P, Q198K; 
 S173P, S175P, Q198K; 
 S173P, S175P, N199K; 
 S173P, S175P, N199K; 
 S27K, S173P, S175P, F180Y, E194*, Q197K, Q198E, N199K, T297P; 
 S27K, E127Q, S173P, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, E127Q, S173P, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, I121V, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, N109S, S111R, S173P, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, S173P, S175P, F180Y, Q197K, Q198E, N199K, T297P; 
 S27K, Q70N, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, I150S, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, S114V, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, Q70N, G107N, I121V, E127Q, S173P, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, Q70A, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, Q70V, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, S173P, G174K, S175P, F180Y, L184F, Q198E, N199K, T297P; 
 S27K, S171D, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, S171R, S173P, G174E, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, S171K, S173P, G174E, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, S171E, S173P, G174K, S175P, F180Y, Q198E, N199K, T297P; 
 S173P, S175P, G200H; 
 S173P, S175P, D206N; 
 S173P, S175P, N212K; 
 S173P, S175P, F180Y, A214R; 
 S173P, S175P, F180Y, A214K; 
 S173P, S175P, T215K; 
 S173P, S175P, D218N; 
 S173P, S175P, D218H; 
 S173P, S175P, F180Y, D218R; 
 S173P, S175P, Y219R; 
 S173P, S175P, Y219H; 
 S173P, S175P, I220K; 
 S173P, S175P, Q222K; 
 S27K, Q97K, E127Q, S173P, S175P, F180Y, Q198E, D225N, I247M, T297P; 
 S173P, S175P, F180Y, A233R; 
 S173P, S175P, F180Y, A233R; 
 S173P, S175P, F180Y, A233R; 
 S173P, S175P, F180Y, A233R; 
 S173P, S175P, F180Y, S234K; 
 T5C, S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, E236C, T297P; 
 Q6C, S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, E236C, T297P; 
 S4C, S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, E236C, T297P; 
 A1*, T5C, S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, E236C, T297P; 
 A1*, Q6C, S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, E236C, T297P; 
 A1*, S4C, S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, E236C, T297P; 
 A1*, P3*, T5C, S27K, S171N, S173P, G174R, S175P, F180Y, Q198E, E236C, T297P; 
 S173P, S175P, T241K; 
 S173P, S175P, N245K; 
 S173P, S175P, T246K; 
 S173P, S175P, S248K; 
 S173P, S175P, F180Y, I264R; 
 S173P, S175P, F180Y, I264R; 
 S173P, S175P, F180Y, I264R; 
 S173P, S175P, F180Y, I264R; 
 S173P, S175P, A267H; 
 S173P, S175P, N268K; 
 S27K, E127Q, S173P, S175P, F180Y, Q198E, N268K, T297P; 
 S173P, S175P, T269K; 
 S173P, S175P, S270K; 
 S173P, S175P, S270K; 
 S173P, S175P, F180Y, Q198E, S270K, T297P; 
 S27K, S173P, S175P, F180Y, Q198E, S270K, T297P; 
 S27K, I137E, S173P, S175P, F180Y, S270K, T297P; 
 S27K, N87K, I121V, S171N, S173P, S175P, F180Y, S270K, T297P; 
 S27K, S173P, S175P, F180Y, S270K, T297P; 
 S27K, S173P, S175P, F180Y, S270K, T297P; 
 S173P, S175P, S272K; 
 S173P, S175P, S274K; 
 S173P, S175P, T278K; 
 S173P, S175P, E279Q; 
 S173P, S175P, E279H; 
 S173P, S175P, V286H; 
 I137E, S173P, S175P, F180Y, V286K, T297P; 
 I137E, S173P, S175P, F180Y, V286K, T297P; 
 S173P, S175P, Y287R; 
 S173P, S175P, Y287H; 
 I137E, S173P, S175P, F180Y, D288N, T297P; 
 S173P, S175P, F180Y, Q198E, D288N, T297P; 
 I137E, S173P, S175P, F180Y, D288K, T297P; 
 S27K, S173P, S175P, F180Y, Q198E, I293R, T297P; 
 S27K, I137E, S173P, S175P, F180Y, I293R, T297P; 
 S173P, S175P, I293K; 
 S173P, S175P, T297K; 
 G28D, S86K, N87R, Q89R, S173P, S175P, K311R; 
 G28N, S86K, N87R, Q89R, S173P, S175P, K311R; 
 E127S, S173P, S175P, F180Y; 
 S173P, S175P, F180Y; 
 S173P, S175P, F180Y, D299R; 
 S27K, D147N, I150S, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q197K, Q198E, S274V, T297P; 
 S27K, D147N, I150S, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S27K, D147N, I150S, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, R224G, T297P; 
 S27K, D147S, I150S, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S27K, G142E, D147N, I150S, S171N, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, G142E, I150S, S171N, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, I150L, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, T297P; 
 S27K, I150L, S171N, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, I150S, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, I150S, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S27K, I150S, S171N, S173P, G174R, S175P, F180Y, Q198E, N199K, R224G, T297P; 
 S27K, N109K, S111D, S171D, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, N109K, S111D, S171E, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, N109K, S111E, S171D, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, N109K, S111E, S171E, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, N109K, S171D, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, N109S, *109aE, S111R, S171E, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, N109S, *109aE, S171E, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, N109S, I121V, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, N109S, I121V, S171 D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S27K, N87K, G132H, V162T, S171 D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S27K, N87K, S171 D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, T297P; 
 S27K, N87K, S171 D, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, N87K, S171N, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, N87K, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, V286R, T297P; 
 S27K, N87K, V162T, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S27K, Q70A, D147N, S171 D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, Q70A, N109K, S111E, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, Q70A, S111E, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, Q70A, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, Q70A, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S27K, S111 D, S171D, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, S111 D, S171E, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, S111E, S171 D, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, S111E, S171E, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, N199K, T297P; 
 S27K, S111R, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S270V, S274V, L280V, T297P; 
 S27K, S114V, I121V, S129K, S173P, G174K, S175P, F180Y, L184F, Q198E, N199K, T297P; 
 S27K, S114V, I121V, S173P, G174K, S175P, F180Y, L184F, Q198E, N199K, T297P; 
 S27K, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S270V, S274V, L280V, T297P; 
 S27K, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, T297P; 
 S27K, S171D, S173P, G174R, S175P, F180Y, Q197K, Q198E, T297P; 
 S27K, S171D, S173P, G174R, S175P, F180Y, Q198E, T297P; 
 S27K, S171N, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, N199K, S274V, T297P; 
 S27K, S173P, G174K, S175P, F180Y, G182A, L184F, Q198E, V286R, T297P; 
 S27K, V162T, S171D, S173P, G174R, S175P, F180Y, G182A, L184F, Q198E, S274V, T297P; 
 S4K, S27K, E127Q, G132H, Y156R, S173P, S175P, F180Y, Q198E, T297P; 
 S4K, S27K, N87K, E127Q, G132H, Y156R, S173P, S175P, F180Y, Q198E, T297P and 
 T24R, S173P, S175P, F180Y 
 
     
     
         4 . The protease variant of  claim 1 , wherein the variant has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99 sequence identity to SEQ ID NO: 3. 
     
     
         5 . The protease variant of  claim 1 , wherein the total number of alterations compared to SEQ ID NO: 3 is 1-20, e.g. 1-10 and 1-5, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 alterations. 
     
     
         6 . A detergent composition comprising a protease variant according to  claim 1 . 
     
     
         7 . The detergent composition of  claim 6 , further comprising one or more detergent components. 
     
     
         8 . The detergent composition according to  claim 6 , further comprising one or more additional enzymes selected from the group comprising of proteases, amylases, lipases, cutinases, cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases, peroxidaes, haloperoxygenases, catalases and mannanases, or any mixture thereof. 
     
     
         9 . The detergent composition according to  claim 6  in form of a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid. 
     
     
         10 . Use of a detergent composition according to  claim 6  in a cleaning process, such as laundry or hard surface cleaning such as dish wash. 
     
     
         11 . A method for obtaining a protease variant, comprising introducing into a parent protease one or more of the following alterations: V2H, S4K, S4H, T5K, Q6K, N16K, N16H, D17N, Q18K, Q18H, S19K, T24K, T24H, T24R, S27K, Y39R, S41K, L43H, G47K, E50Q, E50T, E50L, Q57K, S58K, N59K, D63N, G64H, S65K, T67K, Q70H, G85R, G85H, S86K, N87K, Y92R, Q97K, D108N, D108S, D108A, N109K, S111K, S111R, G112R, S114K, D116S, D116A, A118R, A118K, D126G, D126W, D126K, E127Q, E127K, E127L, E127S, S129K, R130K, T131K, T131H, G132H, G142K, G142R, S143K, S143H, S144K, D147N, D147S, D147*, D147Q, D147H, D147W, S148K, L149K, D155H, Y156R, Y156H, G159H, G174R, G174H, T177K, L184H, V185K, N186K, N186R, E194*, E194S, E194K, E194Q, N195H, N195K, V196H, Q197K, Q197H, Q197R, Q198K, N199K, G200H, D206N, D206H, F207H, N212K, A214R, A214K, T215K, D218N, D218H, D218R, Y219R, Y219H, I220K, I220H, Q222K, D225N, A233R, S234K, E236C, T241K, N245K, T246K, I247K, S248K, I264R, A267H, N268K, N268H, T269K, T269H, S270K, S272K, S274K, T278K, T278H, E279Q, E279H, V286H, V286K, Y287R, Y287H, D288N, D288K, I293R, I293K, T297K, D299R and K311R of SEQ ID NO 3, wherein the variant has an amino acid sequence which is at least 60% identical to SEQ ID NO: 3, and recovering the variant. 
     
     
         12 . The method of  claim 11 , wherein the variant comprises two, three, four or five substitutions corresponding to positions 2, 4, 5, 6, 16, 17, 18, 19, 24, 27, 39, 41, 43, 47, 50, 57, 58, 59, 63, 64, 65, 67, 70, 85, 86, 87, 92, 97, 108, 109, 111, 112, 114, 116, 118, 126, 127, 129, 130, 131, 132, 142, 143, 144, 147, 148, 149, 155, 156, 174, 177, 184, 185, 186, 194, 195, 196, 197, 198, 199, 200, 206, 207, 212, 214, 215, 218, 219, 220, 222, 225, 234, 236, 241, 245, 246, 247, 248, 264, 267, 268, 269, 270, 272, 274, 278, 279, 286, 287, 288, 293, 297, 299 or 311 of the polypeptide of SEQ ID NO: 3. 
     
     
         13 . The method according to  claim 11 , wherein the protease variant has at least 75%, such as at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the mature polypeptide of SEQ ID NO: 3. 
     
     
         14 . The method according to  claim 11 , wherein the parent protease selected from the group consists of:
 a) a polypeptide having at least 60% sequence identity to the mature polypeptide of SEQ ID NO: 2;   b) a polypeptide encoded by a polynucleotide that hybridizes under medium or high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) a sequence encoding the mature polypeptide of SEQ ID NO: 2, or (iii) the full-length complement of (i) or (ii);   c) a polypeptide encoded by a polynucleotide having at least 60% identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or a sequence encoding the mature polypeptide of SEQ ID NO: 2; and   d) a fragment of the mature polypeptide of SEQ ID NO: 2, which has protease activity.   
     
     
         15 . The method according to  claim 11 , wherein the parent protease has at least 75%, such as at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, sequence identity to SEQ ID NO: 3.

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