US2013203124A1PendingUtilityA1
Alpha-Amylase Variant With Altered Properties

Assignee: NOVOZYMES ASPriority: May 15, 2001Filed: Nov 21, 2012Published: Aug 8, 2013
Est. expiryMay 15, 2021(expired)· nominal 20-yr term from priority
Inventors:Allan Svendsen Carsten Andersen Thomas Thisted Claus Von Der Osten
C12P 19/14C12P 7/14C12N 9/2417C12Y 302/01001C11D 3/386C12P 7/06C11D 3/38618Y02E50/10
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References
Claims
Abstract

The present invention relates to variants (mutants) of parent Termamyl-like alpha-amylases, which variant has alpha-amylase activity and exhibits altered properties relative to the parent alpha-amylase.
Claims

exact text as granted — not AI-modified
1 . A variant of a parent Termamyl-like alpha-amylase, comprising an alteration at one or more positions selected from the group of:
 5, 6, 36, 37, 38, 39, 42, 45, 47, 63, 66, 69, 70, 71, 72, 74, 75, 76, 79, 82, 83, 86, 87, 89, 93, 112, 113, 117, 120, 137, 213, 216, 220, 223, 225, 226, 227, 229, 243, 245, 279, 282, 311, 321, 324, 352, 353, 354, 357, 361, 362, 364, 368, 390, 395, 397, 399, 400, 401, 425, 451, 452, 453, 466, 468, 470, 471, 478,   wherein   (a) the alteration(s) are independently
 (i) an insertion of an amino acid downstream of the amino acid which occupies the position, 
 (ii) a deletion of the amino acid which occupies the position, or 
 (iii) a substitution of the amino acid which occupies the position with a different amino acid, 
   (b) the variant has alpha-amylase activity and (c) each position corresponds to a position of the amino acid sequence of the parent Termamyl-like alpha-amylase having the amino acid sequence shown in SEQ ID NO: 8 ( Bacillus licheniformis  alpha-amylase).   
     
     
         2 . A variant of a parent Termamyl-like alpha-amylase, comprising one or more of the following substitutions:
 X1A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y;   X2R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V;   X3A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y;   X4A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X13R,N,D,C,Q,E,G,H,K,M,P,S,T,W;   X14A,R,D,C,G,K,M,P,W;   X16R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V;   X17A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X18A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X20A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X23A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,W,Y,V;   X24A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X26A,D,C,E,G,H,I,L,M,F,P,S,T,W,V;   X34A,R,N,C,Q,E,G,H,I,L,K,M,F,P,T,W,Y,V;   X35A,R,N,D,C,Q,E,G,H,K,M,F,P,S,T,W,Y,V;   X49A,C,G,H,P,T;   X50A,R,N,C,Q,E,G,H,K,M,F,P,S,W;   X51A,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X52A,R,D,C,Q,E,G,H,K,P;   X53A,D,C,G,H,K,M,P;   X61A,R,N,D,C,Q,E,G,H,I,L,K,M,P,S,T,Y;   X62A,R,D,C,G,K,M,P,Y;   X67A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,Y,V;   X68A,R,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X73A,R,N,D,C,Q,E,G,H,K,M,P,S,T,W,Y,V;   X84A,R,N,D,C,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X85A,R,N,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X88A,R,N,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X91A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X92A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X96A,R,N,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X106A,N,D,C,Q,E,G,H,I,L,K,M,P,S,T,Y,V;   X108R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X114A,N,C,Q,E,G,H,K,F,P,S,T,W,Y;   X116A,R,D,C,Q,E,G,H,I,L,M,F,P,S,W,Y,V;   X119A,R,N,D,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X121A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X122R,N,Q,G,H,I,L,M,F,S,T,W,Y,V;   X123N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X124N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X125R,N,Q,E,G,I,K,M,F,S,T,W,Y;   X126N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X127A,R,N,D,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X128A,R,N,D,C,Q,G,H,I,L,K,M,F,P,S,W,Y,V;   X129A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,W,Y,V;   X130A,R,N,D,Q,E,G,H,I,L,K,M,F,P,S,W,Y,V;   X131A,R,N,D,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X132R,N,D,C,Q,E,G,H,I,L,K,M,F,S,W,Y;   X133R,N,D,C,M,T,W,V;   X134A,N,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X136A,R,N,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X138R,N,D,Q,E,G,I,K,M,P,S,T,W,V;   X145A,R,N,D,C,Q,E,G,H,I,L,K,M,P,S,T,Y,V;   X147A,R,N,D,C,Q,E,G,H,I,L,K,M,P,S,T,W,Y,V;   X148A,R,D,C,Q,E,G,H,I,L,K,M,F,P,T,W,Y,V;   X149A,R,N,D,C,Q,E,G,H,L,K,M,F,P,S,T,W,Y,V;   X150A,D,C,G,M,P,W,Y;   X152A,R,N,C,Q,E,G,H,I,L,K,M,F,P,T,W,Y,V;   X153A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X154A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X155A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X156A,C,Q,E,G,I,L,M,F,P,S,T,W,V;   X157R,I,L,M,F,P,S,T,W,Y,V;   X158R,M,P,W,Y;   X164R,I,L,M,F,P,S,T,W,Y,V;   X165A,N,D,C,Q,E,H,I,L,K,M,F,P,S,T,W,Y,V;   X167A,R,N,D,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X168A,R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,V;   X169A,R,N,D,C,Q,E,G,H,M,P,S,W,Y,V;   X170A,R,N,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X171A,R,N,D,C,Q,E,G,H,K,M,P,S,T,W,Y,V;   X172A,N,D,C,Q,E,G,I,L,M,F,P,T,W,Y,V;   X173A,N,D,C,Q,E,G,H,M,P,S,W,Y,V;   X176A,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X179D,C,Q,E,H,I,L,K,M,F,P,W,Y,V;   X180A,G,I,L,M,F,P,W,Y,V;   X181G;   X182A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X184I,L,M,F,P,W,Y,V;   X185R,I,L,M,F,P,S,T,W,Y,V;   X188A,R,N,Q,G,H,L,M,F,W,V;   X189A,R,N,G,H,I,L,M,F,P,S,T,W,Y,V;   X190N;   X191A,R,N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X193A,R,G,M,P,W,Y;   X196A,N,Q,G,H,I,L,M,P,S,T,W,V;   X198A,R,G,M,P,W;   X204R,L,M,F,P,T,W,Y,V;   X205A,G,H,I,L,M,F,P,W,Y,V;   X206R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X209R,P,S,W,Y;   X210A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,W,Y,V;   X211E;   X214A,D,C,Q,E,G,I,L,M,F,P,S,T,Y,V;   X217A,R,N,D,C,Q,G,H,I,L,M,F,P,S,T,W,Y;   X218A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X221A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X222A,R,N,D,C,E,G,H,I,L,K,M,F,P,S,W,Y,V;   X234A,D,C,G,H,I,K,M,F,P,S,T,W,Y,V;   X235A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X237A,G,H,I,L,M,F,W,Y,V;   X239G,H,I,L,M,F,P,S,T,Y,V;   X242G,I,L,M,F,S,T,W,Y,V;   X246A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X247R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,V;   X249A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X250A,R,N,D,C,E,H,I,L,K,M,P,T,W,Y,V;   X251 R,N,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X252A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,W,Y,V;   X253R,D,C,Q,E,H,I,L,K,M,F,P,S,T,W,Y;   X254A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X255A,R,D,C,G,H,I,L,K,M,F,S,T,W,Y,V;   X257A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X261A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X263A,R,N,D,C,Q,E,G,I,L,K,M,F,P,S,T,W,Y,V;   X265C,Q,E,H,I,L,M,F,P,W;   X266A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X267A,R,N,D,C,Q,E,G,H,K,P,S,T,W,Y,V;   X268A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X269A,N,C,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X271A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X272A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X275A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,W,Y,V;   X276A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X278A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X280A,R,D,C,E,G,H,I,L,K,M,F,P,W,Y,V;   X290W,Y;   X291A,R,N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X293A,R,N,G,I,L,M,P,S,T,W,V;   X294R,N,Q,G,H,I,L,M,F,P,S,T,W,Y;   X297A,G,H,I,L,M,F,P,W,Y,V;   X298G,H,I,L,M,F,P,S,T,W,Y,V;   X299A,G,H,M,P,S,T;   X300A,C,G,H,I,L,M,F,P,T,W,Y,V;   X301N,Q,H,I,L,M,F,P,S,T,W,Y,V;   X302R,M,P,W,Y;   X303R,I,L,M,F,P,S,T,W,Y,V;   X305G,I,L,M,F,P,S,T,W,Y,V;   X306Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X308A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X309N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X310A,R,N,C,Q,E,H,I,L,K,M,F,P,S,T,W,Y,V;   X314A,D,C,E,G,I,L,M,F,P,W,Y,V;   X315A,N,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X316A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,V;   X317R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X318A,R,N,D,C,Q,E,G,H,I,K,P,S,W,Y,V;   X319A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X328A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X332A,R,N,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X333A,N,D,C,G,I,M,F,P,S,T,Y,V;   X334R,N,C,Q,E,G,H,K,M,F,P,S,W,Y;   X335R,D,C,Q,E,H,I,L,K,M,F,P,W,Y,V;   X336A,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X337A,R,N,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X338A,R,N,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X340I,L,M,F,P,S,T,W,Y,V;   X341A,R,D,C,G,H,I,L,K,M,F,W,Y,V;   X342R,I,L,M,F,P,S,T,W,Y,V;   X345R,N,D,C,Q,E,G,H,I,L,K,M,F,P,T,W,Y,V;   X355A,R,N,D,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X358A,R,D,C,G,K,M,P,W,Y;   X363A,R,D,C,G,K,M,P,W,Y;   X370A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X371A,N,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X373A,R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V;   X374A,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V;   X375A,R,N,D,C,Q,G,H,I,L,K,M,F,P,S,T,W,V;   X376A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X378R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V;   X379A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,T,W,Y,V;   X381A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X389A,D,C,G,H,M,F,P,S,T,W,Y,V;   X393A,N,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X394A,R,D,C,G,K,M,P,W,Y;   X396A,R,D,C,G,K,M,P,W,Y;   X398A,R,N,D,C,Q,E,G,H,I,L,M,F,W,Y,V;   X402R,C,G,K,M,P,W,Y;   X403A,R,N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X404R,I,L,M,F,P,S,T,W,Y,V;   X405R,G,H,I,L,M,F,P,W,Y,V;   X406A,R,G,H,I,M,F,P,Y,V;   X407A,R,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X408A,R,N,D,C,Q,E,G,H,I,K,M,P,S,T,W,Y,V;   X413A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X414A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X415A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X416A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X417A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,W,Y,V;   X418A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X419A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X420A,N,D,C,E,G,H,I,L,K,M,F,S,T,W,Y,V;   X421A,R,N,D,C,Q,E,H,I,L,K,M,F,P,S,T,W,Y,V;   X422A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X431A,R,N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X432G,H,I,L,M,F,P,S,T,W,Y,V;   X433A,R,N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X434A,R,N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X435Q,G,H,I,L,M,F,P,T,W,Y,V;   X436A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X437A,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X439A,R,D,C,G,K,M,P,W,Y;   X442A,R,N,D,C,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X443A,R,N,D,C,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X444A,C,G,H,I,L,M,F,P,S,T,W,Y,V;   X445A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X446A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X447A,N,D,C,G,H,I,L,M,F,P,S,T,W,Y,V;   X448A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X450A,N,D,C,Q,E,G,I,L,K,M,F,P,S,T,W,V;   X454A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X455A,R,N,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X456A,D,C,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X457A,R,N,D,C,Q,E,G,H,I,L,K,M,F,W,Y,V;   X458A,R,N,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X459R,N,D,C,Q,E,G,H,I,L,K,M,F,S,W,Y,V;   X460A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X461A,R,N,D,C,Q,E,G,H,I,L,M,F,P,S,W,Y,V;   X463A,R,N,D,C,Q,E,H,I,L,K,M,F,P,S,T,W,Y,V;   X464A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X465A,R,N,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X467A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X469A,R,N,D,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X473N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X474A,R,G,H,I,L,M,F,P,W,Y,V;   X475A,N,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X476G,H,I,L,M,F,P,S,T,W,Y,V;   X482A,N,D,C,G,H,I,L,M,F,S,T,W,Y,V;   X483A,N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V   wherein   (a) the variant has alpha-amylase activity and (b) each position corresponds to a position of the amino acid sequence of the parent Termamyl-like alpha-amylase having the amino acid sequence shown in SEQ ID NO: 8 ( Bacillus licheniformis  alpha-amylase).   
     
     
         3 . A variant of a parent Termamyl-like alpha-amylase, comprising one or more of the following substitutions:
 X7A,R,N,D,C,Q,E,G,H,K,M,P,S,Y,V;   X8C,M   X9A,R,N,D,C,Q,G,H,M,P,S,T,W,Y,V;   X11A,N,D,C,Q,G,H,I,L,M,P,S,T,W,Y,V;   X12A,R,N,D,C,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X19A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X21A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X22A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X25A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X32A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X40A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X41A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X46A,R,D,C,G,K,M,P,W,Y;   X48R,N,D,C,Q,E,G,H,K,M,F,P,W,Y;   X55A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X57R,N,D,C,Q,E,G,H,K,M,P,W;   X58A,R,N,D,C,Q,E,G,H,K,M,S,T,W,Y;   X60A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X77A,R,D,C,G,K,M,P,W,Y;   X95A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X97A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X98A,R,D,C,G,K,M,P,W,Y;   X99R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X100A,R,D,C,Q,E,G,H,I,K,M,F,P,S,T,W,Y,V;   X101A,N,C,Q,G,I,L,M,P,S,T,W,Y,V;   X102N,D,C,Q,E,H,I,L,M,F,P,W,Y,V;   X103A,N,D,C,Q,E,G,M,P,S,W,Y;   X105A,N,C,Q,G,H,I,L,M,P,S,T,Y,V;   X107R,N,D,Q,E,H,K,M,F,P,W,Y;   X115R,N,D,C,Q,E,H,I,L,K,M,F,P,S,T,W,Y,V;   X118R,N,C,Q,E,H,I,L,K,M,F,P,S,T,W,Y,V;   X135A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X139A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X141A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X143A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,Y,V;   X151A,R,N,D,C,Q,E,G,H,K,M,P,S,T,Y,V;   X159A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,V;   X160A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X161A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X162A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X163A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X166A,R,N,D,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X175A,R,D,C,G,K,M,P,W,Y;   X177A,N,D,C,Q,E,H,I,L,K,M,P,S,T,W,Y,V;   X183A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X186A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X187A,R,C,Q,E,G,H,I,L,K,M,F,P,W,Y,V;   X192A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X199R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X200A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X202A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X203A,R,D,C,G,K,M,P;   X208A,R,N,D,C,Q,E,G,H,L,K,M,F,P,S,T,W,Y,V;   X212A,N,C,Q,G,H,M,P,S,T,V;   X215A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X219A,R,D,C,G,K,M,P,W,Y;   X228A,R,N,D,C,Q,E,G,H,I,L,K,M,P,S,T,W,Y,V;   X230A,R,N,D,C,Q,E,G,L,M,P,S,T,W,Y,V;   X233R,N,C,Q,E,G,H,I,K,M,P,S,T,W,Y;   X236A,C,Q,G,H,I,M,P,S,T,V;   X238A,R,N,D,C,Q,E,G,H,I,K,M,P,S,T,W,Y,V;   X240A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X241A,N,C,Q,G,H,P,S,T,V;   X244A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X248A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X256C,M;   X258A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X259A,R,N,D,C,Q,E,G,H,K,M,P,S,T,W,Y,V;   X260R,N,D,C,Q,E,H,I,L,K,M,F,P,T,W,Y,V;   X262A,R,D,C,G,K,M,P,W,Y;   X270A,N,C,Q,G,I,L,M,F,P,S,T,W,Y,V;   X273A,R,D,C,G,K,M,P,Y;   X274A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X277A,R,N,D,C,Q,E,G,H,K,M,P,S,W,Y,V;   X281A,R,N,D,C,Q,E,G,K,M,P,S,T,W,Y,V;   X283A,R,N,C,Q,E,G,I,L,K,M,F,P,S,T,W,Y,V;   X284A,R,N,D,C,Q,E,G,I,L,K,M,F,P,S,T,Y,V;   X285A,R,D,C,Q,E,G,H,I,K,M,F,P,S,T,W,Y,V;   X286A,R,D,C,Q,E,G,H,I,K,M,F,P,S,T,W,Y,V; preferably X286N,C,Q,I,L,M,P,T,V,Y,F;   X287R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X288A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X289A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X292A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X295A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X296A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X304C,M;   X307A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X312A,N,C,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X313A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X320R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X322R,N,D,C,Q,E,G,H,L,K,M,F,P,S,T,W,Y,V;   X323A,R,N,D,C,Q,E,G,I,L,K,M,F,P,S,T,W,Y,V;   X325A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X326A,R,N,C,Q,E,G,M,P,S,T,W;   X327A,R,C,G,H,I,L,K,M,P,S,T,W,Y,V;   X329A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,T,W,Y,V;   X331N,D,C,Q,E,G,H,I,L,M,F,P,S,T,W,Y,V;   X339A,R,N,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X343A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,Y,V;   X344A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X346A,R,N,D,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X347A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X349R,N,D,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X350A,R,N,C,Q,G,H,I,L,K,M,F,P,S,T,Y,V;   X359R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X360N,D,Q,G,H,I,L,M,F,P,S,T,W,Y,V;   X369A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X377A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X380A,R,N,D,C,Q,E,G,H,K,P,S,W,Y,V;   X387A,R,N,D,C,Q,E,G,H,L,K,M,P,S,T,W,Y,V;   X409N,C,Q,E,G,H,M,P,S,T,W,Y,V;   X410A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X411A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X412R,N,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X423A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X424A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X426A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X427A,R,N,D,C,Q,E,G,H,K,M,P,S,T,Y,V;   X428A,N,D,Q,E,G,H,I,M,F,P,S,W,Y,V;   X429A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X430A,R,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X438C,M;   X440R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X441A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X449A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X462A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X472A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X477A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V;   X479A,R,N,D,Q,E,G,H,I,L,K,M,F,P,S,W,Y,V;   X480A,R,D,C,G,K,M,P,W,Y;   X481A,R,N,D,C,Q,E,G,H,K,M,P,S,T,Y,V   wherein   (a) the variant has alpha-amylase activity and (b) each position corresponds to a position of the amino acid sequence of the parent Termamyl-like alpha-amylase having the amino acid sequence shown in SEQ ID NO: 8 ( Bacillus licheniformis  alpha-amylase).   
     
     
         4 . A variant of a parent Termamyl-like alpha-amylase, comprising an alteration at one or more positions selected from the group of:
 A1 insertion;   L3 insertion;   N4 insertion;   N17 insertion;   D18 insertion;   Q20 insertion;   R23 insertion;   R24 insertion;   D28 insertion;   Y56 insertion;   L61 insertion or deletion;   Y62 insertion;   F67 insertion or deletion;   H68 insertion;   K80 insertion or deletion;   G81 insertion or deletion;   Q84 insertion;   S85 insertion;   H91 insertion or deletion;   S92 insertion or deletion;   K106 insertion or deletion;   D110 insertion or deletion;   D114 deletion;   E119 insertion or deletion;   D121 insertion;   P122 insertion;   A123 insertion;   D124 insertion;   R125 insertion;   N126 insertion;   R127 insertion;   I129 insertion;   G131 insertion;   L134 insertion;   K136 insertion;   N172 insertion;   E185 insertion;   L196 insertion or deletion;   P206 insertion or deletion;   T217 insertion;   W218 insertion;   D231 insertion or deletion;   A232 insertion or deletion;   H235 insertion or deletion;   N246 insertion;   H247 insertion;   R249 insertion;   K251 insertion;   F257 insertion or deletion;   N278 insertion;   G310 insertion or deletion;   H316 insertion;   P317 insertion;   D328 insertion or deletion;   G332 insertion or deletion;   E355 insertion or deletion;   Y358 insertion;   Y363 insertion;   Y367 insertion;   K370 insertion;   S373 insertion;   R375 insertion;   E376 insertion;   K381 insertion;   H382 insertion;   R391 insertion or deletion;   Y396 insertion;   R413 insertion or deletion;   E414 insertion or deletion;   G415 insertion or deletion;   D416 insertion;   S417 insertion;   S418 insertion;   V419 insertion;   A420 insertion;   N421 insertion;   S422 insertion or deletion;   Y439 insertion;   A445 insertion or deletion;   G446 insertion or deletion;   T448 insertion or deletion;   H450 insertion;   G454 insertion or deletion;   N455 insertion;   E458 insertion;   P459 insertion;   V460 insertion;   V461 insertion;   N463 insertion;   S464 insertion;   E465 insertion;   W467 insertion;   wherein   (a) the alteration(s) are independently (as specified above):
 (i) an insertion of an amino acid downstream of the amino acid which occupies the position, or 
 (ii) a deletion of the amino acid which occupies the position, 
   (b) the variant has alpha-amylase activity and (c) each position corresponds to a position of the amino acid sequence of the parent Termamyl-like alpha-amylase having the amino acid sequence shown in SEQ ID NO: 8 ( Bacillus licheniformis  alpha-amylase).   
     
     
         5 . A variant of a parent Termamyl-like alpha-amylase, comprising an alteration at one or more positions selected from the group of:
 L7 insertion or deletion;   M8 insertion;   Y10 insertion;   F11 insertion;   E12 insertion or deletion;   M15 insertion;   G19 insertion;   H21 insertion;   W22 insertion;   L25 insertion;   V40 insertion or deletion;   W41 insertion;   P43 insertion or deletion;   P44 insertion or deletion;   Y46 insertion;   G55 insertion;   Y59 insertion;   Y77 insertion;   G78 insertion or deletion;   L90 insertion or deletion;   I95 insertion;   V97 insertion;   Y98 insertion;   G99 insertion;   D100 insertion;   V101 insertion;   V102 insertion;   H105 insertion or deletion;   A109 insertion or deletion;   V115 insertion or deletion;   V118 insertion or deletion;   I135 insertion;   T139 insertion or deletion;   F141 insertion or deletion;   Y195 insertion;   V208 insertion or deletion;   W215 insertion;   Y219 insertion;   I236 insertion or deletion;   F238 insertion or deletion;   F240 insertion or deletion;   W244 insertion;   V248 insertion;   M256 insertion;   T258 insertion or deletion;   V259 insertion or deletion;   V312 insertion or deletion;   V313 insertion or deletion;   S320 insertion;   T322 insertion or deletion;   F323 insertion or deletion;   D325 insertion or deletion;   N326 insertion;   H327 insertion or deletion;   Q330 insertion or deletion;   P331 insertion or deletion;   Y348 insertion;   A349 insertion or deletion;   F350 insertion or deletion;   P359 insertion or deletion;   Q360 insertion;   D365 insertion or deletion;   M366 insertion;   T369 insertion;   I377 insertion;   I384 insertion or deletion;   L388 insertion or deletion;   G423 insertion or deletion;   L424 insertion or deletion;   M438 insertion;   G441 insertion or deletion;   W449 insertion;   I462 insertion;   I479 insertion or deletion;   Y480 insertion;   V481 insertion or deletion;   wherein   (a) the alteration(s) are independently (as specified above):
 (i) an insertion of an amino acid downstream of the amino acid which occupies the position, or 
 (ii) a deletion of the amino acid which occupies the position, 
   (b) the variant has alpha-amylase activity and (c) each position corresponds to a position of the amino acid sequence of the parent Termamyl-like alpha-amylase having the amino acid sequence shown in SEQ ID NO: 8 ( Bacillus licheniformis  alpha-amylase).   
     
     
         6 . The variant of any of  claims 1 - 5 , wherein the parent Termamyl-like alpha-amylase is derived from a strain of  B. licheniformis  (SEQ ID NO: 8),  B. amyloliquefaciens  (SEQ ID NO: 10),  B. stearothermophilus  (SEQ ID NO: 6),  Bacillus  sp. (SEQ ID NO: 12 (AA560),  Bacillus  sp. (SEQ ID NO: 2 (SP690));  Bacillus  sp. (SEQ ID NO: 4 (SP722);  Bacillus  sp. #707 alpha-amylase (SEQ ID NO: 13); KSM-AP1378. 
     
     
         7 . The variant of any of  claims 1  to  6 , wherein the parent Termamyl-like alpha-amylase has an amino acid sequence which has a degree of identity to SEQ ID NO: 8 of at least 60%, preferably 70%, more preferably at least 80%, even more preferably at least about 90%, even more preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, and even more preferably at least 99%. 
     
     
         8 . The variant of any of  claims 1 - 7 , wherein the parent Termamyl-like alpha-amylase is encoded by a nucleic acid sequence (DNA), which hydridizes under low, preferably medium, preferred high stringency conditions, with the nucleic acid sequence of SEQ ID NO: 7. 
     
     
         9 . The variant of  claims 1 - 8 , which variant has alpha-amylase activity and exhibits an alteration in at least one of the following properties relative to said parent alpha-amylase: altered substrate specificity, substrate binding, substrate cleavage pattern, thermal stability, pH activity profile, pH stability profile, stability towards oxidation, Ca 2+  dependency, reduced and increased pI and improved wash performance, specific activity, stability under high temperature and/or low pH conditions, in particular at low calcium concentrations and/or in particular at high temperatures from 70-120° C. and/or low pH in the range from pH 4-6 
     
     
         10 . A DNA construct comprising a DNA sequence encoding an alpha-amylase variant according to any one of  claims 1 - 9 . 
     
     
         11 . A recombinant expression vector which carries a nucleic acid sequence (DNA) construct according to  claim 10 . 
     
     
         12 . A cell which is transformed with a DNA construct according to  claim 10  or a vector according to  claim 11 . 
     
     
         13 . A cell according to  claim 12 , which is a microorganism, preferably a bacterium or a fungus, in particular a gram-positive bacterium, such as  Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Bacillus stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus circulans, Bacillus lautus  or  Bacillus thuringiensis.    
     
     
         14 . A composition comprising an alpha-amylase variant of any of  claims 1 - 9 . 
     
     
         15 . The composition of  claim 14 , wherein the composition further comprises glucoamylase, pullulanase and/or phytase. 
     
     
         16 . A detergent composition comprising an alpha-amylase variant according to any of  claims 1 - 9 . 
     
     
         17 . A detergent composition of  claim 16 , which additionally comprises another enzyme such as a protease, a lipase, a peroxidase, another amylolytic enzyme, glucoamylase, maltogenic amylase, CGTase, mannanase, cutinase, laccase and/or a cellulase. 
     
     
         18 . Use of an alpha-amylase variant according to any of  claims 1 - 9  or a composition according to  claim 15  for starch liquefaction, in particular for syrup or ethanol production. 
     
     
         19 . Use of an alpha-amylase variant according to any one of  claims 1 - 9  or a composition according to  claim 15  for washing and/or dishwashing. 
     
     
         20 . Use of an alpha-amylase variant of any one of  claims 1 - 9  or a composition according to  claim 15  for textile desizing.
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