Mannanase variants and polynucleotides encoding same
The present invention relates to mannanase 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.
1 . A variant of a parent mannanase, which comprises
(a) a first substitution at one or more positions corresponding to positions selected from the group consisting of:
(i) 260 with Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Ile, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val;
(ii) 288 with Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val;
(iii) 294 with Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val; and
(iv) 295 with Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val; and
(b) a second substitution at one or more other positions,
wherein
numbering is according to SEQ ID NO: 2,
the variant comprises an amino acid sequence that has at least 80% and less than 100% sequence identity to the amino acid sequence of SEQ ID NO: 2, and
the variant has mannanase activity.
2 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 260 and 288.
3 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 260 and 294.
4 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 260 and 295.
5 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 288 and 294.
6 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 288 and 295.
7 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 294 and 295.
8 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 260, 288, and 294.
9 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 260, 288, and 295.
10 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 288, 294, and 295.
11 . The variant of claim 1 , which comprises a substitution at positions corresponding to positions 260, 288, 294, and 295.
12 . The variant of claim 1 , wherein the first substitution is
260F, 260Y, or 260T,
294P, 294K, 294R, 294V, or 294H, or
295K, 295V, 295P, 295L, 295R, 295N, 295M, or 295I.
13 . The variant of claim 1 , wherein the second substitution is at one or more positions corresponding to the positions selected from the group consisting of: 1, 2, 3, 4, 5, 6, 8, 11, 13, 14, 18, 30, 32, 33, 34, 35, 37, 41, 45, 47, 57, 59, 60, 63, 65, 70, 71, 74, 77, 78, 80, 81, 82, 83, 93, 95, 97, 98, 100, 104, 108, 111, 114, 116, 118, 119, 131, 133, 135, 136, 139, 142, 143, 150, 169, 172, 174, 176, 177, 180, 183, 184, 185, 196, 200, 202, 203, 205, 210, 213, 228, 229, 234, 235, 241, 243, 244, 250, 254, 257, 258, 262, 266, 268, 270, 272, 273, 276, 279, 280, 283, 286, 290, 296, and 298.
14 . The variant of claim 1 , wherein the second substitution is at one or both of positions corresponding to positions 93 and 136.
15 . The variant of claim 1 , wherein the second substitution is selected from the group consisting of
A1G or A1V,
N2E,
S3P,
G4D,
F5H,
Y6H, Y6M, Y6F, Y6W, or Y6H,
S8T, S8P, or S8R,
T11K or T11R,
Y13F,
D14S or D14K,
N18V or N18R,
A30T,
Y32F or Y32W,
K33Q,
D34G,
Q35L,
T37P,
E41V or E41N,
N45G, G47S, or G47A,
D57N,
G59Q,
Q60R,
K63R or K63Q,
D65E,
R70K,
N71S,
S74K,
E77T or E77N,
D78G,
H80K,
V82R, V82I, or V82S,
A83P or A83S,
Y93Q or Y93A,
S95D,
A97R,
S98P or S98D,
N100Y,
D104A or D104G,
E108S,
S111A, S111K, or S111R,
I114Q, 1114M, or I114W,
K116R,
D118K,
T119R,
S131T,
E133R or E133Q,
D135P,
A136P,
D139A or D139R,
K142M, K142V, K142S, or K142R,
Q143R,
N150T, N150R, or N150S,
Q169A, Q169R, or Q169K,
H172R,
Y174R, Y174L, Y174W, or Y174F,
R176Q,
E177S or E177Y,
N180R,
P183T or P183G,
Q184E or Q184K,
R185G,
Y196W or Y196F,
N200T,
S202R,
Q203T,
R205K,
R210L, R210G, or R210M,
N213V or N213D,
T228S,
N229D,
E234F or E234Y,
A235K or A235R,
S241C,
Q243K or Q243E,
R244K or R244V,
A250G,
K254Y,
G257W, G257E, G257A, or G257G,
Y262F,
S266A,
D268N,
A270D,
N272M, or N272T,
N273E or N273D,
A276E, A276W, or A276D,
N279D or N279E,
T280L,
N283W or N283H,
Y286W or Y286F,
E290A,
T296S,
F298Y, or
any combination thereof.
16 . A composition comprising
(a) the variant of claim 1 and
(b) a surfactant, a bleaching system, a chelating agent, stabilizing agents, hydrotopes, builders, co-builders, bleach activators, polymers and/or fabric-hueing agents.
17 . A method of dishwashing, comprising of adding the composition of claim 16 in a detergent composition compartment in an automatic dishwashing machine, and releasing said composition during a main-wash cycle.
18 . A method of laundering, comprising adding the composition of claim 16 in a detergent composition compartment in an automatic laundering machine, and releasing said composition during a main wash cycle.
19 . A variant of a parent mannanase, which comprises
(a) a first substitution at the position corresponding to positions selected from the group consisting of:
(i) 260 with Leu;
(ii) 288 with Ile;
(iii) 294 with Ile; and
(iv) 295 with Ala; and
(b) a second substitution at one or more other positions,
wherein
numbering is according to SEQ ID NO: 2,
the variant comprises an amino acid sequence that has at least 85% and less than 100% sequence identity to the amino acid sequence of SEQ ID NO: 2, and
the variant has mannanase activity.
20 . A composition comprising
(a) the variant of claim 19 and
(b) a surfactant, a bleaching system, a chelating agent, stabilizing agents, hydrotopes, builders, co-builders, bleach activators, polymers and/or fabric-hueing agents.