US2005181446A1PendingUtilityA1
Protein variants having modified immunogenicity
Est. expiryApr 28, 2020(expired)· nominal 20-yr term from priority
A61K 39/00C12N 9/54A21D 2/267A21D 8/042C07K 1/047C07K 5/1008C07K 5/101C07K 5/1013C07K 5/1019C07K 5/1021C07K 14/001C07K 16/4283C11D 3/38627C11D 3/38645C12N 9/0061C12N 9/20C12Y 110/03002C07K 16/40Y10S435/911C12Y 304/21062
57
PatentIndex Score
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Cited by
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References
0
Claims
Abstract
The present invention relates to a method of selecting a protein variant having modified immunogenicity as compared to the parent protein comprising the steps obtaining antibody binding peptide sequences, using the sequences to localise epitope sequences on the 3-dimensional structure of parent protein, defining an epitope area including amino acids situated within 5 Å from the epitope amino acids constituting the epitope sequence, changing one or more of the amino acids defining the epitope area of the parent protein by genetical engineering mutations of a DNA sequence encoding the parent protein, introducing the mutated DNA sequence into a suitable host, culturing said host and expressing the protein variant, and evaluating the immunogenicity of the protein variant using the parent protein as reference. The invention further relates to the protein variant and use thereof, as well as to a method for producing said protein variant.
Claims
exact text as granted — not AI-modified1 . A method of selecting a protein variant having modified immunogenicity as compared to a parent protein, comprising the steps of:
(a) obtaining antibody binding peptide sequences, (b) using the sequences to localise epitope sequences on the 3-dimensional structure of the parent protein, (c) defining an epitope area including amino acids situated within 5 Å from the epitope amino acids constituting the epitope sequence, (d) changing one or more of the amino acids defining the epitope area of the parent protein by genetic engineering mutations of a DNA sequence encoding the parent protein, (e) introducing the mutated DNA sequence into a suitable host, culturing said host and expressing the protein variant, and (f) evaluating the immunogenicity of the protein variant using the parent protein as reference.
2 . The method of claim 1 , wherein the sequences of step a) are obtained by screening a random peptide display package library with antibodies raised against any protein of interest and sequencing the amino acid sequence of the antibody binding peptide, or the DNA sequence encoding the antibody binding peptide.
3 . The method of claim 2 , wherein antibodies for screening the random peptide display package library are raised against the parent protein.
4 . The method of claim 2 , wherein the peptide display package library is a phage display library.
5 . The method of claim 2 , wherein the peptides of the peptide display package library are oligopeptides having from 5 to 25 amino acids.
6 . The method of claim 1 , wherein the antibody binding peptide sequences of step a) are obtained by screening a library of known peptides related to the primary sequence of any protein of interest, with antibodies raised against the protein of interest.
7 . The method of claim 1 , wherein epitope patterns are identified by sequence alignment of ntibody binding peptide sequences and these epitope patterns are used to quide localisation of pitope sequences on the 3-dimensional structure of the parent protein.
8 . The method of claim 1 , wherein the epitope area of step c) equals the epitope sequence.
9 . The method of claim 1 , wherein hot spot amino acids of the parent protein are identified.
10 . The method of claim 1 , wherein the epitope area is changed by substituting, adding and/or deleting at least one amino acid of the epitope area.
11 . The method of claim 10 , wherein the epitope area is changed by substituting, adding and/or deleting at least one hot spot amino acid.
12 . The method of claim 10 , wherein amino acids in the epitope area are changed by substituting and/or inserting at least one amino acid by an amino acid which render the substituted and/or inserted amino acid a target for covalent conjugation to an activated polymer.
13 . The method of claim 12 , wherein the amino acid for substitution and/or insertion is selected from the group consisting of K, C, D, E.
14 . The method of claim 12 , wherein the molecule for covalent conjugation is selected from the group of activated synthetic or natural polymers.
15 . The method of claim 14 , wherein the activated synthetic polymer is a polyethylene glycol.
16 . The method of claim 1 , wherein the immunogenicity is measured by competitive ELISA.
17 . The method of claim 1 , wherein the protein variant has reduced allergenicity.
18 . The method of claim 17 , wherein the allergenicity of the protein variant is below 75%, preferably below 50%, more preferably below 25% of the allergenicity of the parent protein.
19 . The method of claim 1 , wherein the parent protein is an enzyme or an environmental allergen or a pharmaceutical protein.
20 . The method of claim 19 , wherein the enzyme is selected from the group consisting of glycosyl hydrolases, carbohydrases, peroxidases, proteases, lipolytic enzymes, phytases, polysaccharide lyases, oxidoreductases, transglutaminases and glucoseisomerases.
21 . The method of claim 19 , wherein the environmental allergen is selected from the group consisting of pollen, dust mites, mammals, venoms, fungi, food allergens or other plant allergens.
22 . A protein variant obtainable by a method of claim 1 .
23 . A protein variant, wherein the amino acid sequence of the protein variant differs from the amino acid sequence of the parent protein with respect to at least one epitope area of the parent protein.
24 . The protein variant of claim 23 having modified immunogenicity as compared to its parent protein.
25 . A protein variant of claim 23 , wherein the epitope areas are defined on the parent protein structure by being localised less than 5 Å from any of the following epitope patterns: P>S/T D P G; P>>D A G; >P>R D T G; P>S/T D P G; >R Y>K/R; >R S A; >G>>A G; V H>G >; A>I D P R/K; A R>A; Q>Y>D>; >P>>A P>S; R/K R F>N; D/E Q I F F T; A>>>>Y P>; L>G R S; R P P R; >E Y; >P>>P A P>S; >K L>>; K Q S; >K L>>; Y I>K L; R Q>>D/E; N>>E L.
26 . The protein variant of claim 22 , wherein the epitope areas correspond to antibody binding peptide sequences reactive to antibodies raised against the parent protein.
27 . The protein variant of claim 22 , wherein the epitope pattern is a IgE epitope pattern.
28 . The protein variant of claim 22 , wherein at least one hot spot amino acid is substituted or deleted.
29 . The protein variant of claim 22 , wherein the allergenicity of the protein variant is below 75%, preferably below 50%, more preferably below 25% of the allergenicity of the parent protein.
30 . The protein variant of claim 22 , wherein the protein variant is an environmental allergen, preferable an allergen selected from the group consisting of pollen, dust mites, mammals, venoms, fungi, food allergens or other plant allergens.
31 . The protein variant of claim 22 , wherein the protein variant is an antifungal peptide or antimicrobial peptide.
32 . The protein variant of claim 30 , wherein the allergen is pollen allergen comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 6:
Position
T
10
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
V
12
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
P
14
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
A
16
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
17
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
K
20
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
24
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
F
30
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
P
31
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
K
32
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
34
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
35
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
Q
36
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
A
37
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
39
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
S
40
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
E
42
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
57
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, T, V, W, Y;
Position
F
58
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
59
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
E
60
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
61
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
62
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
P
63
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
F
64
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
65
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
77
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
F
79
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
90
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
D
93
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
V
105
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
A
106
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
107
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
D
109
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
110
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
123
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
127
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
129
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
131
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
136
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
G
140
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
143
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
R
145
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
S
149
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
Y
150
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
L
152
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
A
153
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
156
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
158
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W.
33 . The protein variant of claim 32 , wherein the pollen allergen comprises one or more of the following substitutions:
position
P31
to
A, G, L, or S;
position
A34
to
D, E, F, H, K, N, P, Q, R, W, or Y;
position
P35
to
A, G, L, or S;
position
A37
to
D, E, F, H, K, N, P, Q, R, W, or Y;
position
S39
to
D, E, F, H, K, N, P, Q, R, W, or Y;
position
S40
to
D, E, F, H, K, N, P, Q, R, W, or Y;
position
P59
to
A, G, L, or S;
position
L62
to
D, E, F, H, K, N, P, Q, R, W, or Y;
position
P63
to
A, G, L, or S.
34 . The allergen of claim 32 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 6.
35 . The pollen allergen of claim 34 , wherein the allergen has the amino acid sequence of SEQ ID NO: 6.
36 . The protein variant of claim 30 , wherein the allergen is mite allergen comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 7:
Position
D
1
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
2
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
N
11
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
E
12
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
14
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
15
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
19
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
20
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
H
30
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
31
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
G
32
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
34
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
T
36
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
L
37
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
E
38
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
39
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
40
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
D
59
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
61
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
E
62
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
64
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
N
71
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
H
74
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
F
75
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
79
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
Q
85
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
D
87
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
90
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
T
91
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
W
92
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
N
93
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
P
95
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
K
96
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
I
97
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
98
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
99
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
S
101
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
E
102
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
123
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
K
126
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
128
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
D
129
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y.
37 . The mite allergen of claims 36 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 7.
38 . The mite allergen of claim 37 , wherein the allergen has the amino acid sequence of SEQ ID NO: 7.
39 . The protein variant of claim 30 , wherein the allergen is mite allergen comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 8:
Position
L
17
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
P
19
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
G
20
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
26
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
I
28
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
H
30
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
31
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
P
34
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
F
35
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
36
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
K
55
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
56
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
57
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
D
59
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
60
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
61
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
E
62
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
64
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
66
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
D
69
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
89
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
90
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
T
91
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
W
92
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
P
95
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
K
96
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
I
97
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
99
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
K
100
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
102
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
N
103
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
T
123
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
A
125
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
128
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y.
40 . The mite allergen of claim 39 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 8.
41 . The mite allergen of claim 40 , wherein the allergen has the amino acid sequence of SEQ ID NO: 8.
42 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 9:
Position
V
1
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
E
9
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
10
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
N
13
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
E
14
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
15
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
H
16
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
18
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
34
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
H
36
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
37
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
38
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
W
41
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
V
42
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
A
43
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
F
54
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
55
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
56
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
E
57
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
59
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
L
60
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
61
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
P
63
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
R
67
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
L
69
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
D
79
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
84
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
85
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
87
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
P
94
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y.
43 . The allergen of claims 42 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 9.
44 . The allergen of claim 43 , wherein the allergen has the amino acid sequence of SEQ ID NO: 9.
45 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 12:
Position
I
1
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
18
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
41
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
43
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
65
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
70
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
K
113
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
114
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
116
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
P
119
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
E
120
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
122
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
K
124
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
126
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
Q
127
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
S
130
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
R
132
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
I
139
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
I
143
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y.
46 . The allergen of claim 45 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 12.
47 . The allergen of claim 46 , wherein the allergen has the amino acid sequence of SEQ ID NO: 12.
48 . The protein variant of claim 30 , wherein the allergen is a mammal allergen comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 13:
Position
S
9
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
S
12
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
Y
16
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
D
23
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
V
40
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
R
42
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
A
43
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
44
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
50
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
D
69
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
N
80
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
Y
84
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
P
110
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
Q
112
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
E
120
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
121
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
D
122
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
129
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
K
133
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
139
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
142
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
156
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
L
157
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
R
158
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
G
159
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y.
49 . The allergen of claim 48 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 13.
50 . The allergen of claim 49 , wherein the allergen has the amino acid sequence of SEQ ID NO: 13.
51 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 15:
Position
K
1
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
24
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
E
35
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
45
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
T
47
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
D
52
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
53
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
N
59
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
R
61
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
W
62
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
W
63
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
N
65
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
D
66
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
67
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
70
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
S
72
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
R
73
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
L
75
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
I
78
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
79
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
S
81
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
A
82
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
84
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
T
118
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
R
125
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
G
126
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
128
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
L
129
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y.
52 . The allergen of claim 51 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 15.
53 . The allergen of claim 52 , wherein the allergen has the amino acid sequence of SEQ ID NO: 15.
54 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 16:
Position
T
4
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
D
28
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
V
31
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
Q
40
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
F
41
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
42
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
44
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
45
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
47
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
48
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
51
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
54
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
58
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
P
61
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
T
62
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
H
65
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
68
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
70
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
143
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
146
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
148
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
S
173
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
A
178
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
181
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
184
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
185
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
186
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
G
187
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
188
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
A
190
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
192
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
V
203
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
I
204
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
207
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
208
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
G
209
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
213
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
K
215
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
236
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
238
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
T
240
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
P
241
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
G
242
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y.
55 . The allergen of claim 54 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 16.
56 . The allergen of claim 55 , wherein the allergen has the amino acid sequence of SEQ ID NO: 16.
57 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 17:
Position
A
33
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
36
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
38
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
P
54
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
R
56
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
A
57
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
58
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
V
68
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
L
70
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
R
71
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
Y
78
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
K
80
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
81
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
83
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
A
84
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
102
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
103
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
106
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
E
114
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
118
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
119
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
I
121
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y.
58 . The allergen of claim 57 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 17.
59 . The allergen of claim 58 , wherein the allergen has the amino acid sequence of SEQ ID NO: 17.
60 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 18:
Position
W
2
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
D
13
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
15
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
16
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
28
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
V
31
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
Q
34
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
Q
40
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
L
41
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
P
43
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
Q
44
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
D
47
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
50
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
51
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
54
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
57
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
60
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
62
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
G
67
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
68
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
69
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
71
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
I
74
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
75
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
Q
78
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
R
83
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
K
85
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
87
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
88
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
N
97
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
D
106
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
108
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
T
110
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
R
120
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
D
123
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
124
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
E
127
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
129
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y.
61 . The allergen of claim 60 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 18.
62 . The allergen of claim 61 , wherein the allergen has the amino acid sequence of SEQ ID NO: 18.
63 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 19:
Position
T
28
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
T
31
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
A
33
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
34
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
36
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
53
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
A
54
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
56
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
G
64
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
65
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
R
66
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
V
68
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
R
71
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
D
74
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
78
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
S
83
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
A
84
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
N
101
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
E
102
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
103
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
105
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
P
106
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
T
108
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
K
115
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
119
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
T
133
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
V
136
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
G
137
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
150
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
153
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
A
158
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
161
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
169
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
175
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
176
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
181
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
D
199
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
200
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
K
206
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
207
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
208
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
A
209
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
215
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
227
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
228
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
I
229
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
231
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
G
232
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
233
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
N
236
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
E
239
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
243
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
244
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
I
246
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
247
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
248
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
G
249
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y.
64 . The allergen of claim 63 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 19.
65 . The allergen of claim 64 , wherein the allergen has the amino acid sequence of SEQ ID NO: 19.
66 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 20:
Position
S
1
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
Y
5
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
E
8
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
H
9
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
12
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
E
47
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
48
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
70
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
71
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
76
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
K
78
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
80
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
81
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
K
88
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
90
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
91
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
E
99
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
100
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
101
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
V
102
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
T
103
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
P
104
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
G
105
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
106
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
E
112
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
116
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
117
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
I
119
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
120
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
121
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
G
122
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
123
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y.
67 . The allergen of claim 66 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 20.
68 . The allergen of claim 67 , wherein the allergen has the amino acid sequence of SEQ ID NO: 20.
69 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 21:
Position
L
4
to
A, C, D, E, F, G, H, I, K, M, N, P, Q,
R, S, T, V, W, Y;
Position
Y
6
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W;
Position
Y
17
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W;
Position
S
20
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, T, V, W, Y;
Position
S
31
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, T, V, W, Y;
Position
K
32
to
A, C, D, E, F, G, H, I, L, M, N, P, Q,
R, S, T, V, W, Y;
Position
A
33
to
C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W, Y;
Position
K
37
to
A, C, D, E, F, G, H, I, L, M, N, P, Q,
R, S, T, V, W, Y.
70 . The allergen of claim 69 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 21.
71 . The allergen of claim 70 , wherein the allergen has the amino acid sequence of SEQ ID NO: 21.
72 . The protein variant of claim 30 , wherein the allergen comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 22:
Position
N
6
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
C
9
to
A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
23
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
24
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
G
25
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
26
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
L
27
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
K
28
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
29
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
K
34
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
V
35
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
Y
39
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
G
40
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
41
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
K
43
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
45
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
47
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
D
48
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
L
50
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
K
51
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
52
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
55
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Q
58
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
K
59
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
R
62
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
G
71
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
72
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
P
74
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
P
75
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
V
83
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
Position
N
85
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
D
86
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
87
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
90
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
Q
93
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
L
120
to
A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Position
T
121
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
G
122
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
123
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
T
124
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
A
125
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
A
126
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
Y
128
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
D
130
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
D
140
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
P
147
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
K
148
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
150
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
S
152
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
G
153
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
F
156
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
158
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
H
161
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
I
181
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
183
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
K
184
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
W
185
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
H
186
to
A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
N
199
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
K
201
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y
Position
N
202
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
E
203
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
E
204
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
T
208
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y.
73 . The allergen of claim 72 , wherein the allergen has at least 81%, preferably at least 85%, more preferably at least 90%, even more preferably at least 96%, most preferably at least 98% homology to SEQ ID NO: 22.
74 . The allergen of claim 73 , wherein the allergen has the amino acid sequence of SEQ ID NO: 22.
75 . The protein variant of claim 22 , wherein the protein variant is an enzyme.
76 . The protein variant of claim 75 , wherein the enzyme is a protease, a lipolytic enzyme, a carbohydrase or a oxidoreductase.
77 . The protein variant of claim 76 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
−6
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
−5
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
−4
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
−2
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
3a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
28a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
44a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
44b
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
139
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
148
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
149
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion;
Position
264a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion,
deletion.
78 . The protein variant of claim 76 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
−1
to
G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E,
R, H;
Position
1
to
V, L, I, W, M, F, Y, S, T, R;
Position
2
to
G, V, I, M, F, N, Q, Y, S, T, H;
Position
3
to
W, M, F, N, Q, Y, S, D, E, R, H;
Position
4
to
V, L, W, M, F, Y, R;
Position
5
to
V, L, I, W, M, F, N, Q, Y, T, R, H;
Position
6
to
G, V, L, I, W, P, M, N, Q, T, D, E, R, H;
Position
9
to
G, V, L, I, W, P, M, F, Q, Y, S, T, R, H;
Position
10
to
G, A, V, I, W, P, M, N, Q, Y, S, T, D, E, R;
Position
12
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E;
Position
14
to
V, L, I, W, P, M, F, N, Q, Y, T, R, H;
Position
15
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, E, H;
Position
17
to
G, A, V, I, W, P, M, F, Y, H;
Position
18
to
G, A, L, I, W, P, M, F, N, Q, Y, T, D, E, H;
Position
19
to
A, V, I, W, M, F, N, Y, S, T, D, R, H;
Position
20
to
G, V, L, I, W, M, F, N, Q, Y, S, T, D, E;
Position
21
to
G, V, I, W, N, Q, Y, S, T, D, E, R, H;
Position
22
to
G, V, L, I, W, M, F, Y, S, T;
Position
24
to
G, V, L, I, W, M, F, N, Q, Y, S, D, E, R;
Position
25
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position
27
to
G, L, I, W, P, M, F, Y, T, H;
Position
38
to
V, L, I, W, M, F, N, Q, Y, T, H;
Position
39
to
G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H;
Position
40
to
V, L, I, W, M, F, N, Q, Y, T, R, H;
Position
42
to
G, A, L, W, C, M, F, N, Q, Y, S, T, D, E, R, H;
Position
43
to
G, L, H;
Position
44
to
G, V, L, I, W, P, M, F, Y, S, T;
Position
45
to
G, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position
46
to
G, A, L, I, W, P, M, F, Y, H;
Position
47
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E,
R, H;
Position
48
to
A, L, I, P, M, F, N, Y, D, H;
Position
49
to
G, A, V, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position
50
to
G, A, W, M, N, Q, Y, S, T, D, E, H;
Position
51
to
V, L, I, W, M, F, N, Y, R;
Position
52
to
V, L, I, W, M, F, Y, S, T, R;
Position
53
to
A, V, L, I, W, M, F, N, Q, Y, S, D, E, H;
Position
54
to
V, L, I, W, M, F, S, R;
Position
55
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T, D, E, R,
K, H;
Position
56
to
G, V, L, I, W, M, F, N, Q, Y, S, T, H;
Position
57
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
58
to
L, W, M, F, N, Y, R;
Position
59
to
A, V, L, I, C, T, H;
Position
61
to
V, L, I, W, M, F, Y;
Position
62
to
G, A, L, W, M, F, N, Y, R;
Position
64
to
G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
75
to
L;
Position
79
to
I;
Position
80
to
G;
Position
87
to
A, V, L, I, W, M, F, Q, Y, S, T, D, E, H;
Position
89
to
G, V, L, I, W, P, F, N, Y, T, E;
Position
91
to
G, A, V, L, I, W, P, M, N, Y, S, T, D, E,
R, H;
Position
98
to
A;
Position
99
to
V, L, I, W, M, F, Q, Y, H;
Position
100
to
G, V, L, I, W, M, F, Y, R, H;
Position
101
to
V, I, W, M, F, N, Q, Y, H;
Position
102
to
V, L, I, W, M, F, Y, R, H, G;
Position
108
to
I;
Position
109
to
N;
Position
112
to
E
Position
113
to
W;
Position
115
to
I;
Position
117
to
N;
Position
118
to
N;
Position
126
to
L;
Position
127
to
G, A, V, I, W, M, F, Y, R, H, L;
Position
128
to
I, W;
Position
129
to
W;
Position
130
to
W, F, Y, R;
Position
131
to
W, Y, R;
Position
132
to
L, W, M, F, Y, S, H;
Position
133
to
A, L, I, W, M, F, Y, R;
Position
134
to
L, I, W, F, N, Q, Y, R, H;
Position
136
to
G, A, W, P, N, Y, S, T, D, E, H;
Position
137
to
G, A, V, I, W, P, M, N, Y, H;
Position
140
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, H;
Position
141
to
G, V, L, I, W, P, M, F, Q, S, D, E, H;
Position
143
to
V, L, I, P, M, F, N, Y, R;
Position
144
to
L, W, P, M, F, N, Q, Y, S, D, E, R, H;
Position
145
to
G, V, L, I, W, M, F, Q, Y, D, E, R, H;
Position
146
to
G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H;
Position
155
to
V, L, I, W, M, F, Y, R;
Position
156
to
V, I, W, F, R;
Position
157
to
G, A, V, L, I, W, M, F, Y, T, R, H;
Position
158
to
V, L, I, W, M, F, Y;
Position
159
to
A, W, M, Y, T, R, H;
Position
160
to
W, M, F, Y, R, H;
Position
161
to
I, W, M, F, Y, H;
Position
167
to
R, K;
Position
171
to
D;
Position
172
to
G, A, V, L, I, S, T, H;
Position
173
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, H;
Position
181
to
G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H;
Position
182
to
A, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, H;
Position
183
to
G, A, V, L, W, C, M, F, N, Q, Y, S, T, E, R, H;
Position
184
to
A, V, L, I, W, C, M, F, N, Q, Y, T, E, H;
Position
185
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T, E, H;
Position
186
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position
188
to
G, A, V, L, W, F, S, R, K;
Position
189
to
W, F;
Position
191
to
A, V, L, I, W, M, F, Y, T, R, H;
Position
192
to
G, L, I, W, M, N, Q, Y, S, T, D, R, H;
Position
194
to
W, N, Q, Y, D, H;
Position
195
to
W, P, Y;
Position
196
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E,
R, H;
Position
203
to
V, F, Y, R, H;
Position
204
to
I, W, M, Y, H;
Position
206
to
F;
Position
209
to
Y, R;
Position
210
to
W, F, Y;
Position
211
to
L, W, M, F, Y, H;
Position
212
to
V, L, I, W, M, F, Y, T, R, H;
Position
214
to
W, Y, R;
Position
215
to
A, L, I, W, M, F, Y;
Position
216
to
A, L, I, W, M, F, Y, R;
Position
217
to
W, R;
Position
218
to
G, A, L, W, P, M, F, Y, R, H;
Position
221
to
S;
Position
236
to
S;
Position
240
to
N;
Position
241
to
W;
Position
243
to
N;
Position
245
to
Q;
Position
247
to
G, V, I, W, P, F, Y, S, T, R;
Position
248
to
W, P, F, Y, E, R, H;
Position
249
to
L, W, P, F, S, D, E, H;
Position
251
to
G, L, I, W, P, M, F, Y, H;
Position
252
to
G, A, W, P, N, Q, Y, T, E, R, H;
Position
254
to
G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, H;
Position
255
to
G, L, W, M, F, N, Y, T, D, H;
Position
256
to
G, A, V, L, I, W, M, F, Q, Y, S, T, D, H;
Position
257
to
G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H;
Position
258
to
G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, E, K, H;
Position
259
to
A, V, I, W, M, F, N, Q, Y, S, T, E, R;
Position
260
to
L, I, W, M, F, Y, T, H;
Position
261
to
L, N, S, H;
Position
262
to
G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H;
Position
263
to
G, A, V, L, I, P, C, M, N, Q, Y, S, T, R, K;
Position
265
to
V, L, I, W, M, F, Y;
Position
269
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, R, H;
Position
271
to
A, L, I, W, P, M, F, N, Y, S, T, R, H;
Position
272
to
G, A, V, L, I, W, P, M, F, N, Q, Y, T, D, E, H;
Position
275
to
G, A, V, L, I, W, M, F, N, Y, T, D.
79 . The protein variant of claim 76 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
−1
to
Deletion;
Position
9
to
Insertion, deletion;
Position
10
to
Insertion, deletion;
Position
12
to
Insertion, deletion;
Position
14
to
Insertion, deletion;
Position
15
to
Insertion, deletion;
Position
17
to
Insertion, deletion;
Position
18
to
Insertion, deletion;
Position
19
to
Insertion, deletion;
Position
20
to
Insertion, deletion;
Position
21
to
Insertion, deletion;
Position
22
to
Insertion, deletion;
Position
24
to
Insertion, deletion;
Position
25
to
Insertion, deletion;
Position
46
to
Insertion, deletion;
Position
47
to
Insertion, deletion;
Position
48
to
Insertion, deletion;
Position
49
to
Insertion, deletion;
Position
50
to
Insertion, deletion;
Position
51
to
Insertion, deletion;
Position
52
to
Insertion, deletion;
Position
53
to
Insertion, deletion;
Position
54
to
Insertion, deletion;
Position
55
to
Insertion, deletion;
Position
58
to
Insertion, deletion;
Position
59
to
Insertion, deletion;
Position
61
to
Insertion, deletion;
Position
64
to
Insertion, deletion;
Position
78
to
Insertion:
Position
80
to
Insertion;
Position
91
to
Insertion, deletion;
Position
98
to
Deletion;
Position
99
to
Deletion;
Position
102
to
Deletion;
Position
105
to
Insertion;
Position
108
to
Insertion;
Position
109
to
Insertion;
Position
112
to
Insertion;
Position
113
to
Insertion;
Position
115
to
Insertion;
Position
116
to
Insertion;
Position
117
to
Insertion;
Position
118
to
Insertion;
Position
131
to
Deletion;
Position
134
to
Insertion, deletion;
Position
136
to
Insertion, deletion;
Position
137
to
Insertion, deletion;
Position
140
to
Insertion, deletion;
Position
141
to
Insertion, deletion;
Position
143
to
Insertion, deletion;
Position
144
to
Insertion, deletion;
Position
145
to
Insertion, deletion;
Position
146
to
Insertion, deletion;
Position
171
to
Deletion;
Position
172
to
Deletion;
Position
173
to
Deletion;
Position
181
to
Deletion;
Position
182
to
Deletion;
Position
183
to
Deletion;
Position
184
to
Deletion;
Position
185
to
Deletion;
Position
186
to
Deletion;
Position
188
to
Deletion;
Position
189
to
Deletion;
Position
191
to
Deletion;
Position
192
to
Deletion;
Position
195
to
Deletion;
Position
196
to
Insertion, deletion;
Position
221
to
Insertion;
Position
236
to
Insertion;
Position
237
to
Insertion;
Position
238
to
Insertion;
Position
239
to
Insertion;
Position
240
to
Insertion;
Position
241
to
Insertion;
Position
242
to
Insertion;
Position
243
to
Insertion;
Position
244
to
Insertion;
Position
245
to
Insertion;
Position
247
to
Insertion, deletion;
Position
248
to
Insertion, deletion;
Position
249
to
Insertion, deletion;
Position
251
to
Insertion, deletion;
Position
252
to
Insertion, deletion;
Position
254
to
Insertion, deletion;
Position
255
to
Insertion, deletion;
Position
256
to
Insertion, deletion;
Position
257
to
Insertion, deletion;
Position
258
to
Insertion, deletion;
Position
259
to
Insertion, deletion;
Position
260
to
Insertion, deletion;
Position
261
to
Insertion, deletion;
Position
262
to
Insertion, deletion;
Position
263
to
Insertion, deletion;
Position
265
to
Insertion, deletion;
Position
269
to
Insertion, deletion;
Position
271
to
Insertion, deletion;
Position
272
to
Insertion, deletion;
Position
275
to
Insertion, deletion.
80 . The protein variant of claim 76 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
7
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E,
R, H;
Position
8
to
G, A, L, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
13
to
G, L, I, W, P, M, F, N, Q, Y, S, D, E, H;
Position
16
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, D, E, R, H;
Position
23
to
G, A, V, L, I, W, M, F, Y, E, R, H;
Position
26
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position
28
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
29
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
33
to
V, L, I, W, C, M, F, N, Q, Y, R, H;
Position
35
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
36
to
V, L, I, W, P, M, F, N, Y, S, T, R, H;
Position
37
to
L, I, W, M, F, N, Q, Y, S, R, H;
Position
41
to
G, V, L, I, W, M, F, N, Q, Y, S, T, R, H;
Position
60
to
G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H;
Position
63
to
G, A, V, L, I, W, M, F, Y, T, R, H;
Position
73
to
A;
Position
74
to
A;
Position
81
to
V;
Position
82
to
L;
Position
86
to
G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H;
Position
88
to
A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position
92
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
93
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
94
to
G, V, L, I, W, P, M, F, N, Y, T, D, E, K, H;
Position
96
to
L, W, F, Y, R, K;
Position
97
to
V, L, W, C, M, F, Y, H;
Position
111
to
I;
Position
114
to
A;
Position
119
to
M;
Position
124
to
M;
Position
135
to
G, L, P, C, N, Q, T, R, H;
Position
138
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E,
R, H;
Position
142
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
147
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position
151
to
G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
162
to
I, W, F, Y, R;
Position
163
to
V, W, M, F, H;
Position
168
to
G, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
169
to
C, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y;
Position
174
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
176
to
G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E,
R, K, H;
Position
179
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
187
to
A, V, L, I, W, M, F, Y, R;
Position
190
to
G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, R, K, H;
Position
193
to
G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position
197
to
G, V, L, I, W, P, M, F, Q, Y, S, T, H;
Position
198
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
205
to
W, F, Y, R, K;
Position
208
to
A, V, L, I, W, C, M, F, Y, T, R, K, H;
Position
219
to
G, A, V, L, I, W, F, Y, R, H;
Position
222
to
M;
Position
232
to
A;
Position
233
to
L;
Position
234
to
I;
Position
250
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E,
R, H;
Position
267
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position
268
to
G, V, L, I, W, C, M, N, Q, Y, S, T, D, E, R, K, H;
Position
270
to
G, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position
273
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H;
Position
274
to
W, P, M, F, N, Q, Y, T, D, E, R, H.
81 . The protein variant of claim 76 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
13
to
Insertion, deletion;
Position
16
to
Insertion, deletion;
Position
23
to
Insertion, deletion;
Position
26
to
Insertion, deletion;
Position
28
to
Insertion, deletion;
Position
29
to
Insertion, deletion;
Position
35
to
Deletion;
Position
60
to
Insertion, deletion;
Position
63
to
Insertion;
Position
81
to
Insertion;
Position
82
to
Insertion;
Position
92
to
Insertion, deletion;
Position
93
to
Insertion, deletion;
Position
94
to
Insertion, deletion;
Position
96
to
Deletion,
Position
106
to
Insertion,
Position
111
to
Insertion,
Position
114
to
Insertion,
Position
119
to
Insertion,
Position
124
to
Insertion,
Position
138
to
Insertion, deletion;
Position
142
to
Insertion, deletion;
Position
147
to
Insertion, deletion;
Position
151
to
Insertion, deletion;
Position
174
to
Insertion, deletion;
Position
176
to
Insertion, deletion;
Position
179
to
Insertion, deletion;
Position
187
to
Deletion;
Position
190
to
Deletion;
Position
193
to
Deletion;
Position
197
to
Insertion, deletion;
Position
198
to
Insertion, deletion;
Position
232
to
Insertion,
Position
233
to
Insertion,
Position
234
to
Insertion,
Position
246
to
Insertion,
Position
250
to
Insertion, deletion;
Position
267
to
Insertion, deletion;
Position
268
to
Insertion, deletion;
Position
270
to
Insertion, deletion;
Position
273
to
Insertion, deletion.
82 . The protein variant of claim 76 , wherein the protease is a savinase-like subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
2
to
G, V, I, M, F, N, Q, Y, S, T, H,
Position
3
to
W, M, F, N, Q, Y, S, D, E, R, H,
Position
4
to
V, L, W, M, F, Y, R,
Position
6
to
G, V, L, I, W, P, M, N, Q, T, D, E, R, H,
Position
9
to
G, V, L, I, W, P, M, F, Q, Y, S, T, R, H, insertion, deletion,
Position
10
to
G, A, V, I, W, P, M, N, Q, Y, S, T, D, E, R, insertion, deletion,
Position
12
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, insertion, deletion,
Position
14
to
V, L, I, W, P, M, F, N, Q, Y, T, R, H, insertion, deletion,
Position
15
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, E, H, insertion, deletion,
Position
17
to
G, A, V, I, W, P, M, F, Y, H, insertion, deletion,
Position
18
to
G, A, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position
19
to
A, V, I, W, M, F, N, Y, S, T, D, R, H, insertion, deletion,
Position
20
to
G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, insertion, deletion,
Position
21
to
G, V, I, W, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position
22
to
G, V, L, I, W, M, F, Y, S, T, insertion, deletion,
Position
24
to
G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, insertion, deletion,
Position
25
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position
27
to
G, L, I, W, P, M, F, Y, T, H,
Position
37
to
L, I, W, M, F, N, Q, Y, S, R, H,
Position
40
to
V, L, I, W, M, F, N, Q, Y, T, R, H,
Position
42
to
G, A, L, W, C, M, F, N, Q, Y, S, T, D, E, R, H,
Position
43
to
G, L, H,
Position
44
to
G, V, L, I, W, P, M, F, Y, S, T,
Position
45
to
G, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H,
Position
46
to
G, A, L, I, W, P, M, F, Y, H, insertion, deletion,
Position
47
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion,
deletion,
Position
48
to
A, L, I, P, M, F, N, Y, D, H, insertion, deletion,
Position
50
to
G, A, W, M, N, Q, Y, S, T, D, E, H, insertion, deletion,
Position
51
to
V, L, I, W, M, F, N, Y, R, deletion, insertion,
Position
54
to
V, L, I, W, M, F, S, R, deletion, insertion,
Position
55
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T, D, E, R, K, H, deletion,
insertion,
Position
57
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K, H,
Position
58
to
L, W, M, F, N, Y, R, insertion, deletion,
Position
59
to
A, V, L, I, C, T, H, insertion, deletion,
Position
61
to
V, L, I, W, M, F, Y, insertion, deletion,
Position
64
to
G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion,
deletion,
Position
75
to
L,
Position
78
to
insertion,
Position
79
to
I,
Position
87
to
A, V, L, I, W, M, F, Q, Y, S, T, D, E, H,
Position
89
to
G, V, L, I, W, P, F, N, Y, T, E,
Position
91
to
G, A, V, L, I, W, P, M, N, Y, S, T, D, E, R, H, insertion, deletion,
Position
98
to
A, deletion,
Position
100
to
G, V, L, I, W, M, F, Y, R, H,
Position
101
to
V, I, W, M, F, N, Q, Y, H,
Position
102
to
V, L, I, W, M, F, Y, R, H, G, deletion,
Position
109
to
N, insertion,
Position
112
to
E, insertion,
Position
113
to
W, insertion,
Position
116
to
insertion,
Position
117
to
N, insertion,
Position
126
to
L,
Position
127
to
G, A, V, I, W, M, F, Y, R, H, L,
Position
128
to
I, W,
Position
129
to
W,
Position
130
to
W, F, Y, R,
Position
131
to
W, Y, R, deletion,
Position
132
to
L, W, M, F, Y, S, H,
Position
133
to
A, L, I, W, M, F, Y, R,
Position
134
to
L, I, W, F, N, Q, Y, R, H, insertion, deletion,
Position
136
to
G, A, W, P, N, Y, S, T, D, E, H, insertion, deletion,
Position
137
to
G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position
140
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, H, insertion, deletion,
Position
141
to
G, V, L, I, W, P, M, F, Q, S, D, E, H, insertion, deletion,
Position
143
to
V, L, I, P, M, F, N, Y, R, insertion, deletion,
Position
144
to
L, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position
145
to
G, V, L, I, W, M, F, Q, Y, D, E, R, H, insertion, deletion,
Position
146
to
G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position
155
to
V, L, I, W, M, F, Y, R,
Position
156
to
V, I, W, F, R,
Position
157
to
G, A, V, L, I, W, M, F, Y, T, R, H,
Position
158
to
V, L, I, W, M, F, Y,
Position
160
to
W, M, F, Y, R, H,
Position
161
to
I, W, M, F, Y, H,
Position
167
to
R, K,
Position
170
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
171
to
D, deletion,
Position
172
to
G, A, V, L, I, S, T, H, deletion,
Position
173
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, H, deletion,
Position
181
to
G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H, deletion,
Position
183
to
G, A, V, L, W, C, M, F, N, Q, Y, S, T, E, R, H, deletion,
Position
184
to
A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position
185
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position
186
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position
188
to
G, A, V, L, W, F, S, R, K, deletion,
Position
189
to
W, F, deletion,
Position
191
to
A, V, L, I, W, M, F, Y, T, R, H, deletion,
Position
192
to
G, L, I, W, M, N, Q, Y, S, T, D, R, H, deletion,
Position
194
to
W, N, Q, Y, D, H,
Position
195
to
W, P, Y, deletion,
Position
197
to
G, V, L, I, W, P, M, F, Q, Y, S, T, H, insertion, deletion,
Position
203
to
V, F, Y, R, H,
Position
206
to
F,
Position
209
to
Y, R,
Position
210
to
W, F, Y,
Position
212
to
V, L, I, W, M, F, Y, T, R, H,
Position
214
to
W, Y, R,
Position
216
to
A, L, I, W, M, F, Y, R,
Position
217
to
W, R,
Position
218
to
G, A, L, W, P, M, F, Y, R, H,
Position
221
to
S, insertion,
Position
236
to
S, insertion,
Position
237
to
insertion,
Position
239
to
insertion,
Position
240
to
N, insertion,
Position
241
to
W, insertion,
Position
242
to
insertion,
Position
244
to
insertion,
Position
245
to
Q, insertion,
Position
247
to
G, V, I, W, P, F, Y, S, T, R, insertion, deletion,
Position
248
to
W, P, F, Y, E, R, H, insertion, deletion,
Position
251
to
G, L, I, W, P, M, F, Y, H, insertion, deletion,
Position
252
to
G, A, W, P, N, Q, Y, T, E, R, H, insertion, deletion,
Position
255
to
G, L, W, M, F, N, Y, T, D, H, insertion, deletion,
Position
256
to
G, A, V, L, I, W, M, F, Q, Y, S, T, D, H, insertion, deletion,
Position
257
to
G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H, insertion, deletion,
Position
258
to
G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, E, K, H, insertion, deletion,
Position
259
to
A, V, I, W, M, F, N, Q, Y, S, T, E, R, insertion, deletion,
Position
260
to
L, I, W, M, F, Y, T, H, insertion, deletion,
Position
261
to
L, N, S, H, insertion, deletion,
Position
262
to
G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position
263
to
G, A, V, L, I, P, C, M, N, Q, Y, S, T, R, K, insertion, deletion,
Position
265
to
V, L, I, W, M, F, Y, insertion, deletion,
Position
271
to
A, L, I, W, P, M, F, N, Y, S, T, R, H, insertion, deletion,
Position
272
to
G, A, V, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position
275
to
G, A, V, L, I, W, M, F, N, Y, T, D, insertion, deletion.
83 . The protein variant of claim 82 , wherein the savinase-like subtilisin comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
6
to
G, V, L, I, W, P, M, N, Q, T, D, E, R, H,
Position
9
to
G, V, L, I, W, P, M, F, Q, Y, S, T, R, H, insertion, deletion,
Position
10
to
G, A, V, I, W, P, M, N, Q, Y, S, T, D, E, R, insertion, deletion,
Position
14
to
V, L, I, W, P, M, F, N, Q, Y, T, R, H, insertion, deletion,
Position
15
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, E, H, insertion, deletion,
Position
17
to
G, A, V, I, W, P, M, F, Y, H, insertion, deletion,
Position
18
to
G, A, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position
19
to
A, V, I, W, M, F, N, Y, S, T, D, R, H, insertion, deletion,
Position
20
to
G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, insertion, deletion,
Position
21
to
G, V, I, W, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position
37
to
L, I, W, M, F, N, Q, Y, S, R, H,
Position
43
to
G, L, H,
Position
45
to
G, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H,
Position
47
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion,
deletion,
Position
50
to
G, A, W, M, N, Q, Y, S, T, D, E, H, insertion, deletion,
Position
51
to
V, L, I, W, M, F, N, Y, R, deletion, insertion,
Position
54
to
V, L, I, W, M, F, S, R, deletion, insertion,
Position
59
to
A, V, L, I, C, T, H, insertion, deletion,
Position
89
to
G, V, L, I, W, P, F, N, Y, T, E,
Position
91
to
G, A, V, L, I, W, P, M, N, Y, S, T, D, E, R, H, insertion, deletion,
Position
101
to
V, I, W, M, F, N, Q, Y, H,
Position
109
to
N, insertion,
Position
112
to
E, insertion,
Position
113
to
W, insertion,
Position
127
to
G, A, V, I, W, M, F, Y, R, H, L,
Position
128
to
I, W,
Position
129
to
W,
Position
130
to
W, F, Y, R,
Position
131
to
W, Y, R, deletion,
Position
133
to
A, L, I, W, M, F, Y, R,
Position
136
to
G, A, W, P, N, Y, S, T, D, E, H, insertion, deletion,
Position
137
to
G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position
140
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, H, insertion, deletion,
Position
141
to
G, V, L, I, W, P, M, F, Q, S, D, E, H, insertion, deletion,
Position
143
to
V, L, I, P, M, F, N, Y, R, insertion, deletion,
Position
144
to
L, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position
145
to
G, V, L, I, W, M, F, Q, Y, D, E, R, H, insertion, deletion,
Position
146
to
G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position
155
to
V, L, I, W, M, F, Y, R,
Position
157
to
G, A, V, L, I, W, M, F, Y, T, R, H,
Position
158
to
V, L, I, W, M, F, Y,
Position
160
to
W, M, F, Y, R, H,
Position
161
to
I, W, M, F, Y, H,
Position
167
to
R, K,
Position
170
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
171
to
D, deletion,
Position
172
to
G, A, V, L, I, S, T, H, deletion,
Position
173
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, H, deletion,
Position
181
to
G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H, deletion,
Position
184
to
A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position
185
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position
186
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position
188
to
G, A, V, L, W, F, S, R, K, deletion,
Position
189
to
W, F, deletion,
Position
192
to
G, L, I, W, M, N, Q, Y, S, T, D, R, H, deletion,
Position
194
to
W, N, Q, Y, D, H,
Position
195
to
W, P, Y, deletion,
Position
197
to
G, V, L, I, W, P, M, F, Q, Y, S, T, H, insertion, deletion,
Position
203
to
V, F, Y, R, H,
Position
210
to
W, F, Y,
Position
218
to
G, A, L, W, P, M, F, Y, R, H,
Position
236
to
S, insertion,
Position
237
to
insertion,
Position
239
to
insertion,
Position
240
to
N, insertion,
Position
241
to
W, insertion,
Position
242
to
insertion,
Position
244
to
insertion,
Position
245
to
Q, insertion,
Position
247
to
G, V, I, W, P, F, Y, S, T, R, insertion, deletion,
Position
251
to
G, L, I, W, P, M, F, Y, H, insertion, deletion,
Position
255
to
G, L, W, M, F, N, Y, T, D, H, insertion, deletion,
Position
256
to
G, A, V, L, I, W, M, F, Q, Y, S, T, D, H, insertion, deletion,
Position
257
to
G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H, insertion, deletion,
Position
258
to
G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, E, K, H, insertion, deletion,
Position
260
to
L, I, W, M, F, Y, T, H, insertion, deletion,
Position
262
to
G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position
265
to
V, L, I, W, M, F, Y, insertion, deletion,
Position
271
to
A, L, I, W, P, M, F, N, Y, S, T, R, H, insertion, deletion,
Position
272
to
G, A, V, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position
275
to
G, A, V, L, I, W, M, F, N, Y, T, D, insertion, deletion.
84 . The savinase-like subtilisin of claim 82 , wherein the subtilisin has at least 81%, preferably at least 96%, more preferably at least 98%, most preferably at least 99% homology to SEQ ID NO: 24.
85 . The savinase-like subtilisin of claim 84 , wherein the subtilisin has any of the amino acid sequence of SEQ ID NO: 24, 26, 27, 28, 29, 30, 31, 32, 34, 35.
86 . The protein variant of claim 76 , wherein the protease is a savinase-like subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
8
to
G, A, L, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H,
Position
16
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, D, E, R, H,
insertion, deletion,
Position
23
to
G, A, V, L, I, W, M, F, Y, E, R, H, insertion,
deletion,
Position
26
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H,
insertion, deletion,
Position
35
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K,
H, deletion,
Position
38
to
V, L, I, W, M, F, N, Q, Y, T, H,
Position
39
to
G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H,
Position
41
to
G, V, L, I, W, M, F, N, Q, Y, S, T, R, H,
Position
60
to
G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H,
insertion, deletion,
Position
73
to
A,
Position
74
to
A,
Position
80
to
G, insertion,
Position
81
to
V, insertion,
Position
86
to
G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H,
Position
88
to
A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H,
Position
90
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T,
V, W, Y, insertion, deletion,
Position
93
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H, insertion, deletion,
Position
108
to
I, insertion,
Position
111
to
I, insertion,
Position
124
to
M, insertion,
Position
135
to
G, L, P, C, N, Q, T, R, H,
Position
142
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H, insertion, deletion,
Position
147
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, K, H,
insertion, deletion,
Position
148
to
G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E,
R, K, H, insertion, deletion,
Position
149
to
G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E,
R, K, H, insertion, deletion,
Position
151
to
G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H, insertion, deletion,
Position
163
to
V, W, M, F, H,
Position
168
to
G, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, R,
K, H,
Position
169
to
C, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y,
Position
174
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R,
K, H, insertion, deletion,
Position
179
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H, insertion, deletion,
Position
190
to
G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, R, K, H,
deletion,
Position
193
to
G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H,
deletion,
Position
196
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
H, insertion, deletion,
Position
208
to
A, V, L, I, W, C, M, F, Y, T, R, K, H,
Position
213
to
N, oN, E,
Position
215
to
A, L, I, W, M, F, Y,
Position
232
to
A, insertion,
Position
233
to
L, insertion,
Position
234
to
I, insertion,
Position
246
to
insertion,
Position
250
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
H, insertion, deletion,
Position
254
to
G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, H,
insertion, deletion,
Position
267
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H,
insertion, deletion,
Position
268
to
G, V, L, I, W, C, M, N, Q, Y, S, T, D, E, R, K, H,
insertion, deletion,
Position
269
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, R, H,
insertion, deletion,
Position
273
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R,
K, H, insertion, deletion,
87 . The savinase-like subtilisin of claim 86 , wherein the subtilisin has at least 81%, preferably at least 96%, more preferably at least 98%, most preferably at least 99% homology to SEQ ID NO: 24.
88 . The savinase-like subtilisin of claim 87 , wherein the subtilisin has any of the amino acid sequence of SEQ ID NO: 24, 26, 27, 28, 29, 30, 31, 32, 34, 35.
89 . The protein variant of claim 76 havingmodified immunogenicity as compared to its parent protein having at least 81% homolgy to SEQ ID NO: 25 comprising one or more of the following substitions corresponding to any of the following in SEQ ID NO: 25:
Position
21
to
G, V, I, W, N, Q, Y, S, T, D, E, R, H,
insertion, deletion,
Position
27
to
G, L, I, W, P, M, F, Y, T, H,
Position
50
to
G, A, W, M, N, Q, Y, S, T, D, E, H,
insertion, deletion,
Position
52
to
V, L, I, W, M, F, Y, S, T, R, deletion,
insertion,
Position
55
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T,
D, E, R, K, H, deletion, insertion,
Position
129
to
W,
Position
133
to
A, L, I, W, M, F, Y, R,
Position
172
to
G, A, V, L, I, S, T, H, deletion,
Position
186
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D,
E, R, H, deletion,
Position
194
to
W, N, Q, Y, D, H,
Position
195
to
W, P, Y, deletion,
Position
197
to
G, V, L, I, W, P, M, F, Q, Y, S, T, H,
insertion, deletion,
Position
242
to
insertion,
Position
249
to
L, W, P, F, S, D, E, H, insertion,
deletion,
Position
252
to
G, A, W, P, N, Q, Y, T, E, R, H,
insertion, deletion,
Position
254
to
G, V, L, I, W, M, F, N, Q, Y, S, D, E,
R, H, insertion, deletion,
Position
257
to
G, A, L, I, W, C, M, F, N, Q, Y, S, T,
D, E, K, H, insertion, deletion,
Position
260
to
L, I, W, M, F, Y, T, H, insertion,
deletion,
Position
265
to
V, L, I, W, M, F, Y, insertion,
deletion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
90 . The protein variant of claim 76 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 10 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position
4
to
V, L, W, M, F, Y, R,
Position
38
to
V, L, I, W, M, F, N, Q, Y, T, H,
Position
40
to
V, L, I, W, M, F, N, Q, Y, T, R, H,
Position
43
to
G, L, H,
Position
47
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S,
T, D, E, R, H, insertion, deletion,
Position
49
to
G, A, V, I, W, P, M, F, N, Q, Y, S, T,
D, E, R, H, insertion, deletion,
Position
54
to
V, L, I, W, M, F, S, R, deletion,
insertion,
Position
96
to
L, W, F, Y, R, K, deletion,
Position
99
to
V, L, I, W, M, F, Q, Y, H, deletion,
Position
113
to
W, insertion,
Position
131
to
W, Y, R, deletion,
Position
133
to
A, L, I, W, M, F, Y, R,
Position
137
to
G, A, V, I, W, P, M, N, Y, H,
insertion, deletion,
Position
141
to
G, V, L, I, W, P, M, F, Q, S, D, E,
H, insertion, deletion,
Position
144
to
L, W, P, M, F, N, Q, Y, S, D, E, R,
H, insertion, deletion,
Position
170
to
A, C, D, E, F, G, H, I, K, L, M, N,
P, Q, R, S, T, V, W, Y;
Position
173
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T,
E, H, deletion,
Position
181
to
G, A, V, L, I, W, C, M, F, Q, Y, T, D,
R, K, H, deletion,
Position
185
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T,
E, H, deletion,
Position
186
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D,
E, R, H, deletion,
Position
188
to
G, A, V, L, W, F, S, R, K, deletion,
Position
194
to
W, N, Q, Y, D, H,
Position
203
to
V, F, Y, R, H,
Position
210
to
W, F, Y,
Position
211
to
L, W, M, F, Y, H,
Position
257
to
G, A, L, I, W, C, M, F, N, Q, Y, S, T,
D, E, K, H, insertion, deletion,
Position
261
to
L, N, S, H, insertion, deletion,
Position
262
to
G, A, V, L, I, W, P, F, N, Q, Y, T, D,
E, R, H, insertion, deletion,
Position
265
to
V, L, I, W, M, F, Y, insertion, deletion.
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
91 . The protein variant of claim 76 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 11 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 11:
Position
38
to
V, L, I, W, M, F, N, Q, Y, T, H,
Position
40
to
V, L, I, W, M, F, N, Q, Y, T, R, H,
Position
45
to
G, V, L, I, W, P, M, F, N, Q, Y, S, T, D,
E, R, H,
Position
47
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T,
D, E, R, H, insertion, deletion,
Position
49
to
G, A, V, I, W, P, M, F, N, Q, Y, S, T, D,
E, R, H, insertion, deletion,
Position
50
to
G, A, W, M, N, Q, Y, S, T, D, E, H,
insertion, deletion,
Position
52
to
V, L, I, W, M, F, Y, S, T, R, deletion,
insertion,
Position
53
to
A, V, L, I, W, M, F, N, Q, Y, S, D, E,
H, deletion, insertion,
Position
56
to
G, V, L, I, W, M, F, N, Q, Y, S, T, H,
Position
58
to
L, W, M, F, N, Y, R, insertion, deletion,
Position
96
to
L, W, F, Y, R, K, deletion,
Position
97
to
V, L, W, C, M, F, Y, H,
Position
98
to
A, deletion,
Position
105
to
insertion,
Position
109
to
N, insertion,
Position
113
to
W, insertion,
Position
115
to
I, insertion,
Position
133
to
A, L, I, W, M, F, Y, R,
Position
136
to
G, A, W, P, N, Y, S, T, D, E, H,
insertion, deletion,
Position
137
to
G, A, V, I, W, P, M, N, Y, H,
insertion, deletion,
Position
141
to
G, V, L, I, W, P, M, F, Q, S, D, E, H,
insertion, deletion,
Position
158
to
V, L, I, W, M, F, Y,
Position
159
to
A, W, M, Y, T, R, H,
Position
172
to
G, A, V, L, I, S, T, H, deletion,
Position
186
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D,
E, R, H, deletion,
Position
189
to
W, F, deletion,
Position
192
to
G, L, I, W, M, N, Q, Y, S, T, D, R, H,
deletion,
Position
195
to
W, P, Y, deletion,
Position
197
to
G, V, L, I, W, P, M, F, Q, Y, S, T, H,
insertion, deletion,
Position
257
to
G, A, L, I, W, C, M, F, N, Q, Y, S, T,
D, E, K, H, insertion, deletion,
Position
261
to
L, N, S, H, insertion, deletion,
Position
265
to
V, L, I, W, M, F, Y, insertion,
deletion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
92 . The protein variant of claim 76 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 33 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 33:
Position
−6
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, Y, insertion, deletion,
Position
−5
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, T, V, W, Y, insertion, deletion,
Position
−4
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R,
S, T, V, W, Y, insertion, deletion,
Position
−2
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R,
S, T, V, W, Y, insertion, deletion,
Position
−1
to
G, V, L, I, W, C, M, F, N, Q, Y, S, T, D,
E, R, H, deletion,
Position
1
to
V, L, I, W, M, F, Y, S, T, R,
Position
2
to
G, V, I, M, F, N, Q, Y, S, T, H,
Position
3a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W, Y, insertion, deletion,
Position
5
to
V, L, I, W, M, F, N, Q, Y, T, R, H,
Position
6
to
G, V, L, I, W, P, M, N, Q, T, D, E, R, H,
Position
7
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T,
D, E, R, H,
Position
8
to
G, A, L, W, P, C, M, F, N, Q, Y, S, T, D,
E, R, K, H,
Position
10
to
G, A, V, I, W, P, M, N, Q, Y, S, T, D, E,
R, insertion, deletion,
Position
12
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D,
E, insertion, deletion,
Position
13
to
G, L, I, W, P, M, F, N, Q, Y, S, D, E, H,
insertion, deletion,
Position
14
to
V, L, I, W, P, M, F, N, Q, Y, T, R, H,
insertion, deletion,
Position
15
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T,
E, H, insertion, deletion,
Position
16
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, D,
E, R, H, insertion, deletion,
Position
17
to
G, A, V, I, W, P, M, F, Y, H, insertion,
deletion,
Position
18
to
G, A, L, I, W, P, M, F, N, Q, Y, T, D, E,
H, insertion, deletion,
Position
19
to
A, V, I, W, M, F, N, Y, S, T, D, R, H,
insertion, deletion,
Position
21
to
G, V, I, W, N, Q, Y, S, T, D, E, R, H,
insertion, deletion,
Position
22
to
G, V, L, I, W, M, F, Y, S, T, insertion,
deletion,
Position
23
to
G, A, V, L, I, W, M, F, Y, E, R, H, insertion,
deletion,
Position
24
to
G, V, L, I, W, M, F, N, Q, Y, S, D, E, R,
insertion, deletion,
Position
25
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E,
R, H, insertion, deletion,
Position
26
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E,
R, H, insertion, deletion,
Position
27
to
G, L, I, W, P, M, F, Y, T, H,
Position
28
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D,
E, R, K, H, insertion, deletion,
Position
28a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R,
S, T, V, W, Y, insertion, deletion,
Position
29
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D,
E, R, K, H, insertion, deletion,
Position
33
to
V, L, I, W, C, M, F, N, Q, Y, R, H,
Position
35
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E,
R, K, H, deletion,
Position
37
to
L, I, W, M, F, N, Q, Y, S, R, H,
Position
40
to
V, L, I, W, M, F, N, Q, Y, T, R, H,
Position
42
to
G, A, L, W, C, M, F, N, Q, Y, S, T, D, E,
R, H,
Position
43
to
G, L, H,
Position
44
to
G, V, L, I, W, P, M, F, Y, S, T,
Position
44a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R,
S, T, V, W, Y, insertion, deletion,
Position
44b
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R,
S, T, V, W, Y, insertion, deletion,
Position
46
to
G, A, L, I, W, P, M, F, Y, H, insertion,
deletion,
Position
48
to
A, L, I, P, M, F, N, Y, D, H, insertion,
deletion,
Position
51
to
V, L, I, W, M, F, N, Y, R, deletion,
insertion,
Position
52
to
V, L, I, W, M, F, Y, S, T, R, deletion,
insertion,
Position
53
to
A, V, L, I, W, M, F, N, Q, Y, S, D, E, H,
deletion, insertion,
Position
55
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T, D,
E, R, K, H, deletion, insertion,
Position
56
to
G, V, L, I, W, M, F, N, Q, Y, S, T, H,
Position
57
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T, D,
E, R, K, H,
Position
58
to
L, W, M, F, N, Y, R, insertion, deletion,
Position
61
to
V, L, I, W, M, F, Y, insertion, deletion,
Position
64
to
G, V, L, I, W, P, C, M, F, N, Q, Y, S, T,
D, E, R, K, H, insertion, deletion,
Position
75
to
L,
Position
81
to
insertion,
Position
86
to
G, A, V, L, I, W, M, F, N, Q, Y, T, D, E,
R, H,
Position
87
to
A, V, L, I, W, M, F, Q, Y, S, T, D, E, H,
Position
88
to
A, V, L, I, W, M, F, N, Q, Y, S, T, D, E,
R, H,
Position
89
to
G, V, L, I, W, P, F, N, Y, T, E,
Position
91
to
G, A, V, L, I, W, P, M, N, Y, S, T, D, E,
R, H, insertion, deletion,
Position
92
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T,
D, E, R, K, H, insertion, deletion,
Position
94
to
G, V, L, I, W, P, M, F, N, Y, T, D, E, K,
H, insertion, deletion,
Position
96
to
L, W, F, Y, R, K, deletion,
Position
97
to
V, L, W, C, M, F, Y, H,
Position
98
to
deletion,
Position
101
to
V, I, W, M, F, N, Q, Y, H,
Position
102
to
V, L, I, W, M, F, Y, R, H, G, deletion,
Position
108
to
I, insertion,
Position
109
to
N, insertion,
Position
111
to
insertion,
Position
112
to
E, insertion,
Position
113
to
W, insertion,
Position
114
to
insertion,
Position
115
to
I, insertion,
Position
117
to
N, insertion,
Position
118
to
N, insertion,
Position
119
to
M, insertion,
Position
127
to
G, A, V, I, W, M, F, Y, R, H, L,
Position
133
to
A, L, I, W, M, F, Y, R,
Position
134
to
L, I, W, F, N, Q, Y, R, H, insertion,
deletion,
Position
135
to
G, L, P, C, N, Q, T, R, H,
Position
136
to
G, A, W, P, N, Y, S, T, D, E, H,
insertion, deletion,
Position
137
to
G, A, V, I, W, P, M, N, Y, H, insertion,
deletion,
Position
138
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S,
T, D, E, R, H, insertion, deletion,
Position
139
to
G, A, V, L, I, W, P, C, M, F, N, Q, Y,
S, T, D, E, R, K, H, insertion, deletion,
Position
141
to
G, V, L, I, W, P, M, F, Q, S, D, E, H,
insertion, deletion,
Position
142
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S,
T, D, E, R, K, H, insertion, deletion,
Position
144
to
L, W, P, M, F, N, Q, Y, S, D, E, R, H,
insertion, deletion,
Position
145
to
G, V, L, I, W, M, F, Q, Y, D, E, R, H,
insertion, deletion,
Position
146
to
G, A, W, L, I, W, M, F, N, Q, Y, T, D,
E, R, H, insertion, deletion,
Position
147
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D,
E, R, K, H, insertion, deletion,
Position
148
to
G, A, V, L, I, W, P, C, M, F, N, Q, Y,
S, T, D, E, R, K, H, insertion, deletion,
Position
156
to
V, I, W, F, R,
Position
158
to
V, L, I, W, M, F, Y,
Position
160
to
W, M, F, Y, R, H,
Position
161
to
I, W, M, F, Y, H,
Position
162
to
I, W, F, Y, R,
Position
163
to
V, W, M, F, H,
Position
167
to
R, K,
Position
169
to
C, E, F, G, H, I, K, L, M, N, Q, R, T,
V, W, Y,
Position
170
to
A, C, D, E, F, G, H, I, K, L, M, N, P,
Q, R, S, T, V, W, Y, insertion, deletion,
Position
171
to
D, deletion,
Position
174
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S,
T, D, E, R, K, H, insertion, deletion,
Position
176
to
G, A, V, L, I, W, P, C, M, F, N, Q, Y, S,
T, D, E, R, K, H, insertion, deletion,
Position
182
to
A, V, L, I, W, C, M, F, N, Q, Y, S, T, D,
E, H, deletion,
Position
186
to
G, A, V, L, W, M, F, N, Q, Y, S, T, D, E,
R, H, deletion,
Position
188
to
G, A, V, L, W, F, S, R, K, deletion,
Position
191
to
A, V, L, I, W, M, F, Y, T, R, H, deletion,
Position
192
to
G, L, I, W, M, N, Q, Y, S, T, D, R, H,
deletion,
Position
193
to
G, V, L, I, W, M, F, N, Q, Y, S, T, D,
E, R, H, deletion,
Position
194
to
W, N, Q, Y, D, H,
Position
195
to
W, P, Y, deletion,
Position
196
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T,
D, E, R, H, insertion, deletion,
Position
197
to
G, V, L, I, W, P, M, F, Q, Y, S, T, H,
insertion, deletion,
Position
198
to
G, A, L, I, W, P, C, M, F, N, Q, Y, S, T,
D, E, R, K, H, insertion, deletion,
Position
203
to
V, F, Y, R, H,
Position
205
to
W, F, Y, R, K,
Position
215
to
A, L, I, W, M, F, Y,
Position
216
to
A, L, I, W, M, F, Y, R,
Position
217
to
W, R,
Position
219
to
G, A, V, L, I, W, F, Y, R, H,
Position
233
to
insertion,
Position
234
to
I, insertion,
Position
236
to
insertion,
Position
237
to
insertion,
Position
238
to
insertion,
Position
239
to
insertion,
Position
240
to
insertion,
Position
243
to
insertion,
Position
246
to
insertion,
Position
247
to
G, V, I, W, P, F, Y, S, T, R, insertion,
deletion,
Position
249
to
L, W, P, F, S, D, E, H, insertion,
deletion,
Position
252
to
G, A, W, P, N, Q, Y, T, E, R, H, insertion,
deletion,
Position
254
to
G, V, L, I, W, M, F, N, Q, Y, S, D, E, R,
H, insertion, deletion,
Position
262
to
G, A, V, L, I, W, P, F, N, Q, Y, T, D, E,
R, H, insertion, deletion,
Position
264a
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W, Y, insertion, deletion,
Position
270
to
G, L, I, W, P, M, F, N, Q, Y, S, T, D, E,
R, K, H, insertion, deletion,
Position
273
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S, T,
D, E, R, K, H, insertion, deletion,
Position
274
to
W, P, M, F, N, Q, Y, T, D, E, R, H,
Position
275
to
G, A, V, L, I, W, M, F, N, Y, T, D,
insertion, deletion,
Position
276
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W, Y, insertion, deletion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
93 . The protein variant of claim 76 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 33 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 33:
Position
5
to
V, L, I, W, M, F, N, Q, Y, T, R, H,
Position
22
to
G, V, L, I, W, M, F, Y, S, T, insertion,
deletion,
Position
26
to
G, A, V, L, I, W, M, F, N, Q, Y, S, T,
D, E, R, H, insertion, deletion,
Position
28
to
G, A, V, L, I, W, P, M, F, N, Q, Y, S,
T, D, E, R, K, H, insertion, deletion,
Position
37
to
L, I, W, M, F, N, Q, Y, S, R, H,
Position
40
to
V, L, I, W, M, F, N, Q, Y, T, R, H,
Position
44
to
G, V, L, I, W, P, M, F, Y, S, T,
Position
51
to
V, L, I, W, M, F, N, Y, R, deletion,
insertion,
Position
52
to
V, L, I, W, M, F, Y, S, T, R, deletion,
insertion,
Position
55
to
G, A, V, L, I, W, C, M, F, N, Q, Y, T,
D, E, R, K, H, deletion, insertion,
Position
58
to
L, W, M, F, N, Y, R, insertion, deletion,
Position
61
to
V, L, I, W, M, F, Y, insertion, deletion,
Position
64
to
G, V, L, I, W, P, C, M, F, N, Q, Y, S,
T, D, E, R, K, H, insertion, deletion,
Position
87
to
A, V, L, I, W, M, F, Q, Y, S, T, D, E, H,
Position
97
to
V, L, W, C, M, F, Y, H,
Position
98
to
deletion,
Position
101
to
V, I, W, M, F, N, Q, Y, H,
Position
102
to
V, L, I, W, M, F, Y, R, H, G, deletion,
Position
109
to
N, insertion,
Position
112
to
E, insertion,
Position
118
to
N, insertion,
Position
127
to
G, A, V, I, W, M, F, Y, R, H, L,
Position
137
to
G, A, V, I, W, P, M, N, Y, H, insertion,
deletion,
Position
146
to
G, A, W, L, I, W, M, F, N, Q, Y, T, D,
E, R, H, insertion, deletion,
Position
156
to
V, I, W, F, R,
Position
158
to
V, L, I, W, M, F, Y,
Position
161
to
I, W, M, F, Y, H,
Position
188
to
G, A, V, L, W, F, S, R, K, deletion,
Position
192
to
G, L, I, W, M, N, Q, Y, S, T, D, R, H,
deletion,
Position
194
to
W, N, Q, Y, D, H,
Position
195
to
W, P, Y, deletion,
Position
203
to
V, F, Y, R, H,
Position
216
to
A, L, I, W, M, F, Y, R,
Position
236
to
insertion,
Position
237
to
insertion,
Position
262
to
G, A, V, L, I, W, P, F, N, Q, Y, T, D,
E, R, H, insertion, deletion,
Position
264a
to
A, C, D, E, F, G, H, I, K, L, M, N, P,
Q, R, S, T, V, W, Y, insertion,
deletion.
with the provisio that the amino acids of the parent enzyme are substituted to another mino acid.
94 . The protein variant of claim 76 , wherein the lipolytic enzyme comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 1:
Q15 to A, C, D, E, F, G, I, K, L, M, N, P, R, S, T, V, W, Y;
Y16 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
A18 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
A19 to C, D, E, F, G, H, I, K, L, M, N, Q, R, S, V, W, Y;
A20 to C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
N25 to A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
N26 to A, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
E43 to A, C, D, F, G, H, I, K, L, M, N, R, S, T, V, W, Y;
V44 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, W, Y;
K46 to A, C, D, E, F, G, H, I, L, M, N, Q, S, T, V, W, Y;
A47 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
A49 to C, D, E, F, G, H, I, K, L, M, N, Q, R, S, V, W, Y;
L52 to A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
Y53 to A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
S54 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y;
G65 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
L67 to A, C, D, E, F, G, H, I, K, M, N, Q, R, S, T, V, W, Y;
A68 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
L69 to A, C, D, E, F, G, H, I, K, M, N, P, Q, S, T, V, W, Y;
T72 to A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, V, W, Y;
K74 to A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
L75 to A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
V77 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
S79 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
R81 to A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
S83 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, V, W, Y;
S85 to A, D, E, G, H, I, L, M, N, Q, V, W, Y;
W89 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
L97 to A, C, D, E, F, G, H, I, K, N, P, R, S, T, W, Y;
K98 to A, C, G, H, L, M, N, P, Q, S, T, V, W, Y;
E99 to C, F, G, I, M, P, W, Y;
G106 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
C107 to A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
R108 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, Y;
G109 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
T123 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
L124 to A, C, D, E, F, G, H, I, K, M, N, P, Q, R, T, V, W, Y;
K127 to A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
E129 to A, C, D, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
A131 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
V132 to A, C, D, E, F, G, H, I, K, L, N, P, Q, R, S, T, W, Y;
Y138 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
V140 to A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, W, Y;
L147 to A, C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y;
A150 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
T153 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Y164 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
D165 to A, C, E, F, G, H, I, K, L, M, N, Q, S, T, V, W, Y;
D167 to A, C, E, F, H, I, L, M, N, P, Q, S, T, V, W, Y;
S170 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y;
Y171 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
G172 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
A173 to C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
P174 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
R175 to A, C, D, E, F, G, H, I, K, L, M, N, Q, S, T, V, W, Y;
V176 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, W, Y;
G177 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
R179 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
A182 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Y194 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
H198 to A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
N200 to A, C, D, E, F, G, H, I, K, L, M, P, Q, S, T, V, W, Y;
P207 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
P208 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
R209 to C, D, F, G, H, I, K, L, M, N, Q, T, V, W, Y;
G212 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
S214 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
H215 to A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
S216 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y;
S217 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
P218 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
E219 to C, D, F, H, I, M, P, W, Y;
Y220 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
K223 to A, C, D, E, F, G, H, I, L, M, N, Q, S, T, V, W, Y;
S224 to A, C, D, E, F, G, H, I, K, L, M, N, Q, T, V, W, Y;
D234 to C, E, F, H, I, M, W;
I235 to A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
K237 to A, C, D, E, F, G, H, I, L, N, P, Q, S, T, V, W, Y;
I238 to A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, Y;
D242 to C, E, F, G, H, I, M, P, W, Y;
A243 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
P250 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
P253 to A, C, D, E, F, G, H, I, K, L, M, N, Q, S, T, V, W, Y;
D254 to C, E, F, H, I, M, P, Y;
I255 to C, D, E, F, H, L, M, N, Q, W, Y;
P256 to C, E, F, G, H, I, K, L, M, N, Q, R, V, W, Y;
Y261 to A, C, E, F, G, H, L, M, N, P, Q, R, S, T, V.
95 . The protein variant of claim 94 , wherein the lipolytic enzyme comprises one or more of the substitutions corresponding to any of the following in SEQ ID NO: 1:
G65 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
L67 to A, C, D, E, F, G, H, I, K, M, N, Q, R, S, T, V, W, Y;
R81 to A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
S83 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, V, W, Y;
S85 to A, D, E, G, H, I, L, M, N, Q, V, W, Y;
L97 to A, C, D, E, F, G, H, I, K, N, P, R, S, T, W, Y;
L124 to A, C, D, E, F, G, H, I, K, M, N, P, Q, R, T, V, W, Y;
E129 to A, C, D, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Y164 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
R179 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
A182 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
P207 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
P208 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
R209 to C, D, F, G, H, I, K, L, M, N, Q, T, V, W, Y;
G212 to A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
S214 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
H215 to A, C, D, E, F, G, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
S216 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y;
S217 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
P218 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
E219 to C, D, F, H, I, M, P, W, Y;
Y220 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
A243 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
P250 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
P253 to A, C, D, E, F, G, H, I, K, L, M, N, Q, S, T, V, W, Y.
96 . The protein variant of claim 95 , wherein the lipolytic comprises one or more of the following substitutions:
P207 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
P208 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
S214 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
S216 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y;
S217 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
A243 to C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
P250 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
P253 to A, C, D, E, F, G, H, I, K, L, M, N, Q, S, T, V, W, Y.
97 . The protein variant of claim 94 , wherein the parent lipolytic enzyme has at least 80% homology with SEQ ID NO: 1.
98 . The protein variant of claim 76 , wherein the carbohydrase is a glucoamylase comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 36:
Position
68
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W, Y, deletion, insertion,
Position
94
to
insertion,
Position
102
to
insertion,
Position
122
to
insertion,
Position
125
to
insertion,
Position
272
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W, Y, deletion, insertion,
Position
345
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R,
S, T, V, W, Y, deletion, insertion,
Position
348
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q,
R, S, T, V, W, Y, deletion, insertion,
Position
353
to
insertion,
Position
357
to
insertion,
Position
359
to
insertion,
Position
450
to
insertion,
Position
451
to
insertion,
Position
468
to
insertion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
99 . The protein variant of claim 98 , wherein the enzyme is at least 81% homologous, preferably 90% homologous, more preferably 95% homologous, most preferably 99% homologous to Carezyme core (SEQ ID NO: 36).
100 . The protein variant of claim 76 , wherein the carbohydrase is a Thermamyl-like α-amylase comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 2:
Position
TYR
8
to
A, C, D, G, K, M, P, R, W, Y, insertion;
Position
ASP
25
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
insertion;
Position
ASP
26
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
ALA
27
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
SER
28
to
A, C, D, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
ASN
29
to
A, C, D, G, K, M, P, R, W, Y;
Position
ARG
31
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
PRO
41
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
PRO
42
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
TYR
54
to
A, C, D, E, G, K, M, P, R, Y, insertion;
Position
TYR
57
to
A, C, D, G, K, M, P, R, W, Y, insertion;
Position
LEU
62
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
GLY
63
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
GLY
76
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
ARG
78
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
SER
79
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
LEU
88
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
GLY
92
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
ASN
102
to
A, C, E, F, G, H, I, L, M, P, Q, S, T, V, W, Y, insertion;
Position
ALA
107
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
ASP
108
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
ALA
109
to
A, C, D, E, F, G, H, K, M, N, P, Q, R, S, W, Y;
Position
LYS
138
to
A, C, E, F, G, I, L, M, N, P, Q, S, T, V, W, insertion;
Position
ASP
140
to
A, C, E, F, G, I, K, L, M, N, P, Q, S, T, V, W;
Position
PRO
142
to
C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
ARG
144
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
GLN
170
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
ILE
173
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
ASP
195
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
TYR
196
to
A, C, D, G, K, M, P, R, W, Y, insertion;
Position
ASP
232
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
ALA
233
to
A, C, D, E, I, K, L, M, N, P, Q, R, W, Y, deletion, insertion;
Position
GLN
331
to
A, C, D, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
TYR
349
to
A, C, D, G, K, M, P, R, W, Y, insertion;
Position
ILE
352
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
GLN
357
to
C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, deletion,
insertion;
Position
ASP
366
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
TYR
367
to
C, E, F, H, K, M, N, P, Q, R, V, W, insertion;
Position
TYR
368
to
A, C, D, G, K, M, P, R, W, Y, insertion;
Position
ILE
370
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
ALA
380
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
LYS
381
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y,
deletion, insertion;
Position
ILE
382
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
PRO
384
to
C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, deletion,
insertion;
Position
LEU
386
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
ARG
389
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
GLN
390
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, S, T, V, W, Y;
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
101 . The protein variant of claim 100 , wherein the amylase comprises one or more of the following substitutions:
Position
PRO
41
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
PRO
42
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
ALA
109
to
A, C, D, E, F, G, H, K, M, N, P, Q, R, S, W, Y;
Position
LYS
138
to
A, C, E, F, G, I, L, M, N, P, Q, S, T, V, W;
Position
ASP
140
to
A, C, E, F, G, I, K, L, M, N, P, Q, S, T, V, W;
Position
PRO
142
to
C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
ARG
144
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
ASP
366
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
TYR
367
to
C, E, F, H, K, M, N, P, Q, R, V, W, insertion;
Position
TYR
368
to
A, C, D, G, K, M, P, R, W, Y, insertion;
Position
ALA
380
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
LYS
381
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y,
deletion, insertion;
Position
ILE
382
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
Position
PRO
384
to
C, D, E, F, G, H, I, K, M, N, P, Q, R, S, T, V, W, deletion,
insertion;
Position
ARG
389
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y,
deletion, insertion;
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
102 . The protein variant of claim 100 , wherein the enzyme is at least 81% homologous, preferably 90% homologous, more preferably 95% homologous, most preferably 99% homologous to SEQ ID NO: 2.
103 . The protein variant of claim 100 , wherein the enzyme has an amino acid sequence of SEQ ID NO: 2, 4, 5, or 37.
104 . A cellulase variant of a microbial parent cellulase having a catalytically active domain classified in family 45, said variant comprises a substitution of one or more amino acid residues at a position corresponding to a position in SEQ ID NO: 4 from the group consisting of:
Position
1
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
2
to
A, C, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
7
to
A, C, D, E, F, G, H, K, L, M, N, P, Q, S, T, V, Y;
Position
20
to
C, D, F, H, I, L, M, N, P, Q, S, T, V, W, Y;
Position
23
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
27
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
29
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
36
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
37
to
C, D, E, F, G, H, I, K, L, M, P, Q, T, V, W, Y;
Position
38
to
A, C, D, E, G, H, K, M, N, P, R, S, T, V, W, Y;
Position
40
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
41
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
44
to
A, C, D, E, F, H, I, L, M, N, S, T, W, Y;
Position
54
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
59
to
A, C, D, E, F, G, H, I, K, L, M, N, P, R, S, T, V, W, Y;
Position
61
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
62
to
A, C, D, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
83
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
84
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
95
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
96
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, V, W, Y;
Position
97
to
C, D, E, F, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
98
to
C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
100
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
101
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
131
to
C, D, E, F, K, M, P, R, S, W, Y;
Position
133
to
A, C, E, F, G, H, I, L, M, P, R, S, T, V, W, Y;
Position
134
to
C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
136
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
142
to
A, C, E, F, G, H, I, K, M, N, P, Q, R, V, W, Y;
Position
143
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
145
to
C, E, F, G, H, I, K, L, M, P, R, S, T, V, W, Y;
Position
146
to
A, C, D, F, G, H, I, K, L, M, N, P, T, V, W, Y;
Position
151
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
153
to
C, D, E, F, G, H, I, M, N, P, Q, S, T, V, W, Y;
Position
154
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
155
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
157
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
158
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
160
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
162
to
C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
163
to
A, C, D, E, F, G, H, I, K, M, P, Q, R, S, T, Y;
Position
164
to
A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, Y;
Position
165
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
168
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W;
Position
169
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y;
Position
170
to
A, C, D, E, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
174
to
A, C, D, E, G, H, I, K, L, N, P, Q, R, S, T, V, Y;
Position
176
to
A, C, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
177
to
C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
178
to
A, C, E, F, G, H, I, K, L, M, Q, R, S, T, V, W, Y;
Position
180
to
A, C, D, E, F, G, H, I, K, M, N, Q, R, S, T, V, W, Y;
Position
183
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
191
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
195
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
197
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, V, W, Y; and
Position
200
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y.
105 . The protein variant of claim 104 , wherein the carbohydrase comprises one or more of the following substitutions:
Position
20
to
C, D, F, H, I, L, M, N, P, Q, S, T, V, W, Y;
Position
23
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
27
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
83
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
84
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
95
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
96
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, V, W, Y;
Position
97
to
C, D, E, F, H, I, K, L, M, N, P, Q, R, S, V, W, Y;
Position
98
to
C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position ALA
100
to
C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y;
Position
101
to
A, C, D, E, F, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
131
to
C, D, E, F, K, M, P, R, S, W, Y;
Position
142
to
A, C, E, F, G, H, I, K, M, N, P, Q, R, V, W, Y;
Position
143
to
A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y;
Position
145
to
C, E, F, G, H, I, K, L, M, P, R, S, T, V, W, Y;
Position
151
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y;
Position
154
to
A, C, D, E, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, Y;
Position
155
to
A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
157
to
A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y; and
Position
158
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, Y.
106 . The protein variant of claim 104 , wherein the enzyme is at least 81% homologous, preferably 90% homologous, more preferably 95% homologous, most preferably 99% homologous to Carezyme core (SEQ ID NO: 4).
107 . The protein variant of claim 76 , wherein the laccase is a Coprinus -like laccase.
108 . The protein variant of claim 107 , wherein the laccase comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 3:
Position
5
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
8
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
10
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
12
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
22
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
23
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
30
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
39
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
40
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
41
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
42
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
43
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
51
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
53
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
55
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
58
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
59
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
60
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
71
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
72
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
78
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
79
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
80
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
100
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
101
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
102
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
112
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
113
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
114
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
118
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
139
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
142
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
155
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
157
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
165
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
166
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
168
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
175
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
180
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
183
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
186
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
190
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
191
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
192
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
193
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
211
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
213
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
231
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
234
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
236
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
241
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
251
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
257
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
259
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
265
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
275
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
286
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
294
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
295
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
296
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
299
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
300
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
301
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
302
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
306
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
313
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
314
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
315
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
320
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
321
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
322
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
324
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
329
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
332
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
335
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
336
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
339
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
344
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
345
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
348
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
349
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
350
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
366
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
367
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
369
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
370
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
371
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
372
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
375
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
378
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
379
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
389
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
390
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
409
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
410
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
414
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
416
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
418
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
419
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
420
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
430
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
432
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
433
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
434
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
442
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
443
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
445
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
446
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
469
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
473
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
485
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
488
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
490
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
491
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
492
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
493
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
494
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
495
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
496
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
499
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
500
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
501
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
with the proviso that the amino acids of the parent protein is substituted to another amino acid.
109 . The protein variant of claim 108 , wherein the laccase comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 3:
Position
59
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
96
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
100
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
181
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
348
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
369
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
414
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
432
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position
493
to
A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
with the proviso that the amino acids of the parent protein is substituted to another amino acid.
110 . A subtilisin variant comprising one or more of the insertions, substitutions and/or deletions in any of the positions of claim 77 .
111 . The variant of claim 110 , wherein the subtilisin has at least 60%, preferably at least 70%, more preferably at least 80, even more preferably at least 90, still more preferably at least 95%, most preferably at least 99% homology to SEQ ID NO; 10.
112 . A lipolytic enzyme comprising one or more of the insertions, substitutions and/or deletions in any of the positions of claim 94 .
113 . A glycoamylase variant comprising one or more of the insertions, substitutions and/or deletions in any of the positions of claim 98 .
114 . The variant of claim 113 , wherein the variant has at least 60%, preferably at least 70%, more preferably at least 80, even more preferably at least 90, still more preferably at least 95%, most preferably at least 99% homology to SEQ ID NO: 36.
115 . A Termamyl-like alpha-amylase comprising one or more of the insertions, substitutions and/or deletions in any of the positions of claim 100 .
116 . The variant of claim 115 , wherein the variant has at least 60%, preferably at least 70%, more preferably at least 80, even more preferably at least 90, still more preferably at least 95%, most preferably at least 99% homology to SEQ ID NO; 2.
117 . A cellulase variant comprising one or more of the insertions, substitutions and/or deletions in any of the positions of claim 104 .
118 . The variant of claim 117 , wherein the variant has at least 60%, preferably at least 70%, more preferably at least 80, even more preferably at least 90, still more preferably at least 95%, most preferably at least 99% homology to SEQ ID NO: 10.
119 . A Coprinus -like laccase variant comprising one or more of the insertions, substitutions and/or deletions in any of the positions of claim 107 .
120 . A composition comprising a protein variant of claim 22 .
121 . The composition of claim 120 , wherein the composition is in form of a pharmaceutical composition such as a vaccine.
122 . The composition of claim 120 , wherein the compositions is in form of a industrial composition such as, a detergent composition, personal care composition.
123 . The use of the composition as defined in claim 120 for the production of a pharmaceutical.
124 . The use of the composition as defined in claim 120 for industrial application.
125 . A DNA construct comprising a DNA sequence encoding a protein variant of claim 22 .
126 . An expression vector comprising a DNA construct of claim 125 .
127 . A host cell which is capable of expressing a polypeptide and comprising a DNA construct of claim 125 .
128 . A host cell which is capable of expressing a polypeptide and which is transformed by an expression vector of claim 126 .
129 . A host cell of claim 127 , which is a fungal cell, an insect cell, a mammalian cell, or a plant cell.
130 . A method of producing a protein variant having reduced immunogenicity as compared to the parent protein, comprising:
(a) culturing a host of claim 127 in a suitable culture medium to obtain expression and secretion of the protein into the medium, followed by (b) isolation of the protein from the culture medium.
131 . A kit for characterizing specificity of the allergic response of a patient, comprising a set of antibody binding peptide sequences corresponding to at least one epitope on at least one potential allergen.
132 . The kit of claim 131 , for which the antibody binding sequences each are specific for one out of a known range of allergens, such that the characterization of allergic specificity becomes less susceptibility to cross-reactivity interferences.
133 . A kit of claim 131 , which further comprises other diagnostic reagents, which facilitate determination of the serum response to each of the antibody binding sequences.
134 . A kit of claim 131 , further comprising allergen vaccines, which can be administered to the patient according to the test results obtained using the antibody binding sequences.Join the waitlist — get patent alerts
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