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US 2026/0234581 A1Published application
Lipase variant
AssigneeKao Corporation
InventorsTakahiro HIOKI, Akihito KAWAHARA
Life of this patent
Estimating the term…
Abstract
Provided is a lipase mutant with alleviated inhibition of activity against a dispersed substrate in the presence of a surfactant. The lipase mutant consists of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 and has a predetermined amino acid residue at a predetermined position numbered according to SEQ ID NO: 2.
Claims15
1:
- A lipase mutant that consists of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO:
- 2, 4, 6, or 8 and has one or more amino acid residues selected from the group consisting of the following (a) to (e):
- (a) an amino acid residue other than leucine at a position corresponding to position 120 numbered according to SEQ ID NO:
- 2;
- (b) an amino acid residue other than serine at a position corresponding to position 130 numbered according to SEQ ID NO:
- 2;
- (c) an amino acid residue other than alanine at a position corresponding to position 134 numbered according to SEQ ID NO:
- 2;
- (d) an amino acid residue other than leucine at a position corresponding to position 136 numbered according to SEQ ID NO:
- 2;
- and
- (e) an amino acid residue other than serine at a position corresponding to position 137 numbered according to SEQ ID NO:
- 2,
- provided that, when the lipase mutant has only an amino acid residue of (d) in the above (a) to (e) and the amino acid residue is methionine, the lipase mutant consists of an amino acid sequence having an identity of at least 93% with the amino acid sequence of SEQ ID NO:
- 2, 4, or 6.
12 more elements
2:
- The lipase mutant according to claim 1, wherein the (a) to (e) are the following (a′) to (e′), respectively:
- (a′) alanine, phenylalanine, glycine, histidine, isoleucine, lysine, methionine, asparagine, glutamine, arginine, serine, threonine, valine, tryptophan, or tyrosine at a position corresponding to position 120 numbered according to SEQ ID NO:
- 2;
- (b′) alanine, cysteine, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, glutamine, arginine, threonine, valine, tryptophan, or tyrosine at a position corresponding to position 130 numbered according to SEQ ID NO:
- 2;
- (c′) cysteine, aspartic acid, glutamic acid, glycine, isoleucine, threonine, or valine at a position corresponding to position 134 numbered according to SEQ ID NO:
- 2;
- (d′) alanine, cysteine, glutamic acid, phenylalanine, glycine, histidine, isoleucine, methionine, serine, threonine, valine, or tryptophan at a position corresponding to position 136 numbered according to SEQ ID NO:
- 2;
- and
- (e′) cysteine, asparagine, or threonine at a position corresponding to position 137 numbered according to SEQ ID NO:
- 2,
- provided that, when the lipase mutant has only methionine of (d′) in the above (a′) to (e′), the lipase mutant consists of an amino acid sequence having an identity of at least 93% with the amino acid sequence of SEQ ID NO:
- 2, 4, or 6.
11 more elements
3:
- The lipase mutant according to claim 2, wherein the lipase mutant has two or more amino acid residues selected from the group consisting of the (a′) to (e′).
4:
- A polynucleotide encoding the lipase mutant according to claim 1.
5:
- A vector or DNA fragment comprising the polynucleotide according to claim 4.
6:
- A transformed cell comprising the vector or DNA fragment according to claim 5.
7:
- The transformed cell according to claim 6, wherein the transformed cell is a microorganism.
8:
- A cleansing composition comprising the lipase mutant according to claim 1.
(canceled)
10:
- A stain-washing method comprising using the cleansing composition according to claim 8.
11:
- A method for producing a lipid mutant, comprising performing one or more steps selected from the group consisting of the following (i) to (v) in a polypeptide that consists of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO:
- 2, 4, 6, or 8 and has a lipase activity:
- (i) a step of substituting an amino acid residue at a position corresponding to position 120 numbered according to SEQ ID NO:
- 2 with an amino acid residue other than leucine;
- (ii) a step of substituting an amino acid residue at a position corresponding to position 130 numbered according to SEQ ID NO:
- 2 with an amino acid residue other than serine;
- (iii) a step of substituting an amino acid residue at a position corresponding to position 134 numbered according to SEQ ID NO:
- 2 with an amino acid residue other than alanine;
- (iv) a step of substituting an amino acid residue at a position corresponding to position 136 numbered according to SEQ ID NO:
- 2 with an amino acid residue other than leucine;
- and
- (v) a step of substituting an amino acid residue at a position corresponding to position 137 numbered according to SEQ ID NO:
- 2 with an amino acid residue other than serine,
- provided that, when the method comprises performing only the step (iv) in the above (i) to (v) and the amino acid residue is substituted with methionine, the polypeptide consists of an amino acid sequence having an identity of at least 93% with the amino acid sequence of SEQ ID NO:
- 2, 4, or 6.
12 more elements
12:
- A method for evaluating or selecting a lipase having an ability to decompose an oil stain adhering to a hard surface in the presence of a sulfosuccinate or a salt thereof, comprising steps of:
- measuring lipase activity of a test lipase against a dispersed substrate in the presence of a surfactant;
- evaluating a degree of inhibition of lipase activity of the test lipase by the surfactant based on the measured result;
- and
- evaluating or selecting a test lipase which has lower degree of lipase activity inhibition by the surfactant than a standard lipase as a lipase having an ability to decompose an oil stain adhering to a hard surface in the presence of a sulfosuccinate or a salt thereof.
2 more elements
13:
- The lipase mutant according to claim 1, wherein the lipase mutant has any of amino acid residues Nos. 1 to 75 in Table 1 below:
- TABLE 1
- No.
- Amino acid residue
- 1
- 120A
- 2
- 120F
- 3
- 120G
- 4
- 120H
- 5
- 120I
- 6
- 120K
- 7
- 120M
- 8
- 120N
- 9
- 120Q
- 10
- 120R
- 11
- 120S
- 12
- 120T
- 13
- 120V
- 14
- 120W
- 15
- 120Y
- 16
- 130A
- 17
- 130C
- 18
- 130E
- 19
- 130F
- 20
- 130H
- 21
- 130I
- 22
- 130K
- 23
- 130L
- 24
- 130M
- 25
- 130Q
- 26
- 130R
- 27
- 130T
- 28
- 130V
- 29
- 130W
- 30
- 130Y
- 31
- 134C
- 32
- 134D
- 33
- 134E
- 34
- 134G
- 35
- 134I
- 36
- 134T
- 37
- 134V
- 38
- 136A
- 39
- 136C
- 40
- 136E
- 41
- 136F
- 42
- 136G
- 43
- 136H
- 44
- 136I
- 45
- 136M
- 46
- 136T
- 47
- 136V
- 48
- 136W
- 49
- 137C
- 50
- 137N
- 51
- 137T
- 52
- 120G 130A
- 53
- 120G 130L
- 54
- 120H 130A
- 55
- 120H 130L
- 56
- 120R 130A
- 57
- 120R 130K
- 58
- 120R 130L
- 59
- 120S 130A
- 60
- 120S 130K
- 61
- 120S 130L
- 62
- 120S 134T
- 63
- 120S 136A
- 64
- 120S 136S
- 65
- 120S 137N
- 66
- 130A 134T
- 67
- 130A 136M
- 68
- 130A 137T
- 69
- 130K136M
- 70
- 130M 137T
- 71
- 130Y 137T
- 72
- 136M 137N
- 73
- 120S 130A 136M
- 74
- 120R 130K 136M 137N
- 75
- 120R 130A 134T 135V 136M 137N
- wherein the amino acid position is a position numbered according to SEQ ID NO:
- 2.
153 more elements
14:
- The lipase mutant according to claim 13, wherein the lipase mutant is a lipase mutant consisting of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO:
- 2 and having any of amino acid residues Nos. 1 to 74 in the Table 1, a lipase mutant consisting of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO:
- 4 and having any of amino acid residues Nos. 11, 59, and 75 in the Table 1, a lipase mutant consisting of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO:
- 6 and having the amino acid residue No. 59 in the Table 1, or a lipase mutant consisting of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO:
- 8 and having any of amino acid residues Nos. 4, 6, 11, 22, 24, 30, 36, 41, 43, 50, 51, and 59 in the Table 1.
2 more elements
15:
- The method according to claim 12, further comprising a step of measuring the lipase activity of the test lipase against a dispersed substrate in the absence of a surfactant.
Dependencies are inferred from the claim text.