Lipase variant

{
  "publication_number": "US2026103693A1",
  "country": "US",
  "kind": "A1",
  "title": "Lipase variant",
  "abstract": "Provided is a lipase mutant with improved heterologous expression. The lipase mutant consists of an amino acid sequence obtained by substituting an amino acid residue at a predetermined position numbered according to SEQ ID NO: 2 with a predetermined amino acid residue in an amino acid sequence of a parent lipase.",
  "claims": [
    "1. A lipase mutant selected from the group consisting of (a) to (v) and (aa) to (ae): (a) a lipase mutant consisting of an amino acid sequence having an identity of at least 82% with the amino acid sequence of SEQ ID NO: 2 and having aspartic acid at a position corresponding to position 118 of the amino acid sequence of SEQ ID NO: 2; (b) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and having tyrosine at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2; (c) a lipase mutant consisting of an amino acid sequence having an identity of at least 91% with the amino acid sequence of SEQ ID NO: 4 and having glycine at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2; (d) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and having cysteine at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2; (e) a lipase mutant consisting of an amino acid sequence having an identity of at least 94% with the amino acid sequence of SEQ ID NO: 4 and having leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (f) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 6 and having alanine, threonine, histidine, or glycine at a position corresponding to position 222 of the amino acid sequence of SEQ ID NO: 2; (g) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and having lysine at a position corresponding to position 39 of the amino acid sequence of SEQ ID NO: 2; (h) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and having threonine at a position corresponding to position 125 of the amino acid sequence of SEQ ID NO: 2; (i) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and having proline at a position corresponding to position 126 of the amino acid sequence of SEQ ID NO: 2; (j) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and having glycine at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2; (k) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and having tyrosine at a position corresponding to position 29 of the amino acid sequence of SEQ ID NO: 2; (l) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and having tyrosine at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2; (m) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and having glycine at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2; (n) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and having glycine at a position corresponding to position 186 of the amino acid sequence of SEQ ID NO: 2; (o) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 12 and having glutamine at a position corresponding to position 206 of the amino acid sequence of SEQ ID NO: 2; (p) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 14 and having tyrosine at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2; (q) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 14 and having glycine at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2; (r) a lipase mutant consisting of an amino acid sequence having an identity of at least 97% with the amino acid sequence of SEQ ID NO: 14 and having lysine at a position corresponding to position 77 of the amino acid sequence of SEQ ID NO: 2; (s) a lipase mutant consisting of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and having leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (t) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 16 and having cysteine at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2; (u) a lipase mutant consisting of an amino acid sequence having an identity of at least 81% with the amino acid sequence of SEQ ID NO: 16 and having leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (v) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 34 and having tyrosine at a position corresponding to position 29 of the amino acid sequence of SEQ ID NO: 2; (aa) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and having at least two amino acid residues selected from tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, cysteine at a position corresponding to position 90, and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (ab) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and having at least two amino acid residues selected from lysine at a position corresponding to position 39, threonine at a position corresponding to position 125, proline at a position corresponding to position 126 and glycine at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2; (ac) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and having at least two amino acid residues selected from tyrosine at a position corresponding to position 29, tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, and glycine at a position corresponding to position 186 of the amino acid sequence of SEQ ID NO: 2; (ad) a lipase mutant consisting of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and having at least two amino acid residues selected from tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, lysine at a position corresponding to position 77, and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; and (ae) a lipase mutant consisting of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 16 and having cysteine at a position corresponding to position 90 and leucine at a position corresponding to position 181 of the amino acid sequence of SEO ID NO: 2. 2. (canceled) 3. The lipase mutant according to claim 1, wherein the lipase mutant is a lipase mutant selected from the group consisting of (a), (c) to (g), (i) to (k), (n), (o), (r), and (t) to (v). 4. The lipase mutant according to claim 1, wherein the lipase mutant is a lipase mutant selected from the group consisting of (aa-1) to (ae): (aa-1) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has tyrosine at a position corresponding to position 31 and glycine at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2; (aa-2) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has cysteine at a position corresponding to position 90 and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (aa-3) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has glycine at a position corresponding to position 32 and cysteine at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2; (aa-4) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, cysteine at a position corresponding to position 90, and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (ab-1) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has threonine at a position corresponding to position 125 and proline at a position corresponding to position 126 of the amino acid sequence of SEQ ID NO: 2; (ab-2) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lysine at a position corresponding to position 39 and glycine at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2; (ab-3) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has threonine at a position corresponding to position 125, proline at a position corresponding to position 126, and glycine at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2; (ab-4) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lysine at a position corresponding to position 39, threonine at a position corresponding to position 125, and proline at a position corresponding to position 126 of the amino acid sequence of SEQ ID NO: 2; (ab-5) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lysine at a position corresponding to position 39, proline at a position corresponding to position 126, and glycine at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2; (ab-6) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lysine at a position corresponding to position 39, threonine at a position corresponding to position 125, proline at a position corresponding to position 126, and glycine at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2; (ac-1) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has tyrosine at a position corresponding to position 29 and glycine at a position corresponding to position 186 of the amino acid sequence of SEQ ID NO: 2; (ac-2) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has tyrosine at a position corresponding to position 29, tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, and glycine at a position corresponding to position 186 of the amino acid sequence of SEQ ID NO: 2; (ad-1) a lipase mutant that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has tyrosine at a position corresponding to position 31 and glycine at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2; (ad-2) a lipase mutant that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has lysine at a position corresponding to position 77 and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (ad-3) a lipase mutant that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; (ad-4) a lipase mutant that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, lysine at a position corresponding to position 77, and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2; and (ae) a lipase mutant that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 16 and has cysteine at a position corresponding to position 90 and leucine at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2. 5. An enzyme composition comprising the lipase mutant according to claim 1. 6. A polynucleotide encoding the lipase mutant according to claim 1. 7. A vector or DNA fragment comprising the polynucleotide according to claim 6. 8. A transformed cell comprising the vector or DNA fragment according to claim 7. 9. The transformed cell according to claim 8, wherein the transformed cell is a microorganism. 10. The transformed cell according to claim 9, wherein the transformed cell is Escherichia coli or a Bacillus bacterium. 11. A method for producing a lipase mutant, comprising a step of culturing the transformed cell according to claim 8. 12. A method for manufacturing a lipase mutant, comprising a step selected from the group consisting of (i) to (xxvii): (i) a step of substituting an amino acid residue at a position corresponding to position 118 of the amino acid sequence of SEQ ID NO: 2 with aspartic acid in a polypeptide that consists of an amino acid sequence having an identity of at least 82% with the amino acid sequence of SEQ ID NO: 2 and has lipase activity; (ii) a step of substituting an amino acid residue at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (iii) a step of substituting an amino acid residue at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 91% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (iv) a step of substituting an amino acid residue at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2 with cysteine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (v) a step of substituting an amino acid residue at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 94% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (vi) a step of substituting an amino acid residue at a position corresponding to position 222 of the amino acid sequence of SEQ ID NO: 2 with alanine, threonine, histidine, or glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 6 and has lipase activity; (vii) a step of substituting an amino acid residue at a position corresponding to position 39 of the amino acid sequence of SEQ ID NO: 2 with lysine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (viii) a step of substituting an amino acid residue at a position corresponding to position 125 of the amino acid sequence of SEQ ID NO: 2 with threonine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (ix) a step of substituting an amino acid residue at a position corresponding to position 126 of the amino acid sequence of SEQ ID NO: 2 with proline in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (x) a step of substituting an amino acid residue at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (xi) a step of substituting an amino acid residue at a position corresponding to position 29 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xii) a step of substituting an amino acid residue at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xiii) a step of substituting an amino acid residue at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xiv) a step of substituting an amino acid residue at a position corresponding to position 186 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xv) a step of substituting an amino acid residue at a position corresponding to position 206 of the amino acid sequence of SEQ ID NO: 2 with glutamine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 12 and has lipase activity; (xvi) a step of substituting an amino acid residue at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; (xvii) a step of substituting an amino acid residue at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; (xviii) a step of substituting an amino acid residue at a position corresponding to position 77 of the amino acid sequence of SEQ ID NO: 2 with lysine in a polypeptide that consists of an amino acid sequence having an identity of at least 97% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; (xix) a step of substituting an amino acid residue at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; (xx) a step of substituting an amino acid residue at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2 with cysteine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 16 and has lipase activity; (xxi) a step of substituting an amino acid residue at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 81% with the amino acid sequence of SEQ ID NO: 16 and has lipase activity; (xxii) a step of substituting an amino acid residue at a position corresponding to position 29 of the amino acid sequence of SEQ ID NO. 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 34 and has lipase activity; (xxiii) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 31, 32, 90, and 181 of the amino acid sequence of SEQ ID NO: 2 with tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, cysteine at a position corresponding to position 90, and leucine at a position corresponding to position 181 in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (xxiv) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 39, 125, 126, and 293 of the amino acid sequence of SEQ ID NO: 2 with lysine at a position corresponding to position 39, threonine at a position corresponding to position 125, proline at a position corresponding to position 126, and glycine at a position corresponding to position 293 in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (xxv) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 29, 31, 32, and 186 of the amino acid sequence of SEQ ID NO: 2 with tyrosine at a position corresponding to position 29, tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, and glycine at a position corresponding to position 186 in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xxvi) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 31, 32, 77, and 181 of the amino acid sequence of SEQ ID NO: 2 with tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, lysine at a position corresponding to position 77, and leucine at a position corresponding to position 181 in a polypeptide that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; and (xxvii) a step of substituting an amino acid residue at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2 with cysteine and an amino acid residue at a position corresponding to position 181 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 16 and has lipase activity. 13. (canceled) 14. A method for improving heterologous expression of a lipase, comprising a step selected from the group consisting of (i) to (xxvii): (i) a step of substituting an amino acid residue at a position corresponding to position 118 of the amino acid sequence of SEQ ID NO: 2 with aspartic acid in a polypeptide that consists of an amino acid sequence having an identity of at least 82% with the amino acid sequence of SEQ ID NO: 2 and has lipase activity; (ii) a step of substituting an amino acid residue at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (iii) a step of substituting an amino acid residue at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 91% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (iv) a step of substituting an amino acid residue at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2 with cysteine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (v) a step of substituting an amino acid residue at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 94% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (vi) a step of substituting an amino acid residue at a position corresponding to position 222 of the amino acid sequence of SEQ ID NO: 2 with alanine, threonine, histidine, or glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 6 and has lipase activity; (vii) a step of substituting an amino acid residue at a position corresponding to position 39 of the amino acid sequence of SEQ ID NO: 2 with lysine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (viii) a step of substituting an amino acid residue at a position corresponding to position 125 of the amino acid sequence of SEQ ID NO: 2 with threonine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (ix) a step of substituting an amino acid residue at a position corresponding to position 126 of the amino acid sequence of SEQ ID NO: 2 with proline in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (x) a step of substituting an amino acid residue at a position corresponding to position 293 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (xi) a step of substituting an amino acid residue in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity with tyrosine at a position corresponding to position 29 of the amino acid sequence of SEQ ID NO: 2; (xii) a step of substituting an amino acid residue at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xiii) a step of substituting an amino acid residue at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity: (xiv) a step of substituting an amino acid residue at a position corresponding to position 186 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xv) a step of substituting an amino acid residue at a position corresponding to position 206 of the amino acid sequence of SEQ ID NO: 2 with glutamine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 12 and has lipase activity; (xvi) a step of substituting an amino acid residue at a position corresponding to position 31 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; (xvii) a step of substituting an amino acid residue at a position corresponding to position 32 of the amino acid sequence of SEQ ID NO: 2 with glycine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; (xviii) a step of substituting an amino acid residue at a position corresponding to position 77 of the amino acid sequence of SEQ ID NO: 2 with lysine in a polypeptide that consists of an amino acid sequence having an identity of at least 97% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; (xix) a step of substituting an amino acid residue at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity: (xx) a step of substituting an amino acid residue at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2 with cysteine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 16 and has lipase activity; (xxi) a step of substituting an amino acid residue at a position corresponding to position 181 of the amino acid sequence of SEQ ID NO: 2 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 81% with the amino acid sequence of SEQ ID NO: 16 and has lipase activity; (xxii) a step of substituting an amino acid residue at a position corresponding to position 29 of the amino acid sequence of SEQ ID NO: 2 with tyrosine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 34 and has lipase activity; (xxiii) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 31, 32, 90, and 181 of the amino acid sequence of SEQ ID NO: 2 with tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, cysteine at a position corresponding to position 90, and leucine at a position corresponding to position 181 in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 4 and has lipase activity; (xxiv) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 39, 125, 126, and 293 of the amino acid sequence of SEQ ID NO: 2 with lysine at a position corresponding to position 39, threonine at a position corresponding to position 125, proline at a position corresponding to position 126, and glycine at a position corresponding to position 293 in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 8 and has lipase activity; (xxv) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 29, 31, 32, and 186 of the amino acid sequence of SEQ ID NO: 2 with tyrosine at a position corresponding to position 29, tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, and glycine at a position corresponding to position 186 in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 10 and has lipase activity; (xxvi) a step of substituting at least two amino acid residues at positions selected from positions corresponding to positions 31, 32, 77, and 181 of the amino acid sequence of SEQ ID NO: 2 with tyrosine at a position corresponding to position 31, glycine at a position corresponding to position 32, lysine at a position corresponding to position 77, and leucine at a position corresponding to position 181 in a polypeptide that consists of an amino acid sequence having an identity of at least 83% with the amino acid sequence of SEQ ID NO: 14 and has lipase activity; and (xxvii) a step of substituting an amino acid residue at a position corresponding to position 90 of the amino acid sequence of SEQ ID NO: 2 with cysteine and an amino acid residue at a position corresponding to position 181 with leucine in a polypeptide that consists of an amino acid sequence having an identity of at least 80% with the amino acid sequence of SEQ ID NO: 16 and has lipase activity. 15. (canceled)"
  ],
  "description_excerpt": "The present invention relates to a lipase mutant.\n\nLipases contribute to removal of dirt including oil by hydrolyzing ester bonds in lipids to generate fatty acids and are useful in cleansing applications. As a lipase useful for cleansing, a Thermomyces lanuginosus -derived lipase (hereinafter, TLL) is commercially available under the trade name LIPOLASE (registered tradename). Patent Literature 1 discloses that Cedecea sp-16640 strain-derived lipase Lipr139 exhibits excellent cleansing performance compared to TLL. Patent Literature 2 discloses a mutant of a Proteus bacterium-derived lipase (hereinafter, PvLip), having improved cleansing performance compared to one or more reference lipolytic enzymes. Patent Literature 3 discloses that a metagenome-derived lipase Lipr138 exhibits excellent cleansing performance compared to TLL. Lipr139, PvLip, and Lipr138 are lipases belonging to the same clade (Proteus/Yersinia clade) that includes lipases derived from a Proteus bacterium or Yersinia bacterium.\n\nMany lipases derived from gram-negative bacteria require a lipase-specific chaperone for folding into the active form (Non Patent Literature 1) and have a challenge in low-cost manufacturing by heterologous expression. It has been reported that the heterologous expression is improved by coexpression with a specific chaperone (Non Patent Literature 2), but the productivity thereof is low, and there is still a large challenge in low-cost manufacturing by heterologous expression.",
  "cpc": [
    "C12N 9/20",
    "C12Y 301/01003"
  ],
  "ipc": [
    "C12N 9/20"
  ],
  "assignees": [
    "Kao Corp"
  ],
  "inventors": [
    "Takahiro HIOKI",
    "Akihito Kawahara"
  ],
  "filing_date": "2023-09-19",
  "publication_date": "2026-04-16",
  "priority_date": "2022-09-20",
  "application_number": "US-202319113016-A",
  "family_id": "90454563",
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}