US 2012/0058928 A1Published application

Alkaline Protease Variants

AssigneeKao Corporation

InventorsMasatoshi Tohata, Mitsuyoshi Okuda, Tsuyoshi Sato, Keiji Endo, Katsuhisa Saeki

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Abstract

An alkaline protease variant derived from an alkaline protease consisting of an amino acid sequence represented by SEQ ID NO: 2 or consisting of an amino acid sequence having an identity of 90% or more therewith, which variant has mutations wherein one or more amino acid residues at positions selected from (a) position 6, (b) position 15, (c) position 16, (d) position 65, (e) position 66, (f) position 82, (g) position 83, (h) position 204, (i) position 319, and (j) position 337 of the amino acid sequence represented by SEQ ID NO: 2, or at positions corresponding thereto are substituted with the following amino acid residues: (a) or a position corresponding thereto: Typ, Leu, Val, Ile, Met, Tyr, Gln, Lys, Thr, Phe, Arg, Ser, Cys, Ala, or His; (b) or a position corresponding thereto: Glu, Met, Asp, Val, Gln, Arg, Cys, Trp, Ala, or Phe; (c) or a position corresponding thereto: Met, Glu, Arg, Val, Lys, Phe, Tyr, Ile, His, Asp, or Cys; (d) or a position corresponding thereto: Trp; (e) or a position corresponding thereto: His, Trp, Ser, or Leu; (f) or a position corresponding thereto: Ala, Glu, Gln, Ser, Cys, Gly, H is, Lys, Arg, Met, or Asn; (g) or a position corresponding thereto: Ala, Ser, or Cys; (h) or a position corresponding thereto: Glu, Asp, Cys, Val, Thr, Pro, His, Ile, Trp, Ser, Asn, Lys, or Arg; (i) or a position corresponding thereto: Trp, Val, Thr, Leu, Ile, Cys, Glu, Lys, Tyr, Arg, Phe, Gln, Met, Pro, Asp, Asn, H is, or Ser; and (j) or a position corresponding thereto: Arg, Gly, Ser, Lys, Gln, Thr, His, Ala, Cys, or Val.

Claims10

Dependent claims

An alkaline protease variant derived from an alkaline protease consisting of an amino acid sequence represented by SEQ ID NO:

2 or consisting of an amino acid sequence having an identity of 900 or more therewith, wherein one or more amino acid residues at positions selected from (a) position 6, (b) position 15, (c) position 16, (d) position 65, (e) position 66, (f) position 82, (g) position 83, (h) position 204, (i) position 319, and (j) position 337 of the amino acid sequence represented by SEQ ID NO:
2, or at positions corresponding thereto are substituted with the following amino acid residues:
(a) or a position corresponding thereto:
tryptophan, leucine, valine, isoleucine, methionine, tyrosine, glutamine, lysine, threonine, phenylalanine, arginine, serine, cysteine, alanine, or histidine;
(b) or a position corresponding thereto:
glutamic acid, methionine, aspartic acid, valine, glutamine, arginine, cysteine, tryptophan, alanine, or phenylalanine;
(c) or a position corresponding thereto:
methionine, glutamic acid, arginine, valine, lysine, phenylalanine, tyrosine, isoleucine, histidine, aspartic acid, or cysteine;
(d) or a position corresponding thereto:
tryptophan;
(e) or a position corresponding thereto:
histidine, tryptophan, serine, or leucine;
(f) or a position corresponding thereto:
alanine, glutamic acid, glutamine, serine, cysteine, glycine, histidine, lysine, arginine, methionine, or asparagine;
(g) or a position corresponding thereto:
alanine, serine, or cysteine;
(h) or a position corresponding thereto:
glutamic acid, aspartic acid, cysteine, valine, threonine, proline, histidine, isoleucine, tryptophan, serine, asparagine, lysine or, arginine;
(i) or a position corresponding thereto:
tryptophan, valine, threonine, leucine, isoleucine, cysteine, glutamic acid, lysine, tyrosine, arginine, phenylalanine, glutamine, methionine, proline, aspartic acid, asparagine, histidine, or serine;
and
(j) or a position corresponding thereto:
arginine, glycine, serine, lysine, glutamine, threonine, histidine, alanine, cysteine, or valine.

20 more elements

A method for enhancing the stability of an alkaline protease in a liquid detergent, the method comprising, in an alkaline protease consisting of an amino acid sequence represented by SEQ ID NO:

2 or consisting of an amino acid sequence having an identity of 90% or more therewith, substituting one or more amino acid residues at positions selected from (a) position 6, (b) position 15, (c) position 16, (d) position 65, (e) position 66, (f) position 82, (g) position 83, (h) position 204, (i) position 319, and (j) position 337 of the amino acid sequence represented, by SEQ ID NO:
2, or at positions corresponding thereto with the following amino acid residues:
(a) or a position corresponding thereto:
tryptophan, leucine, valine, isoleucine, methionine, tyrosine, glutamine, lysine, threonine, phenylalanine, arginine, serine, cysteine, alanine, or histidine;
(b) or a position corresponding thereto:
glutamic acid, methionine, aspartic acid, valine, glutamine, arginine, cysteine, tryptophan, alanine, or phenylalanine;
(c) or a position corresponding thereto:
methionine, glutamic acid, arginine, valine, lysine, phenylalanine, tyrosine, isoleucine, histidine, aspartic acid, or cysteine;
(d) or a position corresponding thereto:
tryptophan;
(e) or a position corresponding thereto:
histidine, tryptophan, serine, or leucine;
(f) or a position corresponding thereto:
alanine, glutamic acid, glutamine, serine, cysteine, glycine, histidine, lysine, arginine, methionine, or asparagine;
(g) or a position corresponding thereto:
alanine, serine, or cysteine;
(h) or a position corresponding thereto:
glutamic acid, aspartic acid, cysteine, valine, threonine, proline, histidine, isoleucine, tryptophan, serine, asparagine, lysine or, arginine;
(i) or a position corresponding thereto:
tryptophan, valine, threonine, leucine, isoleucine, cysteine, glutamic acid, lysine, tyrosine, arginine, phenylalanine, glutamine, methionine, proline, aspartic acid, asparagine, histidine, or serine;
and
(j) or a position corresponding thereto:
arginine, glycine, serine, lysine, glutamine, threonine, histidine, alanine, cysteine, or valine.

20 more elements

Dependencies are inferred from the claim text.

Facts

Application: 13/266,525Filed 26 Apr 2010
Priority: JP 2009-11079230 Apr 2009
Published: 8 Mar 2012
Term ends: Not estimatedThe source records don’t support an estimate.

Classification6 codes

C11D 3/38618
Preparations containing enzymes , e.g. protease or amylase: protease or amylase in liquid compositions only
C11D 3/386
Products with no well-defined composition , e.g. natural products: preparations containing enzymes , e.g. protease or amylase
C11D 3/38681
Preparations containing enzymes , e.g. protease or amylase: chemically modified or immobilised enzymes
C12N 9/54
Derived from bacteria or Archaea: bacteria being Bacillus

Family

US 8,778,650 B2Related patent
US 2012/0058928 A1This patent

From the US filing history. Foreign family members aren’t listed.

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