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1. WO1999001544 - VARIANTS D'ENDO-1,4-$g(b)-GLUCANASE DE FAMILLE 6 ET COMPOSITIONS NETTOYANTES CONTENANT DE TELS COMPOSES

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

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CLAIMS

1. Cleaning composition comprising one or more enzymes having cellulolytic activity wherein at least 25% of the total weight of cellulolytic active protein derives from the presence of a

Humicola endo-1,4-β-glucanase or Humicola-like cellulase of the glycosyl hydrolase family 6.

2. Cleaning composition according to claim 1, wherein the Humicola-like cellulase comprises a catalytically core domain which has an amino acid sequence being at least 35% homologous to SEQ ID NO: 4.

3. Cleaning composition according to claim 1 or 2, wherein the endo-1,4-β-glucanase comprises one or two cellulose binding domains (CBD) operably linked to the catalytically active domain.

4. Cleaning composition according to any of the claims 1-3 wherein the composition is a detergent composition.

5. Cleaning composition according to claim 4 wherein the detergent composition comprises one or more components selected from anionic, nonionic, cationic, amphoteric, ampholytic and zwitterionic surfactants, bleaching agents, additional enzymes, sud suppressores, dispersants, soil suspension and anti-redeposition agents, smectite clays, and builder components.

6. Cleaning composition according to claim 4 or 5 wherein the detergent composition is a granular detergent composition containing no more than 40% by weight of inorganic filler salt.

7. Cleaning composition according to claim 6 wherein the granular detergent composition contains no more than 15% by weight of inorganic filler salt.

8. Cleaning composition according to claim 5 wherein the detergent composition is a heavy duty liquid composition.

9. Cleaning composition according to any of the claims 4-8 wherein the enzymes are selected from the group consisting of proteases, cellulases, β-glucanases, hemicellulases, lipases, peroxidases, laccases, α-amylase, glucoamylases, cutinases, pectinases, reductases, oxidases, phenoloxidases, ligninases, pullulanases, arabinosidases or mixtures thereof.

10. Cleaning composition according to any of the claims 4-9 which is a laundry detergent additive.

11. Cleaning composition according to any of the claims 1-3 wherein the composition is a fabric softener or fabric conditioning composition for the treatment of fabrics.

12. Cleaning composition according to claim 11 wherein the fabric softener composition comprises from about 1% to about 90% of one or more cationic fabric softening agents, nonionic fabric softening agents, or mixtures thereof.

13. Cleaning composition according to claim 12 comprising from about 2% to about 50% by weight of one or more fabric softening agents.

14. Cleaning composition according to claim 12 or 13 wherein the cationic fabric softening agents comprise quaternary ammonium softening agent or amine precursor thereof.

15. Cleaning composition according to claim 14 wherein the quaternary ammonium softening agent is N,N-di(2-tallowoyl-oxyethyl)-N,N-dimethyl ammonium chloride.

16. Cleaning composition according to any of the claims 1-15 wherein endo-1,4-β-glucanase is derived or derivable from the fungus Orpinomyces sp.

17. Cleaning composition according to any of the claims 1-15 wherein endo-1,4-β-glucanase is derived or derivable from the fungus Neocallimastix patriciarum.

18. Cleaning composition according to any of the claims 1-15 wherein endo-1,4-β-glucanase is derived or derivable from the fungus Trichoderma reesei.

19. Cleaning composition according to any of the claims 1-15 wherein endo-1,4-β-glucanase is derived or derivable from the fungus Fusarium oxysporum.

20. Cleaning composition according to any of the claims 1-19 which comprises the family 6 endo-1,4-β-glucanase in an amount corresponding to from about 1 ECU to about 100000 ECU per liter washing or rinsing solution.

21. Cleaning composition according to any of the claims 1-20 which further comprises an endo-1,4-β-glucanase of the glycosyl hydrolase family 5, or an endo-1,4-β-glucanase of the glycosyl hydrolase family 7, or an endo-1,4-β-glucanase of the glycosyl hydrolase family 8, or an endo-1,4-β-glucanase of the glycosyl hydrolase family 9, or an endo-1,4-β-glucanase of the glycosyl hydrolase family 44, or an endo-1,4-β-glucanase of the glycosyl hydrolase family 45, or an endo-1,4-β-glucanase of the glycosyl hydrolase family 48, or an endo-1,4-β-glucanase of the glycosyl hydrolase family 12 optionally being operably linked to a cellulose binding domain, or any mixture thereof.

22. A process for machine treatment of fabrics which process comprises treating fabric during a rinse cycle of a machine washing process with a rinse solution containing the composition according to any of the claims 11-21.

23. A method of constructing a variant of a parent Humicola family 6 endo-beta-1,4-glucanase, which variant has endo-beta-1,4-glucanase activity and improved detergent compability as compared to the parent endo-beta-1,4-glucanase, which method comprises i) analysing the structure of the parent Humicola family 6 endo-beta-1,4-glucanase to identify at least one amino acid residue or at least one structural part of the Humicola family 6 endo-beta-1,4-glucanase catalytically core domain structure, which amino acid residue or structural part is believed to be of relevance for altering the detergent compatibility of the parent Humicola family 6 endo-beta-1,4-glucanase as evaluated on the basis of structural or functional considerations,

ii) constructing a Humicola family 6 endo-beta-1,4-glucanase variant, which as compared to the parent Humicola family 6 endo-beta-1,4-glucanase has been modified in the amino acid residue or structural part identified in i) so as to alter the detergent compatibility, and, optionally,

iii) testing the resulting Humicola family 6 endo-beta-1,4-glucanase variant with respect to detergent compatibility.

24. The method according to claim 23, wherein the structural part to be modified is the binding cleft, the loop region encompassing the binding cleft, or the side chain of the catalytic acid Aspl39.

25. A variant of a parent Humicola family 6 endo-beta-1,4-glucanase, which comprises a mutation in a position corresponding to at least one of the following positions in SEQ ID NO: 4: K20, S56, S94, A95, K103, A182, N183 and Q318.

26. The variant according to claim 25, which comprises at least one of the mutations:
K20E
S56D
S94D
A95G
K103E
K103Q
A182G
N183E A182G+N183H
Q318E

27. A method of constructing a variant of a parent Humicola-like family 6 cellulase, which variant has endo-beta-1,4-glucanase activity and improved detergent compatibility as compared to the parent cellulase, which method comprises
i) comparing the three-dimensional structure of the Humicola endo-beta-1,4-glucanase with the structure of a Humicola-like
cellulase,
ii) identifying a part of the Humicola-like cellulase structure which is different from the Humicola endo-beta-1,4-glucanase structure and which from structural or functional considerations is contemplated to be responsible for differences in the detergent compatibility of the Humicola endo-beta-1,4-glucanase and
Humicola-like cellulase,
iii) modifying the part of the Humicola-like cellulase identified in ii) whereby a Humicola-like endo-beta-1,4-glucanase variant is obtained, which has an improved detergent compatibility compared to the parent Humicola-like cellulase , and optionally,
iv) testing the resulting Humicola-like endo-beta-1,4-glucanase variant with respect to detergent compatibility.

28. The method according to claim 27, wherein, in step iii), the part of the Humicola-like cellulase is modified so as to resemble the corresponding part of the Humicola family 6 endo-beta-1,4-glucanase.

29. The method according to claim 27 or 28, wherein, in step iii), the modification is accomplished by deleting one or more amino acid residues of the part of the Humicola-like cellulase to be modified; or the modification is accomplished by replacing one or more amino acid residues of the part of the Humicola-like
cellulase to be modified with the amino acid residues occupying corresponding positions in the Humicola endo-beta-1,4-glucanase; or the modification is accomplished by insertion of one or more amino acid residues present in the Humicola endo-beta-1,4-glucanase into a corresponding position in the Humicola-like cellulase.

30. The method according to any of claims 27-29, wherein the
Humicola-like cellulase is selected from the group consisting of Neocallimastix patriciarum endo-beta-1,4-glucanase, Orpinomyces sp. endo-beta-1,4-glucanase, Trichoderma reseei exoglucanase, Agaricus bispora exoglucanase, Phanerochaete chrysosporium
exoglucanse, Penicillium purpurogenum exoglucanase, Acremonium cellulyticus exoglucanase, Fusarium oxysporum exoglucanase and homologues thereof.

31. The method according to claim 23 or 27, wherein the parent Humicola endo-beta-1,4-glucanase is derived from a strain of
Humicola insolens.

32. The method according to claim 31, wherein the parent Humicola endo-beta-1,4-glucanase is derived from Humicola insolens, DSM 1800.

33. An isolated polynucleotide molecule comprising a DNA sequence encoding an endo-beta-1,4-glucanase variant according to claim 26.

34. A recombinant expression vector which carries the
polynucleotide molecule according to claim 33.

35. A cell which is transformed with the polynucleotide molecule according to claim 33 or a vector according to claim 34.

36. The cell according to claim 35, which is a microorganism.

37. The cell according to claim 36, which is a bacterium or a fungus.

38. A cell according to claim 37, which is a Fusarium oxysporum, an Aspergillus niger or an Aspergillus oryzae cell.

39. Cleaning composition comprising an endo-beta-1,4-glucanase which is constructed using the method according to any of the claims 23, 24 and 27-32.