Polypeptides with xylanase activity

Patent metadata

Field	Value
Publication number	US-9249403-B2
Application number	US-201414579980-A
Country	US
Kind code	B2
Filing date	Dec 22, 2014
Priority date	Dec 23, 2008
Publication date	Feb 2, 2016
Grant date	Feb 2, 2016

Abstract

The present invention relates to polypeptides with xylanase activity and uses thereof. The present invention also relates to method of modifying polypeptides with xylanase activity to affect, preferably to increase, xylanase activity and/or bran solubility.

First claim

The invention claimed is:

1. A recombinant polypeptide having xylanase activity and comprising an amino acid sequence, said amino acid sequence having at least 85% identity with SEQ ID No. 2 and an amino acid substitution in position 110, with any one different amino acid residue selected from the group consisting of:
asparagine (N), glutamic acid (E), tryptophan (W), alanine (A) and cysteine (C), wherein said position 110 is determined as the position within said polypeptide that corresponds to position 110 of B. subtilis xylanase sequence shown as SEQ ID No. 1 by alignment.

2. A recombinant polypeptide having xylanase activity and comprising an amino acid sequence, said amino acid sequence having at least 85% identity with SEQ ID No. 3 and an amino acid substitution in position 110, with any one different amino acid residue selected from the group consisting of:
asparagine (N), glutamic acid (E), tryptophan (W), alanine (A) and cysteine (C), wherein said position 110 is determined as the position within said polypeptide that corresponds to position 110 of B. subtilis xylanase sequence shown as SEQ ID No. 1 by alignment.

3. The recombinant polypeptide according claim 1 , wherein said polypeptide has at least 90% identity with SEQ ID No. 2.

4. The recombinant polypeptide according claim 2 , wherein said polypeptide has at least 90% identity with SEQ ID No. 3.

5. The recombinant polypeptide according to claim 1 , having a β-jelly roll fold.

6. The recombinant polypeptide according to claim 1 , wherein the amino acid substitution in position 110 is a substitution to alanine.

7. The recombinant polypeptide according to claim 1 , comprising one or more modification(s) at any one or more of amino acid positions: 11, 12, 13, 34, 54, 77, 81, 99, 104, 113, 114, 118, 122, 141, 154, 159, 162, 164, 166 and 175, the position(s) being determined as the corresponding position of B. subtilis amino acid sequence shown as SEQ ID No. 1.

8. The recombinant polypeptide according claim 1 , comprising one or more amino acid substitutions selected from the group consisting of: 11F, 12F, 122D, 113A, 13Y, 54Q, 54W, 113D, 141Q, 175L, 122F, 34K, 99Y, 104W, 154R, 159D, 175K, 81I , 166F, 162E, 162D, 164F, 114D, 114Y, 114F, 118V, 175K, 77L, 77M, 77S, 77V, and 77Y, the position(s) being determined as the corresponding position of B. subtilis amino acid sequence shown as SEQ ID No. 1.

9. A method of preparing a recombinant polypeptide according to claim 1 , said method comprising expressing a nucleotide sequence encoding said polypeptide; and optionally isolating and/or purifying the polypeptide after expression.

10. A nucleotide sequence encoding a recombinant polypeptide according to claim 1 .

11. A composition comprising a recombinant polypeptide prepared according to claim 9 admixed with a non-toxic component.

12. A composition comprising the nucleotide sequence according to claim 10 admixed with a non-toxic component.

13. The recombinant polypeptide of claim 3 , wherein said polypeptide has at least 95% identity with SEQ ID No. 2.

14. The recombinant polypeptide of claim 4 , wherein said polypeptide has at least 95% identity with SEQ ID No. 3.

15. The recombinant polypeptide according to claim 2 , wherein the amino acid substitution in position 110 is a substitution to alanine.

16. The recombinant polypeptide according to claim 2 , comprising one or more modification(s) at any one or more of amino acid positions: 11, 12, 13, 34, 54, 77, 81, 99, 104, 113, 114, 118, 122, 141, 154, 159, 162, 164, 166 and 175, the position(s) being determined as the corresponding position of B. subtilis amino acid sequence shown as SEQ ID No. 1.

17. The recombinant polypeptide according claim 2 , comprising one or more amino acid substitutions selected from the group consisting of: 11F, 12F, 122D, 113A, 13Y, 54Q, 54W, 113D, 141Q, 175L, 122F, 34K, 99Y, 104W, 154R, 159D, 175K, 81I, 166F, 162E, 162D, 164F, 114D, 114Y, 114F, 118V, 175K, 77L, 77M, 77S, 77V, and 77Y, the position(s) being determined as the corresponding position of B. subtilis amino acid sequence shown as SEQ ID No. 1.

18. A method of preparing a recombinant polypeptide according to claim 2 , said method comprising expressing a nucleotide sequence encoding said polypeptide; and optionally isolating and/or purifying the polypeptide after expression.

19. A nucleotide sequence encoding a recombinant polypeptide according to claim 2 .

20. A composition comprising a recombinant polypeptide prepared according to claim 18 admixed with a non-toxic component.

21. A composition comprising the nucleotide sequence according to claim 19 admixed with a non-toxic component.

Assignees

• Dupont Nutrition Biosci Aps

Inventors

• Sibbesen Ole
• Sorensen Jens Frisbaek

Classifications

• C12N9/248

  Xylanases · CPC title

• C12N9/2482 Primary

  Endo-1,4-beta-xylanase (3.2.1.8) · CPC title

• C12Y302/01008

  Endo-1,4-beta-xylanase (3.2.1.8) · CPC title

• A23L29/06

  Enzymes · CPC title

• A61K38/47

  acting on glycosyl compounds (3.2), e.g. cellulases, lactases · CPC title