Polypeptides with xylanase activity
View Patent ↗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.
1. A polynucleotide encoding a polypeptide having xylanase activity and comprising an amino acid sequence, said amino acid sequence having at least 85% identity with the amino acid sequence set forth in SEQ ID NO: 1, and which polypeptide has
i) one or two amino acid substitutions in a position selected from the group consisting of: 12 and 13; and
ii) one or more further amino acid substitutions in a position selected from the group consisting of: 15, 34, 54, 77, 81, 82, 99, 104, 110, 113, 114, 118, 122, 141, 154, 159, 162, 164, 166, 175, and 179,
wherein said positions are determined as the position corresponding to the position of B. subtilis xylanase sequence shown as SEQ ID NO: 1 by alignment;
wherein the amino acid substitution in position 12 is an amino acid substitution to any one different amino acid residue selected from the group consisting of: isoleucine, alanine, leucine, asparagine, lysine, aspartic acid, methionine, cysteine, glutamic acid, threonine, glutamine, tryptophan, valine, proline, serine, tyrosine, arginine, and histidine; and
wherein the amino acid substitution in position 13 is an amino acid substitution to any one different amino acid residue selected from the group consisting of: alanine, leucine, asparagine, lysine, aspartic acid, methionine, cysteine, glutamic acid, threonine, glutamine, tryptophan, valine, proline, serine, tyrosine, arginine, and histidine.
2. A composition comprising the polynucleotide according to claim 1 admixed with a non-toxic component.
3. The polynucleotide according to claim 1 , wherein said polypeptide comprises one or more amino acid substitutions selected from the group consisting of: 13Y, 15Y, 34K, 77V, 77M, 77Y, 77L, 77S, 81I, 82I, 99Y, 104W, 110A, 113D, 113A, 114F, 114D, 114Y, 118V, 122F, 122D, 154R, 159D, 162E, 162D, 164F, 166F, 175L, 175K, 175E, 175Y, and 179Y, the position(s) being determined as the corresponding position of B. subtilis amino acid sequence shown as SEQ ID NO: 1.
4. The polynucleotide according to claim 1 , wherein said polypeptide has at least 90% identity to SEQ ID NO: 1.
5. The polynucleotide according to claim 1 , wherein said polypeptide has a β-jelly roll fold.
6. The polynucleotide according to claim 1 , wherein the amino acid modification in position 12 of said polypeptide is a substitution to a tyrosine.
7. The polynucleotide according to claim 1 , wherein the amino acid modification in position 13 of said polypeptide is a substitution to a tyrosine.
8. The polynucleotide according to claim 1 , wherein said polypeptide has a total number of amino acids less than 250 amino acids.
9. The polynucleotide according to claim 1 , wherein said polypeptide comprises one or more amino acid substitutions selected from the group consisting of: 13Y, 110A, 122D, 113A, 13Y, 113D, 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.
10. A recombinant expression vector comprising the polynucleotide according to claim 1 .
11. A host cell comprising the expression vector of claim 10 .