Xylanases, nucleic acids encoding them and methods for making and using them

Patent metadata

Field	Value
Publication number	US-RE46733-E
Application number	US-200814732217-A
Country	US
Kind code	E1
Filing date	Aug 1, 2008
Priority date	Oct 3, 2007
Publication date	Feb 27, 2018
Grant date	Feb 27, 2018

Abstract

The invention relates to enzymes having xylanase, mannanase and/or glucanase activity, e.g., catalyzing hydrolysis of internal β-1,4-xylosidic linkages or endo-β-1,4-glucanase linkages; and/or degrading a linear polysaccharide beta-1,4-xylan into xylose. Thus, the invention provides methods and processes for breaking down hemicellulose, which is a major component of the cell wall of plants, including methods and processes for hydrolyzing hemicelluloses in any plant or wood or wood product, wood waste, paper pulp, paper product or paper waste or byproduct. In addition, methods of designing new xylanases, mannanases and/or glucanases and methods of use thereof are also provided. The xylanases, mannanases and/or glucanases have increased activity and stability at increased pH and temperature.

First claim

What is claimed is:

1. An isolated, synthetic, or recombinant polypeptide having a xylanase activity, wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2, and amino acid residue change at amino acid residue 44 such that amino acid residue 44 Serine, Ser (or “S”) is changed to Threonine, “Thr” (or “T”).

2. A protein preparation, or an immobilized polypeptide comprising the polypeptide of claim 1 , wherein the protein preparation comprises a liquid, a solid or a gel.

3. The polypeptide of claim 1 , wherein the polypeptide specifically binds to an antibody and the antibody is a monoclonal, a polyclonal antibody, or a single chained antibody.

4. A method for hydrolyzing, liquefying, breaking up, or disrupting a xylan-, cellulose- or hemicellulose-comprising composition, comprising: contacting the polypeptide of claim 1 with the composition comprising a xylan, a cellulose, or a hemicellulose under conditions wherein the polypeptide hydrolyze, liquefy, breakup, or disrupt the xylan, cellulose, or hemicellulose-comprising composition, wherein optionally the composition comprises a plant cell, or a bacterial cell.

5. A biomass, wood, wood pulp, wood product, paper pulp, paper product, newspaper or paper waste comprising the polypeptide of claim 1 .

6. An enzyme cocktail comprising the polypeptide of claim 1 , and one or more enzymes selected from a group consisting of: a xylanase, a mannanase, a glucanase, a cellulose, a lipase, an esterase, a protease, an endoglycosidase, an endo-beta.-1,4-glucanase, a beta-glucanase, an endo-beta-1,3(4)-glucanase, a cutinase, a peroxidase, a catalase, a laccase, an amylase, a glucoamylase, a pectinase, a reductase, an oxidase, a phenoloxidase, a ligninase, a pullulanase, an arabinanase, a hemicellulase, a mannanase, a xyloglucanase, a xylanase, a mannanase, a glucanase, a pectin acetyl esterase, a rhamnogalacturonan acetyl esterase, a polygalacturonase, a rhamnogalacturonase, a galactanase, a pectin lyase, a pectin methylesterase, a cellobiohydrolase, a transglutaminase, or a combination thereof.

7. A process for hydrolyzing xylans, celluloses, or hemicelluloses in any organic compound, plant or wood or wood product or byproduct, wood waste, paper pulp, paper product or paper waste or byproduct with the polypeptide of claim 1 .

8. A composition comprising the polypeptide of claim 1 .

9. The enzyme cocktail of claim 6 , further comprising at least one additional enzyme selected from: an Endo glucanase, an Oligomerase I (beta glucosidase), a CBHI (GH family 7), a CBH2 (GH family 6), a Xylanase (GH family 11), an Arabinofuranosidase, a Xylanase (GH family 10), and an Oligomerase II (beta-xylosidase).

10. A fabric, yarn, cloth or textile comprising the polypeptide of claim 1 wherein the fabric, yarn, cloth or textile comprises a non-cotton cellulosic fabric, yarn, cloth, or textile.

11. A food, a feed, or a nutritional supplement comprising the polypeptide of claim 1 .

12. The polypeptide of claim 1 , further comprising a heterologous amino acid sequence.

13. The polypeptide of claim 12 , wherein the heterologous amino acid sequence comprises:
(a) a heterologous signal sequence, a heterologous carbohydrate binding module, a heterologous dockerin domain, a heterologous catalytic domain (CD), or a combination thereof;
(b) the sequence of (a), wherein the heterologous signal sequence, carbohydrate binding module or heterologous catalytic domain (CD) is derived from a heterologous enzyme; a tag, an epitope, a targeting peptide, a cleavable sequence, a detectable moiety or an enzyme; or
(c) the sequence of (a), wherein the heterologous carbohydrate binding module (CBM) comprises, a xylan binding module, a cellulose binding module, a lignin binding module, a xylose binding module, a mannanse binding module, a xyloglucan-specific module or a arabinofuranosidase binding module.

14. The polypeptide of claim 1 , wherein the amino acid sequence further comprising a second amino acid change selected from a group consisting of:
(a) a change at amino acid residue 4 such that amino acid residue 4 Threonine, or “Thr” (or “T”) is changed to Asparagine, or Asn (or “N”);
(b) a change at amino acid residue 4 such that amino acid residue 4 Threonine, or “Thr” (or “T”) is changed to Arginine, Arg (or “R”);
(c) a change at amino acid residue 4 such that amino acid residue 4 Threonine, or “Thr” (or “T”) is changed to Histidine, His (or “H”);
(d) a change at amino acid residue 73 such that amino acid residue 73 Glycine, Gly (or “G”) is changed to Tyrosine, “Tyr” (or “Y”);
(e) a change at amino acid residue 63 such that amino acid residue 63 Isoleucine, Ile (or “I”) is changed to Valine, “Val” (or “V”);
(f) a change at amino acid residue 17 such that amino acid residue 17 Phenylalanine, Phe (or “F”) is changed to Valine, “Val” (or “V”);
(g) a change at amino acid residue 38 such that amino acid residue 38 Arginine, Arg (or “R”) is changed to Histidine, His (or “H”);
(h) a change at amino acid residue 33 such that amino acid residue 33 Leucine, Leu (or “L”) is changed to Alaninw, Ala (or “A”);
(i) a change at amino acid residue 73 such that amino acid residue 73 Glycine, Gly (or “G”) is changed to Glutamate, Glu (or “E”);
(j) a change at amino acid residue 73 such that amino acid residue 73 Glycine, Gly (or “G”) is changed to Valine, “Val” (or “V”);
(k) a change at amino acid residue 125 such that amino acid residue 125 Glutamine, Gln (or “Q”) is changed to Tyrosine, “Tyr” (or “Y”):
(l) a change at amino acid residue 188 such that amino acid residue 188 Serine, Ser (or “S”) is changed to Glutamate, Glu (or “E”);
(m) a change at amino acid residue 9, such that amino acid residue 9 Proline, Pro (or “P”) is changed to Aspartate, Asp (or “D”);
(n) a change at amino acid residue 150, such that amino acid residue 150 Valine, Val (or “V”) is changed to Alanine, Ala (or “A”);
(o) a change at amino acid residue 189, such that amino acid residue 189 Serine, Ser (or “S”) is changed to Glutamine, Gln (or “Q”);
(p) a change at amino acid residue 21, such that amino acid residue 21 Phenylalanine, Phe (or “F”) is changed to Tyrosine, “Tyr” (or “Y”); and
(q) any combination of the changes (a) to (m).

15. A method for reducing the amount of lignin (delignification), or solubilizing a lignin, in a paper or paper product, a wood, wood pulp or wood product, or a wood or paper recycling composition, comprising: contacting the paper or paper product, wood, wood pulp or wood product, or wood or paper recycling composition with the polypeptide of claim 1 thereby reducing the amount of lignin.

16. A method for an enzymatic decoloring of paper, hemp, or flax pulp comprising: contacting the paper, hemp, or flax pulp with the polypeptide of claim 1 and a decoloring agent, wherein optionally the decoloring agent comprises oxygen or hydrogen peroxide under conditions suitable for enzymatic decoloring.

17. A method for an enzymatic deinking of paper, paper waste, paper recycled product, deinking toner from non-contact printed wastepaper or mixtures of non-contact and contact printed wastepaper, comprising: contacting the paper, paper waste, paper recycled product, non-contact printed wastepaper, or contact printed wastepaper with the polypeptide of claim 1 under conditions suitable for enzymatic deinking.

18. A method for decoloring a fabric, yarn, cloth, or textile comprising: contacting the fabric, yarn, cloth, or textile with the polypeptide of claim 1 under conditions suitable to produce a whitening of the te

Assignees

• Bp Corp North America Inc

Inventors

• Gray Kevin A
• Dirmeier Richard

Classifications

• A61P3/00

  Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00) · CPC title

• A61P1/00

  Drugs for disorders of the alimentary tract or the digestive system · CPC title

• D21H17/22

  Proteins · CPC title

• C12N9/248 Primary

  Xylanases · CPC title

• Y10T428/249921

  Web or sheet containing structurally defined element or component · CPC title