Polypeptide having xylanase activity

Patent metadata

Field	Value
Publication number	US-10550375-B2
Application number	US-201515116281-A
Country	US
Kind code	B2
Filing date	Jan 30, 2015
Priority date	Jan 31, 2014
Publication date	Feb 4, 2020
Grant date	Feb 4, 2020

Abstract

The present invention relates to an isolated polypeptide having xylanase activity, selected from the group consisting of:
a) a polypeptide comprising an amino acid sequence having at least 87% identity with SEQ ID NO: 1; b) a polypeptide encoded by a polynucleotide having at least 87% identity with SEQ ID NO:2; or c) a fragment of a polypeptide of a) or b) which fragment has xylanase activity.

First claim

The invention claimed is:

1. A polypeptide having xylanase activity, selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence having at least 94% identity with SEQ ID NO: 1; or
(b) a polypeptide encoded by a polynucleotide having at least 94% identity with SEQ ID NO:2.

2. The polypeptide according to claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID No. 1.

3. The polypeptide of claims 1 or 2 wherein the polypeptide is encoded by the polynucleotide of SEQ ID No. 2.

4. The polypeptide of claims 1 or 2 , wherein the polypeptide is an endo-1,4-β-xylanase.

5. The polypeptide of claims 1 or 2 , wherein the polypeptide has at least 50% residual activity of xylanase activity after incubation at 65° C. for 10 minutes at pH 6 and/or has at least 80% residual activity of xylanase activity after incubation at 61° C. for 10 minutes at pH 6.

6. The polypeptide according to claim 1 , wherein the polypeptide has a residual activity of at least 70% when incubated with 0.2 mg/ml pepsin in a buffered solution at pH 3.5 for two hours at a temperature of 40° C.

7. The polypeptide according to claim 1 , wherein the polypeptide solubilizes arabinoxylans without increasing viscosity in a reaction medium.

8. A feed or feedstuff comprising the polypeptide of claims 1 or 2 .

9. A method for producing the polypeptide of claims 1 or 2 comprising (a) cultivating a host cell comprising a polynucleotide having at least 94% identity with SEQ ID No. 2 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

10. The method according to claim 9 wherein the polypeptide produced is isolated and/or purified.

11. A method for solubilizing arabinoxylan in a xylan-containing material by contacting the xylan-containing material with the polypeptide of claims 1 or 2 .

12. The method of claim 11 wherein the arabinoxylan is insoluble arabinoxylan (AXinsol).

13. The method of claim 11 wherein the xylan-containing material is selected from one or more of the group consisting of: a feed or feedstuff; a feed component; a grain-based material; a mash; a wort; a malt; malted barley; an adjunct, a barley mash; and a cereal flour.

14. The method of claim 13 wherein the feed or feedstuff or feed component comprises or consists of corn, Distillers Dried Grain Solubles (DDGS), corn based Distillers Dried Grain Solubles (cDDGS), wheat, wheat bran or a combination thereof.

15. The method of claim 11 wherein the polypeptide is used in combination with one or more of the enzymes selected from the group consisting of endoglucanases (E.C. 3.2.1.4) celliobiohydrolases (E.C. 3.2.1.91), β-glucosidases (E.C. 3.2.1.21), cellulases (E.C. 3.2.1.74), lichenases (E.C. 3.2.1.73), lipases (E.C. 3.1.1.3), lipid acyltransferases (E.C. 2.3.1.x), phospholipases (E.C. 3.1.1.4, E.C. 3.1.1.32 and E.C. 3.1.1.5), 6-phytase (E.C. 3.1.3.26) 3-phytase (E.C. 3.1.3.8), amylases, alpha-amylases (E.C. 3.2.1.1), xylanases (E.C. 3.2.1.8, E.C. 3.2.1.32, E.C. 3.2.1.37, E.C. 3.2.1.72, and E.C. 3.2.1.136), glucoamylases (E.C. 3.2.1.3), hemicellulases, subtilisin (E.C. 3.4.21.62) bacillolysin (E.C. 3.4.24.28) alkaline serine proteases (E.C. 3.4.21.x) akeratinases (E.C. 3.4.x.x), debranching enzymes, cutinases, esterases and β-mannanase (E.C. 3.2.1.78).

16. The method of claim 12 wherein the polypeptide is used in combination with one or more of the enzymes selected from the group consisting of endoglucanases (E.C. 3.2.1.4) celliobiohydrolases (E.C. 3.2.1.91), β-glucosidases (E.C. 3.2.1.21), cellulases (E.C. 3.2.1.74), lichenases (E.C. 3.2.1.73), lipases (E.C. 3.1.1.3), lipid acyltransferases (E.C. 2.3.1.x), phospholipases (E.C. 3.1.1.4, E.C. 3.1.1.32 and E.C. 3.1.1.5), 6-phytase (E.C. 3.1.3.26) 3-phytase (E.C. 3.1.3.8), amylases, alpha-amylases (E.C. 3.2.1.1), xylanases (E.C. 3.2.1.8, E.C. 3.2.1.32, E.C. 3.2.1.37, E.C. 3.2.1.72, and E.C. 3.2.1.136), glucoamylases (E.C. 3.2.1.3), hemicellulases, subtilisin (E.C. 3.4.21.62) bacillolysin (E.C. 3.4.24.28) alkaline serine proteases (E.C. 3.4.21.x) akeratinases (E.C. 3.4.x.x), debranching enzymes, cutinases, esterases and β-mannanase (E.C. 3.2.1.78).

17. The method of claim 13 wherein the polypeptide is used in combination with one or more of the enzymes selected from the group consisting of endoglucanases (E.C. 3.2.1.4) celliobiohydrolases (E.C. 3.2.1.91), β-glucosidases (E.C. 3.2.1.21), cellulases (E.C. 3.2.1.74), lichenases (E.C. 3.2.1.73), lipases (E.C. 3.1.1.3), lipid acyltransferases (E.C. 2.3.1.x), phospholipases (E.C. 3.1.1.4, E.C. 3.1.1.32 and E.C. 3.1.1.5), 6-phytase (E.C. 3.1.3.26) 3-phytase (E.C. 3.1.3.8), amylases, alpha-amylases (E.C. 3.2.1.1), xylanases (E.C. 3.2.1.8, E.C. 3.2.1.32, E.C. 3.2.1.37, E.C. 3.2.1.72, and E.C. 3.2.1.136), glucoamylases (E.C. 3.2.1.3), hemicellulases, subtilisin (E.C. 3.4.21.62) bacillolysin (E.C. 3.4.24.28) alkaline serine proteases (E.C. 3.4.21.x) akeratinases (E.C. 3.4.x.x), debranching enzymes, cutinases, esterases and β-mannanase (E.C. 3.2.1.78).

Assignees

• Dupont Nutrition Biosci Aps

Inventors

• Lorentsen Rikke Hoeegh
• Arent Lund Susan
• Haaning Svend
• Nikolaev Igor
• Barends Sharief
• Van Tuijl Jan Hendrick A
• Koops Bart

Classifications

• C12N9/2485

  Xylan endo-1,3-beta-xylosidase (3.2.1.32), i.e. endo-1,3-beta-xylanase · CPC title

• A23L29/06

  Enzymes · CPC title

• A61P1/14

  Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents · CPC title

• Y02A40/818

  Alternative feeds for fish, e.g. in aquacultures · CPC title

• A23K50/80

  for aquatic animals, e.g. fish, crustaceans or molluscs · CPC title