Xylanases for Solubilising Arabinoxylan-Containing Material

Abstract

The present invention relates to a method for solubilising arabinoxylan-containing material (particularly insoluble arabinoxylan-containing material), comprising admixing a xylan-containing material with a xylanase comprising a polypeptide sequence shown herein as SEQ ID No. 3, SEQ ID No. 2, SEQ ID No. 1, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11 or SEQ ID No. 15, or a variant, homologue, fragment or derivative thereof having at least 75% identity with SEQ ID No. 3 or SEQ ID No. 2 or SEQ ID No. 1 or SEQ ID No. 9 or SEQ ID No. 10 or SEQ ID No. 11 or SEQ ID No. 15; or a polypeptide sequence which comprises SEQ ID No. 3, SEQ ID No. 2, SEQ ID No. 1, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11 or SEQ ID No. 15 with a conservative substitution of at least one of the amino acids; or a xylanase which is encoded by a nucleotide sequence shown herein as SEQ ID No. 6, SEQ ID No. 5, SEQ ID No. 4, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 16, SEQ ID No. 17 or SEQ ID No. 18, or a nucleotide sequence which can hybridize to SEQ ID No. 6, SEQ ID No. 5, SEQ ID No. 4, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 16, SEQ ID No. 17 or SEQ ID No. 18 under high stringency conditions, or a nucleotide sequence which has at least 75% identity with SEQ ID No. 6, SEQ ID No. 5, SEQ ID No. 4, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 16, SEQ ID No. 17 or SEQ ID No. 18, or a nucleotide sequence which differs from SEQ ID No. 6 or SEQ ID No. 5 or SEQ ID No. 4 or SEQ ID No. 12 or SEQ ID No. 13 or SEQ ID No. 14 or SEQ ID No. 16 or SEQ ID No. 17 or SEQ ID No. 18 due to the degeneracy of the genetic code, or a xylanase obtainable (or obtained) from Fusarium verticilloides . The present invention also relates to a novel xylanase comprising (or consisting of) a polypeptide sequence shown herein as SEQ ID No. 3, SEQ ID No. 2 or SEQ ID No. 1, or a variant, homologue, fragment or derivative thereof having at least 99% identity with SEQ ID No. 3 or SEQ ID No. 2 or SEQ ID No. 1; or a xylanase which is encoded by a nucleotide sequence shown herein as SEQ ID No. 6, SEQ ID No. 5 or SEQ ID No. 4, or a nucleotide sequence which can hybridize to SEQ ID No. 4 or SEQ ID No. 5 under high stringency conditions, or a nucleotide sequence which has at least 97.7% identity (preferably 98% identity) with SEQ ID No. 6, SEQ ID No. 5 or SEQ ID No. 4. The present invention yet further relates to methods relating to feedstuffs, malting and brewing, processing of grain-based materials such as during the production of bioethanol or biochemical (e.g. bio-based isopropanol), or wheat gluten-starch separation processes and the like.

1 - 10 . (canceled) 
     
     
         11 . A polypeptide having xylanase activity comprising a polypeptide sequence shown herein as SEQ ID No. 3, SEQ ID No. 2 or SEQ ID No. 1, or a variant, homologue, fragment or derivative thereof having at least 75% identity with SEQ ID No. 3 or SEQ ID No. 2 or SEQ ID No. 1; or a polypeptide which is encoded by a nucleotide sequence shown herein as SEQ ID No. 6, SEQ ID No. 5 or SEQ ID No. 4, or a nucleotide sequence which has at least 75% identity with SEQ ID No. 6, SEQ ID No. 5 or SEQ ID No. 4. 
     
     
         12 . The polypeptide according to  claim 11  wherein the polypeptide is an endo-1,4̂-d˜xylanase. 
     
     
         13 . The polypeptide according to  claim 11  wherein the polypeptide has an optimum temperature in the range of 50-70° C. 
     
     
         14 . The polypeptide according to  claim 11  wherein the polypeptide has a pH optimum in the range of 4.6 to 7. 
     
     
         15 . The polypeptide according to  claim 11  wherein the enzyme is formulated with a coating or is encapsulated. 
     
     
         16 . An isolated or recombinant nucleic acid molecule comprising a polynucleotide sequence selected from the group consisting of:
 a. a polynucleotide sequence which encodes a polypeptide sequence selected from the group consisting of SEQ ID No. 3, SEQ ID No. 2 or SEQ ID No. 1, or a variant, homologue, fragment or derivative thereof having at least 75% identity with SEQ ID No. 3 or SEQ ID No. 2 or SEQ ID No. 1; or   b. a polynucleotide sequence shown herein as SEQ ID No. 6, SEQ ID No. 5 or SEQ ID No. 4; or a nucleotide sequence which has at least 75% identity with SEQ ID No. 6, SEQ ID No. 5 or SEQ ID No. 4.   
     
     
         17 . A vector comprising the nucleic acid molecule of  claim 16 . 
     
     
         18 . A host cell comprising the nucleic acid molecule of  claim 16 . 
     
     
         19 . A feed additive composition or a premix comprising the polypeptide of  claim 11 , and optionally at least one mineral and/or at least one vitamin. 
     
     
         20 . The feed additive composition or premix according to  claim 19  which further comprises one or more of the enzymes selected from the group consisting of a protease (e.g. subtilisin (E.C. 3.4.21.62) or a bacillolysin (E.C. 3.4.24.28) or an alkaline serine protease (E.C. 3.4.21.x) or a keratinase (E.C. 3.4.x.x)) and/or an amylase (including ct-amylases (E.C. 3.2.1.1), G4-forming amylases (E.C. 3.2.1.60), β-amylases (E.C. 3.2.1.2) and γ-amylases (E.C. 3.2.1.3). 
     
     
         21 . A method of preparing a feedstuff comprising admixing a feed component with the polypeptide of  claim 11 . 
     
     
         22 . A feedstuff comprising a feed additive composition or premix according to  claim 19 . 
     
     
         23 . (canceled) 
     
     
         24 . The method according to claim  1  wherein the method is part of a wheat gluten-starch separation process, a biofuel (e.g. bioethanol) or biochemical (e.g. bio-based isoprene) production process, or a malting or brewing process. 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . A method for degrading arabinoxylan-containing material in a xylan-containing material, comprising admixing said xylan-containing material with a xylanase comprising a polypeptide sequence shown herein as SEQ ID No. 3, SEQ ID No. 2, SEQ ID No. 1, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11 or SEQ ID No. 15; or a variant, homologue, fragment or derivative thereof having at least 75% identity with SEQ ID No. 3 or SEQ ID No. 2 or SEQ ID No. 1 or SEQ ID No. 9 or SEQ ID No. 10 or SEQ ID No. 11 or SEQ ID No. 15; or a polypeptide sequence which comprises SEQ ID No. 3, SEQ ID No. 2, SEQ ID No. 1, SEQ ID No. 9, SEQ ID No. 10, SEQ ID No. 11 or SEQ ID No. 15 with a conservative substitution of at least one of the amino acids; or a xylanase which is encoded by a nucleotide sequence shown herein as SEQ ID No. 6, SEQ ID No. 5. SEQ ID No. 4, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 16, SEQ ID No. 17 or SEQ ID No. 18, or a nucleotide sequence which can hybridize to SEQ ID No. 6, SEQ ID No. 5, SEQ ID No. 4, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 16, SEQ ID No. 17 or SEQ ID No. 18 under high stringency conditions, or a nucleotide sequence which has at least 75% identity with SEQ ID No. 6, SEQ ID No. 5, SEQ ID No. 4, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 16, SEQ ID No. 17 or SEQ ID No. 18, or a nucleotide sequence which differs from SEQ ID No. 6 or SEQ ID No. 5 or SEQ ID No. 4 or SEQ ID No. 12 or SEQ ID No. 13 or SEQ ID No. 14 or SEQ ID No. 16 or SEQ ID No. 17 or SEQ ID No. 18 due to the degeneracy of the genetic code, or a xylanase obtainable (or obtained) from  Fusarium verticilloides.    
     
     
         28 . The method according to  claim 27  wherein the arabinoxylan is insoluble arabinoxylan (AXinsol). 
     
     
         29 . The method according to  claim 27  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 barely; an adjunct, a barley mash; and a cereal flour. 
     
     
         30 . The method according to  claim 27  wherein the arabinoxylans are solubilized without increasing viscosity in the reaction medium. 
     
     
         31 . The method according to  claim 29  wherein the feed or feedstuff or feed component comprises or consists of corn, DDGS (such as cDDGS), wheat, wheat bran or a combination thereof. 
     
     
         32 . The method according to  claim 31  wherein the feed or feedstuff is a corn-based feedstuff. 
     
     
         33 . The method according to  claim 27  wherein the xylanase is used in combination with one or more of the enzymes selected from the group consisting of a protease (e.g. subtilisin (E.C. 3.4.21.62) or a bacillolysin (E.C. 3.4.24.28) or an alkaline serine protease (E.C. 3.4.21.x) or a keratinase (E.C. 3.4.x.x)) and/or an amylase (including α-amylases (E.C. 3.2.1.1), G4-forming amylases (E.C. 3.2.1.60), β-amylases (E.C. 3.2.1.2) and γ-amylases (E.C. 3.2.1.3); and/or a protease (e.g. subtilisin (E.C. 3.4.21.62) or a bacillolysin (E.C. 3.4.24.28) or an alkaline serine protease (E.C. 3.4.21.x) or a keratinase (E.C. 3.4.x.x)).