Carbohydrate degrading polypeptide and uses thereof
The invention relates to a polypeptide having hemicellulase activity. The invention features the full length coding sequence of the novel gene as well as the amino acid sequence of the full-length functional protein and functional equivalents of the gene or the amino acid sequence. The invention also relates to methods for using the polypeptide in industrial processes. Also included in the invention are cells transformed with a polynucleotide according to the invention suitable for producing these proteins.
1. A method for producing a sugar from a lignocellulosic material, comprising:
(a) pretreatment of the lignocellulosic material; and
(b) contacting the pretreated lignocellulosic material with a polypeptide having hemicellulase activity, comprising:
(i) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:32;
(ii) an amino acid sequence encoded by a nucleic acid sequence having at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 31, 34, and 35;
(iii) an amino acid sequence encoded by a nucleic acid sequence hybridizing with the complement of a nucleic acid sequence having at least 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 31, 34, and 35; or
(iv) an amino acid sequence encoded by a nucleic acid sequence which is the reverse complement of a nucleic acid sequence as defined in (i), (ii) or (iii).
2. The method of claim 1 , further comprising contacting the lignocellulosic material with a cellulase and/or an additional hemicellulase and/or a pectinase.
3. The method of claim 2 , wherein the cellulase is selected from the group consisting of a GH61, cellobiohydrolase, cellobiohydrolase I, cellobiohydrolase II, endo-β-1,4-glucanase, β-glucosidase, and β-(1,3)(1,4)-glucanase.
4. The method of claim 2 , wherein the additional hemicellulase is selected from the group consisting of endoxylanase, β-xylosidase, α-L-arabinofuranosidase, α-D-glucuronidase, cellobiohydrolase, feruloyl esterase, coumaroyl esterase, α-galactosidase, β-galactosidase, β-mannanase, and β-mannosidase.
5. The method of claim 2 , wherein the pectinase is selected from the group consisting of endo-polygalacturonase, pectin methyl esterase, endo-galactanase, β-galactosidase, pectin acetyl esterase, endo-pectin lyase, pectate lyase, α-rhamnosidase, exo-galacturonase, exo-polygalacturonate lyase, rhamnogalacturonan hydrolase, rhamnogalacturonan lyase, rhamnogalacturonan acetyl esterase, rhamnogalacturonan galacturonohydrolase, xylogalacturonase, and α-arabinofuranosidase.
6. The method of claim 1 , wherein the lignocellulosic material comprises fiber from corn kernels.
7. A method for preparing a fermentation product, comprising:
(a) performing the method according to claim 1 ; and
(b) fermentation of the resulting material, to thereby prepare the fermentation product.
8. The method of claim 1 , wherein said polypeptide has alpha-arabinofuranosidase activity.
9. A method for treating a substrate comprising hemicellulose, comprising:
(a) pretreatment of the substrate; and
(b) contacting the pretreated substrate with a polypeptide having hemicellulase activity, comprising:
(i) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:32;
(ii) an amino acid sequence encoded by a nucleic acid sequence having at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 31, 34, and 35;
(iii) an amino acid sequence encoded by a nucleic acid sequence hybridizing with the complement of a nucleic acid sequence having at least 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 31, 34, and 35; or
(iv) an amino acid sequence encoded by a nucleic acid sequence which is the reverse complement of a nucleic acid sequence as defined in (i), (ii) or (iii).
10. The method of claim 9 , further comprising contacting the lignocellulosic material with a cellulase and/or an additional hemicellulase and/or a pectinase.
11. The method of claim 10 , wherein the cellulase is selected from the group consisting of a GH61, cellobiohydrolase, cellobiohydrolase I, cellobiohydrolase II, endo-β-1,4-glucanase, β-glucosidase, and β-(1,3)(1,4)-glucanase.
12. The method of claim 10 , wherein the additional hemicellulase is selected from the group consisting of endoxylanase, β-xylosidase, α-L-arabinofuranosidase, α-D-glucuronidase, cellobiohydrolase, feruloyl esterase, coumaroyl esterase, α-galactosidase, β-galactosidase, β-mannanase, and β-mannosidase.
13. The method of claim 10 , wherein the pectinase is selected from the group consisting of endo-polygalacturonase, pectin methyl esterase, endo-galactanase, β-galactosidase, pectin acetyl esterase, endo-pectin lyase, pectate lyase, α-rhamnosidase, exo-galacturonase, exo-polygalacturonate lyase, rhamnogalacturonan hydrolase, rhamnogalacturonan lyase, rhamnogalacturonan acetyl esterase, rhamnogalacturonan galacturonohydrolase, xylogalacturonase, and α-arabinofuranosidase.
14. The method of claim 9 , wherein said substrate comprises a plant material.
15. The method of claim 9 , wherein said substrate comprises fiber from corn kernels.
16. A method for the preparation of a fermentation product, which method comprises:
(a) performing the method according to claim 9 ; and
(b) fermentation of the resulting material, to thereby prepare the fermentation product.
17. A method for preparing a polypeptide having hemicellulase activity, comprising cultivating a cell under conditions which allow for expression of said polypeptide and, optionally, recovering the expressed polypeptide, wherein said polypeptide comprises:
(i) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:32;
(ii) an amino acid sequence encoded by a nucleic acid sequence having at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 31, 34, and 35;
(iii) an amino acid sequence encoded by a nucleic acid sequence hybridizing with the complement of a nucleic acid sequence having at least 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 31, 34, and 35; or
(iv) an amino acid sequence encoded by a nucleic acid sequence which is the reverse complement of a nucleic acid sequence as defined in (i), (ii) or (iii).