Polypeptides having glucuronyl esterase activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9394556-B2
Application number	US-201314395945-A
Country	US
Kind code	B2
Filing date	Apr 22, 2013
Priority date	Apr 23, 2012
Publication date	Jul 19, 2016
Grant date	Jul 19, 2016

Abstract

The present invention relates to isolated polypeptides having glucuronyl esterase activity, catalytic domains and polynucleotides encoding the polypeptides or catalytic domains. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides or catalytic domains.

First claim

The invention claimed is:

1. A nucleic acid construct comprising a polynucleotide encoding a polypeptide having glucuronyl esterase activity operably linked to one or more heterologous control sequences that direct the expression of the polypeptide in an expression host, wherein the polypeptide having glucuronyl esterase activity is selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1, wherein the high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.;
(c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1; and
(d) a fragment of the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2, wherein the fragment has glucuronyl esterase activity.

2. An isolated recombinant host cell comprising the nucleic acid construct of claim 1 .

3. A method of producing a polypeptide having glucuronyl esterase activity, comprising:
(a) cultivating the isolated recombinant host cell of claim 2 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.

4. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

5. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 96% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

6. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 97% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

7. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 98% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

8. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 99% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

9. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of SEQ ID NO: 2.

10. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity consists of the amino acid sequence of SEQ ID NO: 2.

11. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

12. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity consists of the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

13. The nucleic acid construct of claim 1 , wherein the polypeptide is a fragment of the amino acid sequence of SEQ ID NO:2, wherein the fragment has glucuronyl esterase activity.

14. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1, wherein the very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.

15. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1.

16. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 96% sequence identity to the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1.

17. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 97% sequence identity to the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1.

18. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 98% sequence identity to the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1.

19. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 99% sequence identity to the nucleotide sequence of nucleotides 33 to 1457 of SEQ ID NO: 1.

20. An expression vector comprising the nucleic acid construct of claim 1 .

21. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 91% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

22. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 92% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

23. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 93% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

24. The nucleic acid construct of claim 1 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 94% sequence identity to the amino acid sequence of amino acids 101 to 474 of SEQ ID NO: 2.

Assignees

• Novoyzmes As
• Novozymes As

Inventors

• Börjesson Johan
• Viksø-Nielsen Anders
• Spodsberg Nikolaj
• Krogh Kristian

Classifications

• C12N9/24

  acting on glycosyl compounds (3.2) · CPC title

• C12P7/14

  Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms · CPC title

• C12N9/2434

  acting on beta-1,4-glucosidic bonds · CPC title

• C12P19/14 Primary

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title