Polypeptides having feruloyl esterase activity and nucleic acids encoding the same
The present invention relates to isolated polypeptides having esterase activity and isolated polynucleotides encoding the polypeptides. 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.
1. An isolated polypeptide having feruloyl esterase activity, wherein the polypeptide is selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of amino acids 21 to 520 of SEQ ID NO: 2; and
(b) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence that hybridizes under at least very high stringency conditions with (i) the nucleotide sequence of nucleotides 61 to 1560 of SEQ ID NO: 1, (ii) a cDNA sequence contained in the nucleotide sequence of nucleotides 61 to 1560 of SEQ ID NO: 1, or (iii) a full-length complementary strand of (i) or (ii), wherein 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 following standard Southern blotting procedures for 12 to 24 hours optimally, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.
2. The isolated polypeptide of claim 1 , which has at least 97% identity to the mature polypeptide of amino acids 21 to 520 of SEQ ID NO: 2.
3. The isolated polypeptide of claim 1 , which has at least 98% identity to the mature polypeptide of amino acids 21 to 520 of SEQ ID NO: 2.
4. The isolated polypeptide of claim 1 , which has at least 99% identity to the mature polypeptide of amino acids 21 to 520 of SEQ ID NO: 2.
5. The isolated polypeptide of claim 1 , comprising or consisting of the amino acid sequence of SEQ ID NO: 2.
6. The isolated polypeptide of claim 1 , comprising or consisting of the mature polypeptide of amino acids 21 to 520 of SEQ ID NO: 2.
7. A nucleic acid construct comprising a nucleotide sequence that encodes the polypeptide of claim 1 , wherein the nucleotide sequence is operably linked to one or more heterologous control sequences that direct the production of the polypeptide in a suitable expression host.
8. A recombinant expression vector comprising the nucleic acid construct of claim 7 .
9. A recombinant host cell comprising the nucleic acid construct of claim 7 .
10. A method for producing a polypeptide, the method comprising (a) cultivating the recombinant host cell of claim 9 to produce a supernatant comprising the polypeptide; and (b) recovering the polypeptide.
11. A method for improving the nutritional value of an animal feed, wherein at least one polypeptide of claim 1 is added to the animal feed.
12. A composition comprising at least one polypeptide of claim 1 , and
(a) at least one fat soluble vitamin, and/or
(b) at least one water soluble vitamin, and/or
(c) at least one trace mineral.
13. The composition of claim 12 , which further comprises amylase, phytase, xylanase, galactanase, alpha-galactosidase, protease, phospholipase, beta-glucanase, and/or arabinofuranosidase.
14. An animal feed composition comprising the polypeptide of claim 1 .
15. A method for improving the nutritional value of an animal feed, the method comprising adding the polypeptide of claim 1 to the feed.