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1. (WO2010077775) POLYPEPTIDES HAVING ALPHA-MANNOSIDASE ACTIVITY AND POLYNUCLEOTIDES ENCODING SAME
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Claims

What is claimed is:

1. An isolated polypeptide having alpha-mannosidase activity, selected from the group consisting of:

(a) a polypeptide comprising an amino acid sequence having at least 60% identity to SEQ ID NO: 2;

(b) a polypeptide encoded by a polynucleotide that hybridizes under at least medium stringency conditions with (i) SEQ ID NO: 1 , (ii) the genomic DNA sequence comprising SEQ ID NO: 1 , or (iii) a full-length complementary strand of (i) or (ii);

(c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 60% identity to SEQ ID NO: 1 ; and

(d) a variant comprising a substitution, deletion, and/or insertion of one or more (several) amino acids of SEQ ID NO: 2.

2. The polypeptide of claim 1 , comprising or consisting of the amino acid sequence of SEQ ID NO: 2; or a fragment thereof having alpha-mannosidase activity.

3. The polypeptide of claim 1 , comprising or consisting of the amino acid sequence of SEQ ID NO: 2.

4. The polypeptide of claim 1 , which is encoded by a polynucleotide comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 ; or a subsequence thereof encoding a fragment having alpha-mannosidase activity.

5. The polypeptide of claim 1 , which is encoded by a polynucleotide comprising or consisting of the nucleotide sequence of SEQ ID NO: 1.

6. The polypeptide of claim 1 , which is encoded by the polynucleotide contained in plasmid pTterO8D4 which is contained in E. coli NRRL B-50205.

7. An isolated polynucleotide comprising a nucleotide sequence that encodes the polypeptide of any of claims 1-6.

8. A nucleic acid construct comprising the polynucleotide of claim 7 operably linked to one or more (several) control sequences that direct the production of the polypeptide in an expression host.

9. A recombinant host cell comprising the nucleic acid construct of claim 8.

10. A method of producing the polypeptide of any of claims 1-6, comprising: (a) cultivating a cell, which in its wild-type form produces the polypeptide, under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

11 . A method of producing the polypeptide of any of claims 1-6, comprising: (a) cultivating a host cell comprising a nucleic acid construct comprising a nucleotide sequence encoding the polypeptide under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

12. A method of producing a mutant of a parent cell, comprising disrupting or deleting a polynucleotide encoding the polypeptide, or a portion thereof, of any of claims 1 -6, which results in the mutant producing less of the polypeptide than the parent cell.

13. A mutant cell produced by the method of claim 12.

14. The mutant cell of claim 13, further comprising a gene encoding a native or heterologous protein.

15. A method of producing a protein, comprising: (a) cultivating the mutant cell of claim 14 under conditions conducive for production of the protein; and (b) recovering the protein.

16. A method of producing the polypeptide of any of claims 1-6, comprising: (a) cultivating a transgenic plant or a plant cell comprising a polynucleotide encoding the polypeptide under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

17. A transgenic plant, plant part or plant cell transformed with a polynucleotide encoding the polypeptide of any of claims 1-6.

18. A double-stranded inhibitory RNA (dsRNA) molecule comprising a subsequence of the polynucleotide of claim 7, wherein optionally the dsRNA is a siRNA or a miRNA molecule.

19. A method of inhibiting the expression of a polypeptide having alpha-mannosidase activity in a cell, comprising administering to the cell or expressing in the cell a double-stranded RNA (dsRNA) molecule, wherein the dsRNA comprises a subsequence of the polynucleotide of claim 7.

20. A method for producing a demannosylated protein, comprising: treating a protein comprising O-linked mannose residues with the polypeptide of any of claims 1-6, wherein the O-linked mannose residues on the protein are removed.