Polypeptides having Alpha-Galactosidase Activity and Polynucleotides Encoding Same
AssigneeNovozymes A/S
InventorsLone Carstensen, Nikolaj Spodsberg, Morten Gjermansen, Jesper Salomon, Kristian Bertel Roemer M. Krogh
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Abstract
The present invention relates to methods of releasing galactose from legumes using polypeptides having alpha-galactosidase activity. The invention also relates to polypeptides having alpha-galactosidase activity, polynucleotides encoding the polypeptides, nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of 5 producing the polypeptides. The invention also relates to compositions comprising the polypeptides of the invention and the use of the polypeptides in animal feed.
Claims18
An animal feed additive comprising one or more GH36 polypeptides having alpha-galactosidase activity selected from the group consisting of:
- (a) a polypeptide having at least 80% 100% sequence identity to the polypeptide of SEQ ID NO:
- 3;
- (b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions, or very high stringency conditions with
- (i) the mature polypeptide coding sequence of SEQ ID NO:
- 1,
- (ii) the cDNA sequence thereof, or
- (iii) the full-length complementary strand of (i) or (ii);
- (c) a polypeptide encoded by a polynucleotide having at least 80% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:
- 1;
- (d) a variant of the polypeptide selected from the group consisting of SEQ ID NO:
- 3, wherein the variant has alpha-galactosidase activity and comprises one or more amino acid substitutions, and/or one or more amino acid deletions, and/or one or more amino acid insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 positions;
- (e) a polypeptide comprising the polypeptide of (a), (b), (c) or (d) and a N-terminal and/or C-terminal His-tag and/or HQ-tag;
- (f) a polypeptide comprising the polypeptide of (a), (b), (c) or (d) and a N-terminal and/or C-terminal extension of up to 10 amino acids, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids;
- and
- (g) a fragment of the polypeptide of (a), (b), (c) or (d) having alpha-galactosidase activity and having at least 90% of the length of the mature polypeptide.
A liquid formulation comprising one or more GH36 polypeptides having alpha-galactosidase activity, wherein the liquid formulation comprises:
- (A) 0.001% to 25% w/w of polypeptide having alpha-galactosidase activity wherein the polypeptide having alpha-galactosidase activity is selected from the group consisting of:
- (a) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO:
- 3;
- (b) a variant of SEQ ID NO:
- 3, wherein the variant has alpha-galactosidase activity and comprises one or more amino acid substitutions, and/or one or more amino acid deletions, and/or one or more amino acid insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 positions;
- (c) a polypeptide comprising the polypeptide of (a) or (b) and a N-terminal and/or C-terminal His-tag and/or HQ-tag;
- (d) a polypeptide comprising the polypeptide of (a) or (b) and a N-terminal and/or C-terminal extension of up to 10 amino acids, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids;
- and
- (e) a fragment of the polypeptide of (a) or (b) having alpha-galactosidase activity and having at least 90% of the length of the mature polypeptide;
- and
- (B) water.
An isolated polypeptide having alpha-galactosidase activity, selected from the group consisting of:
- (a) a polypeptide having at least 96.5% sequence identity to the polypeptide of SEQ ID NO:
- 3;
- (b) a polypeptide encoded by a polynucleotide having at least 96.5% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:
- 1;
- (c) a variant of the polypeptide selected from the group consisting of SEQ ID NO:
- 3, wherein the variant has alpha-galactosidase activity and comprises one or more amino acid substitutions, and/or one or more amino acid deletions, and/or one or more amino acid insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27 positions;
- (d) a polypeptide comprising the polypeptide of (a), (b) or (c) and a N-terminal and/or C-terminal His-tag and/or HQ-tag;
- (e) a polypeptide comprising the polypeptide of (a), (b) or (c) and a N-terminal and/or C-terminal extension of up to 10 amino acids, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids;
- and
- (f) a fragment of the polypeptide of (a), (b) or (c) having alpha-galactosidase activity and having at least 90% of the length of the mature polypeptide.
(canceled)
Dependencies are inferred from the claim text.