Compositions comprising polypeptides having galactanase activity and polypeptides having beta-galactosidase activity

Patent metadata

Field	Value
Publication number	US-10927359-B2
Application number	US-201716300978-A
Country	US
Kind code	B2
Filing date	May 24, 2017
Priority date	May 24, 2016
Publication date	Feb 23, 2021
Grant date	Feb 23, 2021

Abstract

The present invention relates to compositions comprising polypeptides having galactanase activity and polypeptides having beta-galactosidase activity for use in e.g. animal feed. The present invention further relates to polypeptides having beta-galactosidase activity, polypeptides having galactanase activity and 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.

First claim

What is claimed is:

1. A method of improving one or more performance parameters of an animal, comprising feeding the animal with plant-based material and a composition comprising one or more GH53 polypeptides having galactanase activity and one or more GH35 polypeptides having beta-galactosidase activity, wherein the plant-based material comprises soybean or soybean meal and the one or more performance parameters are selected from the group consisting of body weight gain (BWG), European Production Efficiency Factor (EPEF) and feed conversion ratio (FCR).

2. The method of claim 1 , wherein the composition is a granule.

3. The method of claim 1 , wherein the plant-based material and the composition are a pelleted animal feed.

4. The method of claim 1 , wherein the composition further comprises one or more components selected from the group consisting of:
one or more vitamins;
one or more minerals;
one or more amino acids;
one or more prebiotics;
one or more phytogenics;
one or more organic acids; and
one or more other feed ingredients.

5. The method of claim 1 , wherein the composition is a liquid composition.

6. The method of claim 1 , wherein the GH53 polypeptide comprises the motif GV[T/M]PD[W/M]VQ[I/V]GNE (SEQ ID NO: 65) and/or the motif WADP[A/G]xQxKPxAW (SEQ ID NO: 66).

7. The method of claim 1 , wherein the GH53 polypeptide 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 polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 7;
(c) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 11;
(d) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 15;
(e) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 19;
(f) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 23;
(g) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 27;
(h) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 31;
(i) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 35; and
(j) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 39.

8. The method of claim 1 , wherein the GH35 polypeptide is selected from the group consisting of:
(a) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 43;
(b) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 46;
(c) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 49;
(d) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 52;
(e) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 55;
(f) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 58;
(g) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 61;
(h) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 70;
(i) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 73;
(j) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 76; and
(k) a polypeptide having at least 80% sequence identity to the polypeptide of SEQ ID NO: 79.

9. The method of claim 1 , wherein the GH53 polypeptide has at least 80% sequence identity to the polypeptide of SEQ ID NO: 3 and the GH35 polypeptide has at least 80% sequence identity to the polypeptide of SEQ ID NO: 58.

10. The method of claim 1 , wherein the GH53 polypeptide has at least 85% sequence identity to the polypeptide of SEQ ID NO: 3 and the GH35 polypeptide has at least 85% sequence identity to the polypeptide of SEQ ID NO: 58.

11. The method of claim 1 , wherein the GH53 polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO: 3 and the GH35 polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO: 58.

12. The method of claim 1 , wherein the GH53 polypeptide has at least 95% sequence identity to the polypeptide of SEQ ID NO: 3 and the GH35 polypeptide has at least 95% sequence identity to the polypeptide of SEQ ID NO: 58.

13. The method of claim 1 , wherein the GH53 is the polypeptide of SEQ ID NO: 3 or 4 and the GH35 polypeptide is the polypeptide of SEQ ID NO: 58.

14. The method of claim 9 , wherein the GH53 polypeptide:
comprises an N-terminal and/or C-terminal His-tag and/or HQ-tag;
comprises an N-terminal and/or C-terminal extension of up to 10 amino acids; or
is a fragment having galactanase activity and having at least 90% of the length of the mature polypeptide.

15. The method of claim 9 , wherein the GH35 polypeptide:
comprises an N-terminal and/or C-terminal His-tag and/or HQ-tag;
comprises an N-terminal and/or C-terminal extension of up to 10 amino acids; or
is a fragment having beta-galactosidase activity and having at least 90% of the length of the mature polypeptide.

16. The method of claim 9 , wherein the GH53 polypeptide is a variant of SEQ ID NO: 3, wherein the variant has galactanase activity and comprises one or more amino acid substitutions, one or more amino acid deletions, and/or one or more amino acid insertions.

17. The method of claim 9 , wherein the GH35 polypeptide is a variant of SEQ ID NO: 58, wherein the variant has beta-galactosidase activity and comprises one or more amino acid substitutions, one or more amino acid deletions, and/or one or more amino acid insertions.

18. The method of claim 1 , wherein the composition releases at least 12 g galactose per kg soybean meal when performed under the reaction conditions 20 mg galactanase and 20 mg beta-galactosidase per kg soybean meal in 10% w/v 0.1M citric acid-phosphate buffer pH 6.5 incubating at 40° C. for 2 hours.

Assignees

• Novozymes As

Inventors

• Carstensen Lone
• Spodsberg Nikolaj
• Gjermansen Morten
• Salomon Jesper
• Krogh Kristian B R M
• Peng Wei
• Liu Cui
• Della Pia Eduardo Antonio

Classifications

• C12N9/2402

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• A23K20/20

  Inorganic substances, e.g. oligoelements · CPC title

• A23K10/30

  from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms (obtained by microbiological or biochemical processes, e.g. using yeasts or enzymes, A23K10/10) · CPC title

• C12Y302/01023

  Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase · CPC title

• C12N9/2471 Primary

  Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase · CPC title