US 11,896,034 B2Granted

Polypeptides having lysozyme activity, polynucleotides encoding same and uses and compositions thereof

AssigneeNovozymes A/S

InventorsMikkel Klausen, Kirk Matthew Schnorr, Lars Kobberoee Skov, Soeren Nymand-Grarup, Marianne Thorup Cohn, Peter Bjarke Olsen, Ming Li, Ye Liu

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Abstract

The present invention relates animal feed or animal feed additives comprising one or more polypeptides having lysozyme activity. The invention also relates to polypeptides having lysozyme activity, polynucleotides encoding the polypeptides nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Claims7

Dependent claims

A granule comprising one or more Glycoside Hydrolase Family 25 (GH25) polypeptides having lysozyme activity, wherein the polypeptide has improved activity compared to the activity of the polypeptide of SEQ ID NO:

39 and wherein the polypeptide is selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO:
3;
(b) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
6;
(c) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
9;
(d) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
12;
(e) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
18;
(f) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
21;
(g) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
24;
(h) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
30;
(i) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
33;
(j) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
38;
(k) a variant of the polypeptide selected from the group consisting of SEQ ID NO:
3, SEQ ID NO:
6, SEQ ID NO:
9, SEQ ID NO:
12, SEQ ID NO:
18, SEQ ID NO:
21, SEQ ID NO:
24, SEQ ID NO:
30, SEQ ID NO:
33 and SEQ ID NO:
38, wherein the variant has lysozyme 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 1-20 positions;
(l) a polypeptide comprising the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k), and a N-terminal and/or C-terminal histidine tag (His-tag) and/or histidine-glutamine tag (HQ-tag);
(m) a polypeptide comprising the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k) and a N-terminal and/or C-terminal extension of between 1 and 10 amino acids;
and
(n) a fragment of the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k) or having lysozyme activity and having at least 90% of the length of the mature polypeptide.

33 more elements

An isolated polypeptide having lysozyme activity, selected from the group consisting of:

(a) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO:
3;
(b) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
6;
(c) a polypeptide having at least 91% sequence identity to the polypeptide of SEQ ID NO:
9;
(d) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
12;
(e) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
21;
(f) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
24;
(g) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
30;
(h) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
33;
(i) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
38;
(j) a variant of the polypeptide of SEQ ID NO:
3, wherein the variant has lysozyme 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 1-20 positions;
(k) a variant of the polypeptide of SEQ ID NO:
6, wherein the variant has lysozyme 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 1-20 positions;
(l) a variant of the polypeptide of SEQ ID NO:
9, wherein the variant has lysozyme 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 1-18 positions;
(m) a variant of the polypeptide selected from the group consisting of SEQ ID NO:
12, SEQ ID NO:
21, SEQ ID NO:
24, SEQ ID NO:
30 and SEQ ID NO:
33, wherein the variant has lysozyme 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 thereof in 1-20 positions;
(n) a variant of the polypeptide of SEQ ID NO:
38, wherein the variant has lysozyme 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 1-20 positions;
(o) a polypeptide comprising the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (l), (m), or (n) and a N-terminal and/or C-terminal histidine tag (His-tag) and/or histidine-glutamine tag (HQ-tag);
(p) a polypeptide comprising the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (l), (m), or (n) and a N-terminal and/or C-terminal extension of between 1 and 10 amino acids;
and
(q) a fragment of the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (l), (m), or (n) having lysozyme activity and having at least 90% of the length of the mature polypeptide.

33 more elements

A method of hydrolysing peptidoglycan in a bacterial cell wall, comprising treating the bacterial cell wall with one or more Glycoside Hydrolase Family 25 (GH25) polypeptides having lysozyme activity, wherein the polypeptide has improved activity compared to the activity of the polypeptide of SEQ ID NO:

39 and wherein the polypeptide is selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO:
3;
(b) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
6;
(c) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
9;
(d) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
12;
(e) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
18;
(f) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
21;
(g) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
24;
(h) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
30;
(i) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
33;
(j) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO:
38;
(k) a variant of the polypeptide selected from the group consisting of SEQ ID NO:
3, SEQ ID NO:
6, SEQ ID NO:
9, SEQ ID NO:
12, SEQ ID NO:
18, SEQ ID NO:
21, SEQ ID NO:
24, SEQ ID NO:
30, SEQ ID NO:
33 and SEQ ID NO:
38, wherein the variant has lysozyme 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 1-20 positions;
(l) a polypeptide comprising the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k), and a N-terminal and/or C-terminal histidine tag (His-tag) and/or histidine-glutamine tag (HQ-tag);
(m) a polypeptide comprising the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k) and a N-terminal and/or C-terminal extension of between 1 and 10 amino acids;
and
(n) a fragment of the polypeptide of (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), or (k) having lysozyme activity and having at least 90% of the length of the mature polypeptide.

33 more elements

Dependencies are inferred from the claim text.

Facts

Application: 17/212,019Filed 25 Mar 2021 · division
Priority: WO PCT/CN2016/11132021 Dec 2016
Published: US 2021/0212343 A115 Jul 2021
Granted: 13 Feb 2024
Term ends: Not estimatedThe source records don’t support an estimate.

Classification16 codes

A23K 20/147
Amino acids; Derivatives thereof: polymeric derivatives, e.g. peptides or proteins
A23K 20/189
Organic substances: enzymes
A23K 10/30
Animal feeding-stuffs: from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
A23K 20/174
Organic substances: vitamins

Family

PCT/CN2017/117753Parent of this division · filed 21 Dec 2017
US 2021/0212343 A1Pre-grant publication · published 15 Jul 2021
Application16/463,205Parent of this division
US 10,986,850 B2Parent of this division
US 2019/0328005 A1Published application
US 11,896,034 B2This patent

From the US filing history. Foreign family members aren’t listed.

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