WO2020099490A1

WO2020099490A1 - Oral care composition comprising enzymes

Info

Publication number: WO2020099490A1
Application number: PCT/EP2019/081184
Authority: WO
Inventors: Thomas Thomasen DURHUUS; Lorena González PALMÉN; Anna Verena REISER; Werner W STREICHER
Original assignee: Novozymes A/S
Priority date: 2018-11-14
Filing date: 2019-11-13
Publication date: 2020-05-22
Also published as: WO2020099491A1

Abstract

The invention provides relates to oral care compositions comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component.

Description

ORAL CARE COMPOSITION COMPRISING ENZYMES

REFERENCE TO SEQUENCE LISTING

This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to oral care compositions comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component.

BACKGROUND OF THE INVENTION:

Biofilms are communities of bacteria that are found on solid surfaces in many different environments, including surfaces of the oral cavity. Oral biofilm, or dental plague, contains many of the bacteria that are associated with oral health issues such as oral malodor, demineralization, dental caries, tooth decay, potential loss of teeth and gum disease (gingivitis and periodontitis).

The formation of oral biofilm occurs in three stages known as the lag phase, growth phase, and steady state, respectively. In the lag phase, glycoproteins from saliva bind to an oral surface such as teeth and create a structure termed the pellicle that functions as attachment site for bacteria. In the growth phase, co-aggregation occurs, i.e., secondary bacterial colonizers attach to the primary bacterial colonizers, causing the diversity of the biofilm to increase and the biofilm to grow and mature. In the steady state, the biofilm growth slows down and eventually stops. This stage-based formation cycle causes biofilms to exist in several consecutive layers, which makes physical abrasion of biofilm more difficult.

Within a biofilm, the residing bacterial cells are distributed in an extracellular polymeric matrix that consists primarily of water, proteins, exopolysaccharides, lipopolysaccharides, lipids, surfactants, and extracellular DNA (eDNA). eDNA is an important component of the extracellular polymeric matrix and plays a vital role in both the formation and stability of biofilm and in the antimicrobial properties of its embedded bacteria. eDNA in biofilm is known to influence the initial attachment and adhesion of biofilm to surfaces as well as the subsequent buildup, and eDNA also has a stabilizing effect on biofilms as it coats the surface of the biofilm. Moreover, eDNA derived from lysed bacteria may contain genes conferring resistance to anti-microbial agents. In case such DNA fragments are transferred within the biofilm and integrated into the chromosome of living bacterial cells, this may lead to new phenotypes with improved anti-microbial resistance profiles.

Because of the increased resistance to anti-microbial agents as well as the mechanical properties of biofilm, many current oral care products are rather inefficient in addressing biofilm formation and alleviating the associated oral health issues. The main focus for biofilm removal has been on mechanical abrasion. However, this approach is difficult due to the multilayered nature of biofilms and is further compromised by the fact that mechanical removal of biofilm, e.g., by brushing the teeth, expands and deepens the areas in the oral cavity where biofilms attach and expand, thus potentially increasing the severity of the problem rather than reducing it.

In view of the important role of biofilm in oral disease, there is a need in the art for oral care compositions that can effectively target oral biofilm. In particular, there is a need for oral care formulations that can target biofilm by removing the eDNA. However, this would require agents such as enzymes that at the same time are effective against biofilm as well as stable under conditions suitable for oral care, including in oral care formulations.

WO 201 1/098579 relates to NucA/NucB-type deoxyribonucleases and methods for biofilm disruption and prevention.

SUMMARY OF THE INVENTION

In a first aspect, the present invention relates to an oral care composition comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component.

In a second aspect, the present invention relates to a composition according to the first aspect for use as a medicament.

In a third aspect, the present invention relates to a composition according to the first aspect for use in the treatment of oral disease.

In a fourth aspect, the present invention relates to use of a composition according to the first aspect for treatment or prophylactic treatment of a human or animal subject.

In a fifth aspect, the present invention relates to a method of treatment of a human or animal subject, the method comprising administering a composition according to the first aspect to a human or animal subject.

In a sixth aspect, the present invention relates to a kit of parts comprising:

a) a composition according to the first aspect; and

b) instructions for use.

BRIEF DESCRIPTION OF DRAWINGS

Fig. 1 shows an example of the thermal stability data generated using the nanoDSF instrument. Panel A is an example of the data obtained (the ratio of the fluorescence emission at 350 nm to 330 nm) in triplicate for SEQ ID NO: 32 (for clarity only every fifth data point is shown) as a function of temperature. Panel B shows the first derivative of the raw data in Panel A. The peak maximum in the first derivative plot corresponds to the mid-point of the thermal unfolding transition, referred to as Td. In this example the Td corresponds to 58°C and is highly reproducible within the three replicates. Fig. 2 shows the average thermal stability of DNases in 1 mM EDTA (black bars) or 5 mM EDTA (grey bars). The dotted line represents the average for all Td values measured in EDTA.

Fig. 3 shows thermal stability of DNases in the presence of 1 or 5% sodium dodecyl sulphate (SDS). The dotted line represents the average for all Td values measured in SDS.

Fig. 4 shows thermal stability of DNases in the presence of 45 or 90% Plax COOL MINT (Colgate®) mouthwash. The dotted line represents the average for all Td values measured in mouthwash Plax COOL MINT (Colgate®).

Fig. 5 shows thermal stability of DNases in the presence of 1 or 10% (w/v) Cavity Protection Caries toothpaste (Colgate®, Colgate-Palmolive). The dotted line represents the average for all Td values measured in Cavity Protection Caries toothpaste (Colgate ®, Colgate- Palmolive).

DEFINITIONS:

Clade: The term“clade” means a group of polypeptides clustered together on the basis of homologous features traced to a common ancestor. Polypeptide clades can be visualized as phylogenetic trees and a clade is a group of polypeptides that consists of a common ancestor and all its lineal descendants. Polypeptides forming a group within the clade (a subclade) of the phylogenetic tree can also share common properties and are more closely related than other polypeptides in the clade.

Dentures: The term“dentures” is meant to cover dentures as such as well as braces, aligners, retainers, and the like.

Polypeptide having DNase activity: The term“polypeptide having DNase activity” means a polypeptide with DNase (deoxyribonuclease) activity that catalyzes the hydrolytic cleavage of phosphodiester linkages in a DNA backbone, thus degrading DNA. Exodeoxyribonuclease cut or cleaves residues at the end of the DNA backbone where endo-deoxyribonucleases cleaves or cut within the DNA backbone. A DNase may cleave only double-stranded DNA or may cleave double stranded and single stranded DNA. The term “DNases” and the expression “a polypeptide with DNase activity” are used interchangeably throughout this application. For purposes of the present invention, DNase activity may be determined according to the procedure described in Assay I or Assay II ( vide infra).

Polypeptide having mutanase activity: The term“polypeptide having mutanase activity” means a polypeptide having mutanase activity that catalyzes the hydrolytic cleavage of -1 ,3- glycosidic linkages in mutan, thereby degrading mutan. which is a polysaccharide found in oral biofilm. The term“mutanase” and the expression“a polypeptide with mutanase activity” are used interchangeably throughout this application. For purposes of the present invention, mutanase activity may be determined according to the procedure described in WO 2017/083228 or in A. Wiater et a!., Mycological Research, vol. 105, pp. 1357-1363, 2001. Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter“sequence identity”.

For purposes of the present invention, the sequence identity between two amino acid sequences is determined as the output of “longest identity” using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et ai, 2000, Trends Genet. 16: 276-277), preferably version 6.6.0 or later. The parameters used are a gap open penalty of 10, a gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. In order for the Needle program to report the longest identity, the -nobrief option must be specified in the command line. The output of Needle labeled“longest identity” is calculated as follows:

(Identical Residues x 100)/(Length of Alignment - Total Number of Gaps in Alignment)

For purposes of the present invention, the sequence identity between two polynucleotide sequences is determined as the output of “longest identity” using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, supra) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, supra), preferably version 6.6.0 or later. The parameters used are a gap open penalty of 10, a gap extension penalty of 0.5, and the EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. In order for the Needle program to report the longest identity, the nobrief option must be specified in the command line. The output of Needle labeled“longest identity” is calculated as follows:

(Identical Deoxyribonucleotides x 100)/(Length of Alignment - Total Number of Gaps in Alignment)

Nomenclature

For purposes of the present invention, the nomenclature [G/N] means that the amino acid at this position may be a glycine (Gly, G) or an asparagine (Asn, N). Likewise, the nomenclature [T/D/S] means that the amino acid at this position may be a threonine (Thr, T), aspartic acid (Asp, D), or serine (Ser, S), and so forth for other combinations as described herein. Unless otherwise limited further, the amino acid X is defined such that it may be any of the natural amino acids.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based on the surprising and inventive finding that the combination of a DNase and a mutanase exhibits an improved and synergistic effect on removal and/or prevention of oral biofilm when compared to treatment with DNase or mutanase alone. The combination of DNase and mutanase may be formulated in a composition suitable for oral care so that they retain sufficient stability and efficacy for degradation of eDNA and removal and/or prevention of oral biofilm.

Thus, in a first aspect, the present invention relates to oral care compositions comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component.

Advantages of oral care compositions of the invention include improved degradation of eDNA, improved removal of oral biofilm, improved prevention of oral biofilm, improved oral health, improved denture cleaning, improved stability, prolonged shelf-life, reduced side-effects, lower toxicity, as well as other advantages.

Polypeptides having DNase activity

The term“DNase” means a polypeptide with DNase (deoxyribonuclease) activity that catalyzes the hydrolytic cleavage of phosphodiester linkages in a DNA backbone, thus degrading DNA. Exodeoxyribonuclease cut or cleaves residues at the end of the DNA back bone where endo-deoxyribonucleases cleaves or cut within the DNA backbone. A DNase may cleave only double-stranded DNA or may cleave double stranded and single stranded DNA. The term “DNases” and the expression“a polypeptide with DNase activity” are used interchangeably throughout this application. For purposes of the present invention, DNase activity is determined according to the procedure described in the Assay I or Assay II.

Preferably the DNase is selected from any of the enzyme classes E.C.3.1 , preferably E.C.3.1.21 , e.g., such as E.C.3.1.21 .X, where X = 1 , 2, 3, 4, 5, 6, 7, 8 or 9, or, e.g., Deoxyribonuclease I, Deoxyribonuclease IV, Type I site-specific deoxyribonuclease, Type II site- specific deoxyribonuclease, Type III site-specific deoxyribonuclease, CC-preferring endo- deoxyribonuclease, Deoxyribonuclease V, T(4) deoxyribonuclease II, T(4) deoxyribonuclease IV or E.C. 3.1.22.Y where Y = 1 , 2, 4 or 5, e.g., Deoxyribonuclease II, Aspergillus deoxyribonuclease K(1 ), Crossover junction endo-deoxyribonuclease, Deoxyribonuclease X.

Preferably, the polypeptide having DNase activity is obtained from a microorganism and the DNase is a microbial enzyme. The DNase is preferably of fungal or bacterial origin.

The DNase may be obtainable from Bacillus, e.g, Bacillus licheniformis, Bacillus subtilis, Bacillus sp-62451, Bacillus horikoshii, Bacillus sp-62451, Bacillus sp-16840, Bacillus sp-62668, Bacillus sp-13395, Bacillus horneckiae, Bacillus sp-11238, Bacillus cibi, Bacillus idriensis, Bacillus sp-62520, Bacillus sp-16840, Bacillus sp-62668, Bacillus algicola, Bacillus vietnamensis, Bacillus hwajinpoensis, Bacillus indicus, Bacillus marisflavi, Bacillus luciferensis, and Bacillus sp. SA2-6.

The DNase may also be obtained from any of the following: Pyrenochaetopsis sp., Vibrissea flavovirens, Setosphaeria rostrate, Endophragmiella valdina, Corynespora cassiicola, Paraphoma sp. XZ1965, Monilinia fructicola, Curvularia lunata, Penicillium reticulisporum, Penicillium quercetorum, Setophaeosphaeria sp., Alternaria, Alternaria sp. XZ2545, Trichoderma reesei, Chaetomium thermophilum, Scytalidium thermophilum, Metapochonia suchlasporia, Daldinia fissa, Acremonium sp. XZ2007, Acremonium sp. XZ2414, Acremonium dichromosporum, Sarocladium sp. XZ2014, Metarhizium sp. HNA15-2, Isaria tenuipes Scytalidium circinatum, Metarhizium lepidiotae, Thermobispora bispora, Sporormia fimetaria, Pycnidiophora cf. dispera, Enviromental sample D, Enviromental sample O, Clavicipitaceae sp-70249, Westerdykella sp. AS85-2, Humicolopsis cephalosporioides, Neosartorya massa, Roussoella intermedia, Pleosporales, Phaeosphaeria or Didymosphaeria futilis.

DNases comprising the DUF1524 domain

In addition to the DNases mentioned above, examples of polypeptides having DNase activity also include polypeptides comprising the PFAM domain DUF1524 (http://pfam.xfam.org/),“The Pfam protein families database: towards a more sustainable fu ture", R.D. Finn, et.al. Nucleic Acids Research (2016) Database Issue 44, D279- D285". The DUF1524 domain contains a conserved HXXP sequence motif commonly found in nucleases (M.A. Machnicka, et. al. Phylogenomics and sequence-structure-function relationships in the GmrSD family of Type IV restriction enzymes, BMC Bioinformatics, 2015, 16, 336).

DUF means domain of unknown function, and the polypeptide families comprising, e.g., DUF1524 have been collected together in the Pfam database (the Pfam ID for DUF1524 is PF07510). The Pfam data base provides sequence alignments and hidden Markov models that define the collected protein domains. A protein domain is a conserved part of a given protein sequence and (tertiary) structure that can evolve, function, and exist independently of the rest of the protein chain. Each domain forms a compact three-dimensional structure and often can be independently stable and folded. Many proteins consist of several structural domains. One domain may appear in a variety of different proteins.

A particular DUF may be identified using the prefix DUF followed by a number, e.g., 1524. The DUF1524 is a family of proteins all comprising the HXXP motif, where H is the amino acid histidine, P is the amino acid proline and X is any amino acid. In one aspect of the invention the polypeptides of the present invention having DNase activity comprise the DUF1524 domain.

The DNases to be used in a composition of the invention preferably belong to the NUC1 group of DNases, which is a sub-group of the DUF1524 domain. The NUC1 group of DNases comprises polypeptides which, in addition to having DNase activity, may comprise one or more of the motifs [T/D/S][G/N]PQL (SEQ ID NO: 1 ), [F/L/Y/I]A[N/R]D[L/I/P/V] (SEQ ID NO: 2), or C[D/N]T[A R] (SEQ ID NO: 3). In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, wherein the polypeptide comprises one or more of the motifs [T/D/S][G/N]PQL (SEQ ID NO: 1 ), [F/L/Y/I]A[N/R]D[L/I/PA/] (SEQ ID NO: 2) or C[D/N]T[A/R] (SEQ ID NO: 3).

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity and at least one oral care component, wherein the polypeptide comprises one or more of the motifs [T/D/S][G/N]PQL (SEQ ID NO: 1 ), [F/L/Y/I]A[N/R]D[L/I/P/V] (SEQ ID NO: 2) or C[D/N]T[A/R] (SEQ ID NO: 3), and wherein the polypeptide is selected from the group consisting of:

a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 7;

b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 8;

c) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 9;

d) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 10;

e) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 1 1 ;

f) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 12;

g) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 13;

h) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 14;

i) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 15;

j) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 16;

k) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 17; L) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 18;

m) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 19;

n) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 20;

o) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 21 ;

p) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 22;

q) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 23;

r) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 24;

s) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 25;

t) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 26;

u) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 27;

v) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 28;

w) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 29; x) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 30;

y) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 31 ;

z) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 32;

aa) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 33;

ab) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 34; and

ac) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 45.

The DNases preferably comprise a NUC1_A domain [D/Q][I V]DH (SEQ ID NO: 4). In addition to comprising any of the domain motifs [T/D/S][G/N]PQL, [F/L/Y/I]A[N/R]D[L/I/P V], and C[D/N]T[A R], the polypeptides having DNase activity may comprise the NUC1_A domain and may share the common motif [D/Q][I/V]DH (SEQ ID NO: 4). A preferred embodiment of the invention relates to oral care compositions comprising a polypeptide having DNase activity, wherein the polypeptide comprises one or more motifs selected from the group of [T/D/S][G/N]PQL, [F/L/Y/I]A[N/R]D[L/I/P V], C[D/N]T[A R], and [D/Q][I V]DH, and wherein the polypeptide has DNase activity.

The DNases to be added to a composition of the invention preferably belong to the group of DNases comprised in the GYS-clade, which are a group of DNases on the same branch of a phylogenetic tree having both structural and functional similarities. These NUC1 and/or NUC1_A DNases comprise the conservative motifs [D/M/L][S/T]GYSR[D/N] (SEQ ID NO: 5) or ASXNRSKG (SEQ ID NO: 6) and share similar structural and functional properties. The DNases of the GYS-clade are preferably obtained from the Bacillus genus.

A preferred embodiment of the invention relates to an oral care composition comprising a polypeptide of the GYS clade having DNase activity, optionally wherein the polypeptide comprises one or both of the motifs [D/M/L][S/T]GYSR[D/N] (SEQ ID NO: 5) and ASXNRSKG (SEQ ID NO: 6), and wherein the polypeptide is selected from the group consisting of: a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 7;

k) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 17;

L) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 18; m) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 19;

w) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 29;

x) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 30; y) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 31 ;

z) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 32; and

aa) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 45.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide having DNase activity obtainable from Bacillus, e.g., obtainable from Bacillus sp- 62451 , and having a sequence identity to the polypeptide shown in SEQ ID NO: 7 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 7.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide having DNase activity obtainable from Bacillus, e.g., obtainable from Bacillus horikoshii, and having a sequence identity to the polypeptide shown in SEQ ID NO: 8 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 8.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide having DNase activity obtainable from Bacillus, e.g., obtainable from Bacillus sp- 62520, and having a sequence identity to the polypeptide shown in SEQ ID NO: 9 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 9.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp-62520, and having a sequence identity to the polypeptide shown in SEQ ID NO: 10 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 10.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus horikoshii, and having a sequence identity to the polypeptide shown in SEQ ID NO: 11 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 11.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus horikoshii, and having a sequence identity to the polypeptide shown in SEQ ID NO: 12 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 12.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp-16840, and having a sequence identity to the polypeptide shown in SEQ ID NO: 13 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 13.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp-16840, and having a sequence identity to the polypeptide shown in SEQ ID NO: 14 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 14.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp-62668, and having a sequence identity to the polypeptide shown in SEQ ID NO: 15 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 15.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp-13395, and having a sequence identity to the polypeptide shown in SEQ ID NO: 16 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 16.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus horneckiae, and having a sequence identity to the polypeptide shown in SEQ ID NO: 17 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 17.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp-1 1238, and having a sequence identity to the polypeptide shown in SEQ ID NO: 18 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 18.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus cibi, and having a sequence identity to the polypeptide shown in SEQ ID NO: 19 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 19.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp-18318, and having a sequence identity to the polypeptide shown in SEQ ID NO: 20 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 20.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus idriensis, and having a sequence identity to the polypeptide shown in SEQ ID NO: 21 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 21.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus algicola, and having a sequence identity to the polypeptide shown in SEQ ID NO: 22 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 22.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Enviromental sample J and having a sequence identity to the polypeptide shown in SEQ ID NO: 23 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 23.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus vietnamensis, and having a sequence identity to the polypeptide shown in SEQ ID NO: 24 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 24.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus hwajinpoensis, and having a sequence identity to the polypeptide shown in SEQ ID NO: 25 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 25.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Paenibacillus mucilaginosus and having a sequence identity to the polypeptide shown in SEQ ID NO: 26 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or

100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 26.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus indicus, and having a sequence identity to the polypeptide shown in SEQ ID NO: 27 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 27.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus marisflavi, and having a sequence identity to the polypeptide shown in SEQ ID NO: 28 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 28.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus luciferensis, and having a sequence identity to the polypeptide shown in SEQ ID NO: 29 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 29.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus marisflavi, and having a sequence identity to the polypeptide shown in SEQ ID NO: 30 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 30.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus sp. SA2-6, and having a sequence identity to the polypeptide shown in SEQ ID NO: 31 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 31.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus indicus, and having a sequence identity to the polypeptide shown in SEQ ID NO: 32 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 32.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Cladosponum, e.g., obtainable from Cladosporium cladosporioides , and having a sequence identity to the polypeptide shown in SEQ ID NO: 33 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least

89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least

96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 33.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Deconica, e.g., obtainable from Deconica coprophila, and having a sequence identity to the polypeptide shown in SEQ ID NO: 34 of at least 60%, e.g., at least

65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least

84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least

91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least

98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 34.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus cibi, and having a sequence identity to the polypeptide shown in SEQ ID NO: 45 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 45.

Other DNases comprising the DUF1524 domain, e.g., NUC1 nucleases and GYS-clade nucleases, are also contemplated to be within the scope of the present invention and are readily identified by the skilled person, e.g., according to WO 2017/059802 and WO 2017/060475.

The polypeptides having DNase activity may also be selected from the DNases belonging to the endonuclease/exonuclease/phosphatase family (Pfam ID PF03372). The DNases to be used in a composition of the invention preferably belong to the NUC2 group of DNases, which is a sub-group of the endonuclease/exonuclease/phosphatase family. The NUC2 group of DNases comprises polypeptides which, in addition to having DNase activity, may comprise the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V] (SEQ ID NO: 35). Polypeptides having DNase activity and comprising this motif have shown to effectively prevent, remove or reduce biofilm. The NUC2 group of DNases has been further described in WO 2017/059801.

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity and at least one oral care component, wherein the polypeptide comprises the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V] (SEQ ID NO: 35), and wherein the polypeptide is selected from the group consisting of:

a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 36;

b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 37;

c) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 38;

d) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 39;

e) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 40;

f) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 46; g) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 47;

h) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 48; and

i) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 49.

In a particularly preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity and at least one oral care component, wherein the polypeptide comprises the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V] (SEQ ID NO: 35), and wherein the polypeptide is selected from the group consisting of:

f) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 46;

g) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 47;

h) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 48; and i) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 49.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Pseudoplectania, e.g., obtainable from Pseudoplectania vogesiaca, and having a sequence identity to the polypeptide shown in SEQ ID NO: 36 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 36.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Aspergillus, e.g., obtainable from Aspergillus niger, and having a sequence identity to the polypeptide shown in SEQ ID NO: 37 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 37.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Phialophora, e.g., obtainable from Phialophora geniculata, and having a sequence identity to the polypeptide shown in SEQ ID NO: 38 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 38.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Paradendryphiella, e.g., obtainable from Paradendryphiella salina, and having a sequence identity to the polypeptide shown in SEQ ID NO: 39 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 39.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Simplicillium, e.g., obtainable from Simplicillium obclavatum, and having a sequence identity to the polypeptide shown in SEQ ID NO: 40 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 40.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Acrophialophora, e.g., obtainable from Acrophialophora fusispora, and having a sequence identity to the polypeptide shown in SEQ ID NO: 46 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 46.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Aspergillus, e.g., obtainable from Aspergillus aculeatus, and having a sequence identity to the polypeptide shown in SEQ ID NO: 47 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 47.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Achaetomium, e.g., obtainable from Achaetomium luteum, and having a sequence identity to the polypeptide shown in SEQ ID NO: 48 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 48. In some embodiments, the present invention relates to oral care compositions comprising a polypeptide obtainable from Warcupiella, e.g., obtainable from Warcupiella spinulosa, and having a sequence identity to the polypeptide shown in SEQ ID NO: 49 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 49.

Other DNases belonging to the endonuclease/exonuclease/phosphatase family, e.g., NUC2 nucleases, are also contemplated to be within the scope of the present invention and are readily identified by the skilled person, e.g., according to WO 2017/059801 and WO 2017/060493.

Other DNases

Yet other DNases are also contemplated to be suitable for oral care compositions of the present invention. In a preferred embodiment, the present invention relates to oral care compositions comprising a polypeptide having DNase activity and at least one oral care component, wherein the polypeptide is selected from the group consisting of:

a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least

80%, at least 85%, at least 90%, at least 91 %, at least 952%, at least 93%, at least

94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide shown in SEQ ID NO: 41 ,

b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least

94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide shown in SEQ ID NO: 42,

c) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least

94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide shown in SEQ ID NO: 43, and

d) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least

94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the polypeptide shown in SEQ ID NO: 44.

In some embodiments, the present invention relates compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus licheniformis, and having a sequence identity to the polypeptide shown in SEQ ID NO: 41 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 41.

In some embodiments, the present invention relates compositions comprising a polypeptide obtainable from Bacillus, e.g., obtainable from Bacillus subtilis, and having a sequence identity to the polypeptide shown in SEQ ID NO: 42 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 42.

In some embodiments, the present invention relates compositions comprising a polypeptide obtainable from Aspergillus, e.g., obtainable from Aspergillus oryzae, and having a sequence identity to the polypeptide shown in SEQ ID NO: 43 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 43.

In some embodiments, the present invention relates compositions comprising a polypeptide obtainable from Trichoderma, e.g., obtainable from Trichoderma harzianum, and having a sequence identity to the polypeptide shown in SEQ ID NO: 44 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have DNase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 44.

Polypeptides having mutanase activity

Mutanases are -1 ,3-glucanases (also known as -1 ,3-glucanohydrolases) that degrade the -1 ,3-glycosidic linkages in mutan, which is a polysaccharide found in oral biofilm. Mutanases are particularly useful against biofilm generated by the cariogenic microorganism Streptococcus mutans.

Suitable mutanases have been derived from Trichoderma (Hasegawa et al., (1969), Journal of Biological Chemistry 244, p. 5460-5470; Guggenheim and Haller, (1972), Journal of Dental Research 51 , p. 394-402) and from strains of Streptomyces (Takehara et al., (1981 ), Journal of Bacteriology 145, p. 729-735), Cladosporium resinae (Hare et al. (1978), Carbohydrate Research 66, p. 245-264), Pseudomonas sp. (US patent no. 4,438,093), Flavobacterium sp. (JP 77038113), Bacillus circulans (JP 63301788) and Aspergillus sp.. A mutanase gene from Trichoderma harzianum has been cloned and sequenced (Japanese Patent No. 4-58889/A).

A mutanase suitable for the use in an oral care composition of the invention may be produced by filamentous fungi from the group including Trichoderma, in particular from a strain of Trichoderma harzianum, such as Trichoderma harzianum CBS 243.71 , or Penicillium, in particular a strain of Penicillium funiculosum, such as Penicillium funiculosum NRRL 1768, or a strain of Penicillium lilacinum, such as Penicillium lilacinum NRRL 896, or a strain of Penicillium purpurogenum, such as the strain of Penicillium purpurogenum CBS 238.95, or a strain of the genus Pseudomonas, or a strain of Flavobacterium sp., or a strain of Bacillus circulans or a strain of Aspergillus sp., or a strain of Streptomyces sp..

The mutanase may also be derived from Penicillium purpurogenum.

US patent 4,353,981 (Guggenheim et al.) discloses the use of the Trichoderma harzianum CBS 243.71 mutanase, the Penicillium funiculosum NRRL 1768 mutanase and the Penicillium lilacinum NRRL 896 mutanase for the removal of dental plaque.

In some embodiments, the present invention relates to oral care compositions comprising a polypeptide having mutanase activity obtainable from Trichoderma, e.g., obtainable from Trichoderma harzianum, and having a sequence identity to the polypeptide shown in SEQ ID NO: 50 of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, and which have mutanase activity. In one aspect, the polypeptides differ by up to 10 amino acids, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide shown in SEQ ID NO: 50.

In a preferred embodiment, the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50, and which have mutanase activity. In a preferred embodiment, the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50. In a preferred embodiment, the polypeptide having DNase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 39, and which have DNase activity; and the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50, and which have mutanase activity. In a preferred embodiment, the polypeptide having DNase activity comprises, consists essentially of, or consists of SEQ ID NO: 39, and the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

In a preferred embodiment, the polypeptide having DNase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 40, and which have DNase activity; and the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50, and which have mutanase activity. In a preferred embodiment, the polypeptide having DNase activity comprises, consists essentially of, or consists of SEQ ID NO: 40, and the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

In a preferred embodiment, the polypeptide having DNase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 45, and which have DNase activity; and the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50, and which have mutanase activity. In a preferred embodiment, the polypeptide having DNase activity comprises, consists essentially of, or consists of SEQ ID NO: 45, and the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

In an aspect, the oral care compositions of the invention comprise a polypeptide having DNase activity is selected from the group consisting of:

a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 39, which have DNA activity;

a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 40, which have DNase activity; and

a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 45, which have DNase activity;

and the polypeptide having mutanase activity is a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50, which have mutanase activity.

a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 39, which have DNA activity; and

a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 40, which have DNase activity;

Other enzymes with beneficial effects in oral care

The compositions of the invention may also comprise additional enzymes that have beneficial effect for oral care.

Thus, in a preferred embodiment, the oral care compositions of the inventions comprise a polypeptide having DNase activity, at least one oral care component, and at least one enzyme; preferably the at least one enzyme is selected from the group consisting of dispersin, protease, lipase, carbohydrase, dextranase, oxidoreductase, laccase, peroxidase, oxidase, and lysozyme. In general, the properties of the selected enzyme(s) should be compatible with the selected oral care composition, ( i.e ., pH-optimum, compatibility with other enzymatic and non-enzymatic ingredients, etc.), and the enzyme(s) should be present in effective amounts.

Examples of dispersins, proteases, lipases, carbohydrases, dextranases, oxidoreductases, laccases, peroxidases, oxidases, and lysozyme suitable for use in the compositions of the invention include those that are described below as well as others available in the art, which may be readily identified by the skilled artisan.

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and a dispersin.

Dispersins are polypeptides having hexosaminidase activity, preferably poly-N- acetylglucosamine (PNAG) degrading activity. One example is dispersin B (DspB), which is a beta-N-acetylglucosamininidases belonging to the Glycoside Hydrolase 20 family. Dispersins are produced by the periodontal pathogen Aggregatibacter actinomycetemcomitans, a Gram negative oral bacterium. Dispersin B is a beta-hexosaminidase that specifically hydrolyzes beta- 1 ,6-glycosidic linkages of acetylglucosamine polymers found in biofilm. Suitable dispersin B and variants thereof are described in WO 2014/0611 17 and WO 2017/186936.

Other suitable dispersins include dispersin 2 and variants thereof (WO 2017/186936), dispersin 5 and variants thereof, and dispersin 8 and variants thereof.

In an embodiment, the compositions of the invention comprise a dispersin selected from dispersin B, dispersin 2, dispersin 5, and dispersin 8.

In an embodiment, the compositions of the invention comprise a polypeptide having hexosaminidase activity or a polypeptide comprising a catalytic domain belonging to the Glycoside Hydrolase family 20 (GH20, www.cazy.org).

Proteases

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and a protease.

Within oral care, proteases break down salivary proteins, which are adsorbed onto the tooth surface and form the pellicle that acts as attachment point for oral biofilm. Proteases may also degrade proteins that form part of the structural components of bacterial cell walls and membranes.

Proteases suitable for compositions of the invention are enzymes classified under the Enzyme Classification number (E.C.) 3.4 in accordance with the Recommendations (1992) of the International Union of Biochemistry and Molecular Biology (IUBMB)). Examples include proteases selected from those classified under the Enzyme Classification (E.C.) numbers: 3.4.11 (i.e. so-called aminopeptidases), including 3.4.11 .5 (Prolyl aminopeptidase), 3.4.1 1 .9 (X-pro aminopeptidase), 3.4.11.10 (Bacterial leucyl aminopeptidase), 3.4.1 1.12 (Thermophilic aminopeptidase), 3.4.1 1.15 (Lysyl aminopeptidase), 3.4.11.17 (Tryptophanyl aminopeptidase), 3.4.1 1.18 (Methionyl aminopeptidase);

3.4.21 (i.e. so-called serine endopeptidases), including 3.4.21.1 (Chymotrypsin), 3.4.21 .4 (Trypsin), 3.4.21.25 (Cucumisin), 3.4.21 .32 (Brachyurin), 3.4.21.48 (Cerevisin) and 3.4.21.62 (Subtilisin);

3.4.22 (i.e. so-called cysteine endopeptidases), including 3.4.22.2 (Papain), 3.4.22.3 (Ficain), 3.4.22.6 (Chymopapain), 3.4.22.7 (Asclepain), 3.4.22.14 (Actinidain), 3.4.22.30 (Caricain) and 3.4.22.31 (Ananain);

3.4.23 (i.e. so-called aspartic endopeptidases), including 3.4.23.1 (Pepsin A), 3.4.23.18 (Aspergillopepsin I), 3.4.23.20 (Penicillopepsin) and 3.4.23.25 (Saccharopepsin); and

3.4.24 (i.e. so-called metalloendopeptidases), including 3.4.24.28 (Bacillolysin).

Examples of relevant subtilisins comprise subtilisin BPN', subtilisin amylosacchariticus, subtilisin 168, subtilisin mesentericopeptidase, subtilisin Carlsberg, subtilisin DY, subtilisin 309, subtilisin 147, thermitase, aqualysin, Bacillus PB92 protease, proteinase K, Protease TW7, and Protease TW3.

Specific examples of such readily available commercial proteases include those sold under the tradenames Esperase™, Alcalase™, Neutrase™, Dyrazym™, Savinase™, Pyrase™, Pancreatic Trypsin NOVO™ (PTN), Bio-FeedC Pro™, Clear-Lens Pro™, Maxtase™, Maxacal™, Maxapem™, Opticlean™, and Purafect™.

Proteases included in compositions of the invention are further contemplated to also include variants of the above-mentioned proteases. Examples of such protease variants are disclosed in EP 130 756; EP 214 435; WO 87/04461 ; WO 87/05050; EP 251 446; EP 260 105; Thomas et al., (1985), Nature 318, pp. 375-376; Thomas et al., (1987), J. Mol. Biol. 193, pp. 803-81 ; Russel et al., (1987), Nature 328, p. 496-500; WO 88/08028; WO 88/08033; WO 89/06279; WO 91/00345; EP 525 610); and WO 94/02618.

The activity of proteases can be determined as described in "Methods of Enzymatic Analysis", third edition, vol. 5, 1984, Verlag Chemie, Weinheim.

Lipases

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, an oral care component, and a lipase.

Within oral care, lipases target oral bacteria by degrading the lipids that form part of the structural components of bacterial cell walls and membranes.

Lipases suitable for compositions of the invention include enzymes classified under the Enzyme Classification number (E.C.) 3.1.1 (Carboxylic Ester Hydrolases) in accordance with the Recommendations (1992) of the International Union of Biochemistry and Molecular Biology (IUBMB)). Examples include lipases selected from those classified under the Enzyme Classification (E.C.) numbers 3.1.1.3 (Triacylglycerol lipases) and 3.1 .1 .4 (Phospholipase A2).

Examples of suitable lipases include lipases derived from the following microorganisms: Humicola, e.g., H. brevispora, H. lanuginosa, H. brevis var. thermoidea and H. insolens (US 4,810,414); Pseudomonas, e.g., Ps. fragi, Ps. stutzeri, Ps. cepacia and Ps. fluorescens (WO 89/04361 ), or Ps. plantarii or Ps. gladioli (US patent no. 4,950,417) or Ps. alcaligenes and Ps. pseudoalcaligenes (EP 218 272) or Ps. mendocina (WO 88/09367; US 5,389,536); Fusarium, e.g., F. oxysporum (EP 130,064) or F. solani pisi (WO 90/09446); Mucor (also called Rhizomucor), e.g., M. miehei (EP 238 023); Chromobacterium (especially C. viscosum)·, Aspergillus (especially A. niger) Candida, e.g., C. cylindracea (also called C. rugosa) or C. Antarctica (WO 88/02775) or C. antarctica lipase A or B (WO 94/01541 and WO 89/02916); Geotricum, e.g., G. candidum (Schimada et al. , (1989), J. Biochem., 106, 383-388); Penicillium, e.g., P. camembertii (Yamaguchi et al., (1991 ), Gene 103, 61 -67); Rhizopus, e.g., R. delemar (Hass et al., (1991 ), Gene 109, 107-1 13) or R. niveus (Kugimiya et al., (1992) Biosci. Biotech. Biochem. 56, 716-719) or R. oryzae Bacillus, e.g., B. subtilis (Dartois et al., (1993) Biochemica et Biophysica Acta 1131 , 253-260) or B. stearothermophilus (JP 64/7744992) or B. pumilus (WO 91/16422).

Specific examples of readily available commercial lipases include those sold under the tradenames Lipolase™, Lipolase C Ultra™, Lipozyme™, Palatase™, Novozym 435™, and Lecitase™ (Novozymes).

Examples of other lipases are Lumafast™, Ps. mendocian lipase from Genencor Int. Inc.; Lipomax™, Ps. pseudoalcaligenes lipase from Gist Brocades/Genencor Int. Inc.; Fusarium solani lipase (cutinase) from Unilever; Bacillus sp. lipase from Solvay Enzymes. Other lipases are available from other companies.

It is to be understood that also lipase variants are contemplated as suitable lipases for the present invention. Examples of such are described in, e.g., WO 93/01285 and WO 95/2261.

The activity of the lipase can be determined as described in "Methods of Enzymatic Analysis", Third Edition, 1984, Verlag Chemie, Weinhein, vol. 4.

Carbohydrases

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and a carbohydrase; preferably the carbohydrase is an alpha-amylase.

Carbohydrases may be defined as all enzymes capable of breaking down carbohydrate chains (e.g., starches) of especially five and six membered ring structures (i.e. enzymes classified under the Enzyme Classification number (E.C.) 3.2 (glycosidases) in accordance with the Recommendations (1992) of the International Union of Biochemistry and Molecular Biology (IUBMB)). Such carbohydrate structures are an important structural component of oral biolfilm.

Examples include carbohydrases selected from those classified under the Enzyme Classification (E.C.) numbers:

alpha-amylase (3.2.1.1 ) beta-amylase (3.2.1.2), glucan 1 ,4-alpha-glucosidase (3.2.1.3), cellulase (3.2.1 .4), endo-1 ,3(4)-beta-glucanase (3.2.1.6), endo-1 ,4-beta-xylanase (3.2.1.8), dextranase (3.2.1.1 1 ), chitinase (3.2.1.14), polygalacturonase (3.2.1.15), beta-glucosidase (3.2.1.21 ), alpha-galactosidase (3.2.1.22), beta-galactosidase (3.2.1.23), amylo-1 ,6- glucosidase (3.2.1.33), xylan 1 ,4-beta-xylosidase (3.2.1.37), glucan endo-1 , 3-beta-D- glucosidase (3.2.1 .39), alpha-dextrin endo-1 ,6-glucosidase (3.2.1.41 ), sucrose alpha- glucosidase (3.2.1.48), glucan endo-1 , 3-alpha-glucosidase (3.2.1.59), glucan 1 ,4-beta- glucosidase (3.2.1.74), glucan endo-1 , 6-beta-glucosidase (3.2.1 .75), arabinan endo-1 ,5-alpha- arabinosidase (3.2.1 .99), lactase (3.2.1.108), chitonanase (3.2.1 .132) and xylose isomerase (5.3.1.5).

Examples of relevant carbohydrases include alpha-1 , 3-glucanases derived from Trichoderma harzianurrr, alpha-1 ,6-glucanases derived from a strain of Paecilomyces, beta- glucanases derived from Bacillus subtilis, beta-glucanases derived from Humicola insolens, beta-glucanases derived from Aspergillus niger, beta-glucanases derived from a strain of Trichoderma·, beta-glucanases derived from a strain of Oerskovia xanthineolytica, exo-1 ,4- alpha-D-glucosidases (glucoamylases) derived from Aspergillus niger, alpha-amylases derived from Bacillus subtilis, alpha-amylases derived from Bacillus amyloliquefaciens, alpha-amylases derived from Bacillus stearothermophilus, alpha-amylases derived from Aspergillus oryzae alpha-amylases derived from non-pathogenic microorganisms; alpha-galactosidases derived from Aspergillus niger, Pentosanases, xylanases, cellobiases, cellulases, and hemi-cellulases derived from Humicola insolens, cellulases derived from Trichoderma reeser, cellulases derived from non-pathogenic mold; pectinases, cellulases, arabinases, and hemi-celluloses derived from Aspergillus niger, dextranases derived from Penicillium lilacinum·, endo-glucanase derived from non-pathogenic mold; pullulanases derived from Bacillus acidopullyticus, beta-galactosidases derived from Kluyveromyces fragilis ; xylanases derived from Trichoderma reesei.

In an embodiment of the invention the starch-modifying enzyme is a CGTase (E.C. 2.4.1.19) or a transglucosidase (2.4.1.18).

When the starch-modifying enzyme is a CGTase, it may be derived from a strain of Bacillus autolyticus, a strain of Bacillus cereus, a strain of Bacillus circulars, a strain of Bacillus circulars var. alkalophilus, a strain of Bacillus coagulans, a strain of Bacillus firmus, a strain of Bacillus halophilus, a strain of Bacillus macerans, a strain of Bacillus megaterium, a strain of Bacillus ohbensis, a strain of Bacillus stearothermophilus, a strain of Bacillus subtilis, a strain of Klebsiella pneumoniae, a strain of Thermoanaerobacter sp., a strain of Thermoanaerobacter ethanolicus, a strain of Thermoanaerobacter finnii, a strain of Clostridium thermoamylolyticum , a strain of Clostridium thermosaccharolyticum, or a strain of Thermoanaerobacterium thermosuifurigenes.

When the starch-modifying enzyme is a transglucosidase, it may be derived from Aspergillus niger.

In another embodiment of the invention, the oral care composition comprises a starch hydrolysing enzyme. This will typically be an alpha-amylase, such as a bacterial alpha-amylase, such as BAN™ or Maltogenase™, or an alpha-amylase derived from Bacillus subtilis, an alpha- amylase derived from Bacillus amyloliquefaciens, an alpha-amylase derived from Bacillus stearothermophilus, an alpha-amylase derived from Aspergillus oryzae\ or an alpha-amylase derived from a non-pathogenic microorganism.

The alpha-amylase may also be a fungal alpha-amylase, such as Fungamyl™.

The starch-hydrolysing enzyme may in another embodiment of the invention be a debranching enzyme, in particular a pullulanase (E. C. 3.2.1.41 ), such as Promozyme™.

In a preferred embodiment the oral care composition comprises at least one starch modifying enzyme as defined above, in particular a CGTase and/or a dextranase.

In another preferred embodiment the oral care composition of the invention comprises at least one starch-hydrolysing enzyme as defined above, in particular a bacterial alpha-amylase and/or a dextranase.

The dextranase may be derived from a strain of Paecilomyces sp., in particular Paecilomyces Hlacinus.

Dextranase

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and a dextranase.

Dextranases degrade carbohydrate molecules that are important structural components of oral biofilm.

Dextranases are alpha-1 ,6-glucanases (also known as 1 ,6-alpha-D-glucan-6- glucanohydrolases) which degrade the alpha-1 ,6-glycosidic linkages in dextran. Several microorganisms are capable of producing dextranases, among them fungi of Penicillium, Paecilomyces, Aspergillus, Fusarium, Spicaria, Verticillium, Helminthosporium and the Chaetomium genera; bacteria of the genera Lactobacillus, Streptococcus, Cellvibrio, Cytophaga, Brevibacterium, Pseudomonas, Corynebacterium, Arthrobacter and Flavobacterium, and yeasts such as Lipomyces starkeyi.

Commercially available products include Dextranase™ 50 L from Novozymes produced by fermentation of strains of Penicillium lilacium. Dextranase 50 L is used in the sugar industry to break down dextran in raw sugar juice or syrup. Oxidoreductases

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and an oxidoreductase; preferably the oxidoreductase is a laccase or a related enzyme, an oxidase, or a peroxidase.

Oxidoreductases are enzymes catalysing oxidoreductions, and they have been found to bleach teeth. They are classified under the Enzyme Classification number (E.C.) 1 (Oxidoreductases) in accordance with the Recommendations (1992) of the International Union of Biochemistry and Molecular Biology (IUBMB).

According to the present invention, three types of oxidoreductases are especially contemplated: 1 ) laccases or related enzymes such as tyrosinase that act on molecular oxygen (O2) and yield water (H₂0) without any need for peroxide (e.g. H2O2); 2) oxidases that act on molecular oxygen (O2) and yield peroxide (H2O2); and 3) peroxidases that act on peroxide (e.g., H2O2) and yield water (H₂0).

Preferred oxidoreductases are of microbial origin, especially recombinant and/or substantially purified enzymes without any side activity. Microbial enzyme means in the context of the present invention enzymes derived from bacteria, filamentous fungi or yeasts.

In the case of an enzyme acting on oxygen (O2) as the acceptor, said oxygen may be molecular oxygen supplied by the air.

Enzyme systems which comprise a combination of the three types of enzymes are also contemplated as being suitable for compositions of the invention. The enzyme systems may, e.g., consist of a laccase or a related enzyme and an oxidase; a laccase or a related enzyme and a peroxidase; a laccase or a related enzyme and an oxidase and a peroxidase; or an oxidase and a peroxidase.

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and a laccase or a related enzyme.

Examples of suitable enzymes within the group of laccases and related enzymes are those capable of oxidizing volatile sulphur compounds (VSCs) and nitrogen compounds in question are mono-and diphenolicoxidases, such as catechol oxidase (1.10.3.1 ), laccase (E.C. 1.10.3.2), tyrosinase (E.C. 1.14.18.1 ), and bilirubin oxidase (E.C. 1.3.3.5).

Laccase oxidizes o-diphenol as well as p-diphenol forming their corresponding quinones. Tyrosinase and catechol oxidase catalyses the hydroxylation of monophenols in o-diphenols and the oxidation of o-diphenols in o-quinones.

Laccase is usually applied together with a suitable donor, preferably chlorogenic acid. Laccases suitable for compositions of the invention may be derived from a strain of Polyporus sp., in particular a strain of Polyporus pinsitus (also called Trametes villosa) or Polyporus versicolor, or a strain of Myceliophthora sp., e.g., M. thermophila or a strain of Rhizoctonia sp., in particular a strain of Rhizoctonia praticola or Rhizoctonia solani, or a strain of Scytalidium sp., in particular S. thermophilium, or a strain of Pyricularia sp., in particular Pyricularia oryzae, or a strain of Coprinus sp., such as a C. cinereus. The laccase may also be derived from a fungus such as Collybia, Fornes, Lentinus, Pleurotus, Aspergillus, Neurospora, Podospora, Phlebia, e.g., P. radiata (WO 92/01046), Coriolus sp., e.g., C. hirsitus (JP 2-238885), and Botrytis.

In a preferred embodiment of the invention the laccase is derived from a strain of Myceliophthora sp., especially the Myceliophthora thermophila laccase described in WO 95/33836.

Bilirubin oxidase may be derived from a strain of Myrothecium sp., such as a strain of M. verrucaria.

Peroxidases

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and a peroxidase.

Peroxidases must be used in combination with either H2O2 or an oxidase to obtain the desired result, i.e., removal or at least reduction of malodour.

Suitable peroxidases can be found within the group of enzymes acting on peroxide as acceptor, e.g., E.C. 1.1 1.1 , especially peroxidase (E.C. 1.1 1.1.7).

Specific examples of suitable enzymes acting on peroxide as acceptor include peroxidases derived from a strain of the fungus species Coprinus, in particular a strain of Coprinus cinereus or Coprinus macrorhizus, or derived from a strain of the bacteria genus Bacillus, in particular a strain of Bacillus pumilus.

Haloperoxidases are also suitable according to the invention. Haloperoxidases form a class of enzymes which are able to oxidise halides, i.e., chloride (CI-), bromide (Br-), and iodide (I-), in the presence of hydrogen peroxide to the corresponding hypohalous acids. A suitable haloperoxidase is derivable from Curvularia sp., in particular C. verruculosa.

Oxidases

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and an oxidase.

Oxidases yielding peroxide (H2O2) must be used in combination with a peroxidase to be able to remove or at least reduce malodour. Suitable oxidases include glucose oxidase (E.C.

1 .1 .3.4), hexose oxidase (E.C. 1.1 .3.5), L-amino-acid oxidase (E.C. 1.4.3.2), xylitol oxidase, galactose oxidase (E.C. 1.1.3.9), pyranose oxidase (E.C. 1.1.3.10), alcohol oxidase (E.C. 1 .1 .3.13).

If a L-amino acid oxidase is used, it may be derived from a Trichoderma sp. such as Trichoderma harzianum, such as the L-amino acid oxidase described in WO 1994/25574, or Trichoderma viride.

A suitable glucose oxidase may originate from Aspergillus sp., such as a strain of Aspergillus niger, or from a strain of Cladosporium sp. in particular Cladosporium oxysporum.

Hexose oxidases from the red sea-weed Chondrus crispus (commonly known as Irish moss) (Sullivan and Ikawa, (1973), Biochim. Biophys. Acta 309, p. 1 1-22; Ikawa, (1982), Meth. in Enzymol. 89, Carbohydrate Metabolism Part D, 145-149) oxidises a broad spectrum of carbohydrates, such as D-glucose, D-galactose, maltose, cellobiose, lactose, D-glucose 6- phasphate, D-mannose, 2-deoxy-D-glucole, 2-deoxy-D-galactose, D-fucase, D-glucuronic acid, and D-xylose.

The red sea-weed Iridophycus flaccidum produces easily extractable hexose oxidases, which oxidise several different mono- and disaccharides (Bean and Hassid, (1956), J. Biol. Chem 218, p. 425; Rand et al. (1972), J. Food Science 37, p. 698-710).

Another suitable group of enzymes is xylitol oxidase (disclosed in JP 80892242) which oxidises xylitol, D-sorbitol, D-galactitol, D-mannitol and D-arabinitol in the presence of oxygen. A xylitol oxidase can be obtained from strains of Streptomyces sp. (e.g., Streptomyces IKD472, FERM P-14339). Said enzyme has a pH optimum at 7.5 and is stable at pH 5.5 to 10.5 and at temperatures up to 65°C.

Lysozymes

In a preferred embodiment, the oral care compositions of the invention comprise a polypeptide having DNase activity, a polypeptide having mutanase activity, at least one oral care component, and a lysozyme.

Lysozymes suitable for compositions of the present invention include those classified under EC 3.2.1 .17. Lysozyme is also known as muramidase and occurs naturally in many organisms such as viruses, plants, insects, birds, reptiles, and mammals. In mammals, lysozyme has been isolated from nasal secretions, saliva, tears, intestinal content, urine and milk. The enzyme cleaves the glycosidic bond between carbon number 1 of /V-acetylmuramic acid and carbon number 4 of /V-acetyl-D-glucosamine. In vivo, these two carbohydrates are polymerized to form the cell wall polysaccharide of many microorganisms. Due to its ability to degrade bacterial peptidoglycans, lysozyme functions as an antibacterial agent.

Lysozyme has been classified into five different glycoside hydrolase (GH) families (CAZy, www.cazy.org): hen egg-white lysozyme (GH22), goose egg-white lysozyme (GH23), bacteriophage T4 lysozyme (GH24), Sphingomonas flagellar protein (GH73) and Chalaropsis lysozymes (GH25). Lysozymes from the families GH23 and GH24 are primarily known from bacteriophages and have recently been identified in fungi. The lysozyme family GH25 has been found to be structurally unrelated to the other lysozyme families. Lysozyme extracted from hen egg white is the primary product available on the commercial market.

For the compositions of the present invention, preferred lysozymes may be selected from GH22 lysozymes, GH23 lysozymes, GH24 lysozymes, GH73 lysozymes, and GH25 lysozymes. Preferably, the lysozyme is a GH25 lysozyme. Examples of GH25 lysozymes can be found in, e.g., WO 2013/076253, WO 2005/080559, PCT/CN2017/1 17753, and PCT/CN2017/1 17765.

Oral care compositions

The oral care compositions of the invention comprise a polypeptide having DNase activity and at least one oral care component. The oral care components are ingredients that may be varied according to the type of oral care composition as well as the desired characteristics and/or activities of the oral care compositions. For the purpose of the present invention, the terms “component” and“ingredient” are used interchangeably in relation to oral care compositions.

An oral care composition of the invention may be an internal oral care composition such as toothpaste, dental cream, mouthwash, mouth rinse, lozenges, pastilles, chewing gum, confectionary, candy, and the like, which is designed to remove biofilm inside the oral cavity, e.g., biofilm residing on teeth, on soft tissues of the oral cavity, and on dentures residing in the oral cavity.

An oral care composition of the invention may also be an external oral care composition such as denture cleaning solution, denture cleaning tablet, denture cleaning powder, and the like, which is designed to remove biofilm from dentures that have been removed from the oral cavity for cleaning.

In a preferred embodiment, the oral care composition is an internal oral care composition, and the at least one oral care component is selected from the group consisting of abrasives, humectants, solvents, thickening agents, binding agents, buffering agents, foaming agents, foaming modulators, sweetening agents, softening agents, plasticizing agents, flavoring agents, coloring agents, therapeutic agents, anti-microbial agents, tartar-controlling agents, fluoride sources, preservatives, detergents, surfactants, coloring agents, buffering agents, softeners, plastizisers, whitening agents, bleaching agents, gum-base ingredients, and bulking agents.

Although the oral care components mentioned herein are categorized by a general header according to a functionality, this is not to be construed as a limitation, as a component may comprise additional functionalities as will be appreciated by the skilled person.

Toothpaste, dental cream, mouthwash, and mouth rinse

Internal oral care compositions of the invention in the form of toothpaste, dental cream, mouthwash, and mouth rinse may include substances and/or ingredients selected from the following categories:

Toothpaste

Toothpastes and dental creams/gels typically include abrasives, solvents, humectants, detergents/surfactants, thickening and binding agents, buffering agents, flavoring agents, sweetening agents, fluoride sources, therapeutic agents, enzymes, coloring agents, and preservatives.

In a preferred embodiment, the present invention relates to oral care compositions in the form of a toothpaste or dental cream comprising a polypeptide having DNase activity and at least one oral care component, wherein the at least one oral care component is selected from the following components:

An oral care composition of the invention may be a toothpaste comprising the following ingredients (in weight % of the final toothpaste composition):

Abrasive: 10 to 70%

Humectant: 0 to 80%

Thickening agent: 0.1 to 20%

Binding agent: 0.01 to 10%

Sweetening agent: 0.1 to 5%

Foaming agent: 0 to 15%

Polypeptide(s) having DNase activity: 0.01 to 10%

Enzyme(s) with action(s) on oral cavity: 0.01 to 20%

Abrasive: 10 to 70%

Humectant: 0 to 80%

Thickening agent: 0.1 to 20%

Binding agent: 0.01 to 10%

Sweetening agent: 0.1 to 5%

Foaming agent: 0 to 15%

Polypeptide(s) having DNase activity: 0.01 to 10%

Enzyme(s) with action(s) on oral cavity: 0.01 to 20%

Mouthwash

Mouthwashes and mouth rinses of the invention, including plaque removing liquids, typically comprise a polypeptide having DNase activity, a carrier liquid, detergents/surfactants, buffering agents, flavoring agents, humectants, sweetening agents, therapeutic agents, fluoride sources, coloring agents, preservatives, and enzymes.

In a preferred embodiment, the present invention relates to oral care compositions in the form of a mouthwash or mouth rinse comprising a polypeptide having DNase activity and at least one oral care component, wherein the at least one oral care component is selected from the following components:

An oral care composition of the invention may be a mouthwash comprising the following ingredients (in weight % of the final mouthwash composition):

Water: 0 to 70%

Ethanol: 0 to 20%

Humectant: 0 to 20%

Surfactant: 0 to 2%

Polypeptide(s) having DNase acitivty: 0.01 to 10%

Enzyme(s) with action(s) on oral cavity: 0.01 to 20%

Other ingredients: 0 to 2% ( e.g ., flavors, sweeteners, fluoride sources).

The mouthwash composition may be buffered with an appropriate buffer, e.g., sodium citrate or phosphate in the pH-range 6-7.5.

Relevant oral care components suitable for toothpastes, dental creams, mouthwashes, and mouth rinses is further detailed below. The skilled person is capable of varying the oral care components according to the type of oral care composition as well as the desired characteristics and/or activities of the specific oral care composition. An oral care composition may not necessarily comprise all of the mentioned ingredients.

Abrasives

Abrasive polishing material might be incorporated into the oral care composition of the invention. According to the invention said abrasive polishing material includes alumina and hydrates thereof, such as alpha alumina trihydrate, magnesium trisilicate, magnesium carbonate, kaolin, aluminosilicates, such as calcined aluminum silicate and aluminum silicate, calcium carbonate, zirconium silicate, bentonite, silicium dioxide, sodium bicarbonate, and also powdered plastics, such as polyvinyl chloride, polyamides, polymethyl methacrylate, polystyrene, phenol-formaldehyde resins, melamine-formaldehyde resins, urea-formaldehyde resins, epoxy resins, powdered polyethylene, silica xerogels, hydrogels and aerogels, and the like.

Also suitable as abrasive agents are calcium pyrophosphate, water-insoluble alkali metaphosphates, poly-metaphosphates, dicalcium phosphate and/or its dihydrate, dicalcium orthophosphate, tricalcium phosphate, particulate hydroxyapatite, and the like. It is also possible to employ mixtures of these substances.

Silica dental abrasives of various types are preferred because of their unique benefits of exceptional dental cleaning and polishing performance without unduly abrading tooth enamel or dentine and which have a good compatibility with other possible ingredients, like metal ions and fluoride.

Dependent on the oral care composition, the abrasive product may be present in from 0 to 70% by weight, preferably from 1 % to 70%.

For toothpastes the abrasive material content typically lies in the range of from 10% to 70% by weight of the final tooth-paste product.

Humectants

Humectants are employed to prevent loss of water from, e.g., toothpastes and to avoid hardening of toothpastes upon exposure to air. Some humectants also give a desirable sweetness of flavor to toothpaste and mouthwash compositions. Suitable humectants for use in oral care compositions according to the invention include the following compounds and mixtures thereof: glycerol, polyol, sorbitol, xylitol, maltitol, lactitol, polyoxyethylene, polyethylene glycols (PEG), polypropylene glycols, propylene glycol, 1 ,3-propanediol, 1 ,4-butanediol, hydrogenated partially hydrolysed polysaccharides and the like, coconut fatty acid, amide of N-methyl-taurine, and Pluronic®.

Humectants are in generally present in from 0% to 80%, preferably 5 to 70% by weight. Thickeninq/bindinq agents

Suitable thickening and/or binding agents include silica, starch, tragacanth gum, xanthan gum, karaya gum, carrageenans (extracts of Irish moss), gum arabic, alginates, pectin, cellulose derivatives, such as hydroxyethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose and hydroxyethylpropyl cellulose, polyacrylic acid and its salts, polyvinylpyrrolidone and carboxyvinylpolymers, as well as inorganic thickeners such as amorphous silica compounds. These agents stabilize the oral care compositions of the invention.

Thickeners may be present in toothpaste, dental creams and gels as well as in mouthwashes in an amount of from 0.1 to 20% by weight, and binders to the extent of from 0.01 to 10% by weight of the final product.

Foaming agents and foaming modulators

As foaming agent soap, anionic, cationic, non-ionic, amphoteric and/or zwitterionic surfactants can be used, either alone or in combinations. These may be present at levels of from 0% to 15%, preferably from 0.1 % to 13%, more preferably from 0.25% to 10% by weight of the final product. Surfactants are only suitable to the extent that they do not exert an inactivation effect on the enzymes and other components included in the oral care composition. Useful surface-active agents include anionic, nonionic, and ampholytic compounds, with anionic compounds being preferred.

Examples of suitable surfactants include salts of the higher alkyl sulfates, such as sodium lauryl sulfate or other suitable alkyl sulfates having 8 to 18 carbon atoms in the alkyl group; sodium lauryl sulfoacetate, salts of sulfonated monoglycerides of higher fatty acids, such as sodium coconut monoglyceride sulfonate or other suitable sulfonated monoglycerides of fatty acids of 10 to 18 carbon atoms; salts of amides of higher fatty acid, e.g., 12 to 16 carbon atom acids, with lower aliphatic amino acids, such as sodium-N-methyl-N-palmitoyl tauride, sodium N-lauroyl-, N-myristoyl- and N-palmitoyl sarcosinates; salts of the esters of such fatty acids with isotopic acid or with glycerol monosulfate; such as the sodium salt of monosulfated monoglyceride of hydrogenated coconut oil fatty acids; salts of olefin sulfonates, e.g., alkene sulfonates or alkene sulfonates or mixtures thereof having 12 to 16 carbon atoms in the carbon chain of the molecule; and soaps of higher fatty acids, such as those of 12 to 18 carbon atoms, e.g., coconut fatty acids.

The cation of the salt may be sodium, potassium or mono-, di or triethanol amine. The nonionic surfactants include sucrose/fatty acid esters, maltose/fatty acid esters, maltitol/fatty acid esters, maltotriitol/fatty acid esters, maltotetraitol/fatty acid esters, maltopentaitol/fatty acid esters, maltohexaitol/fatty acid esters, mahoheptaitol/fatty acid esters, sorbitan/fatty acid esters, lactose/fatty acid esters, lactinose/fatty acid esters, polyoxyethylene/polyoxypropylene copolymers, polyoxyethylene alkyl ethers, polyoxyethylene/fatty acid esters, fatty acid alkanolamides, polyoxyethylene sorbitan/fatty acid esters, polyoxyethylene/hydrogenated castor oil, and polyglycerin/fatty acid esters.

Most preferred are sodium lauryl sulphate, sodium dodecylbenzene sulphonate and sodium lauryl sarcosinate.

Preferred foaming modulators include polyethylene glycols.

Foaming agents and foaming modulators may be present from in an amount of from 0% to 15% by weight, preferably from 0.01 % to 10% by weight.

Sweetening agents

Suitable sweeteners include, but are not limited to, saccharin and water-soluble salts thereof, dextrose, sucrose, lactose, maltose, levulose, aspartame, cyclamate salts, D- tryptophan, dihydrochalchones, acesulphame, stevioside, levaudioside, glycyrrhizins, pellartine, thaumatin, p-methoxycinnamic aldehyde, hydrogenated starch hydrolysates, xylitol, sorbitol, erythritol, mannitol, and mixtures thereof.

Sweeteners may be present from in an amount of from 0.001 % to 60% by weight, preferably from 0.01 % to 50% by weight.

Flavoring agents

Flavoring agents are usually present in low amounts, such as from 0.01 % to about 5% by weight, especially from 0.1 % to 5%. The flavors that may be used in the invention include, but are not limited to, wintergreen oil, peppermint oil, spearmint oil, clove bud oil, menthol, anethole, methyl salicvlate, eucalyptol, cassia, 1-inenthvl acetate, sage, eugenol, parsley oil, oxanone, alpha-irisone, marjoram, lemon, orange, cranberry, propenyl guaethol, cinnamon, vanillin, ethyl vanillin, heliotropine, 4-cis-heptenal, diacetyl, methylpara-tert-butyl phenyl acetate, carvone, cineole, menthone, cinnamic aldehyde, limonene, ocimene, n-decyl alcohol, citronellol, alpha- terpineol, methyl acetate, citronellyl acetate, methyl eugenol, linalool, thymol, rosemary oil, pimento oil, diatomaceous oil, eucalyptus oil, and mixtures thereof.

Coolants may also be part of the flavor system or added separately to the composition. Preferred coolants in the present compositions are the paramenthan carboxyamide agents such as N-ethyl-p-menthan-3-carboxamide (known commercially as WS-3"), menthol, 3-1- menthoxypropanc-1 ,2-diol ("TK-10"), menthone glycerol acetal ("MGA"), menthyl lactate and mixtures thereof.

Whitening/bleaching agents

Whitening/bleaching agents include H2O2 and may be added in amounts less than 5%, preferably from 0.05 to 4%, calculated on the basis of the weight of the final composition.

Other bleaching components which might be comprised by the present invention include, peroxydiphosphate, urea, peroxide, metal peroxides such as calcium peroxide, sodium peroxide, stronthium peroxide, magnesium peroxide, hypochlorite salts such as sodium hypochlorite, and the salts of perborate, persilicate, perphosphate and percarbonate such as sodium perborate, potassium persilicate and sodium percarbonate. The peroxide compounds can be stabilised by addition of a triphenylmethane dye, a chelating agent or antioxidants such as butylated hydroxy anisol (BHA) or butylated hydroxy toluene (BHT).

Solvent

A solvent is usually added to compositions of the invention in an amount sufficient for giving the compositions a flowable form in case the compositions is; e.g., a tooth paste, dental cream or gel, or to dissolve the other components of a compositions, in case of, e.g., a mouthwash or mouth rinse.

Suitable solvents include water, ethanol and water/ethanol mixtures, which may be present in an amount of from 0.1 % to 70%.

Anti-microbial agents

The present invention also includes water-soluble anti-microbial agents, such as chlorhexidine, triclosan, digluconate, hexetidine, alexidine, quaternary ammonium antibacterial compounds, and water-soluble sources of certain metal ions such as zinc, copper, silver and stannous (e.g., zinc, copper and stannous chloride, and silver nitrate) may also be included.

Sparingly soluble zinc salts such as zinc citrate, zinc C14-alkyl maleate, zinc benzoate, zinc caproate, zinc carbonate might also be included used in the compositions of the present invention to prolong the anti-microbial effectiveness of zinc ions due to the slow dissolution of these zinc salts in saliva.

Anti-microbial agents may be present in an amount of from 0% to 50% by weight, preferably from 0.01 % to 40% by weight, most preferably from 0.1% to 30% by weight.

Tartar-controlling agent

Compositions of the invention may comprise a tartar-controlling agent such as inorganic phosphorous tartar-controlling agents including any of the pyrophosphates such as disodium pyrophosphate, dipotassium pyrophosphate, tetrapotassium pyrophosphate, tetrasodium pyrophosphate, and mixtures thereof.

Organic phosphorous compounds that may serve as tartar-controlling agents include polyphosphonates such as disodium ethane-1- hydroxy-1 , 1-diphosphonate (EHDP), methanediphosphonic acid, and 2-phosphonobutane-1 2,4-tricarboxylic acid.

Tartar-controlling agents may be present in an amount of from 0% to 10% by weight, preferably from 0.1 % to 5% by weight.

Preservatives

Suitable preservatives include sodium benzoate, potassium sorbate, p-hydroxybenzoate esters, methyl paraben, ethyl paraben, propyl paraben, citric acid, calcium citrate, and mixtures thereof. Preservatives may be present in an amount of from 0% to 40% by weight, preferably from 0.01 % to 30% by weight.

Fluoride sources

Compositions of the invention may also comprise compounds that can be used as fluoride source. Preferred soluble fluoride sources include sodium fluoride, potassium fluoride, stannous fluoride, indium fluoride, sodium monofluorophosphate, sodium heexafluorosilicate, zinc fluoride, lithium fluoride, aluminium fluoride, acidulated phosphate fluoride, ammonium bifluoride, titatium tetrafluoride, and amine fluoride.

Especially preferred are sodium fluoride and sodium monofluorophosphate.

Fluoride sources may be present in an amount of from 0% to 20% by weight, preferably from 0.01 % to 15% by weight, most preferably from 0.1 % to 10% by weight.

Coloring agents

Coloring agents or pigments suitable for oral care compositions of the invention include non-toxic, water-insoluble inorganic pigments such as titanium dioxide and chromium oxide greens, ultramarine blues and pinks and ferric oxides as well as water insoluble dye lakes prepared by extending calcium or aluminum salts of FD&C dyes on alumina such as FD&C Green No.1 lake, FD&C Blue No.2 lake, FD&C Red No. 30 lake, FD&C Yellow No. 16 lake, and FD&C Yellow No. 10.

A preferred opacifier is titanium dioxide.

Coloring agents may be present in an amount of from 0% to 20% by weight, preferably from 0.01 % to 15% by weight, most preferably from 0.1% to 10% by weight.

Buffering agents

The oral care compositions of present invention may also include buffering agents, i.e., pH-adjusting agents, such as alkali metal hydroxides, carbonates, sesquicarbonates, borates, silicates, phosphates, imidazole, and mixtures thereof.

Specific buffering agents include monosodium phosphate, trisodium phosphate, sodium hydroxide, potassium hydroxide, alkali metal carbonate salts, sodium carbonate, imidazole, pyrophosphate salts, sodium citrate, hydrochloric acid, sodium hydroxide, triethanolamine, triethylamine, lactic acid, malic acid, fumaric acid, tartaric acid, phosphoric acid and mixtures of these.

Buffering agents may be present in an amount of from 0% to 10% by weight, preferably from 0.01 % to 5% by weight.

Chewing Gum

When the oral composition according to the invention is a chewing gum, it can be any known type of chewing gum, such as chewing gum pieces optionally coated, as well as sticks or chewing gum provided with an arbitrary desired shape in response to the intended use. The chewing gum preparation can be of any quality including the bubble gum quality.

In a preferred embodiment, the present invention relates to oral care compositions in the form of a chewing gum comprising a polypeptide having DNase activity and at least one oral care component, wherein the at least one oral care component is selected from elastomer, softening agent, plasticizing agent, emulsifier, wax, coloring agent, sweetening agent, flavoring agent, bulking agent, thickening agent.

Gum base ingredients

Chewing gum is traditionally considered as being comprised of a water-insoluble or base portion and a water-soluble portion that contains flavoring agents, sweetening agents, and coloring agents. The gum base part of the gum is a masticatory substance which imparts the chew characteristics to the final product. It defines the release profile of flavors and the sweeteners and plays a significant role in the gum product. The flavors, sweeteners and colors can be thought of as providing the sensory appeal aspects of the chewing gum. No limitations as to the chewing gum bases used in a chewing gum preparation according to the invention exist. Conventional chewing gum bases available for instance from Dansk Tyggegummi Fabrik A/S, L.A. Dreyfus or Cafasa Gum SIA, are usually suitable, but specially made formulations can also be used. The formulation depends on the desired type of chewing gum or the desired type of structure. Suitable raw materials for gum bases include the substances according to the U.S. Chewing Gum Base Regulations - Code of Federal Regulations, Title 21 , Section 172,615 and in accordance with other national and international lists (or positive lists) and include elastomers, resins, waxes, polyvinyl acetates, oils, fats, emulsifiers, fillers and antioxidants.

The gum base usually comprises from 15 to 90% by weight, preferably from 30 to 40% by weight, more preferably from 5 to 25% of the final product.

Elastomers provide the chew, springiness or bounce to the base and control bubble and flavor release in the final chewing gum. They may be any water-insoluble polymer known in the art. They include styrene butadiene copolymers (SBR) and non-SBR types, both natural and synthetic. Examples of natural elastomers include, without limitation, rubbers such as rubber latex (natural rubber) and guayule, and gums such as chicle, jelutong, balata, guttapercha, lechi capsi, sorva, crown gum, nispero, rosidinha, perillo, niger gutta, tunu, gutta kay, pendare, leche de vaca, chiquibul, crown gum, and the like, and mixtures thereof. Examples of synthetic elastomers include, without limitation, polyisobutylene, isobutylene-isoprene copolymers (butylrubber), polyethylene, polybutadiene, styrenebutadiene copolymers, polyisoprene, and the like, and mixtures thereof.

The amounts of elastomer (rubbers) employed in the gum base composition will vary greatly depending upon various factors such as the, type of gum base used (adhesive, or conventional, bubble or standard) the consistency of the gum base composition desired, and the other components used in the composition to make the final chewing gum product. In general, the elastomer is present in the gum base composition in an amount of from about 15% to about 60%, preferably from about 25% to about 30%, by weight based on the total weight of the gum base composition.

Elastomer solvents aid in softening or plasticizing the elastomer component. In doing so they provide a bulkiness to the chew.

Elastomer solvents include, but are not limited to, natural rosin esters and synthetic derivatives of, e.g., terpenes. Examples of elastomer solvents suitable for use herein include tall oil rosin ester; partially hydrogenated wood and gum rosin; the glycerol esters of wood and gum rosin, partially hydrogenated wood/gum rosin, partially dimerized wood and gum rosin, polymerized wood and gum rosin, and tall oil rosin; the deodorized glycerol ester of wood rosin; the pentaerythritol esters of wood and gum rosin; partially hydrogenated wood and gum rosin; the methyl ester of partially hydrogenated wood rosin; methyl, glycerol and pentaerythritol esters of rosins and modified rosins such as hydrogenated, dimerized and polymerized rosins; terpene resins such as polymers of alpha-pinene or beta-pinene, terpene hydrocarbon resins; polyterpene; and the like, and mixtures thereof. The elastomer solvent may be employed in the gum base composition in an amount of from about 2% to about 40%, and preferably from about 7% to about 15% by weight of the gum base composition.

Polyvinyl acetates provide stretch or elasticity to the gum base. They also affect chew bulkiness, softness and bubble, hydrophilic character and flavor release.

The amounts of the different molecular weight polyvinyl acetates present in the gum base composition should be effective to provide the finished chewing gum with the desired chew properties, such as integrity, softness, chew bulkiness, film-forming characteristic, hydrophilic character, and flavor release. The total amount of polyvinyl acetate used in the gum base composition is usually from about 45% to about 92% by weight based on the total gum base composition. The vinyl polymers may possess a molecular weight ranging from about 2000 Da up to about 95,000 Da.

Typically, the low molecular weight polyvinyl acetate has a weight average molecular weight of from about 2,000 Da to about 14,000 Da. The medium molecular weight polyvinyl acetate typically has a weight average molecular weight of from about 15,000 Da to 55,000 Da. The high molecular weight polyvinyl acetate typically has a weight average molecular weight of from 55,000 Da to about 95,000 Da but may range as high as 500,000 Da.

Waxes, fats, and oils plasticize the elastomer mixture and improve the elasticity of the gum base. Waxes can provide a soft or firm chew, affect the flavor release and provide bulkiness and smoothness to the gum base. Fats and oils provide a soft chew. The fats, oils and waxes may be use individually or in combination or the gum base may be a wax free gum base.

Waxes when used, may be of mineral, animal vegetable or synthetic origin. Non-limiting examples of mineral waxes include petroleum waxes such as paraffin and microcrystalline waxes, animal waxes include beeswax, vegetable waxes include carnauba, candellila, rice bran, esparto, flax and sugarcane, and synthetic waxes include those produced by the Fischer- Tropsch synthesis, and mixtures thereof.

Suitable oils and fats usable in gum compositions include hydrogenated or partially hydrogenated vegetable or animal fats, such as cottonseed oil, soybean oil, coconut oil, palm kernel oil, beef tallow, hydrogenated tallow, lard, cocoa butter, lanolin and the like; fatty acids such as palmitic, oleic, stearic, linoleic, lauric, myristic, caproic, caprylic, decanoic or esters and salts as sodium stearate and potassium stearate. These ingredients when used are generally present in amounts up to about 7% by weight of the gum composition, and preferably up to about 3.5% by weight of the gum composition.

Preferred as softeners are the hydrogenated vegetable oils and include soybean oil and cottonseed oil which may be employed alone or in combination. These softeners provide the gum base composition with good texture and soft chew characteristics. These softeners are generally employed in an amount from about 5% to about 14% by weight of the gum base composition.

Emulsifiers aid in dispersing the immiscible components of the gum base composition into a single stable system. They provide hydrophilic character to a gum base and aid in plasticizing the resins and polyvinyl acetates. They also affect the softness of the base and the, bubble character of the base. Typical emulsifiers include acetylated monoglyceride, glyceryl monostearate, lecithin, fatty acid monoglycerides, diglycerides, propylene glycol monostearate, lecithin, triacetin, glyceryl triacetate and the like, and mixtures thereof.

Preferred emulsifiers are glyceryl monostearate and acetylated monogylcerides. These serve as plasticizing agents. The emulsifiers may be employed in an amount of from about 2% to about 15% by weight of the gum base composition, and preferably from about 7% to about 1 1% by weight of the gum base composition.

The fats, oils, waxes, emulsifiers and certain sugar bulking agents are often grouped together and referred to as softening agents. Because of the low molecular weight of these ingredients, the softeners are able to penetrate the fundamental structure of the gum base making it plastic and less viscous. Useful plasticizers and softeners of the above include lanolin, palmitic acid, oleic acid, stearic acid, sodium stearate, potassium stearate, glyceryl triacetate, glyceryl lecithin, glyceryl monostearate, propylene glycol nonastearate, acetylated monoglyceride, glycerin, fully unsaturated vegetable oils such as nonhydrogenated cottonseed oil, hydrogenated vegetable oils, petroleum waxes, sorbitan monostearate, tallow, and the like, and mixtures thereof and also include high fructose corn syrup, corn syrup, sorbitol solution, hydrogenated starch hydrolysate, and the like, and mixtures thereof.

The amount of softener present should he an effective amount to provide a finished chewing gum with the desired chew bulkiness and softness. When used as softeners these materials are generally employed in the gum base composition in an amount of up to about 25%, and preferably in an amount of from about 1 % to about 17%, by weight of the gum base composition.

The gum base may further contain a surfactant. Examples of suitable surfactants include polyoxyethylene (20) sorbitan monoleate, polyoxyethylene (20) sorbitan monolaurate, polyethylene (4) sorbitan monolaurate, polyoxyethylene (20) sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene, (4) sorbitan monostearate, polyoxyethylene (20) sorbitan tristearate, polyoxyethylene (5) sorbitan monooleate, polyoxyethylene (20) sorbitan trioleate, sorbitan monolaurate, and the like. The amount of surfactant present should be effective to provide the finished chewing gum with the desired softness. Typically, the surfactant is employed in the base in an amount of from about 0.5% to about 3.0% by weight based on the total weight of the gum base.

The gum base composition of this invention may also include effective amounts of fillers sometimes referred to as bulking agents. These materials add firmness and bulk and affect the texture and the flavor release of the chewing gum. Useful fillers include organic and inorganic compounds (mineral adjuvants) such as calcium carbonate, magnesium carbonate, ground limestone, magnesium silicate, calcium phosphate, cellulose polymers, clay, alumina, aluminium hydroxide, aluminum silicate, tale, tricalcium phosphate, dicalcium phosphate, and the like, and mixtures thereof. These fillers or adjuvants may be used in the gum base compositions in various amounts. The amount of the filler present should be effective to provide a finished chewing gum with the desired flavor release and integrity. Typically, the filler is employed in the gum base composition in an amount from about 1 % to about 40%, and preferably from about 5% to about 20%, by weight of the gum base composition.

The gum base may also comprise an antioxidant to provide improved stability, lessen any oil-taste and provide longer shelf life. Typical non-limiting examples of antioxidants are butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate. Mixtures thereof may also be used.

Other gum ingredients

The remaining ingredients in chewing gum compositions are conventional and usually comprise from 10 to 85% by weight of the final product.

Examples thereof are sweetening agents, softeners, coloring agents, bulking agents, thickening agents, and flavoring agents of the type and in the amounts conventionally used for chewing gum.

Suitable flavoring agents those flavors known to the skilled artisan such as natural and artificial flavors. These flavorings may be chosen from synthetic flavor oils and flavoring aromatics and/or oils, oleoresins and extracts derived from plants, leaves, flowers, fruits, and so forth, and combinations thereof. Non-limiting representative flavor oils include spearmint oil, cinnamon oil, wintergreen oil (methyl salicylate), peppermint oil, clove oil, bay oil, anise oil, eucalyptus oil, thyme oil, cedar leaf oil, oil of nutmeg, allspice, oil of sage, mace, oil of bitter almonds, and cassia oil. Other useful flavorings are artificial, natural and synthetic fruit flavors such as vanilla, and citrus oils including lemon, orange, lime, grapefruit, and fruit essences including apple, pear, peach, grape, strawberry, raspberry, cherry, plum, pineapple, apricot and so forth. These flavoring agents may be used in liquid or solid form and may be used individually or in admixtures. Commonly used flavors include mints such as peppermint, menthol, artificial vanilla, cinnamon derivatives, and various fruit flavors, whether employed individually or in admixture.

Other useful flavoring agents include aldehydes and esters such as cinnamyl acetate, cinnamaldehyde, citral diethylacetal, dihydrocarvyl acetate, eugenyl formate, pmethylamisol, and so forth may be used. Generally, any flavoring or food additive may be used.

Further examples of aldehyde flavorings include, but are not limited to, acetaldehyde (apple), benzaldehyde (cherry, almond), anisic aldehyde (licorice, anise), cinnamic aldehyde (cinnamon), citral, i.e., alpha-citral (lemon lime), neral, i.e., beta-citral (lemon, lime), decanal (orange, lemon), ethyl vanillin (vanilla, cream), heliotrope, i.e., piperonal (vanilla, cream), vanillin (vanilla, cream), alpha-amyl cinnamaldehyde (spicy fruity flavors), butyraldehyde (butter, cheese), valeraldehyde (butter, cheese), citronellal (many types), decanal (citrus fruits), aldehyde C-8 (citrus fruits), aldehyde C-9 (citrus fruits), aldehyde C-12 (citrus fruits), 2-ethyl butyraldehyde (berry fruits), hexenal, i.e., trans-2-hexenal (berry fruits), tolyl aldehyde (cherry, almond), veratraldehyde (vanilla), 2,6-dimethyl-5-heptenal, i.e., melonal (melon), 2,6- dimethyloctanal (green fruit), and 2-dodecenal (citrus, mandarin), cherry, grape, strawberry shortcake, mixtures thereof and the like.

The amount of flavoring agent employed herein is normally a matter of preference subject to such factors as the type of final chewing gum composition, the individual flavor, the gum employed, and the strength of flavor desired. Thus, the amount of flavoring may be varied in order to obtain the result desired in the final product and such variations are within the capabilities of those skilled in the art without the need for undue experimentation. In gum compositions, the flavoring agent is generally present in amounts from about 0.02% to about 5% by weight of the chewing gum composition.

The chewing gum compositions generally include bulking agents. These bulking agents (carders, extenders) may be water-soluble and include bulking agents selected from the group consisting of, but not limited to, monosaccharides, disaccharides, polysaccharides, sugar alcohols, and mixtures thereof; sorbitol, xylitol, maltitol, mannitol, isomalt (a racemic mixture of alpha-D-glucopyranosyl-1 ,6-mannitol and alpha-D-glucopyranosyl-1 , 6-sorbitol manufactured under the tradename Palatinit™ by Suddeutsche Zucker), glycerol, aspartame, Lycasin® glycerol, galactitol acesulphame K, saccharine and salts thereof, cyclamate and salts thereof, neohesperidine dihydrochalcone, glycyrrhizinic acid and salts thereof, thaumantine and sucralose as well as mixtures thereof or mixtures thereof with other suitable sweeteners, maltodextrins; hydrogenated starch hydrolysates; hydrogenated hexoses; hydrogenated disaccharides; minerals, such as calcium carbonate, talc, titanium dioxide, dicalcium phosphate, celluloses and the and the like, and mixtures thereof. Bulking agents may be used in amounts up to about 60%, and preferably in amounts from about 25% to about 60%, by weight of the chewing gum composition.

The chewing gum compositions may also include a high intensity sweetening agent (sweeteners). High intensity sweetening agents have a sweetness intensity substantially greater than that of sucrose. Examples of suitable intense sweeteners include:

a) water-soluble naturally-occurring intense sweeteners such as dihydrochalcones, monellin, steviosides, glycyrrhizin, dihydroflavenol, and L-aminodicarboxylic acid aminoalkonoic acid ester amides, such as those disclosed in in United States patent no. 4,619,834, and mixtures thereof;

b) water-soluble artificial sweeteners including the soluble saccharin salts such as sodium or calcium saccharin salts, cyclamate salts, the sodium, ammonium or calcium salts of 3,4- dihydro-6-methyl-1 ,2, 3-oxathiazine-4-one-2, 2-dioxide, the potassium salt of 3,4-dihydro-6- methyl-1 , 2, 3-oxathiazine-4-one-2, 2-dioxide (Acesulfam-K), the free acid form of saccharin, and the like, and mixtures thereof;

c) dipeptide based sweeteners including L-aspartic acid derived sweeteners such as 1 - aspartyl-L-phenylalanine methyl ester (Aspartame) and materials described in United States patent no. 3,492,131 , L-alpha-aspartyl-N-(2,2,4,4-tetramethyl-3-thietanyl)-D-alaninamide hydrate (Alitame), methyl esters of L-aspartyl-L-phenylglycerine and L-aspartyl-L-2,5- dihydrophenyl-glycine, L-aspartyl-2.5-dihydro-L-phenylalanine, L-aspartyl-L-(l-cyclohexen)- alanine, and the like, and mixtures thereof;

d) water-soluble intense, sweeteners derived from naturally-occurring water-soluble sweeteners, such as chlorinated derivatives of ordinary sugar (sucrose), e.g., chlorodeoxysugar derivatives such as derivatives of chlorodeoxysucrose or chlorodeoxygalactosucrose, known, for example, under the product designation of Sucralose®; examples of chlorodeoxysucrose and chlorodeoxygalactosucrose derivatives include but are not limited to: to 1 -chloro-T- deoxysucrose; 4-chloro-4-deoxy-alpha-D-galactopyranosyl-alpha-D-fructofuranoside, or 4- chloro-4-deoxygalactosucrose; 4-chloro-4-deoxy-alpha-D-galactopyranosyl-1 -chloro-ldeoxy- beta-D-fructo-furanoside, or 4,T-dichloro-4,T-dideoxygalactosucrose; 1 ',6'-dichloro-T,6'- dideoxysucrose; 4-chloro-4-deoxy-alpha-D-galactopyranosy1-1 ,6-dichloro-1 ,6-dideoxy-beta-D- fructofuranoside, or 4,1 ',6'-trichloro-4,T,6'-trideoxygalactosucrose; 4,6-dichloro-4,6-dideoxy- alpha-D-galactopyranosyl-6-chloro-6-deoxy-beta-D-fructofuranoside, or 4,6,6'-trichloro-4,6,6'- trideoxygalactosucrose; 6,T,6'-trichloro-6,T,6'-trideoxysucrose; 4,6-dichloro-4,6-dideoxy-alpha- D-galactopyranosyl-1 ,6-dichloro-1 ,6-dideoxy-beta-D-fluctofuranoside, or 4,6,1 ',6'-tetrachloro- 4,6,T,6'-tetradeoxygalacto-sucrose; and 4,6,1 ',6'-tetradeoxy-sucrose, and mixtures thereof; and e) protein based intense sweeteners such as Thaumaoccous daniclii (Thaumatin I and II). The amount of sweetener employed in the chewing gum composition will vary with the sweetener selected for a particular chewing gum. Thus, for any given sweetener a sufficient amount of sweetener is used to provide the level of sweetness desired. The saccharide sweeteners and sugar alcohols described above are usually used in an amount of from about 1 % to about 70% and preferably in an amount of from about 40% to about 50%, by weight based on the total weight of the chewing gum composition. The intense sweeteners described above are usually used in an amount of up to about 1 %, preferably from about 0.05% to about 0.4%, by weight based on the total weight of the chewing gum composition.

The coloring agents useful in the present invention are used in amounts effective to produce the desired color. These coloring agents include pigments, which may be incorporated in amounts up to about 6%, by weight of the gum composition. A preferred pigment, titanium dioxide, may be incorporated in amounts up to abou t2%, and preferably less than about 1 %, by weight of the gum composition. The colorants may also include natural food colors and dyes suitable for food, drug and cosmetic applications. These colorants are known as F.D.& C. dyes and lakes. The materials acceptable for the foregoing uses are preferably water-soluble. Illustrative non-limiting examples include the indigoid dye known as F.D.& C. Blue No.2, which is the disodium salt of 5,5-indigotindisulfonic acid. Similarly, the dye known as F.D.& C. Green No.1 comprises a triphenylmethane dye and is the monosodium salt of 4-[4-(N-ethyl-N-p- sulfoniumbenzylamino)diphenylmethylene]-[1-(N-ethyl-N-p-sulfoniumbenzyl)-delta-2,5-cyclo- hexadieneimine].

Examples of thickening agents include methyl cellulose, alginates, carrageenan, xanthan gum, gelatin, carob, tragacanth, and locust bean, emulsifiers, such as lecithin and glyceryl monostearate, acidulants such as malic acid, adipic acid, citric acid, tartaric acid, fumaric acid, and mixtures thereof.

The plasticizers, softening agents, emulsifiers, waxes, and antioxidants discussed above as being suitable for use in the gum base may also be used in the chewing gum composition.

Active gum ingredients

Oral care compositions of the invention in the form of a chewing gum may also contain various active ingredients such as antimicrobial agents, Zn salts, fluorides, and urea.

Moreover, the oral composition according to the invention may, if desired, include any other active ingredients, such as anti-caries agents, anti-calculus agents, anti-plaque agents, anti-periodontal agents, anti-fungal agents, anti-smoking agents, anti-cold agents, agents against gingivitis, etc.

The antimicrobials used in the compositions can be any of a wide of cationic antimicrobial agents such as quaternary ammonium compounds (e.g., cetyl pyridinium chloride) and substituted guanidines such as chlorhexidine and the corresponding compound alexidine. Mixtures of cationic anti-microbials may also be used in the present invention.

Antimicrobial quaternary ammonium compounds include those in which one or two of the substituents on the quaternary nitrogen has a carbon chain length (typically alkyl group) of some 8 to 20, typically 10 to 18 carbon atoms while the remaining substituents (typically alkyl or benzyl group) have a lower number of carbon atoms, such as 1 to 7 carbon atoms, typically methyl or ethyl groups. Dodecyl trimethyl ammonium bromide, tetradecyl pyridinium chloride, tetradecyl ethyl pyridinium chloride, dodecyl dimethyl (2-phenoxyethyl) ammonium bromide, benzyl dimethylstearyl ammonium chloride, cetyl pyridinium chloride, quaternized 5-aminc-1 ,3-bis 2- ethyl-hexyl)-5-methyl hexa hydropyrimidine and benzethonium chloride are exemplary of typical quaternary ammonium antibacterial agents. Other compounds are the bis[4-(R-amino)-1- pyridinium] alkanes as disclosed in U.S. Patent 4,206,215, June 3, 1980 to Bailey incorporated herein by reference. The pyridinium compounds are the preferred quaternary ammonium compounds.

The cationic antimicrobial is generally used in the present compositions at a level of from about 0.02% to about 1 %, preferably from about 0.3% to about 0.7% most preferably from about 0.3% to about 0.5%.

As easily soluble zinc salt it is in principle possible to use any physiologically acceptable, easily soluble zinc salt of an inorganic or organic acid, said salt being able to release zinc ions and being approved for the intended use, such as in foodstuffs, cosmetics or pharmaceutical products. Non-limiting examples are for instance zinc citrate, zinc sulphate, zinc lactate, zinc chloride, zinc acetate as well as mixtures thereof. Among these salts zinc acetate is preferred.

The zinc salt used must be easily soluble such that a release is ensured in the oral cavity of an amount of zinc ions efficient for the purpose aimed at within a suitable period of time.

Advantageously, the zinc salt is present in the oral composition in an amount of from 0.001 to 1.25% by weight. The amount used depends on the administration form and the intended use and is adapted such that an amount of zinc ions efficient for the intended use is released.

As taste-masking salt is used at least one salt selected among sodium chloride, ammonium chloride and physiologically acceptable alkali metal, alkaline earth metal and/or ammonium carbonates.

The alkali metal is in particular sodium or potassium, whereas the alkaline earth metal advantageously is calcium or magnesium. Particularly preferred taste-masking salts are sodium, potassium and magnesium carbonates, sodium chloride, ammonium chloride as well as mixtures thereof.

The taste-masking salt is advantageously used in the oral composition in an amount of from 0.05 to 6.25% by weight, more preferred from 0.25 to 3.50% by weight, such as from 0.50 to 2.50% by weight. The amount used of taste-masking salt for masking the taste of zinc can in each case be determined by a person skilled in the art and depends on the particular zinc salt in question and the selected administration form.

Urea is used as an anticariogenic product for neutralizing the acid produced in dental plaque subsequent to eating or drinking. Beyond urea the composition also can contain pharmacologically acceptable substances capable of releasing urea under the conditions prevailing in the mouth. Examples thereof are: Salts and addition compounds between urea and inorganic compounds such as magnesium sulphate, calcium phosphate, sodium chloride, etc.

The urea content of the composition according to the invention varies between 0.05% by weight and 80% by weight, preferably between 0.2% by weight and 25% by weight.

The chewing gum compositions may be prepared using standard techniques and equipment known to those skilled in the art. The apparatus useful in accordance with the present invention comprises mixing and beating apparatus as well.

Lozenges and pastilles

Lozenges are flavored medicated dosage forms intended to be sucked and held in the mouth or pharynx. They may contain vitamins, antibiotics, antiseptics, local anesthetics, antihistamines, decongestants, corticosteroids, astringents, analgesics, aromatics, demulcents, or combinations of these ingredients. Lozenges may take various shapes, the most common being the flat, circular, octagonal, and biconvex forms. Another type, called bacilli, are in the form of short rods or cylinders. A soft variety of lozenge, called a pastille, consists of medicament in a gelatin or glycerogelatin base or in base of acacia, sucrose, and water (H. A. Lieberman, Pharmaceutical Dosage Forms: Tablets, Volume 1 (1980), Marcel Dekker, Inc., New York, N.Y.).

In a preferred embodiment, the present invention relates to oral care compositions in the form of a lozenge or pastille comprising a polypeptide having DNase activity and at least one oral care component, wherein the at least one oral care component is selected from lubricant, bulking agent, sweetening agent, and flavoring agent.

Lubricants

The use of a lubricant in the manufacture of compressed lozenges is to facilitate the release of the lozenge from the die in which it is formed. The lubricant used in the present invention is a solid material which is not charged and which will not interfere ( e.g complex) with the cationic antimicrobial. The material should preferably be water insoluble. One type of suitable material meeting these requirements is a non-toxic hydrocarbon fat or derivative. Examples include hydrogenated tallow and hydrogenated vegetable oil. Polyethylene glycols may also be used as a lubricant so long as they are solid materials which generally means having a molecular weight in the 4000 Da to 6000 Da range. These materials can also be used as a filler as noted below. Mixtures of lubricants may also be used in the present invention. The lubricant is used at level of from about 0.1 % to about 4.0% preferably from about 0.5% to about 2%.

Lozenge vehicle

The term“lozenge vehicle” is used herein to denote the material(s) which carries the active ingredients, such as polypeptide(s) having DNase activity, other enzymes, and therapeutic agents, as well as the lubricant. These materials are also known as bulking agents or fillers. Since the vehicle is non-cariogenic, the vehicle should be free of sucrose and similar materials.

Acceptable filler materials include mannitol, sorbitol, xylitol, polyethylene glycol and non- cariogenic dextrans. The fillers may be used alone or in combination.

Mannitol is a naturally occurring sugar alcohol and is available as a fine powder. It has a sweetness of only about 50% of that of sucrose. However, mannitol's negative heat of solution enables it to impart a pleasant, cooling sensation in the mouth as the lozenge dissolves.

Sorbitol is a chemical isomer of mannitol and possesses a similar degree of sweetness. Its heat of solution, being negative, also provides for a pleasant, cooling sensation in the mouth. Sorbitol is available either as free flowing granules or as a crystalline powder. Polyethylene glycols (PEG's) can also be used in the present compositions. These materials are polymers of ethylene oxide with the generalized formula HOCH2 (CH₂0CH₂)_nCH₂0H. The use of PEG'S alone is not favored but their use in combination with other fillers is acceptable. The molecular weights found most desirable are between 4000 Da and 6000 Da.

Fillers are generally used in the present invention at a level of from about 85% to about 99.8%, preferably from about 90% to about 98%, most preferably from about 94% to about 97%.

Other lozenge components

Acceptable lozenges may be manufactured using just an active ingredient, the lubricant and the filler material as outlined above. However, in order to make the lozenges more acceptable from an aesthetic viewpoint, generally included are materials such as spray-dried or encapsulated flavors or liquid flavors adsorbed onto a suitable diluent. Spray-dried or encapsulated flavors are preferred. Suitable flavors include oil of peppermint, oil of wintergreen, oil of sassafras, oil of spearmint and oil of clove. Sweetening agents are also acceptable for use in the present compositions. Suitable agents include aspartame, acesulfame, saccharin, dextrose and levulose. Sweetening and flavoring agents are generally used in the compositions of this invention at levels of from about 0.1 % to about 2%, preferably from about 0.25% to about 1 .5%.

It is also acceptable to have a solid form of a water-soluble fluoride compound present in the present lozenges in an amount sufficient to give a fluoride concentration of from about 0.0025% to about 5.0% by weight, preferably from about 0.005% to about 2.0% by weight, to provide additionally anticaries effectiveness. Preferred fluorides are sodium fluoride, stannous fluoride, indium fluoride and sodium monofluorophosphate. The lozenges may also contain various active ingredients such as anti-microbial agents, Zn salts, fluorides, and urea (supra).

Confectionaries and candy

In a preferred embodiment, the present invention relates to oral care compositions in the form of a confectionary or candy comprising a polypeptide having DNase activity and at least one oral care component, wherein the at least one oral care component is selected from coloring agent, sweetening agent, flavoring agent, and oil-modifying agent.

The preparation of confectionery formulations is historically well known and has changed little through the years. Confectionery items have been classified as either "hard" confectionery or "soft" confectionery. The volatile oil-modifying agent of the present invention can be incorporated by admixing the modifying agent into conventional hard and soft confections.

Hard confectionery may be processed and formulated by conventional means. In general, a hard confectionery has a base composed of a mixture of sugar and other carbohydrate bulking agents kept in an amorphous or glassy condition. This form is considered a solid syrup of sugars generally having from about 0.5% to about 1 .5% moisture. Such materials normally contain up to about 92% corn syrup, up to about 55% sugar and from about 0.1 % to about 5% water, by weight of the final composition. The syrup component is generally prepared from corn syrups high in fructose, but may include other materials. Further ingredients such as flavorings, sweeteners, acidulants, colorants and so forth may also be added.

Such confectionery may be routinely prepared by conventional methods such as those involving fire cookers, vacuum cookers, and scraped-surface cookers also referred to as high speed atmospheric cookers.

Fire cookers involve the traditional method of making a candy base. In this method, the desired quantity of carbohydrate bulking agent is dissolved in water by heating the agent in a kettle until the bulking agent dissolves. Additional bulking agent may then be added and cooking continued until a final temperature of 145° to 156° C. is achieved. The batch is then cooled and worked as a plastic-like mass to incorporate additives such as flavor, colorants and the like.

A high-speed atmospheric cooker uses a beat-exchanger surface, which involves spreading a film of candy on a heat exchange surface, the candy is heated to 165° to 170° C. in a few minutes. The candy is then rapidly cooled to 100° to 120° C. and worked as a plastic-like mass enabling incorporation of the additives, such as flavors, colorants and the like.

In vacuum cookers, the carbohydrate bulking agent is boiled to 125° to 132° C, vacuum is applied and additional water is boiled off without extra heating. When cooking is complete, the mass is a semi-solid and has a plastic-like consistency. At this point, flavors, colorants, and other additives are admixed in the mass by routine mechanical mixing operations.

The optimum mixing required to uniformly mix the flavors, colorants and other additives during conventional manufacturing of hard confectionery is determined by the time needed to obtain a uniform distribution of the materials. Normally, mixing times of from 4 to 10 minutes have been found to be acceptable.

Once the candy mass has been properly tempered, it may be cut into workable portions or formed into desired shapes. A variety of forming techniques may be utilized depending upon the shape and size of the final product desired. A general discussion of the composition and preparation of hard confections may be found in H. A. Lieberman, Pharmaceutical Dosage Forms: Tablets, Volume 1 (1980), Marcel Dekker, Inc., New York, N.Y.

The apparatus useful in accordance with the present invention comprises cooking and mixing apparatus well known in the confectionery manufacturing arts, and election of the specific apparatus will be apparent to the artisan. In contrast, compressed tablet confections contain particular materials and are formed into structures under pressure.

These confections generally contain sugars in amounts up to about 95%, by weight of the composition, and typical tablet excipients such as binders and lubricants as well as flavoring agent, colorants and so forth. Similar to hard confectionery, soft confectionery may be utilized in this invention. The preparation of soft confections, such as nougat, involves conventional methods, such as the combination of two primary components, namely (1 ) a high boiling syrup such as corn syrup, hydrogenated starch hydrolysate or the like, and (2) a relatively light textured frappe, generally prepared from egg albumin, gelatin, vegetable proteins, such as soy derived compounds, sugarless milk derived compounds such as milk proteins, and mixtures thereof. The frappe is generally relatively light, and may, for example, range in density from about 0.5 to about 0.7 grams/cc.

The flavoring components of the confection are flavors having an associated bitter taste or other unpleasant after taste. These flavoring components may be chosen from natural and synthetic flavoring liquids such as volatile oils, synthetic flavor oils, flavoring aromatic and oils, liquids, oleoresins or extracts derived from plants, leaves, flowers, fruits, stew and combinations thereof. Non-limiting representative examples of volatile oils include spearmint oil, cinnamon oil, oil of wintergreen (methyl saliclate), peppermint oil, menthol, clove oil, bay oil, anise oil, eucalyptus oil, thyme oil, cedar leaf oil, oil of nutmeg, allspice oil, oil of sage, mace extract, oil of bitter almonds, and cassia oil. In addition, the confection may also contain artificial, natural or synthetic flavors including fruit flavors such as vanilla, and citrus oils including lemon, orange, grape, lime and grapefruit and fruit essences including apple, pear, peach, grape, strawberry, raspberry, cherry, plum, pineapple, apricot and so forth individual and mixed.

Other useful flavorings include aldehydes and esters such as benzaldehyde (cherry, almond), citral, i.e., alpha-citral (lemon, lime), neral, i.e., beta-citral (lemon, lime), decanal (orange, lemon), aldehyde C-8 (citrus fruits), aldehyde C-9 (citrus fruits), aldehyde C-12 (citrus fruits), tolyl aldehyde (cherry, almond), 2,6-dimethyl-octanal (green fruit), and 2-dodecenal (citrus, mandarin), mixtures thereof and the like. In the instance where sweeteners are utilized, the present invention contemplates the inclusion of those sweeteners well known in the art, including both natural and artificial sweeteners. The sweeteners may be chosen from the following non-limiting list: sugars such as sucrose, glucose (corn syrup), dextrose, invert sugar, fructose, and mixtures thereof, saccharin and its various salts such as the sodium or calcium salt; cyclamic acid and its various salts such as the sodium salt; the dipeptide sweeteners such as aspartame, dihydrachalcone compounds, glycyrrhizin; Stevia Rebaudiana (Stevioside); chloro-derivatives of sucrose; dihydroflavinol; hydroxyguaiacol esters; L-amino dicarboxylic acid gem-diamines; L-aminodicarboxylic acid aminoalkenoic acid ester amides; and sugar alcohols such as sorbitol, sorbitol syrup, mannitol, xylitol, and the like. Also contemplated is the synthetic sweetener 3,6-dihydro-6-methyl-1 ,2,3- oxathiazin-4-one-2, 2-dioxide, particularly the potassium (acesulfame-K), sodium and calcium salts thereof.

The confection may also include a colorant. The colorants may be selected from any of the numerous dyes suitable for food, drug and cosmetic applications, and known as FD&C dyes and the like. The materials acceptable for the foregoing spectrum of use are preferably water- soluble. Illustrative examples include indigoid dye, known as FD&C Blue No. 2, which is the disodium salt of 5,5'-indigotindisulfonic acid. Similarly, the dye known as FD&C Green No. 1 comprises a triphenylmethane dye and is the monosodium salts of 4-[ 4-N-ethyl-p- sulfobenzylami no) d iphenylmethylane ]-[ 1-(N-ethyl-N-p-sulfonium benzyl)-2-5-cyclohexadiene imine]. A full recitation of all FD&C and D&C dyes and their corresponding chemical structures may be found in the Kirk-Othmer Encyclopedia of Chemical Technology, in Volume 5.

The confectionary may also include a volatile oil-modifying agent such as capsicum oleoresin. An oil-modifying agent is present in an amount, which is undetected as a separate ingredient in the oral cavity, but nevertheless has the ability to modify sensory perception of the volatile oil.

The oil-modifying agent is present in an amount of from about 1 to about 150 ppm of the confection. The capsicum is available from Capsicum minimum, Capsicum frutescens, Capsicum annuum, and similar varieties. Commercially, the fruits of capsicum are referred to as chilies or as peppers. These fruits are known for their extreme potency of bite, pungency and characteristic odor.

With respect to confectionery compressed tablet formulations, such will contain a tablet granulation base and various additives such as sweeteners and flavors. The tablet granulation base employed will vary depending upon factors such as the type of base used, friability desired and other components used to make the final product. These confections generally contain sugars in amounts up to 95% by weight of the composition.

The confectionery compressed tablet may additionally include tablet excipients such as binders or lubricants, as well as flavoring agents, coloring agents, and volatile oils and volatile oil-modifying agents. The variations that one may practice with regard to these confections are wide ranging and within the ability of those skilled in the art particularly with regard to the use of additional composition fillers, flavoring agents, the use of coloring agents, etc.

External oral care compositions

An external oral care formulation, e.g., denture cleaning solution, denture cleaning tablet, denture cleaning powder, and the like, may include substances and/or ingredients selected from the following categories:

Other oral care compositions

Oral compositions of the invention may also be included in filaments suitable for use in dental cleaning, e.g., filaments useful as dental floss. Preferably, the oral care composition is coated onto the exterior of the filament. Thus, in a preferred embodiment, the present invention relates to a filament comprising an oral care composition comprising a polypeptide having DNase activity and at least one oral care component, wherein the filament is suitable for dental cleaning.

Oral care compositions of the invention may also be included in an animal treat and thereby be used for improving oral health of an animal. Thus, in a preferred embodiment, the present invention relates to an oral care composition of the invention in the form of an animal treat.

Preferably, the animal treat is a pet treat. Most preferably, the animal treat is a dog treat. The oral composition may be coated onto the outer surface of the animal treat, mixed in with the other treat ingredients, or comprised in an inner compartment of the treat. Preferably, the oral care composition is comprised in an inner compartment of the treat.

Suitable types of animal treats as well as methods for making such treats are well-known to the skilled person and are described in, e.g., EP 0 258 037 A2, US 4,892,748 B2, and US 8,496,985 B2.

In an alternative aspect, the present invention relates to an animal treat comprising an oral care composition comprising a polypeptide having DNase activity and at least one oral care component.

Preferably, the animal treat is a pet treat. Most preferably, the animal treat is a dog treat.

The oral composition may be coated onto the outer surface of the animal treat, mixed in with the other treat ingredients, or comprised in an inner compartment of the treat. Preferably, the oral care composition is comprised in an inner compartment of the treat.

Treatment of oral disease

The oral care compositions of the invention are suitable for use in the treatment of oral disease, wherein degradation of eDNA and removal of biofilm is desired. The compositions of the invention are particularly suitable for treating periodontal diseases and dental caries.

Periodontal disease, also known as gum disease, is a set of inflammatory conditions caused by bacterial infection and subsequent biofilm build-up on the test and the tissues surrounding the teeth. Periodontal disease may be divided in terms of severity into the following categories: gingivitis (including plaque-induced gingivitis), chronic periodontitis, aggressive periodontitis, periodontitis as a manifestation of systemic disease, necrotizing ulcerative gingivitis/periodontitis, abscesses of the periodontium, and combined periodontic-endodontic lesions. Periodontal disease may further be considered either localized or generalized depending on the extent of the affected area.

Dental caries, also known as tooth decay or cavities, is caused by organic acids, such as lactic acid, being released by certain biofilm-forming bacteria residing in the oral cavity, including Streptococcus mutans and some Lactobacillus species. Dental caries may be associated with further complications such as inflammation of the tissue around the teeth, tooth loss, and infection or abscess formation. Dental caries may be classified by location, etiology, rate of progression, and affected hard tissues, for instance according to the G.V. Black classification (class I, II, III, IV, V, and VI).

PREFERRED EMBODIMENTS

[1] An oral care composition comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component. [2] The composition according to embodiment 1 , wherein the polypeptide having DNase activity comprises the DUF1524 domain; preferably the polypeptide having DNase activity comprises a NUC1 nuclease; most preferably the polypeptide having DNase activity comprises a member of the GYS clade.

[3] The composition according to any of embodiments 1-2, wherein the polypeptide having DNase activity comprises a NUC1 nuclease having DNase activity, wherein the polypeptide comprises one or more of the motifs [T/D/S][G/N]PQL, [F/L/Y/I]A[N/R]D[L/I/P/V], and C[D/N]T[A/R], and wherein the polypeptide is selected from the group consisting of:

i) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 15; j) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 16;

L) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 18;

u) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 27; v) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 28;

x) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 30;

ac) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 45.

[4] The composition according to any of embodiments 1-3, wherein the polypeptide having DNase activity comprises a polypeptide of the GYS clade having DNase activity, wherein the polypeptide comprises one or both of the motifs [D/M/L][S/T]GYSR[D/N] and ASXNRSKG, and wherein the polypeptide is selected from the group consisting of:

c) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 9; d) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 10;

o) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 21 ; p) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 22;

aa) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 45. [5] The composition according to embodiment 1 , wherein the polypeptide having DNase activity comprises a member of the endonuclease/exonuclease/phosphatase family; preferably the polypeptide having DNase activity comprises a NUC2 nuclease.

[6] The composition according to embodiment 5, wherein the polypeptide having DNase activity comprises a NUC2 nuclease having DNase activity, wherein the polypeptide comprises the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V], and wherein the polypeptide having DNase activity is selected from the group consisting of:

g) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 47;

h) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 48; and

i) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 49.

[7] The composition according to claim 1 , wherein the polypeptide having DNase activity is selected from the group consisting of: a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 41 ;

b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 42;

c) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 43;

d) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 44;

[8] The composition according to any of the preceding embodiments, wherein the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50; preferably the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

[9] The composition according to any of the preceding embodiments, which further comprises at least one enzyme; preferably the at least one enzyme is selected from the group consisting of dispersin, protease, lipase, carbohydrase, dextranase, oxidoreductase, laccase, peroxidase, oxidase, and lysozyme.

[10] The composition according to any of embodiments 1 -9 in the form of an internal oral care composition; preferably in the form of a toothpaste, dental cream, mouthwash, mouth rinse, lozenge, pastille, chewing gum, confectionary, or candy.

[1 1] The composition according to embodiment 10, wherein the composition is a toothpaste or dental cream and the at least one oral care component is selected from the group consisting of abrasive, solvent, humectant, detergent/surfactant, thickening/binding agent, buffering agent, flavoring agent, sweetening agent, fluoride source, therapeutic agent, coloring agent, and preservative.

[12] The composition according to embodiment 10, wherein the composition is a mouthwash or mouth rinse and the at least one oral care component is selected from the group consisting of carrier liquid, detergent/surfactant, buffering agent, flavoring agent, humectant, sweetening agent, therapeutic agent, fluoride source, coloring agent, and preservative.

[13] The composition according to any of embodiment 11 -12, wherein the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, maltitol, lactitol, polyoxyethylene, polyethylene glycol, polypropylene glycols, propylene glycol, 1 ,3-propanediol, and 1 ,4-butanediol; preferably the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, and polyethylene glycol.

[14] The composition according to any of embodiments 1 1-12, wherein the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, lactose, maltose, levulose, aspartame, cyclamate salts, D-tryptophan, dihydrochalchones, acesulphame, stevioside, levaudioside, glycyrrhizins, pellartine, thaumatin, p-methoxycinnamic aldehyde, hydrogenated starch hydrolysates, corn syrup, xylitol, sorbitol, erythritol, mannitol, and mixtures thereof; preferably the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, stevioside, xylitol, sorbitol, erythritol, mannitol, and corn syrup.

[15] The composition according to any of embodiments 11-12, wherein the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, clove bud oil, menthol, anethole, methyl salicylate, eucalyptol, cassia, 1 -inenthyl acetate, sage, eugenol, parsley oil, oxanone, alpha-irisone, marjoram, lemon, orange, cranberry, propenyl guaethol, cinnamon, vanillin, ethyl vanillin, heliotropine, 4-cis-heptenal, diacetyl, methylpara-tert-butyl phenyl acetate, carvone, cineole, menthone, cinnamic aldehyde, limonene, ocimene, n-decyl alcohol, citronellol, alpha-terpineol, methyl acetate, citronellyl acetate, methyl eugenol, linalool, thymol, rosemary oil, pimento oil, diatomaceous oil, and eucalyptus oil; preferably the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, and menthone.

[16] The composition according to any of embodiments 11-12, wherein the fluoride source is selected from the group consisting of sodium fluoride, potassium fluoride, stannous fluoride, indium fluoride, sodium monofluorophosphate, sodium heexafluorosilicate, zinc fluoride, lithium fluoride, aluminium fluoride, acidulated phosphate fluoride, ammonium bifluoride, titatium tetrafluoride, and amine fluoride; preferably the fluoride source is sodium fluoride, stannous fluoride, or sodium monofluorophosphate.

[17] The composition according to embodiment 16, which comprises a fluoride source in an effective concentration of from 0.001 % to 20% by weight; preferably from 0.01 % to 15% by weight; most preferably from 0.1% to 10% by weight. [18] The composition according to embodiment 10, wherein the composition is a chewing gum and the at least one oral care component is selected from the group consisting of elastomer, softening agent, plasticizing agent, emulsifier, wax, coloring agent, sweetening agent, flavoring agent, bulking agent, and thickening agent.

[19] The composition according to embodiment 10, wherein the composition is a lozenge or pastille and the at least one oral care component is selected from lubricant, bulking agent, sweetening agent, and flavoring agent.

[20] The composition according to embodiment 10, wherein the composition is a confectionary or candy and the at least one oral care component is selected from the group consisting of coloring agent, sweetening agent, flavoring agent, and oil-modifying agent.

[21] The composition according to any of embodiments 1 -9 in the form of an external oral care composition; preferably in the form of denture cleaning solution, denture cleaning tablet, or denture cleaning powder.

[22] The composition according to any of embodiments 1-9 in the form of an animal treat; preferably in the form of a pet treat; most preferably in the form of a dog treat.

[23] The composition according to any of embodiments 1-22 for use as a medicament.

[24] The composition according to any of embodiments 1-22 for use in the treatment of oral disease; preferably for use in the treatment of periodontal disease and/or dental caries.

[25] Use of an oral care composition comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component for treatment or prophylactic treatment of a human or animal subject, in particular for removal and/or prevention of oral biofilm.

[26] The use according to embodiment 25, wherein the polypeptide having DNase activity com prises the DUF1524 domain; preferably the polypeptide having DNase activity comprises a NUC1 nuclease; most preferably the polypeptide having DNase activity comprises a member of the GYS clade.

[27] The use according to any of embodiments 25-26, wherein the polypeptide having DNase activity comprises a NUC1 nuclease having DNase activity, wherein the polypeptide comprises one or more of the motifs [T/D/S][G/N]PQL, [F/L/Y/I]A[N/R]D[L/I/P/V], and C[D/N]T[A/R], and wherein the polypeptide is selected from the group consisting of:

[28] The use according to any of embodiments 25-27, wherein the polypeptide having DNase activity comprises a polypeptide of the GYS clade having DNase activity, wherein the polypep tide comprises one or both of the motifs [D/M/L][S/T]GYSR[D/N] and ASXNRSKG, and wherein the polypeptide is selected from the group consisting of:

f) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 12; g) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 13;

r) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 24; s) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 25;

[29] The use according to embodiment 25, wherein the polypeptide having DNase activity com prises a member of the endonuclease/exonuclease/phosphatase family; preferably the polypep tide having DNase activity comprises a NUC2 nuclease.

[30] The use according to embodiment 29, wherein the polypeptide having DNase activity comprises a NUC2 nuclease having DNase activity, wherein the polypeptide comprises the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V], and wherein the polypeptide having DNase activity is selected from the group consisting of: a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 36;

[31] The use according to claim 25, wherein the polypeptide having DNase activity is selected from the group consisting of:

a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 41 ;

b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 42; c) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 43;

[32] The composition according to any of embodiments 25-31 , wherein the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50; preferably the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

[33] The use according to any of embodiments 25-32, which further comprises at least one enzyme; preferably the at least one enzyme is selected from the group consisting of dispersin, protease, lipase, carbohydrase, dextranase, oxidoreductase, laccase, peroxidase, oxidase, and lysozyme.

[34] The use according to any of embodiments 25-33, wherein the composition is an internal oral care composition; preferably in the form of a toothpaste, dental cream, mouthwash, mouth rinse, lozenge, pastille, chewing gum, confectionary, or candy.

[35] The use according to embodiment 34, wherein the composition is a toothpaste or dental cream and the at least one oral care component is selected from the group consisting of abrasive, solvent, humectant, detergent/surfactant, thickening/binding agent, buffering agent, flavoring agent, sweetening agent, fluoride source, therapeutic agent, coloring agent, and preservative.

[36] The use according to embodiment 34, wherein the composition is a mouthwash or mouth rinse and the at least one oral care component is selected from the group consisting of carrier liquid, detergent/surfactant, buffering agent, flavoring agent, humectant, sweetening agent, therapeutic agent, fluoride source, coloring agent, and preservative.

[37] The use according to any of embodiments 35-36, wherein the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, maltitol, lactitol, polyoxyethylene, polyethylene glycol, polypropylene glycols, propylene glycol, 1 ,3-propanediol, and 1 ,4-butanediol; preferably the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, and polyethylene glycol.

[38] The use according to any of embodiments 35-36, wherein the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, lactose, maltose, levulose, aspartame, cyclamate salts, D-tryptophan, dihydrochalchones, acesulphame, stevioside, levaudioside, glycyrrhizins, pellartine, thaumatin, p-methoxycinnamic aldehyde, hydrogenated starch hydrolysates, corn syrup, xylitol, sorbitol, erythritol, mannitol, and mixtures thereof; preferably the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, stevioside, xylitol, sorbitol, erythritol, mannitol, and corn syrup.

[39] The use according to any of embodiments 35-36, wherein the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, clove bud oil, menthol, anethole, methyl salicylate, eucalyptol, cassia, 1 -inenthyl acetate, sage, eugenol, parsley oil, oxanone, alpha-irisone, marjoram, lemon, orange, cranberry, propenyl guaethol, cinnamon, van illin, ethyl vanillin, heliotropine, 4-cis-heptenal, diacetyl, methylpara-tert-butyl phenyl acetate, carvone, cineole, menthone, cinnamic aldehyde, limonene, ocimene, n-decyl alcohol, citronellol, alpha-terpineol, methyl acetate, citronellyl acetate, methyl eugenol, linalool, thymol, rosemary oil, pimento oil, diatomaceous oil, and eucalyptus oil; preferably the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, and menthone.

[40] The use according to any of embodiments 35-36, wherein the fluoride source is selected from the group consisting of sodium fluoride, potassium fluoride, stannous fluoride, indium fluoride, sodium monofluorophosphate, sodium heexafluorosilicate, zinc fluoride, lithium fluoride, aluminium fluoride, acidulated phosphate fluoride, ammonium bifluoride, titatium tetrafluoride, and amine fluoride; preferably the fluoride source is sodium fluoride, stannous fluoride, or sodium monofluorophosphate.

[41] The use according to embodiment 40, wherein the fluoride source is present in an effective concentration of from 0.001% to 20% by weight; preferably from 0.01% to 15% by weight; most preferably from 0.1 % to 10% by weight.

[42] The use according to embodiment 34, wherein the composition is a chewing gum and the at least one oral care component is selected from the group consisting of elastomer, softening agent, plasticizing agent, emulsifier, wax, coloring agent, sweetening agent, flavoring agent, bulking agent, and thickening agent. [43] The use according to embodiment 34, wherein the composition is a lozenge or pastille and the at least one oral care component is selected from lubricant, bulking agent, sweetening agent, and flavoring agent.

[44] The use according to embodiment 34, wherein the composition is a confectionary or candy and the at least one oral care component is selected from the group consisting of coloring agent, sweetening agent, flavoring agent, and oil-modifying agent.

[45] The use according to any of embodiments 25-33, wherein the composition is an external oral care composition; preferably in the form of denture cleaning solution, denture cleaning tablet, or denture cleaning powder.

[46] The use according to any of embodiments 25-33, wherein the composition is in the form of an animal treat; preferably in the form of a pet treat; most preferably in the form of a dog treat.

[47] The composition according to any of embodiments 1-22 for use as a medicament.

[48] The composition according to any of embodiments 1-22 for use in the treatment of oral disease; preferably for use in the treatment of periodontal disease, dental caries, and/or dental plaque.

[49] A method of treatment of a human or animal subject, the method comprising administering a composition comprising a polypeptide having DNase activity, a polypeptide having DNase acitivity, and at least one oral care component to a human or animal subject.

[50] The method according to embodiment 49, wherein the polypeptide having DNase activity comprises the DUF1524 domain; preferably the polypeptide having DNase activity comprises a NUC1 nuclease; most preferably the polypeptide having DNase activity comprises a member of the GYS clade.

[51] The method according to any of embodiments 49-50, wherein the polypeptide having DNase activity comprises a NUC1 nuclease having DNase activity, wherein the polypeptide comprises one or more of the motifs [T/D/S][G/N]PQL, [F/L/Y/I]A[N/R]D[L/I/P V], and C[D/N]T[A/R], and wherein the polypeptide is selected from the group consisting of:

a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 7; b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 8;

m) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 19; n) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 20;

y) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 31 ; z) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 32;

[52] The method according to any of embodiments 49-51 , wherein the polypeptide having DNase activity comprises a polypeptide of the GYS clade having DNase activity, wherein the polypep tide comprises one or both of the motifs [D/IWL][S/T]GYSR[D/N] and ASXNRSKG, and wherein the polypeptide is selected from the group consisting of:

g) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 13; h) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 14;

s) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 25; t) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 26;

[53] The method according to embodiment 49, wherein the polypeptide having DNase activity comprises a member of the endonuclease/exonuclease/phosphatase family; preferably the polypeptide having DNase activity comprises a NUC2 nuclease.

[54] The method according to embodiment 53, wherein the polypeptide having DNase activity comprises a NUC2 nuclease having DNase activity, wherein the polypeptide comprises the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V], and wherein the polypeptide having DNase activity is selected from the group consisting of:

a) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 36; b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 37;

e) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 46;

f) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 47;

g) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 48; and

h) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 49.

[55] The method according to claim 49, wherein the polypeptide having DNase activity is se lected from the group consisting of:

d) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 44; [56] The composition according to any of embodiments 49-55, wherein the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50; preferably the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

[57] The method according to any of embodiments 49-56, which further comprises at least one enzyme; preferably the at least one enzyme is selected from the group consisting of dispersin, protease, lipase, carbohydrase, dextranase, oxidoreductase, laccase, peroxidase, oxidase, and lysozyme.

[58] The method according to any of embodiments 49-57, wherein the composition is an internal oral care composition; preferably in the form of a toothpaste, dental cream, mouthwash, mouth rinse, lozenge, pastille, chewing gum, confectionary, or candy.

[59] The method according to embodiment 58, wherein the composition is a toothpaste or dental cream and the at least one oral care component is selected from the group consisting of abrasive, solvent, humectant, detergent/surfactant, thickening/binding agent, buffering agent, flavoring agent, sweetening agent, fluoride source, therapeutic agent, coloring agent, and preservative.

[60] The method according to embodiment 58, wherein the composition is a mouthwash or mouth rinse and the at least one oral care component is selected from the group consisting of carrier liquid, detergent/surfactant, buffering agent, flavoring agent, humectant, sweetening agent, therapeutic agent, fluoride source, coloring agent, and preservative.

[61] The method according to any of embodiments 59-60, wherein the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, maltitol, lactitol, polyoxyethylene, polyethylene glycol, polypropylene glycols, propylene glycol, 1 ,3-propanediol, and 1 ,4-butanediol; preferably the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, and polyethylene glycol.

[62] The method according to any of embodiments 59-60, wherein the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, lactose, maltose, levulose, aspartame, cyclamate salts, D-tryptophan, dihydrochalchones, acesulphame, stevioside, levaudioside, glycyrrhizins, pellartine, thaumatin, p-methoxycinnamic aldehyde, hydrogenated starch hydrolysates, corn syrup, xylitol, sorbitol, erythritol, mannitol, and mixtures thereof; preferably the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, stevioside, xylitol, sorbitol, erythritol, mannitol, and corn syrup.

[63] The method according to any of embodiments 59-60, wherein the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, clove bud oil, menthol, anethole, methyl salicylate, eucalyptol, cassia, 1 -inenthyl acetate, sage, eugenol, parsley oil, oxanone, alpha-irisone, marjoram, lemon, orange, cranberry, propenyl guaethol, cinnamon, van illin, ethyl vanillin, heliotropine, 4-cis-heptenal, diacetyl, methylpara-tert-butyl phenyl acetate, carvone, cineole, menthone, cinnamic aldehyde, limonene, ocimene, n-decyl alcohol, citronellol, alpha-terpineol, methyl acetate, citronellyl acetate, methyl eugenol, linalool, thymol, rosemary oil, pimento oil, diatomaceous oil, and eucalyptus oil; preferably the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, and menthone.

[64] The method according to any of embodiments 59-60, wherein the fluoride source is selected from the group consisting of sodium fluoride, potassium fluoride, stannous fluoride, indium fluoride, sodium monofluorophosphate, sodium heexafluorosilicate, zinc fluoride, lithium fluoride, aluminium fluoride, acidulated phosphate fluoride, ammonium bifluoride, titatium tetrafluoride, and amine fluoride; preferably the fluoride source is sodium fluoride, stannous fluoride, or sodium monofluorophosphate.

[65] The method according to embodiment 64, wherein the fluoride source is present in an effective concentration of from 0.001 % to 20% by weight; preferably from 0.01 % to 15% by weight; most preferably from 0.1% to 10% by weight.

[66] The method according to embodiment 58, wherein the composition is a chewing gum and the at least one oral care component is selected from the group consisting of elastomer, softening agent, plasticizing agent, emulsifier, wax, coloring agent, sweetening agent, flavoring agent, bulking agent, and thickening agent.

[67] The method according to embodiment 58, wherein the composition is a lozenge or pastille and the at least one oral care component is selected from lubricant, bulking agent, sweetening agent, and flavoring agent.

[68] The method according to embodiment 58, wherein the composition is a confectionary or candy and the at least one oral care component is selected from the group consisting of coloring agent, sweetening agent, flavoring agent, and oil-modifying agent. [69] The method according to any of embodiments 49-57, wherein the composition is an external oral care composition; preferably in the form of denture cleaning solution, denture cleaning tablet, or denture cleaning powder.

[70] The method according to any of embodiments 49-57, wherein the composition is an animal treat; preferably in the form of a pet treat; most preferably in the form of a dog treat.

[71] A kit of parts comprising:

a) a composition comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component; and

b) instructions for use.

[72] The kit of parts according to embodiment 71 , wherein the polypeptide having DNase activity comprises the DUF1524 domain; preferably the polypeptide having DNase activity comprises a NUC1 nuclease; most preferably the polypeptide having DNase activity comprises a member of the GYS clade.

[73] The kit of parts according to any of embodiments 71 -72, wherein the polypeptide having DNase activity comprises a NUC1 nuclease having DNase activity, wherein the polypeptide comprises one or more of the motifs [T/D/S][G/N]PQL, [F/L/Y/I]A[N/R]D[L/I/P/V], and C[D/N]T[A/R], and wherein the polypeptide is selected from the group consisting of:

e) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 1 1 ; f) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 12;

q) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 23; r) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 24;

ac) polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 45. [74] The kit of parts according to any of embodiments 71 -73, wherein the polypeptide having DNase activity comprises a polypeptide of the GYS clade having DNase activity, wherein the polypeptide comprises one or both of the motifs [D/M/L][S/T]GYSR[D/N] and ASXNRSKG, and wherein the polypeptide is selected from the group consisting of:

[75] The kit of parts according to embodiment 71 , wherein the polypeptide having DNase activity comprises a member of the endonuclease/exonuclease/phosphatase family; preferably the pol ypeptide having DNase activity comprises a NUC2 nuclease.

[76] The kit of parts according to embodiment 75, wherein the polypeptide having DNase activity comprises a NUC2 nuclease having DNase activity, wherein the polypeptide comprises the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V], and wherein the polypeptide having DNase activity is selected from the group consisting of:

e) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 40; f) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 46;

[77] The kit of parts according to claim 71 , wherein the polypeptide having DNase activity is selected from the group consisting of:

[78] The composition according to any of embodiments 71-77, wherein the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50; preferably the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

[79] The kit of parts according to any of embodiments 71-78, which further comprises at least one enzyme; preferably the at least one enzyme is selected from the group consisting of dispersin, protease, lipase, carbohydrase, dextranase, oxidoreductase, laccase, peroxidase, oxidase, and lysozyme.

[80] The kit of parts according to any of embodiments 71 -79, wherein the composition is an internal oral care composition; preferably in the form of a toothpaste, dental cream, mouthwash, mouth rinse, lozenge, pastille, chewing gum, confectionary, or candy.

[81] The kit of parts according to embodiment 80, wherein the composition is a toothpaste or dental cream and the at least one oral care component is selected from the group consisting of abrasive, solvent, humectant, detergent/surfactant, thickening/binding agent, buffering agent, flavoring agent, sweetening agent, fluoride source, therapeutic agent, coloring agent, and preservative.

[82] The kit of parts according to embodiment 80, wherein the composition is a mouthwash or mouth rinse and the at least one oral care component is selected from the group consisting of carrier liquid, detergent/surfactant, buffering agent, flavoring agent, humectant, sweetening agent, therapeutic agent, fluoride source, coloring agent, and preservative.

[83] The kit of parts according to any of embodiments 81 -82, wherein the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, maltitol, lactitol, polyoxyethylene, polyethylene glycol, polypropylene glycols, propylene glycol, 1 ,3-propanediol, and 1 ,4-butanediol; preferably the humectant is selected from the group consisting of glycerol, sorbitol, xylitol, and polyethylene glycol.

[84] The kit of parts according to any of embodiments 81 -82, wherein the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, lactose, maltose, levulose, aspartame, cyclamate salts, D-tryptophan, dihydrochalchones, acesulphame, stevioside, levaudioside, glycyrrhizins, pellartine, thaumatin, p-methoxycinnamic aldehyde, hydrogenated starch hydrolysates, corn syrup, xylitol, sorbitol, erythritol, mannitol, and mixtures thereof; preferably the sweetening agent is selected from the group consisting of saccharin, dextrose, sucrose, stevioside, xylitol, sorbitol, erythritol, mannitol, and corn syrup.

[85] The kit of parts according to any of embodiments 81-82, wherein the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, clove bud oil, menthol, anethole, methyl salicylate, eucalyptol, cassia, 1 -inenthyl acetate, sage, eugenol, parsley oil, oxanone, alpha-irisone, marjoram, lemon, orange, cranberry, propenyl guaethol, cinnamon, vanillin, ethyl vanillin, heliotropine, 4-cis-heptenal, diacetyl, methylpara-tert-butyl phenyl acetate, carvone, cineole, menthone, cinnamic aldehyde, limonene, ocimene, n-decyl alcohol, citronellol, alpha-terpineol, methyl acetate, citronellyl acetate, methyl eugenol, linalool, thymol, rosemary oil, pimento oil, diatomaceous oil, and eucalyptus oil; preferably the flavoring agent is selected from the group consisting of wintergreen oil, peppermint oil, spearmint oil, and men- thone.

[86] The kit of parts according to any of embodiments 801-82, wherein the fluoride source is selected from the group consisting of sodium fluoride, potassium fluoride, stannous fluoride, indium fluoride, sodium monofluorophosphate, sodium heexafluorosilicate, zinc fluoride, lithium fluoride, aluminium fluoride, acidulated phosphate fluoride, ammonium bifluoride, titatium tetrafluoride, and amine fluoride; preferably the fluoride source is sodium fluoride, stannous fluoride, or sodium monofluorophosphate.

[87] The kit of parts according to embodiment 86, wherein the fluoride source is present in an effective concentration of from 0.001 % to 20% by weight; preferably from 0.01 % to 15% by weight; most preferably from 0.1% to 10% by weight.

[88] The kit of parts according to embodiment 80, wherein the composition is a chewing gum and the at least one oral care component is selected from the group consisting of elastomer, softening agent, plasticizing agent, emulsifier, wax, coloring agent, sweetening agent, flavoring agent, bulking agent, and thickening agent.

[89] The kit of parts according to embodiment 80, wherein the composition is a lozenge or pastille and the at least one oral care component is selected from lubricant, bulking agent, sweetening agent, and flavoring agent.

[90] The kit of parts according to embodiment 80, wherein the composition is a confectionary or candy and the at least one oral care component is selected from the group consisting of coloring agent, sweetening agent, flavoring agent, and oil-modifying agent.

[91] The kit of parts according to any of embodiments 71 -79, wherein the composition is an external oral care composition; preferably in the form of denture cleaning solution, denture cleaning tablet, or denture cleaning powder.

[92] The kit of parts according to any of embodiments 71 -79, wherein the composition is an animal treat; preferably in the form of a pet treat; most preferably in the form of a dog treat. EXAMPLES

Assays

Assay I: testing of DNase activity

DNase activity was determined on DNase Test Agar with Methyl Green (BD, Franklin Lakes, NJ, USA), which was prepared according to the manual from supplier. Briefly, 21 g of agar was dissolved in 500 ml water and then autoclaved for 15 min at 121 °C. Autoclaved agar was temperated to 48°C in water bath, and 20 ml of agar was poured into petridishes with and allowed to solidify by incubation o/n at room temperature. On solidified agar plates, 5 pi of enzyme solutions are added and DNase activity is observed as colorless zones around the spotted enzyme solutions.

Assay II: testing of DNase activity

DNase activity was determined by using the DNaseAlert Kit (1 1 -02-01-04, IDT Integrated DNA Technologies) according to the supplier’s manual. Briefly, 95 mI DNase sample was mixed with 5 mI substrate in a microtiter plate, and fluorescence was immediately measured using a Clariostar microtiter reader from BMG Labtech (536 nm excitation, 556 nm emission).

Thermal Stability

Methods

Preparation of oral care formulations for thermal stability measurements

Plax COOL MINT (Colgate®, Colgate-Palmolive) mouthwash was used at two concentrations: 45 and 90% (v/v) with 10% of the volume originating from the protein in buffer and the balance of the volume made up with MilliQ water.

Cavity Protection Caries (Colgate®, Colgate-Palmolive) toothpaste was used at two concentrations: 1 and 10% (w/v) in MilliQ water with 10% of the volume originating from the protein in buffer. The toothpaste stock solution was made by resuspending 2.2 or 22 g of toothpaste in 200 ml MilliQ water and mixing the suspensions for 15 min with stirring at 1000 rpm using a magnetic stirrer at room temperature. The insoluble material was removed by filtering the stock solutions through a 0.2 pm asymmetric Polyethersulfone (aPES) membrane (Thermo Scientific) under vacuum. This was necessary as the thermal stability was measured using a spectrophotometric method and insoluble particles will scatter/obscure the light rendering the method ineffective.

Biotechnology grade Sodium Dodecyl Sulfonate (SDS) from Amresco, Ohio, USA, is supplied as a 20% (w/v) solution. SDS concentrations of 2 and 5% (w/v) were used at pH 7 (50mM MES, 50mM Glycine, 50mM Sodium acetate) with 10% of the volume originating from the protein in buffer. Proteomics grade EDTA disodium salt dihydrate from Amresco, Ohio, USA, was used to make a 100 mM stock solution with MilliQ water. The stock was diluted to 1 or 5 mM using MilliQ water with 10% of the volume originating from the protein in buffer.

Protein samples were diluted to a concentration of 1 mg/ml using 50 mM HEPES, 150 mM NaCI, pH 7 prior to a further 10 times dilution in the oral care formulations and corresponds to a final protein concentration of 0.1 mg/ml.

All dilutions were made in a 384 well small volume deep well plate (Greiner Bio-One International, item number 784201 ) with a final volume of 50 pi and used for thermal stability measurements.

Thermal stability in oral care formulations

Thermal stability measurements were performed using a capillary-based nano differential scanning fluorescence instrument (nanoDSF); Prometheus NT.Plex (NanoTemper Technologies GmbH, Munchen, Germany). Standard nanoDSF grade capillary chips were used (Cat#: PR-AC002) from NanoTemper Technologies.

The protein samples were loaded into the capillaries (each sample in triplicate) by capillary action. The emission intensities at 330 and 350 nm were optimized by altering the LED power on the instrument to ensure sufficient signal. The fluorescence signals at 330 and 350 nm were monitored continuously as a function of temperature (heating rate used for thermal unfolding was 3.3°C per minute from 20 to 95°C).

The data was analyzed using the PR.ThermControl_2.1.2.6031 software provided by the manufacturer. The analysis is model independent and simply takes the peak maximum of the first derivative which corresponds to the approximate thermal unfolding transition midpoint, defined as Td (see Fig. 1 ).

Example 1 : Robustness of thermal stability data

Fig. 1 shows an example of the thermal stability data generated using the nanoDSF instrument. Panel A is an example of the data obtained (the ratio of the fluorescence emission at 350 nm to 330 nm) in triplicate for SEQ ID NO: 5 (for clarity only every fifth data point is shown) as a function of temperature. Panel B shows the first derivative of the raw data in Panel A. The peak maximum in the first derivative plot corresponds to the mid-point of the thermal unfolding transition, referred to as Td. In this example the Td corresponds to 58°C and is highly reproducible within the three replicates.

The data shown in Fig. 1 is an example of the type of data that was generated for the nucleases in the different formulations using nanoDSF. In all cases, the data showed a clear unfolding transition, and a clearly defined peak in the first derivative, and were highly reproducible. The thermal stabilities of the nucleases were measured in the single components commonly used in oral care product formulations and selected commercial products. The Td parameter was used to evaluate the thermal stabilities as this is the temperature at which there are equal populations of folded and unfolded protein molecules and is the widely accepted parameter to use when evaluating thermal stability.

Example 2: Thermal stability of DNases in EDTA

Fig. 2 shows the average thermal stabilities measured in the presence of 1 or 5 mM EDTA. EDTA is a strong chelator, and DNases are known to bind divalent cations as they are required as cofactors for activity. In addition, the removal of the divalent cations could lead to a loss in thermal stability as bound ligands generally increase protein thermal stability.

In all cases, there is no significant difference in thermal stability at the different EDTA concentrations which suggests that the divalent cations have been removed at 1 mM EDTA and that there will be no further destabilization of the DNAses if higher levels of EDTA are used. In addition, EDTA is one of the strongest chelators known, so if other weaker chelators are used, their effect would be less than that of EDTA.

It is clear from the data in Fig. 2 that the DNases have reasonable thermal stabilities in the presence of EDTA and in some cases the thermal stability is pronounced. This shows that the DNases will not be unfolded at higher temperatures in the presence of EDTA or other chelators.

Example 3: Thermal stability of DNases in SDS

SDS is one of the most widely used surfactants in oral care formulations as it allows for a more uniform distribution of product and increases the cleaning power. SDS is known to be a strong denaturant of proteins and can unfold proteins completely at elevated temperatures, rendering them inactive. In addition, proteins unfolded by SDS would not regain their activities when the temperature is decreased.

Despite the harsh effects of SDS, Fig. 3 shows that the DNAses exhibit good thermal stability at both high and low SDS concentrations. Overall, the average Td for the DNases is approximately 58°C which is the lowest average for the conditions tested, which can be expected due to the harsh nature of SDS, but it is still relatively high as one would not expect an oral care product comprising a composition of the present invention to be exposed to, or stored at, such high temperatures.

Example 4: Thermal stability of DNases in Plax COOL MINT (Colgate ®) mouthwash.

To test the thermal stabilities of DNases in mouthwash, it was decided use concentrations of mouthwash that would closely resemble concentrations the DNases would encounter in such formulations.

Fig. 4 shows the thermal stabilities of the DNases in 45 or 90% (w/v) concentrations of Plax COOL MINT (Colgate ®) mouthwash. All DNases are stable under these conditions, and there is very little difference in the thermal stabilities at either 45 or 90% (w/v) of mouthwash. Example 5: Thermal stability of DNase in Cavity Protection Caries (Colgate®) toothpaste

The thermal stabilities of DNases in Cavity Protection Caries (Colgate ®) toothpaste (Fig.

5) overall show a relatively high thermal stability with an average Td of all DNases at both 1 and 10% (w/v) toothpaste concentrations of approximately 66°C, which is the highest average Td for all formulations tested.

Conclusion on thermal stability experiments

Taken together, the data presented in Example 5 (thermal stability in toothpaste), Example 3 (thermal stability in SDS) and Example 2 (thermal stability in EDTA) clearly shows that the evaluated DNases have sufficiently high thermal stabilities in these formulations to be used in toothpaste formulations/dental gels containing EDTA and/or formulations with high SDS concentrations.

Since Example 4 (thermal stability in mouthwash) illustrates that it was indeed possible to measure the thermal stabilities of DNases in a mouthwash formulation that very closely resembles a final mouthwash product, it is evident that DNases exhibit sufficiently high thermal stability in order to be used in mouthwash.

Biofilm Removal and Prevention

Materials

Phosphate buffered saline (PBS): Sigma Aldrich catalog number P4417.

Glycerol (99%): Sigma Aldrich catalog number G5516.

Brain Heart Infusion (BHI) broth: Sigma (Merck) catalog number 53286.

1 % Crystal Violet solution: Sigma (Merck) catalog number 548-62-9.

Glacial acetic acid: Sigma (catalog number: A6283).

Nunc MicroWell Plates with Nunclon Delta Surface (sterile): ThermoFisher Scientific (catalog number 163320).

Artificial tap water was made as follows:

1 ) Add 0.145 g CaCI₂ to 1 ml milli-Q water to form 0.713 M CaCI₂.

2) Add 0.0524 g MgCI₂ to 1 ml milli-Q water to form 0.357 M MgCI₂.

3) Add 0.04503 g NaHC0₃ to 1 ml milli-Q water to form 0.536 M NaHC0₃.

4) Add 750 pi 0.713 M CaCI₂, 375 pi 0.357 M MgCI₂ and 1875 mI 0.536 M NaHCOs to 250 ml Milli-Q water followed by sterile filtration.

Saliva collection, preparation and storage

Saliva was collected from 6 different volunteers in sterile tubes. Saliva production was stimulated by chewing paraffin tablets (Ivoclar Vivadent AG). To protect the bacterial cells from lysis upon freezing, 99% Glycerol (Sigma Aldrich) was diluted 1 :1 with phosphate buffered saline (PBS, Sigma Aldrich), and was used to dilute the pooled saliva 1 :1 before storage at -20 °C. The effect of enzymatic treatment on the in vitro biofilm model was measured in two different assays. The first was the prevention of biofilm formation/growth by the addition of enzyme(s) and the second was to measure the extent of established biofilm removal by the addition of enzyme(s).

Prevention of oral biofilm growth in vitro

To grow the biofilms in the presence of enzymes, sterile 96 well-plates were used (ThermoFisher Scientific). To each well, 20 pi non-sterilized saliva (inoculum), 20 pi sterilized saliva (0.22 pm filtered), 16 pi sterile 50% sucrose and 50 pi sterile BHI broth was added. To test the effect of enzyme(s) on the prevention of oral biofilm formation, 20 pi of enzyme(s) were added to yield a final concentration of 20 ppm. In the control experiment, enzyme was replaced with artificial tap water. To each well, 74 pi of artificial tap water was added to reach a total volume of 200 pL.

The plates were incubated for 24 h at 37 °C and 50 rpm after which the liquid was removed from the wells and the plates were dried for 1 hour at 37 °C and 50 rpm.

After drying, 200 pi 1 % Crystal Violet solution was added to each well and incubated for 10 min at room temperature. After removing the Crystal Violet solution, unbound dye was washed away by adding 200 pi deionized water and removing the wash solution (this step was performed 3 times).

After staining, the biofilms were dried for 30 min at 37 °C and 50 rpm. After drying, 200 pi 33% acetic acid was added to each well to extract the dye (for 30 min). After dye extraction, each sample was diluted 4 times with 33% acetic acid followed by measuring the absorbance at 590 nm using a BioTek PowerWave X52 plate reader.

For the data processing, the absorbance was taken to be proportional to the extent of biofilm growth. The biofilm prevention percentage was calculated as follows:

100-((A590nm enzyme treated sample)/(A590nm untreated sample)),

where A590nm refers to the measured absorbance at 590 nm.

Removal of oral biofilm in vitro

To grow the oral biofilm, 20 pi non-sterilized saliva was added to each well, along with 20 pi sterilized saliva, 16 pi sterile 50% sucrose, sterile 50 pi BHI broth and 94 sterile pi deionized water.

The plates were incubated for 24 h at 37 °C and 50 rpm after which the liquid was removed and replaced by 200 pi of artificial tap water containing 20 ppm enzyme(s), or artificial tap water only without enzyme as a control, and the plates were incubated for 2 hr at 37 °C and 50 rpm. The enzyme/control solutions were removed and the biofilms washed three times by adding and removing 200 pL artificial tap water. The plates were dried for 1 hr at 37 °C and 50 rpm. After drying, 200 mI Crystal Violet solution was added to each well and incubated for 10 min at room temperature. After removing the Crystal Violet solution, unbound dye was washed away by adding 200 pi deionized water and removing the wash solution (this step was performed 3 times).

After staining, the biofilms were dried for 30 min at 37 °C and 50 rpm. After drying, 200 mI 33% acetic acid was added to each well to extract the dye (for 30 min). After dye extraction, each sample was diluted 4 times with 33% acetic acid followed by measuring the absorbance at 590 nm using a BioTek PowerWave X52 plate reader.

The absorbance was measured at 590 nm using a BioTek PowerWave X52 plate reader. For the data processing, the absorbance was taken to be proportional to the extent of remaining biofilm. The biofilm removal percentage was calculated as follows:

100-((A590nm enzyme treated sample)/(A590nm untreated sample)),

where A590nm refers to the measured absorbance at 590 nm.

Removal of oral biofilm ex vivo

Experimental setup

Ex vivo biofilms are grown on glass slabs in the oral cavity of healthy volunteers. The glass slabs are 4x4 mm and have a roughness approximating that of the dental surface. The glass slabs are placed in custom-made splints, which the volunteers can wear. In this study, we recruited nine volunteers to wear the splints. The patient wears the splint for 48 hours and dips the splint in a 4% sucrose solution three times a day. The test person removes the splint during eating. Seven slabs can fit on each side of the splint, leaving 14 samples in total. One sample in each side is a control sample, so that 12 enzyme formulations can be tested in each experiment.

Each person wore the splint over three runs, allowing us to test 36 different enzyme formulations on each person. One test person only had space for six slabs in each side, necessitating that person to wear the splint four times.

A large variation may exist between the amount of biofilm on each slab, depending on their location in the mouth. Therefore, the enzyme formulations were randomized with respect to the experiment run number and samples position in the mouth, to prevent any bias from being introduced, due to variation. Likewise, the position of the control in each side of the mouth was randomized. After 48 hours, the slabs were removed from the splint and stored in PBS until enzyme treatment.

Dental biofilm enzyme treatment

Slabs are added to round-bottomed Nunclon microtiter plates with the biofilm facing up. 200 ul of the desired enzyme solution is added. The plates are incubated for 2 hr with enzyme at 37 °C and 50 rpm. Subsequently the biofilms are washed three times with artificial tap water with drying on KimWipes (Sigma/Merck) between each washing. The treated biofilms were fixed with 4% paraformaldehyde for 2 hr at room temperature. Subsequently, the slabs are washed as previously, but in PBS. The slabs are stored in 50% ethanol/50% PBS at -20 °C until analysis.

Dental biofilm data collection

The majority of the liquid on the glass slabs was removed with KimWipes (Sigma/Merck), and the slabs were placed biofilm-down in glass bottom microtiter plates for microscopy, where 50 pi 20 mM Syto12 (ThermoFisher Scientific, S7574) has been placed.

An LSM 700 Confocal Microscope (Zeiss) is used to analyze the samples. A40X objective is used to scan a 160 pm x 160 pm area, with a 488 nm laser at 4% intensity. The images were obtained with the following settings: Detector gain: 620; Detector offset: -15; Detector digital gain: 1.2. The pinhole is set to allow a 1 pm thick scanning layer.

The center of the glass slab is found in the microscopy with the Stage window. All images are taken as 3-layered z-stacks with the first and last stacks at the top and bottom of the biofilm respectively. The center of the glass slabs is used as the reference point, and z-stacks are also collected at all the 8 positions that are located at ±1 mm in both the z and y direction from the center. This results in 9 z-stacks of varying height in the central 1 mm2 of the glass slab. The stacks are saved as czi files and the biovolumes are analyzed in the data treatment.

Data treatment

The czi files are batch-converted to tif files with the ZEN 3.0 Blue software (Zeiss). To each czi files, an XML data file is also created, containing the metadata, including the stack height.

The biovolumes of each tif file is quantified using the Daime software (Daime, version 2.1 ). After the tif files have been batch imported, the scale bar is set to (160x160x1 ). The images are then segmented to define areas with bacteria. 2D Segmentation is used and the threshold set to: Low: 45 and High: 255. The individual objects on each picture are merged, and the biovolume is measured and exported as a text file. The object editor is used to identify any pictures with no objects, and these pictures are noted. The biovolumes can be opened in Excel. The height information is obtained by batch opening the XLM files in Excel’s Query function, and selectively showing the height information in the same column. The height column in the copied into the dataset next to the quantified biovolumes. The height is divided by three, and this value is multiplied with the 2D biovolumes from Daime to achieve the actual 3D biovolumes of each slice. The biovolumes of the three slices are summed to give the total biovolume of the scan area. The average biovolumes are calculated from the nine stacks taken from the same sample. The average biovolume of each sample is normalized to the control sample from the same side of the mouth. The sample batches are discarded if the controls do not live up to the two following criteria: (1 ) the control biovolumes have to exceed 50,000 pm³, and (2) there cannot be more than a 2x difference between the controls from each side of the mouth. An average is calculated of the normalized retained biovolumes of all samples treated with the same enzyme formulation. Example 6: removal of oral biofilm in vitro

A selection of DNases and mutanases was assessed alone and in combination with re spect to removal of oral biofilm using the in vitro procedures described above. The results are indicated in the table below. As can be seen, the combinations DNase1+Mutanase1 and DNase2+Mutanase1 provides a synergistic effect with respect to biofilm removal.

Example 7: removal of oral biofilm ex vivo

A combination of a DNase and a mutanases was assessed with respect to removal of oral biofilm using the ex vivo procedures described above. The results are indicated in the table below. As can be seen, the combination DNase3+Mutanase1 provides a very potent effect with respect to biofilm removal in a setting closely mimicking the oral cavity

Claims

1. An oral care composition comprising a polypeptide having DNase activity, a polypeptide having mutanase activity, and at least one oral care component

2. The composition according to claim 1 , wherein the polypeptide having DNase activity comprises the DUF1524 domain; preferably the polypeptide having DNase activity comprises a NUC1 nuclease; most preferably the polypeptide having DNase activity comprises a member of the GYS clade.

3. The composition according to any of claims 1 -2, wherein the polypeptide having DNase ac tivity comprises a NUC1 nuclease having DNase activity, wherein the polypeptide comprises one or more of the motifs [T/D/S][G/N]PQL, [F/L/Y/I]A[N/R]D[L/I/P/V], and C[D/N]T[A/R], and wherein the polypeptide is selected from the group consisting of:

e) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 11 ;

h) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 14; i) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 15;

t) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 26; u) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 27;

4. The composition according to any of claims 1-3, wherein the polypeptide having DNase ac tivity comprises a polypeptide of the GYS clade having DNase activity, wherein the polypeptide comprises one or both of the motifs [D/M/L][S/T]GYSR[D/N] and ASXNRSKG, and wherein the polypeptide is selected from the group consisting of:

b) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 8; c) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 9;

n) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 20; o) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 21 ;

z) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 32; and aa) a polypeptide having at least 60%, at least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% sequence identity to the polypeptide shown in SEQ ID NO: 45.

5. The composition according to claim 1 , wherein the polypeptide having DNase activity comprises a member of the endonuclease/exonuclease/phosphatase family; preferably the polypeptide having DNase activity comprises a NUC2 nuclease.

6. The composition according to claim 5, wherein the polypeptide having DNase activity comprises a NUC2 nuclease having DNase activity, wherein the polypeptide comprises the motif [G/Y/W/F/A/H]NI[R/Q/D/E/V], and wherein the polypeptide having DNase activity is selected from the group consisting of:

7. The composition according to claim 1 , wherein the polypeptide having DNase activity is se lected from the group consisting of:

8. The composition according to any of the preceding claims, wherein the polypeptide having mutanase activity comprises a polypeptide having a sequence identity of at least 70%, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, to SEQ ID NO: 50; preferably the polypeptide having mutanase activity comprises, consists essentially of, or consists of SEQ ID NO: 50.

9. The composition according to any of the preceding claims, which further comprises at least one enzyme; preferably the at least one enzyme is selected from the group consisting of dispersin, protease, lipase, carbohydrase, dextranase, oxidoreductase, laccase, peroxidase, oxidase, and lysozyme.

10. The composition according to any of claims 1-9 in the form of an internal oral care composition; preferably in the form of a toothpaste, dental cream, mouthwash, mouth rinse, lozenge, pastille, chewing gum, confectionary, or candy.

1 1. The composition according to any of claims 1-9 in the form of an external oral care composition; preferably in the form of denture cleaning solution, denture cleaning tablet, or denture cleaning powder.

12. The composition according to any of claims 1-11 for use as a medicament.

13. The composition according to any of claims 1-1 1 for use in the treatment of oral disease; preferably for use in the treatment of periodontal disease and/or dental caries.

14. Use of a composition according to any of claims 1-11 for treatment or prophylactic treatment of a human or animal subject, in particular for removal and/or prevention of oral biofilm.

15. A method of treatment of a human or animal subject, the method comprising administering a composition according to any of claims 1-12 to a human or animal subject.

16. A kit of parts comprising:

a) a composition according to any of claims 1-11 ; and

b) instructions for use.