Methods and compositions for degrading deoxynivalenol

Abstract

The present invention relates to means and methods for degrading DON and / or DON derivatives comprising a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence set forth in SEQ ID NO: 1.

Description

[0001] The instant application contains a Sequence Listing in computer readable form which is incorporated herein by reference. TECHNICAL FIELD

[0002] The present invention relates to means and methods for degrading DON and / or DON derivatives, comprising a polypeptide having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1. BACKGROUND

[0003] Mycotoxins are secondary metabolites produced by filamentous fungi. These fungi grow, inter alia, on various types of grains and cereals. Typically, crops are infested with fungi before harvest and mycotoxins are produced before and after harvest and during storage. According to the Food and Agriculture Organization, about 25% of agricultural products are contaminated with mycotoxins (e.g. Eskola et al. (2020), Crit. Rev. Food Sci. Nutr. 60(16), 2773-2789), which causes considerable economic losses. Of 19,757 samples analyzed from January 2004 to December 2011, 72% of the samples tested positive for at least one mycotoxin, and 39% were co-contaminated (Schatzmayr and Streit. 2013. World Mycotoxin Journal 6(3):213-222). Mycotoxins are known to cause serious adverse health effects in humans and animals, including mutagenic, carcinogenic, neurotoxic, immunosuppressive or teratogenic effects. One particularly large class of mycotoxins are trichothecene or trichothecenes. Trichothecenes are a class of sesquiterpenes produced, inter alia, by Fusarium, Myrothecium, Podostroma, Trichoderma or Trichothecium species and can comprise a 12,13-epoxy ring (which can be referred to herein interchangeably as “12,13-epoxytrichothec-9-ene” or “trichothecene- ring”) as a common structural feature.

[0004] The trichothecene mycotoxin family comprises, inter alia, deoxynivalenol (DON, CAS no. 51481-10-8), T-2 toxin (CAS no. 21259-20-1), HT-2 toxin (CAS no. 26934-87-2), nivalenol (CAS no. 23282-20-4), fuseranon-X (CAS no. 23255-69-8), scripentriol, 15-acetoxyscirpenol (CAS no. 2623-22-5), 4,15-diacetoxyscirpenol (CAS no. 2270-40-8), trichodermol (CAS no. 2198-93-8), verrucarin A (CAS no. 3148-09-2), verrucarin J (CAS no. 4643-58-7), isotrichodermin (CAS no. 91423-90-4), hydroxyisotrichodermin (CAS no. 344781-02-8), calonectrin (CAS no. 38818-51-8), T-2 tetrol (CAS no. 34114-99-3), deacetylneosolaniol (CAS no. 74833-39-9), neosolaniol (CAS no. 36519-25-2), acetylneosolaniol (CAS no. 65041-92-1), sporotrichiol (CAS no. 101401-89-2) and trichotriol (CAS no. 109890-37-1).

[0005] B-type trichothecenes as referred to herein can refer to trichothecene compounds having a carbonyl group (e.g. C=0) at position 8 of the carbon chain (according to the atom numbering of Yoshizawa and Morooka (1973, Agric. Biol. Chem. 37, 2933-2934)) (e.g. see below) (or in other words, having a double bond to an oxygen atom (=0) on the C8 atom of the carbon chain) and can include DON-derivatives. DON-derivatives as referred to herein can refer to trichothecene compounds having a carbonyl group (e.g. C=0) at position 8 of the carbon chain (or in other words, having a double bond to an oxygen atom (=0) on the C8 atom of the carbon chain) and having a hydroxyl group (-OH) at the C7 atom of the carbon chain and having an unsubstituted C10 (e.g. having a hydrogen (-H) bound thereto, e.g. see below for the DON chemical structure). Exemplary DON-derivatives as referred to herein can include:

[0006]

[0007]

[0008] DON-derivatives as referred to herein do not include 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxytrichothec-9-en-8-one.

[0009] In animals, the consumption of trichothecene mycotoxins leads to reduced feed intake, impaired nutrient absorption and growth, vomiting, diarrhea, immune dysfunction and reduced milk production. In humans, adverse health effects include nausea, vomiting, diarrhea, abdominal pain, headache and fever. Especially DON (chemical name e.g. (3a,7a)-3,7,15-trihydroxy-12,13-epoxytrichothec-9-en-8-one) crop contamination can be found worldwide and several additional toxic DON derivatives have been described, e.g. acetylated DON (like 15-acetyl-DON, 15-ADON), glycosylated DON, DON sulfonates (like DONS-1 or DONS-2) or DON sulfate (like DON-15-sulfate). Therefore, national and regional authorities have established maximum levels of DON in food (EC no. 1881 / 2006, EC no. 1126 / 2007) or feed (2006 / 576 / EC) to avoid mycotoxin poisoning in humans and animals after ingestion of DON, leading to adverse effects like inhibition of protein biosynthesis, interaction with serotonin and dopamine receptors or upregulation of pro-inflammatory cytokines (EFSA Journal 2004, 73, 1-41).

[0010] The risk of some mycotoxin contamination can be reduced by implementing "good agricultural practices". Despite these preventive measures, DON contamination cannot be avoided and therefore additional strategies are needed to avoid DON mycotoxin intoxication. One feasible strategy to prevent mycotoxin intoxication is the addition of binders such as clay, yeast or yeast products to feed and / or food. However, due to the hydrophilic nature of DON, it does not interact with the commonly employed binders and thus cannot be removed by this strategy. Similarly, DON cannot be removed from food or feed by chemical or physical treatment. As an alternative strategy to avoid mycotoxin intoxication, DON can be detoxified by conversion into less toxic molecules.

[0011] EP 1042449 A1 or US 2012 / 0263827 A1 relate to microorganisms capable of detoxifying trichothecenes by cleaving the epoxide ring on the C12 and C13 atoms, respectively, or by bioconversion into 3-epi-DON and 3-keto-DON. Detoxification strategies of DON based on the C3 atom using microorganisms are known in the art. Generally, the use of such microorganisms is often challenged by the specific cultivation requirements of these microorganisms, making their production economically disadvantageous. Furthermore, the mixing of microorganisms into feed or food is not feasible in many food or feed processing processes. Therefore, there is a need for a rapid and safe means of DON detoxification by DON bioconversion using enzyme preparations free of microorganisms. In this regard, WO 2016 / 154640 A1 describes an alcohol dehydrogenase containing a metal ion and a quinone cofactor for the conversion of trichothecenes having a hydroxyl group at the C3 atom. CN 107916266 A describes a multi-enzyme process for the bioconversion of DON into 3-epi-DON. In 2012, a three-enzyme system comprising the cytochrome P450 enzyme DdnA derived from Sphingomonas sp. KSM1 was described for the hydroxylation of DON into 16-hydroxy-DON (Ito et al., 2012. Appl Environ Microbiol 79(5): 1619-1628).

[0012] The present invention was made in view of the above-mentioned prior art. In particular, the object of the present invention can be formulated as providing a new means and method for detoxifying deoxynivalenol (DON). The solution of the present invention is described below, exemplified in the examples, illustrated in the figures and reflected in the claims.

[0013] This object has been achieved by providing an enzyme preparation capable of modifying (e.g. detoxifying and / or degrading) DON into 7-keto-8-hydroxy-8-en-DON (e.g. Figure 2also known as 3,8,15-trihydroxy-12,13-epoxytrichothec-8-en-7-one) is achieved. DON is a trichothecene mycotoxin having a carbonyl group (C=0) at position 8 of the carbon chain (or in other words, having a double bond bonded to an oxygen atom (=0) on the C8 atom of the carbon chain) and a hydroxyl group (-OH) at the C7 atom of the carbon chain, which has the following structural formula:

[0014] SUMMARY

[0015] The present invention relates to methods and compositions for degrading and / or altering the toxicity of DON and / or DON derivatives, the methods / compositions comprising: (i) providing one or more of the following polypeptides selected from the group consisting of: (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 ; (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g. 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1 ; (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, deletion and / or insertion at one or more positions; and / or (d) a fragment of the polypeptide of (a), (b) or (c), wherein the fragment is capable of degrading and / or altering the toxicity of DON and / or DON derivatives, preferably the alteration of toxicity comprises one or more of: detoxifying DON and / or DON derivatives and / or altering the intermolecular rearrangement of DON and / or DON derivatives; (ii) optionally applying one or more of (a)-(d) to DON and / or DON derivatives; preferably the one or more polypeptides is one or more recombinant and / or isolated polypeptides; further preferably the alteration of toxicity comprises one or more of: detoxifying DON and / or DON derivatives and / or intramolecular rearranging DON and / or DON derivatives; most preferably the one or more polypeptides is capable of modifying (e.g. modifying the bond at C7 and / or binding partners and / or functional groups and / or causing a chemical change, such as breaking a bond and / or forming a new bond, etc.) the C7 atom of DON (e.g. producing 7-keto-8-hydroxy-8-ene-DON from DON) and / or DON derivatives. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1: SDS-PAGE gel (12% Mini-PROTEAN (Bio-Rad) TGX unstained precast gels) with fractions corresponding to 10 μl of E. coli gene expression culture lysate (T: total lysate; S: soluble fraction; I: insoluble fraction). SEQ ID NO: 1 : 20.5 kDa. Reference protein of 35.2 kDa included as gene expression control.

[0017] Figure 2 : Molecular structures of DON and 7-keto-8-hydroxy-8-en-DON.

[0018] Figure 3 : Concentrations of DON and 7-keto-8-hydroxy-8-en-DON after incubation of crude E. coli cell lysate in reaction buffer with 100 μM DON at 30 °C for 3 h.

[0019] Figure 4 : Time course of DON and 7-keto-8-hydroxy-8-en-DON concentrations upon incubation with clarified lysate of E. coli BL21(DE3) biomass from gene expression culture in reaction buffer with initially nominally 100 μM DON. DETAILED DESCRIPTION

[0020] DEFINITIONS

[0021] As described herein, “EC number” (Enzyme Commission number) can be used to refer to an enzyme activity according to the Enzyme Nomenclature database published on February 26, 2020 (e.g., available at https: / / enzyme.expasy.org / ). The EC number refers to Enzyme Nomenclature 1992 from NC-IUBMB, Academic Press, San Diego, Calif., including supplements 1-5 published in Eur. J. Biochem. 1994, 223, 1-5; Eur. J. Biochem. 1995, 232, 1-6; Eur. J. Biochem. 1996, 237, 1-5; Eur. J. Biochem. 1997, 250, 1-6; and Eur. J. Biochem. 1999, 264, 610-650, respectively.

[0022] The term "polypeptide" is used herein as equivalent to the term "protein". A protein (including fragments, preferably biologically active fragments thereof, and peptides, which typically have less than 30 amino acids) comprises one or more amino acids coupled to each other by covalent peptide bonds (yielding an amino acid chain). The term "polypeptide" as used herein describes a class of molecules which, for example, consist of more than 30 amino acids. Polypeptides can also form multimers, e.g. dimers, trimers and higher oligomers, i.e. consist of more than one polypeptide molecule. The polypeptide molecules forming such dimers, trimers, etc. can be identical or different. The corresponding higher order structures of these multimers are thus called homo- or heterodimers, homo- or heterotrimers, etc. An example of a heteromultimer is an antibody molecule which in its naturally occurring form consists of two identical light polypeptide chains and two identical heavy polypeptide chains. The terms "polypeptide" and "protein" also refer to naturally modified polypeptides / proteins, wherein the modifications are effected, for example, by post-translational modifications such as glycosylation, acylation, phosphorylation, etc. Such modifications are well known in the art.

[0023] Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter "sequence identity". For the purposes of the present application, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle

[0024] (identical residues x 100) / (length of alignment - total number of gaps in alignment).

[0025] Alternatively, the parameters used can be a gap open penalty of 10, a gap extension penalty of 0.5, and the EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. The Needle output, labeled "longest identity", (obtained using the no-brief option) is used as the percent identity and is calculated as follows:

[0026] (Identical deoxyribonucleotides x 100) / (Aligned Length - Total number of gaps in alignment).

[0027] Expression: The term "expression" includes any step involved in the production of a variant (polypeptide) including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0028] Expression vector: The term "expression vector" can refer to a linear or circular DNA molecule that comprises a polynucleotide encoding a variant (polypeptide) and is operably linked to control sequences to provide for its expression, in particular for its transcription.

[0029] Fragment: The term "fragment" can refer to a polypeptide that is missing one or more (e.g., several, e.g., 5, 10, 20, 30, 40, etc.) amino acids from the amino and / or carboxyl terminus of a mature polypeptide; wherein the fragment has an activity as described elsewhere herein.

[0030] Host cell: The term "host cell" can refer to any cell type that is susceptible to transformation, transfection, transduction, or the like with a nucleic acid construct or expression vector comprising a polynucleotide of the present application. The term "host cell" encompasses any progeny of the parent cell which is not identical to the parent cell (and thus can not contain the same metabolic activity or selectability) due to mutations which occur during replication.

[0031] Nucleic acid construct: The term "nucleic acid construct" can refer to a nucleic acid molecule, either single- or double-stranded, which is isolated from a naturally occurring gene or which is modified from a naturally occurring gene in a manner known to one of skill in the art, or which is synthetic, that comprises one or more control sequences.

[0032] Operably linked: The term "operably linked" can refer to a configuration in which a control sequence is positioned at a place relative to the coding sequence of a polynucleotide such that the control sequence directs expression of the coding sequence.

[0033] Control sequences: The term "control sequences" as used herein can refer to nucleic acid sequences necessary for expression of a polynucleotide encoding a variant of the present application (polypeptide). Each control sequence can be native (i.e., from the same gene) or foreign (i.e., from a different gene) to the polynucleotide encoding the variant or foreign to each other. Such control sequences include, but are not limited to, a leader, polyadenylation sequence, pre-pro peptide sequence, promoter, signal peptide sequence, and transcription terminator. At a minimum, the control sequences include a promoter, and transcriptional and translational stop signals. The control sequences can be provided with linkers for ease of cleavage from the polynucleotide of the present application, as will be understood by those of ordinary skill in the art.

[0034] As used herein, the term "corresponding to" can refer to a way of determining a particular amino acid in a sequence where reference is made to a particular amino acid sequence (such as US2020071638). For example, for the purposes of the present application, when reference is made to a particular amino acid position, the skilled person will be able to align another amino acid sequence with the amino acid sequence to which reference has been made, to thereby determine which particular amino acid can be of interest in the other amino acid sequence. Alignment of another amino acid sequence with, for example, the sequence as set forth in SEQ ID NOs: 1, 2, 3 or any other sequence set forth herein has been described elsewhere herein. Alternative alignment methods can be used and these are well known to the person skilled in the art.

[0035] The term "position" when used in accordance with the present application can refer to the position of an amino acid in an amino acid sequence depicted herein. The term "corresponding to" in this context can include cases where the position is not determined solely by the number of preceding nucleotides / amino acids.

[0036] As used herein, a "silent" mutation means a base substitution in a nucleic acid sequence that does not change the amino acid sequence encoded by the nucleic acid sequence. A "conservative or equivalent" substitution (or mutation) means a substitution as listed in Table 1 below as "Exemplary Substitutions". A "highly conservative" substitution as used herein means a substitution as shown below in Table I under the heading "Preferred Substitutions".

[0037] Table I Amino Acid Substitutions

[0038] Original Exemplary substitution Preferred substitution Ala (A) val; leu; ile Val Arg (R) lys; gin; asn lys Asn (N) gin; his; asp, lys; arg gin Asp (D) glu; asn glu Cys (C) ser; ala ser Gln (Q) asn; glu asn Glu (E) asp; gin asp Gly (G) ala ala His (H) asn; gin; lys; arg arg Ile (I) leu; val; met; ala; phe; leu Leu (L) norleucine; ile; val; met; ala; ile Lys (K) arg; gin; asn arg Met (M) leu; phe; ile leu Phe (F) leu; val; ile; ala; tyr tyr Pro (P) ala ala Ser (S) thr thr Thr (T) ser ser Trp (W) tyr; phe tyr Tyr (Y) trp; phe; thr; ser Phe Val (V) ile; leu; met; phe; ala; leu

[0039] Variant: The term "variant" can refer to a polypeptide having specific activity as described herein comprising an alteration (i.e., a substitution, insertion, and / or deletion) at one or more (e.g., several) positions. A substitution means replacing an amino acid occupying a position with a different amino acid; a deletion means removing an amino acid occupying a position; an insertion means adding an amino acid after the amino acid occupying a position and adjacent and following.

[0040] As used herein, the term "transgenic" can refer to an organism whose genome has been altered, e.g., by introduction of heterologous genetic material or additional copies of homologous genetic material, through transformation or recombination (e.g., US7410800B2). A transgenic organism can be a plant, mammal, fungus, bacterium, or virus. As used herein, a "transgenic plant, seed, or pollen grain" can refer to a plant, seed, or pollen grain or a progeny plant, seed, or pollen grain derived from any offspring of the same, wherein the DNA of the plant, seed, or pollen grain or its progeny comprises introduced foreign DNA that was not initially present in a non-transgenic plant, seed, or pollen grain of the same plant. A transgenic plant, seed, or pollen grain can additionally comprise sequences that are native to the plant that was transformed, but wherein the foreign DNA has been altered to change the expression level or pattern of the encoded sequence.

[0041] The term "modification of the C7 atom" can refer to a process of intramolecular rearrangement and / or bond and / or binding partner and / or functional group alteration at the C7 atom (e.g., position 7 of the carbon chain) of, e.g., DON and / or a DON derivative, (e.g., including altering (e.g., oxidizing) the hydroxyl moiety (e.g., -OH) at the C7 atom (e.g., position 7 of the carbon chain) to a carbonyl moiety (e.g., =0) and / or causing a chemical change, e.g., a bond breakage and / or a new bond formation.

[0042] The term "food" can refer to a substance having edible value.

[0043] The term "forage" can refer to a substance that is fed to livestock.

[0044] The term "feed" can refer to a substance used as food for livestock.

[0045] The term "additive" can refer to a compound or substance that is, e.g., added in small amounts to another product or substance, e.g., for technical purposes in the manufacture, processing, preparation, treatment, packaging, transport, or storage of a food, forage, or feed product, to achieve a desired property and / or characteristic. Exemplary additives of the present invention can include: processing aids (i.e., substances used in food production but not consumed as food), reactants (i.e., substances consumed in the course of a chemical reaction), and catalysts (substances that increase the rate of a reaction without changing the overall standard Gibbs energy change in the reaction), e.g., for the food or feed industry, starch production, e.g., citric acid, bioethanol, etc.

[0046] The term "prebiotic" can refer to a compound or substance that is capable of inducing the growth and / or activity of a microorganism (e.g., a beneficial microorganism).

[0047] The term "detoxifying agent" can refer to a compound or substance that is capable of reducing and / or inhibiting the toxicity of, e.g., a mycotoxin (e.g., DON and / or a DON derivative).

[0048] The term "nutritional supplement" can refer to a compound or substance capable of supporting a nutritional component in a diet, such as vitamins and minerals.

[0049] The term "intermediate" can refer to a compound or substance produced in the course of obtaining a final product of the present application, such as a food, forage, feed, additive (such as a food-, forage- or feed additive), antidote, nutritional supplement or prebiotic of the present application, for example at an intermediate stage of the course.

[0050] The term "nutritional source" can refer to any substance that can be used for nutrition and / or bioenergy production (e.g. comprising carbohydrates and / or proteins and / or fats).

[0051] The term "material" can refer to a raw material of plant origin, such as citric acid or bioethanol vegetable tubers or roots, such as but not limited to potato, carrot, sugar beet, parsnip, parsley root, celery root, sweet potato, yam, arrowroot, radish, turnip, chicory root and cassava; cereals, such as but not limited to wheat, rice, maize, corn, rye, barley, buckwheat, sorghum, oat and millets (ragi); coffee; cocoa; chicory; olives; prunes or raisins.

[0052] The term "intramolecular rearrangement" or "intermolecular rearrangement" can refer to any process involving a transfer (e.g. of atoms, groups, electrons, bonds, etc.) or interaction between different parts of the same molecular entity or between two or more molecular entities.

[0053] It is noted that, as used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a reagent" includes one or more of such different reagents, and reference to "the method" includes reference to equivalent steps and methods known to one of ordinary skill in the art that could be substituted or modified for the methods described herein.

[0054] The term "at least" preceding a series of elements is to be understood to refer to every element in the series unless otherwise indicated. Those skilled in the art will recognize or be able to ascertain using not more than routine experimentation many equivalents of the specific embodiments of the application described herein. Such equivalents are intended to be encompassed by the present application.

[0055] The term "and / or" wherever used herein includes the meaning of "and", "or" and "any or all of the elements linked to the said term".

[0056] The term "about" or "approximately" as used herein when used in connection with a given value or range means within 20%, preferably within 10% and more preferably within 5% of the given value or range.

[0057] Throughout this specification and the claims that follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. As used herein, the term "comprising" can be replaced by the term "containing" or "including" or, as used herein, sometimes replaced by the term "having".

[0058] As used herein, "consisting of excludes any element, step, or ingredient not specified as well as any intangible step or ingredient. As used herein, "consisting essentially of does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claimed invention.

[0059] In each instance herein, any of the terms "including", "consisting essentially of and "consisting of can be replaced by any of the other two terms.

[0060] In the course of the present invention, it was found that the polypeptide SEQ ID NO: 1 is able to convert DON into 7-keto-8-hydroxy-8-en-DON (quantified in Figure 3 ), while the related SEQ ID NO: 2 and SEQ ID NO: 3 did not show any DON conversion or any DON concentration reduction. It is well known in the art that 7-keto-8-hydroxy-8-en-DON is less toxic than DON (e.g. Stadler et al. (2019), Food Chem. 279, 303-311; Stadler et al. (2019), Toxins (Basel) 11, 317). Thus, it was surprisingly found that SEQ ID NO: 1 is able to detoxify DON. Therefore, the object of the present invention has been achieved by providing means, methods and products based on SEQ ID NO: 1 as described herein below.

[0061] In some embodiments, the present application provides a method for producing a food, an intermediate food; a forage, an intermediate forage; a feed, an intermediate feed; an additive (e.g., a food, forage, or feed additive, a processing aid, a reactant, or a catalyst, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.), an intermediate additive (e.g., an intermediate food, forage, or feed additive, an intermediate processing aid, a reactant, or a catalyst, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.); a detoxifying agent, an intermediate detoxifying agent; a nutritional supplement, an intermediate nutritional supplement; a prebiotic, an intermediate prebiotic, and / or a mixture thereof, the method comprising: (i) providing one or more of the following polypeptides: (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1; (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g., 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1; preferably, the polypeptide does not have 100% identity to the amino acid sequence set forth in SEQ ID NO: 2 or 3; (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, a deletion, and / or an insertion at one or more positions; preferably, the variant does not have 100% identity to the amino acid sequence set forth in SEQ ID NO: 2 or 3; and / or (d) a fragment of the polypeptide of (a), (b), or (c), wherein the fragment is capable of degrading DON (e.g., to 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (e.g., the DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom of the carbon chain, a carbonyl moiety (C=0) at position 8; the C10 atom is unsubstituted) and / or altering the toxicity of DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (e.g., the DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom, a carbonyl moiety (=0) at C8; C10 is unsubstituted), preferably, the altering the toxicity comprises one or more of the following: detoxifying DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or altering the intermolecular rearrangement of DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative; (ii) applying one or more of (a)-(d) to a nutritional source (e.g., comprising a carbohydrate and / or protein source) or a material (e.g., a raw material, a crop, a grain, citric acid, bioethanol, etc.) suitable for producing a food, an intermediate food; a forage, an intermediate forage; a feed, an intermediate feed;additives (e.g., food, forage or feed additives, processing aids, reactants or catalysts, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.), intermediate additives (e.g., food, forage or feed intermediate additives, intermediate processing aids, reactants or catalysts, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.); antidotes, intermediate antidotes; nutritional supplements, intermediate nutritional supplements; prebiotics, intermediate prebiotics and / or mixtures thereof; preferably, the one or more polypeptides are one or more recombinant and / or isolated polypeptides; further preferably, the one or more polypeptides are capable of modifying DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative; most preferably, the modification comprises changing a hydroxyl moiety (e.g., can be referred to as -OH or C-OH) to a carbonyl moiety (e.g., can be referred to as =0 or C=0) at the C7 atom (e.g., position 7); further most preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearalenone).

[0062] In other embodiments, the methods of the application further comprise: incubating a nutrient source or substance with one or more of (a)-(d) under conditions suitable for degrading DON and / or a DON derivative and / or altering the toxicity of DON and / or a DON derivative; preferably, the methods further comprise heat treating and / or fractionating and / or drying the incubated product; further preferably, the altering the toxicity comprises one or more of: detoxifying DON and / or a DON derivative (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or intramolecularly rearranging DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearalenone).

[0063] In other embodiments, the application relates to food, intermediate food; forage, intermediate forage; feed, intermediate feed; additives (e.g., food, forage or feed additives, processing aids, reactants or catalysts, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.), intermediate additives (e.g., food, forage or feed intermediate additives, intermediate processing aids, reactants or catalysts, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.); antidotes, intermediate antidotes; nutritional supplements, intermediate nutritional supplements; prebiotics, intermediate prebiotics and / or mixtures thereof produced by the methods of the application.

[0064] In some embodiments, the present application provides a method for degrading DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative (e.g., the DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom of the carbon chain, a carbonyl moiety (C=0) at position 8; the C10 atom is unsubstituted); the degrading and / or altering the toxicity comprises: modifying the C7 atom of the DON and / or the DON derivative; preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy- scirpene-9-ene-8-one), the method comprising: (i) providing one or more of the following polypeptides: (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1; (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g., 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1, preferably, the polypeptide does not have 100% identity to SEQ ID NO: 2-3; (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, a deletion, and / or an insertion at one or more positions; preferably, the variant does not have 100% identity to SEQ ID NO: 2-3; and / or (d) a fragment of the polypeptide of (a), (b), or (c), wherein the fragment is capable of degrading DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative, preferably, the altering the toxicity comprises one or more of the following: detoxifying DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative and / or altering the intermolecular rearrangement of DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative; (ii) applying one or more of (a)-(d) to the DON and / or the DON derivative; preferably, the one or more polypeptides is one or more recombinant and / or isolated polypeptides;Further preferably, said altering toxicity comprises one or more of: detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, said DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one) and / or intramolecular rearranging DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, said DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one); most preferably, said one or more polypeptides are capable of modifying the C7 atom of DON and / or a DON derivative (e.g. said modification comprises changing a C-OH to a C=0 moiety, e.g. at position 7; preferably, said DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one).

[0065] In some embodiments, the present application provides a composition or kit comprising: (i) a polypeptide capable of degrading DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative (e.g., a DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom of the carbon chain, a carbonyl moiety (C=0) at position 8; C10 unsubstituted; preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one) and / or altering the toxicity of DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative, wherein the polypeptide is one or more of: (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1; (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g., 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1; preferably, the polypeptide does not have 100% identity to SEQ ID NO: 2-3; (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, deletion, and / or insertion at one or more positions; preferably, the variant does not have 100% identity to SEQ ID NO: 2-3; and / or (d) a fragment of the polypeptide of (a), (b), or (c), wherein the fragment is capable of degrading DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative, preferably, the altering the toxicity comprises one or more of: detoxifying DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one) and / or altering the intermolecular rearrangement of DON (e.g., by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one); (ii) a nutritional source (e.g., comprising one or more of: a carbohydrate, a protein, citric acid, an ethanol, e.g., bioethanol; preferably, the nutritional source is not 3-Ac-DON) and / or a substrate for at least one of the polypeptides of (a)-(d), preferably, the substrate comprises: DON and / or a DON derivative (preferably, the substrate is not 3-Ac-DON).Further preferably, the composition further comprises the product of a reaction (e.g. enzymatic reaction) of the one or more polypeptides with DON (e.g. 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative, i.e. a reaction product comprising a modification at the C7 atom of DON and / or a DON derivative); preferably, the polypeptide is a recombinant and / or isolated polypeptide; further preferably, the alteration of toxicity comprises one or more of the following: detoxification of DON and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON) and / or intramolecular rearrangement of DON and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON); most preferably, the polypeptide is capable of modifying the C7 atom of DON and / or a DON derivative (preferably, the modification comprises changing a C-OH to a C=0 moiety at position 7; further preferably, the DON derivative is not 3-Ac-DON).

[0066] In other embodiments, the one or more polypeptides of the application are encoded by one or more nucleotide sequences.

[0067] In other embodiments, the one or more nucleotide sequences of the application are comprised by one or more nucleic acids comprised by a host cell.

[0068] In other embodiments, the composition or kit of the application is one or more of the following: a cell-free and / or non-naturally occurring and / or fractionated composition or kit.

[0069] In other embodiments, the composition or kit of the application is a pharmaceutical or veterinary composition or kit.

[0070] In other embodiments, the composition, kit, food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g. food, forage or feed additive), intermediate additive (e.g. food, forage or feed intermediate additive); detoxicant, intermediate detoxicant; nutritional supplement, intermediate nutritional supplement; prebiotic, intermediate prebiotic or mixtures thereof of the application can be used as a medicament (e.g. including veterinary use) and / or in therapy.

[0071] In other embodiments, the composition, kit, food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g. food, forage or feed additive), intermediate additive (e.g. food, forage or feed intermediate additive); detoxicant, intermediate detoxicant; nutritional supplement, intermediate nutritional supplement; prebiotic, intermediate prebiotic or mixtures thereof of the application can be used in the treatment, amelioration, prevention and / or diagnosis of DON mycotoxin poisoning.

[0072] In some embodiments, the present application provides a method for modifying a C7 atom of DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the modification comprises changing a hydroxyl moiety (e.g., -OH or C-OH) to a carbonyl moiety (e.g., can refer to =0 or C=0) at position 7 of the carbon chain; further preferably, the DON derivative is not 3-Ac-DON), the method comprising: (i) providing one or more of the following polypeptides: (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1; (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g., 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1; preferably, the polypeptide does not have 100% identity to SEQ ID NO: 2-3; (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, a deletion, and / or an insertion at one or more positions; preferably, the variant does not have 100% identity to SEQ ID NO: 2-3; and / or (d) a fragment of the polypeptide of (a), (b), or (c), wherein the fragment is capable of degrading DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative, preferably, the altering the toxicity comprises one or more of: detoxifying DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or altering the intermolecular rearrangement of DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON); (ii) applying one or more of (a)-(d) to DON and / or a DON derivative; preferably, the one or more polypeptides is one or more recombinant and / or isolated polypeptides;

[0073] In some embodiments, the method of the present application is an in vitro, ex vivo, or in vivo method.

[0074] In some embodiments, the present application relates to one or more of the following of the present application: (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 ; (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g., 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1 ; preferably, the polypeptide does not have 100% identity to SEQ ID NOs: 2-3; (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, deletion, and / or insertion at one or more positions; and / or (d) a fragment of the polypeptide of (a), (b), or (c); (e) a composition, kit, food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g., a food, forage, or feed additive, a processing aid, a reactant, or a catalyst, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.), intermediate additive (e.g., an intermediate food, forage, or feed additive, an intermediate processing aid, a reactant, or a catalyst, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.); antidote, intermediate antidote; nutritional supplement, intermediate nutritional supplement; prebiotic, intermediate prebiotic, and / or mixtures thereof, for use in / for use in one or more of: (i) modifying a C7 atom of a DON (e.g., by producing a 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative (preferably, the modification comprises changing a hydroxyl moiety (-OH) to a carbonyl moiety (=0) at C7; further preferably, the DON derivative is not 3-Ac-DON); (ii) producing a food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g., a food, forage, or feed additive, a processing aid, a reactant, or a catalyst, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.), intermediate additive (e.g., an intermediate food, forage, or feed additive, an intermediate processing aid, a reactant, or a catalyst, e.g., in the food or feed industry, citric acid, bioethanol, starch production, etc.); antidote, intermediate antidote; nutritional supplement, intermediate nutritional supplement; prebiotic, intermediate prebiotic, and / or mixtures thereof; (iii) degrading a DON and / or a DON derivative and / or altering the toxicity of a DON and / or a DON derivative, preferably, the one or more polypeptides are recombinant and / or isolated polypeptides;Further preferably, said altering toxicity comprises one or more of: detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or intramolecularly rearranging DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, said modification comprises altering a hydroxyl moiety (-OH) to a carbonyl moiety (=0) at C7; further preferably, said DON derivative is not 3-Ac-DON); most preferably, said one or more polypeptides are capable of modifying a C7 atom of DON and / or a DON derivative (preferably, said modification comprises altering a hydroxyl moiety (-OH) to a carbonyl moiety (=0) at C7; further preferably, said DON derivative is not 3-Ac-DON); (iv) any combination of (i)-(iii); (v) the use according to any one of (i)-(iv), wherein said use is an in vitro, ex vivo or in vivo use.

[0075] It is to be understood that the application is not limited to the particular methodology, protocols, and reagents, etc. described herein as these can vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present application which is limited only by the claims.

[0076] All publications and patents cited herein, including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc., are hereby incorporated by reference in their entirety herein. Nothing herein is to be construed as an admission that the application is not entitled to antedate such disclosure by virtue of prior application. To the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern.

[0077] The present application is further characterized by the following items:

[0078] 1. A method for producing a food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g. a food, forage or feed additive, a processing aid, a reactant or catalyst, e.g. in the food or feed industry, in citric acid, bioethanol, starch production, etc.), intermediate additive (e.g. an intermediate food, forage or feed additive, an intermediate processing aid, a reactant or catalyst, e.g. in the food or feed industry, in citric acid, bioethanol, starch production, etc.); a detoxicant, intermediate detoxicant; a nutritional supplement, intermediate nutritional supplement; a prebiotic, intermediate prebiotic and / or mixtures thereof, said method comprising:

[0079] i) providing one or more of the following polypeptides:

[0080] (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 ;

[0081] (b) a polypeptide comprising an amino acid sequence which has at least 70% (e.g. 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%) identity to the amino acid sequence shown in SEQ ID NO: 1 ; preferably the polypeptide does not have 100% identity to the amino acid sequence shown in SEQ ID NO: 2 or 3;

[0082] (c) a variant of the polypeptide shown in SEQ ID NO: 1, wherein the variant comprises: a substitution, deletion and / or insertion at one or more positions; preferably the variant does not have 100% identity to the amino acid sequence shown in SEQ ID NO: 2 or 3; and / or

[0083] (d) a fragment of the polypeptide of (a), (b) or (c), wherein the fragment is capable of degrading DON (e.g. to 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (e.g. a DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom, a carbonyl moiety (C=0) at the C8; C10 is unsubstituted) and / or altering the toxicity of DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (e.g. a DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom, a carbonyl moiety (e.g. =0) at the C8; C10 is unsubstituted), preferably the altering the toxicity comprises one or more of: detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or altering the intermolecular rearrangement of DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative;

[0084] ii) applying one or more of (a)-(d) to a nutrient source (e.g. comprising a carbohydrate and / or protein source) or material (e.g. a raw material, a crop, a cereal, citric acid, bioethanol, etc.) suitable for producing a food, an intermediate food; a forage, an intermediate forage; a feed, an intermediate feed; an additive (e.g. a food, forage or feed additive, a processing aid, a reactant or catalyst, e.g. in the food or feed industry, citric acid, bioethanol, starch production, etc.), an intermediate additive (e.g. an intermediate food, forage or feed additive, an intermediate processing aid, a reactant or catalyst, e.g. in the food or feed industry, citric acid, bioethanol, starch production, etc.); a detoxificant, an intermediate detoxificant; a nutritional supplement, an intermediate nutritional supplement; a prebiotic, an intermediate prebiotic and / or a mixture thereof;

[0085] Preferably, the one or more polypeptides are one or more recombinant and / or isolated polypeptides; further preferably, the one or more polypeptides are capable of modifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative; most preferably, the modification comprises changing a hydroxyl moiety (e.g. C-OH) to a carbonyl moiety (e.g. C=0) at the C7 atom; further most preferably, the DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zeylan-8-one).

[0086] 2. The method according to any one of the preceding items, further comprising incubating the nutrient source or substance with one or more of (a)-(d) under conditions suitable for degrading DON and / or a DON derivative and / or changing the toxicity of DON and / or a DON derivative; preferably, the method further comprises heat-treating and / or fractionating and / or drying the incubated product; further preferably, the changing the toxicity comprises one or more of detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or intramolecularly rearranging DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zeylan-8-one).

[0087] 3. A food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g. food, forage or feed additive, processing aid, reactant or catalyst, e.g. in the food or feed industry, citric acid, bioethanol, starch production, etc.), intermediate additive (e.g. intermediate food, forage or feed additive, intermediate processing aid, reactant or catalyst, e.g. in the food or feed industry, citric acid, bioethanol, starch production, etc.); detoxifying agent, intermediate detoxifying agent; nutritional supplement, intermediate nutritional supplement; prebiotic, intermediate prebiotic and / or mixtures thereof produced by the method according to any one of the preceding items.

[0088] 4. A method for degrading DON (e.g. by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g. by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative (e.g. the DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom, a carbonyl moiety (C=0) at C8; C10 is unsubstituted); the degrading and / or altering toxicity comprising: modifying the C7 atom of the DON and / or DON derivative; preferably, the DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy- scirpene-9-ene-8-one), the method comprising:

[0089] i) providing one or more of the following polypeptides:

[0090] (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 ;

[0091] (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g. 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1 ; preferably, the polypeptide does not have 100% identity to SEQ ID NO: 2-3;

[0092] (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, deletion and / or insertion at one or more positions; preferably, the variant does not have 100% identity to SEQ ID NO: 2-3; and / or

[0093] (d) a fragment of the polypeptide of (a), (b) or (c), wherein the fragment is capable of degrading DON (e.g. by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g. by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative, preferably the altering toxicity comprises one or more of: detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative and / or altering the intermolecular rearrangement of DON (e.g. by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative;

[0094] ii) applying one or more of (a)-(d) to DON and / or a DON derivative;

[0095] Preferably, the one or more polypeptides is one or more recombinant and / or isolated polypeptide; further preferably, the altering toxicity comprises one or more of: detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one) and / or intramolecular rearranging DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one); most preferably, the one or more polypeptides is capable of modifying a bond at the C7 atom of DON and / or a DON derivative (e.g. the modification comprises changing a C-OH to a C=0 moiety at position 7; preferably, the DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one).

[0096] 5. A composition or kit-of-parts comprising:

[0097] (i) a polypeptide capable of degrading DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (e.g. the DON derivative can comprise a hydroxyl moiety (-OH) at the C7 atom, a carbonyl moiety (C=0) at C8; C10 is unsubstituted; preferably, the DON derivative is not 3-Ac-DON, i.e. 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one) and / or altering the toxicity of DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative, wherein the polypeptide is one or more of:

[0098] (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 ;

[0099] (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g. 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1 ; preferably, the polypeptide does not have 100% identity to SEQ ID NO: 2-3;

[0100] (c) a variant of the polypeptide of SEQ ID NO: 1, wherein the variant comprises: a substitution, a deletion, and / or an insertion at one or more positions; preferably, the variant does not have 100% identity to SEQ ID NO: 2-3; and / or

[0101] (d) a fragment of the polypeptide of (a), (b), or (c), wherein the fragment is capable of degrading DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative, preferably, the altering the toxicity comprises one or more of: detoxifying DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one) and / or altering the intermolecular rearrangement of DON (e.g., by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON, i.e., 3-acetyloxy-3a,7a-dihydroxy-12,13-epoxy-9-oxa-zearlan-8-one);

[0102] (ii) a nutrient source (e.g., comprising one or more of: a carbohydrate, a protein, citric acid, ethanol, e.g., bioethanol; preferably, the nutrient source is not 3-Ac-DON) and / or a substrate for at least one of the polypeptides of (a)-(d), preferably, the substrate comprises: DON and / or a DON derivative (preferably, the substrate is not 3-Ac-DON; further preferably, the composition further comprises a reaction product, e.g., an enzymatic reaction, of the one or more polypeptides with DON (e.g., 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative, i.e., a reaction product comprising a modification at the C7 atom of DON and / or a DON derivative);

[0103] preferably, the polypeptide is a recombinant and / or isolated polypeptide; further preferably, the altering the toxicity comprises one or more of: detoxifying DON and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON) and / or intramolecular rearrangement of DON and / or a DON derivative (preferably, the DON derivative is not 3-Ac-DON); most preferably, the polypeptide is capable of modifying the C7 atom of DON and / or a DON derivative (preferably, the modification comprises changing a C-OH to a C=O moiety; further preferably, the DON derivative is not 3-Ac-DON).

[0104] 6. The composition or kit according to any one of the preceding items, wherein the one or more polypeptides are encoded by one or more nucleotide sequences.

[0105] 7. The composition or kit according to any one of the preceding items, wherein the one or more nucleotide sequences are comprised by one or more nucleic acids comprised by a host cell.

[0106] 8. The composition or kit according to any one of the preceding items, wherein the composition or kit is one or more of: a cell-free and / or non-naturally occurring and / or fractionated composition or kit.

[0107] 9. The composition or kit according to any one of the preceding items, wherein the composition or kit is a pharmaceutical and / or veterinary composition or kit.

[0108] 10. The composition, kit, food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g. food, forage or feed additive), intermediate additive (e.g. food, forage or feed intermediate additive); antidote, intermediate antidote; nutritional supplement, intermediate nutritional supplement; prebiotic, intermediate prebiotic or mixtures thereof according to any one of the preceding items for use as a medicament (e.g. for veterinary use) and / or in therapy.

[0109] 11. The composition, kit, food, intermediate food; forage, intermediate forage; feed, intermediate feed; additive (e.g. food, forage or feed additive), intermediate additive (e.g. food, forage or feed intermediate additive); antidote, intermediate antidote; nutritional supplement, intermediate nutritional supplement; prebiotic, intermediate prebiotic or mixtures thereof according to any one of the preceding items for use in the treatment, amelioration, prevention and / or diagnosis of DON mycotoxin poisoning.

[0110] 12. A method for modifying the C7 atom of DON (e.g. by producing 7-keto-8-hydroxy-8- en-DON) and / or a DON derivative (preferably, the modification comprises changing a hydroxyl moiety (-OH) to a carbonyl moiety (=0) at C7; further preferably, the DON derivative is not 3-Ac-DON), the method comprising:

[0111] i) providing one or more of the following polypeptides:

[0112] (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 ;

[0113] (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g. 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1 ; preferably the polypeptide does not have 100% identity to SEQ ID NOs: 2-3;

[0114] (c) a variant of the polypeptide set forth in SEQ ID NO: 1, wherein the variant comprises: a substitution, deletion, and / or insertion at one or more positions; preferably the variant does not have 100% identity to SEQ ID NOs: 2-3; and / or

[0115] (d) a fragment of the polypeptide of (a), (b) or (c), wherein the fragment is capable of degrading DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or altering the toxicity of DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative, preferably the altering the toxicity comprises one or more of: detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or altering the intermolecular rearrangement of DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably the DON derivative is not 3-Ac-DON);

[0116] ii) applying one or more of (a)-(d) to DON and / or a DON derivative;

[0117] Preferably the one or more polypeptides are one or more recombinant and / or isolated polypeptides.

[0118] 13. The method of any one of the preceding items, wherein the method is an in vitro, ex vivo, or in vivo method.

[0119] 14. One or more of the following according to any one of the preceding items:

[0120] (a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 ;

[0121] (b) a polypeptide comprising an amino acid sequence that is at least 70% (e.g. 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%) identical to the amino acid sequence set forth in SEQ ID NO: 1 ; preferably the polypeptide does not have 100% identity to SEQ ID NOs: 2-3;

[0122] (c) a variant of the polypeptide as shown in SEQ ID NO: 1, wherein said variant comprises: a substitution, a deletion, and / or an insertion at one or more positions; and / or

[0123] (d) a fragment of the polypeptide of (a), (b) or (c);

[0124] (e) a composition, a kit, a food, an intermediate food; a forage, an intermediate forage; a feed, an intermediate feed; an additive (e.g. a food, forage or feed additive, a processing aid, a reactant or catalyst, e.g. in the food or feed industry, citric acid, bioethanol, starch production etc.), an intermediate additive (e.g. an intermediate food, forage or feed additive, an intermediate processing aid, a reactant or catalyst, e.g. in the food or feed industry, citric acid, bioethanol, starch production etc.); a detoxifying agent, an intermediate detoxifying agent; a nutritional supplement, an intermediate nutritional supplement; a prebiotic, an intermediate prebiotic or mixtures thereof;

[0125] for use in / in one or more of:

[0126] i) modifying the C7 atom of DON (e.g. by producing 7-keto-8-hydroxy-8-ene-DON) and / or a DON derivative (preferably, said modification comprises changing a hydroxyl moiety (-OH) to a carbonyl moiety (=0) at C7; further preferably, said DON derivative is not 3-Ac-DON);

[0127] ii) producing a food, an intermediate food; a forage, an intermediate forage; a feed, an intermediate feed; an additive (e.g. a food, forage or feed additive), an intermediate additive (e.g. a food, forage or feed intermediate additive); a detoxifying agent, an intermediate detoxifying agent; a nutritional supplement, an intermediate nutritional supplement; a prebiotic, an intermediate prebiotic and / or mixtures thereof;

[0128] iii) degrading and / or altering the toxicity of DON and / or a DON derivative, preferably the one or more polypeptides are recombinant and / or isolated polypeptides; further preferably the altering the toxicity comprises one or more of detoxifying DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative and / or intramolecularly rearranging DON (e.g. by producing 7-keto-8-hydroxy-8-en-DON) and / or a DON derivative (preferably the modification comprises changing a hydroxyl moiety (-OH) to a carbonyl moiety (=0) at C7; further preferably the DON derivative is not 3-Ac-DON); most preferably the one or more polypeptides are capable of modifying the C7 atom of DON and / or a DON derivative (preferably the modification comprises changing a hydroxyl moiety (-OH) to a carbonyl moiety (=0) at C7; further preferably the DON derivative is not 3-Ac-DON);

[0129] iv) any combination of (i)-(iii);

[0130] v) the use according to any one of (i)-(iv), wherein the use is an in vitro, ex vivo or in vivo use.

[0131] The application is further illustrated by the following examples, however the application is not limited to the examples or restricted by any specific embodiment of the examples.

[0132] Embodiments of the invention

[0133] Example 1 : Recombinant production of polypeptides

[0134] Genes for the polypeptide sequences (SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3) were synthesized by the commercial vendor TWIST Bioscience (https: / / www.twistbioscience.com / ) with codons optimized for expression in E. coli. The genes were inserted into the plasmid vector pE2 for expression under the control of a T7lac promoter (Dubendorf and Studier (1991), J. Mol. Biol. 219, 45-59). The resulting plasmids were transformed into E. coli BL21 (DE3) by using a modification of the heat shock procedure (Cohen et al. (1972), Proc. Natl. Acad. Sci. 69, 2110-2114) and by selection for plasmid-mediated kanamycin resistance. E. coli BL21 (DE3) carrying the plasmids were cultured in overnight expression transient TB medium (Merck) at 30 °C overnight for recombinant gene expression by autoinduction (Studier (2005), Protein Expr. Purif. 41, 207-234). The biomass was harvested by centrifugation, resuspended in 1.1x reaction buffer (reaction buffer: 25 mM Hepes pH 7.5; 10 mM ZnCl2) and lysed on ice with an ultrasonic disruption system (QSonica). The crude lysate was clarified by centrifugation (18 min 21 130 rcf 4 °C) and the presence of the polypeptides (enzymes) in the soluble fraction of the lysate was confirmed by SDS-PAGE (Laemmli (1970), Nature 227, 680-685) with a Bio-Rad 12% mini-PROTEAN TGX unstained precast gel.

[0135] Example 2: Analytical quantification of DON and 7-keto-8-hydroxy-8-enyl-DON

[0136] DON and the reaction products of the enzymatic activity of SEQ ID NO: 1 were separated by HPLC on a Phenomenex Kinetex C18 column (150 x 2.1 mm, 2.6 pm particle size) at 30 °C with a gradient of 5.9% to 95% acetonitrile with 0.1% acetic acid. The reaction products were identified by comparison to purified reference material and named 7-keto-8-hydroxy-8-ene-DON, following the atom numbering proposed by Yosizawa and Moroka in the first description of DON (Yosizama and Moroka (1973), Agric. Biol. Chem. 37, 2933-2934). Quantification was based on MS / MS using a Sciex QTRAP 5500 system in negative multiple reaction monitoring (MRM) mode and comparison to pure reference preparations of DON and 7-keto-8-hydroxy-8-ene-DON. The mass transition from the precursor ion [M+Ac - to the product ion 59.1 with m / z 355.1 was used as quantifier and the transition from m / z 355.1 to 265.1 as qualifier.

[0137] Example 3: DON modification / detoxification activity of polypeptides

[0138] A 1 mM aqueous solution of DON was added to the crude E. coli cell lysate prepared as described in Example 1 to start the reaction in a buffer containing 25 mM Hepes pH 7.5, 10 mM ZnCl2 and 100 mM DON. After incubation for 3 h at 30 °C, the reaction was terminated by freezing at -20 °C. To process the reaction end-point samples for HPLC analysis, the reaction was thawed, vortexed and 10 pl / reaction was added to 40 pl water and 200 pl acetonitrile. These samples were incubated with 80% acetonitrile and nominally 4 mM DON for 10 min at room temperature, centrifuged for 10 min at 21 130 rcf and 250 pl supernatant was added to 750 pl water. The samples diluted with 20% acetonitrile and nominally 1 mM DON were centrifuged again for 10 min at 21 130 rcf and 500 pl was transferred to HPLC vials for analysis. The polypeptide SEQ ID NO: 1 was able to convert DON to 7-keto-8-hydroxy-8-ene-DON (quantification shown in Table 1) Figure 3), whereas polypeptides SEQ ID NO:2 and SEQ ID NO:3 did not show any DON conversion or any DON concentration reduction. It is well known in the art that 7-keto-8-hydroxy-8-en-DON is less toxic than DON (Stadler et al. (2019), Food Chem. 279, 303-311; Stadler et al. (2019), Toxins (Basel) 11, 317). Thus, it was surprisingly found that SEQ ID NO: 1 is able to detoxify DON.

[0139] Example 4: Enzymatic kinetics of DON conversion

[0140] The polypeptide SEQ ID NO: 1 and the 35.2 kDa reference protein (negative control) were prepared as described in Example 1 by gene expression in E. coli BL21(DE3) in an overnight expression transient TB medium (Merck). Biomass from the same culture volume or 10 times the culture volume was resuspended in 1.1x reaction buffer (reaction buffer: 25 mM Hepes pH 7.5; 10 mM ZnCI2) and lysed with an ultrasonic disruption system (QSonica) on ice. The lysate was clarified by centrifugation (20 min 30 272 rcf 4°C) and the presence of the recombinant protein in the clarified lysate was confirmed by SDS-PAGE. The reaction was started by adding 1 mM DON to a final concentration of 100 mM DON in 25 mM Hepes pH 7.5 with 10 mM ZnCI2 and incubated at 30°C. Time point samples were taken and inactivated by adding 10 pl sample to 40 pl water and 200 pl acetonitrile. These stopped samples were centrifuged (20 min 30 272 rcf 4°C) and 250 pl supernatant was transferred to 750 pl water to reach a final nominal DON concentration of 1 mM in 20% acetonitrile. After centrifugation (1 min 21 130 rcf 4°C), 500 pl was transferred to HPLC vials for analysis. The time course of DON and 7-keto-8-hydroxy-8-en-DON concentration is shown in Figure 4 . SEQUENCE LISTING <110> Ecolab Inc. <120> Methods and compositions for degrading deoxynivalenol <130> LC21310018 <150> EP 20203359.3 <151> 22 October 2020 <160> 3 <170> PatentIn version 3.5 <210> 1 <211> 182 <212> PRT <213> Gossypium spp. <400> 1 Met Gly Ser Leu Asp Ser Lys Glu Ser Pro Ala Asn Asn Pro Gly Leu 1 5 10 15 His Ser Pro Pro Asp Glu Ala Thr Lys Gly Tyr Ile Met Gln Gln Thr 20 25 30 Met Phe Arg Ile Lys Asp Pro Lys Pro Thr Leu Glu Phe Tyr Ser Arg 35 40 45 Val Leu Gly Met Ser Leu Leu Asn Lys Val Asp Val Pro Tyr Met Lys 50 55 60 Met Thr Leu Tyr Met Met Gly Tyr Glu Asp Val Ser Ser Ala Pro Ser 65 70 75 80 Asp Pro Val Glu Lys Thr Ile Trp Thr Phe Gly Arg Pro Ala Thr Met 85 90 95 Glu Leu Thr His Phe Trp Gly Thr Glu Asn Asp Pro Glu Phe Lys Gly 100 105 110 Tyr His Asp Gly Asn Ser Glu Pro Thr Gly Phe Gly His Ile Gly Ile 115 120 125 Thr Val Asp Asp Met Tyr Lys Ala Cys Glu Arg Phe Glu Ser Leu Gly 130 135 140 Val Glu Phe Val Lys Lys Pro Gly Asp Gly Tyr Ala Phe Ile Lys Asp 145 150 155 160 Pro Asp Gly Tyr Trp Ile Glu Ile Phe Asp Leu Asn Gly Ile Arg Ala 165 170 175 Ile Val Asn Asn Leu Ala 180 <210> 2 <211> 182 <212> PRT <213> Gossypium spp. <400> 2 Met Ala Ser Leu Asp Ser Lys Glu Ser Pro Ala Asn Asn Pro Gly Leu 1 5 10 15 His Phe Pro Pro Asp Glu Ala Thr Lys Gly Tyr Ile Met Gln Gln Thr 20 25 30 Met Phe Arg Ile Lys Asp Pro Lys Arg Thr Leu Glu Phe Tyr Ser Arg 35 40 45 Val Leu Gly Met Thr Leu Leu Asn Lys Val Asp Val Pro Tyr Met Lys 50 55 60 Met Thr Leu Tyr Met Met Gly Tyr Glu Asp Val Ser Ser Ala Pro Thr 65 70 75 80 Asp Pro Val Glu Lys Thr Ile Trp Thr Phe Gly Arg Pro Ala Thr Met 85 90 95 Glu Leu Thr His Phe Trp Gly Thr Glu Asn Asp Pro Glu Phe Lys Gly 100 105 110 Tyr His Asp Gly Asn Ser Glu Pro Ile Gly Phe Gly His Ile Gly Leu 115 120 125 Thr Val Asp Asp Leu Tyr Lys Ala Cys Glu Arg Phe Glu Ser Leu Gly 130 135 140 Val Glu Phe Val Lys Lys Pro Ser Asp Gly Phe Ala Phe Ile Lys Asp 145 150 155 160 Pro Asp Gly Tyr Trp Ile Glu Ile Phe Asp Leu Lys Gly Ile Arg Gln 165 170 175 Ile Val Asn Ser Leu Ala 180 <210> 3 <211> 182 <212> PRT <213> Gossypium ssp. <400> 3 Met Gly Ser Met Asp Ser Lys Glu Ser Pro Ala Asn Asn Pro Gly Leu 1 5 10 15 His Thr Pro Pro Asp Glu Ala Thr Lys Gly Tyr Ile Met Gln Gln Thr 20 25 30 Met Phe Arg Ile Lys Asp Pro Lys Arg Thr Leu Glu Phe Tyr Ser Arg 35 40 45 Val Leu Gly Met Thr Leu Leu Asn Lys Val Asp Val Pro Tyr Met Lys 50 55 60 Met Thr Leu Tyr Met Met Gly Tyr Glu Asp Val Ser Ser Ala Pro Asn 65 70 75 80 Asp Pro Val Glu Lys Thr Ile Trp Thr Phe Gly Arg Pro Ala Thr Met 85 90 95 Glu Leu Thr His Phe Trp Gly Thr Glu Asn Asp Pro Glu Phe Lys Gly 100 105 110 Tyr His Asp Gly Asn Ser Glu Pro Ile Gly Phe Gly His Ile Gly Ile 115 120 125 Thr Val Asp Asp Met Tyr Lys Ala Cys Glu Arg Phe Glu Ser Leu Gly 130 135 140 Val Glu Phe Val Lys Lys Pro Ser Asp Gly Phe Ala Phe Ile Lys Asp 145 150 155 160 Pro Asp Gly Tyr Trp Ile Glu Ile Phe Asp Leu Lys Gly Ile Arg Gln 165 170 175 Ile Val Asn Ser Leu Ala 180

Claims

1. A method for producing food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxificant, intermediate detoxificant, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic, and / or mixtures thereof, the method comprising: i) providing a polypeptide, wherein the amino acid sequence of the polypeptide is set forth in SEQ ID NO: 1 ; ii) applying the polypeptide to a nutritional source or material suitable for producing food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxificant, intermediate detoxificant, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic, and / or mixtures thereof.

2. The method of claim 1, wherein the polypeptide is a recombinant and / or isolated polypeptide.

3. The method of claim 1, wherein the polypeptide is capable of modifying the C7 atom of deoxynivalenol (DON) and / or a DON derivative.

4. The method of claim 1, further comprising: incubating the nutritional source or material with the polypeptide under conditions suitable for degrading and / or altering the toxicity of DON and / or a DON derivative.

5. The method of claim 4, further comprising heat treating and / or fractionating and / or drying the incubated product.

6. The method of claim 4, wherein the altering the toxicity of DON and / or a DON derivative comprises one or more of detoxifying DON and / or the DON derivative and / or intramolecularly rearranging DON and / or the DON derivative.

7. The method of claim 5, wherein the altering the toxicity of DON and / or a DON derivative comprises one or more of detoxifying DON and / or the DON derivative and / or intramolecularly rearranging DON and / or the DON derivative.

8. The method of any one of claims 1 to 7, wherein the DON derivative is selected from the group consisting of: 3-keto deoxynivalenol, 3-epi deoxynivalenol, 15-acetyl deoxynivalenol, nivalenol, fusarenon-X, 3-amino deoxynivalenol.

9. The method of claim 8, wherein fusarenon-X is 4-acetyl nivalenol.

10. A food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxificant, intermediate detoxificant, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic, and / or mixtures thereof produced by the method of any one of claims 1 to 9, wherein the food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxificant, intermediate detoxificant, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic, and / or mixtures thereof comprises the polypeptide as defined in claim 1.

11. A method for degrading deoxynivalenol (DON) and / or a DON derivative and / or altering the toxicity of DON and / or the DON derivative, the DON derivative selected from the group consisting of: 3-ketodeoxynivalenol, 3-epideoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, fusarenon-X, 3-amino deoxynivalenol, the method comprising: i) providing a polypeptide, wherein the amino acid sequence of the polypeptide is set forth in SEQ ID NO: 1 ; ii) applying the polypeptide to DON and / or the DON derivative; and the method is not for preventing, diagnosing or treating a disease.

12. The method of claim 11, wherein the polypeptide is a recombinant and / or isolated polypeptide.

13. The method of claim 11, the altering the toxicity of DON and / or the DON derivative comprises one or more of: detoxifying DON and / or a DON derivative and / or intramolecularly rearranging DON and / or a DON derivative.

14. The method of claim 11, wherein the polypeptide is capable of modifying the C7 atom of DON and / or a DON derivative.

15. The method of any one of claims 11 to 14, wherein the method is an in vitro or ex vivo method.

16. A composition or kit comprising: (i) a polypeptide, wherein the amino acid sequence of the polypeptide is set forth in SEQ ID NO: 1 ; (ii) a nutritional source and / or substrate for the polypeptide, the nutritional source and / or substrate comprising DON and / or a DON derivative.

17. The composition or kit of claim 16, wherein the polypeptide is a recombinant and / or isolated polypeptide.

18. The composition or kit of claim 16, wherein the polypeptide is capable of detoxifying DON and / or a DON derivative and / or intramolecularly rearranging DON and / or a DON derivative.

19. The composition or kit of claim 16, wherein the polypeptide is capable of modifying the C7 atom of DON and / or a DON derivative.

20. The composition or kit of claim 16, wherein the polypeptide is encoded by a nucleotide sequence.

21. The composition or kit of claim 20, wherein the nucleotide sequence is comprised by a nucleic acid comprised by a host cell.

22. The composition or kit of claim 16, wherein the composition or kit is one or more of: a cell-free and / or non-naturally occurring and / or fractionated composition or kit.

23. The composition or kit of claim 16, wherein the composition or kit is a pharmaceutical and / or veterinary composition or kit.

24. The composition or kit of claim 22, wherein the composition or kit is a pharmaceutical and / or veterinary composition or kit.

25. The composition or kit of any one of claims 16 to 24, wherein the DON derivative is selected from the group consisting of: 3-keto-deoxynivalenol, 3-epi-deoxynivalenol, 15-acetyl-deoxynivalenol, deoxynivalenol, fusarenon-X, 3-amino-deoxynivalenol.

26. The composition or kit of claim 25, wherein fusarenon-X is 4-acetyl-deoxynivalenol.

27. Use of the composition or kit of any one of claims 16 to 26 or the food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxifying agent, intermediate detoxifying agent, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic or mixture thereof of claim 10 for the manufacture of a medicament.

28. The use of claim 27, wherein the medicament is for veterinary use.

29. Use of the composition or kit of any one of claims 16 to 26 or the food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxifying agent, intermediate detoxifying agent, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic or mixture thereof of claim 10 for the manufacture of a medicament or agent for the treatment, amelioration, prevention and / or diagnosis of DON mycotoxin poisoning.

30. A method for modifying the C7 atom of deoxynivalenol (DON) and / or a DON derivative selected from the group consisting of: 3-keto-deoxynivalenol, 3-epi-deoxynivalenol, 15-acetyl-deoxynivalenol, deoxynivalenol, fusarenon-X, 3-amino-deoxynivalenol, the method comprising: i) providing a polypeptide, wherein the amino acid sequence of the polypeptide is set forth in SEQ ID NO: 1 ; ii) applying the polypeptide to the DON and / or the DON derivative; and the method is not for the prevention, diagnosis or treatment of a disease.

31. The method of claim 30, wherein the polypeptide is a recombinant and / or isolated polypeptide.

32. The method of claim 30 or 31, wherein the method is an in vitro or ex vivo method.

33. One or more of: (a) a polypeptide, wherein the amino acid sequence of the polypeptide is set forth in SEQ ID NO: 1 ; (b) the composition or kit of any one of claims 16 to 25 or the food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxifying agent, intermediate detoxifying agent, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic and / or mixture thereof of claim 10; for use in: i) modifying the C7 atom of DON and / or a DON derivative, the use not being for the prevention, diagnosis or treatment of a disease; ii) producing food, intermediate food, forage, intermediate forage, feed, intermediate feed, additive, intermediate additive, detoxifying agent, intermediate detoxifying agent, nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic, and / or mixtures thereof; iii) degrading DON and / or DON derivatives and / or altering the toxicity of DON and / or DON derivatives, the use not being for the prevention, diagnosis or treatment of a disease; or iv) any combination of (i) - (iii); and the use according to any one of (i), (iii) and (iv), wherein the use is an in vitro or ex vivo use.

34. The use according to claim 33, wherein the polypeptide is a recombinant and / or isolated polypeptide.

35. The use according to claim 33, the altering the toxicity of DON and / or DON derivatives comprising one or more of detoxifying DON and / or the DON derivatives and / or intramolecularly rearranging DON and / or the DON derivatives.

36. The use according to claim 33, wherein the polypeptide is capable of modifying the C7 atom of DON and / or DON derivatives.

37. The use according to any one of claims 33 to 36, wherein the DON derivative is selected from the group consisting of 3-keto-deoxynivalenol, 3-epi-deoxynivalenol, 15-acetyl- deoxynivalenol, deoxynivalenol, fusarenon-X, 3-amino-deoxynivalenol.

38. The use according to claim 37, wherein fusarenon-X is 4-acetyl-deoxynivalenol.

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