Means and methods for modifying fumonisins

Abstract

The present invention relates to a method for improving the enzyme kinetics of a fumonisins esterase, to an improved fumonisins esterase and the use thereof, to a composition comprising said fumonisins esterase and to a method for modifying fumonisins.

Description

[0001] The present invention relates to a method for improving the enzyme kinetics of fumonisin esterase, an improved fumonisin esterase and uses thereof, a composition comprising said fumonisin esterase and a method for modifying fumonisins.

[0002] Mycotoxins are secondary metabolites produced by fungal species. Fungi not only infect food, but also feed materials. In particular, all types of cereals and forage crops as well as other commodities such as cotton, coffee, peanuts, dates, spices, etc. are affected. After fungal growth, mycotoxins such as aflatoxins, ochratoxins, ergot alkaloids, fumonisins, zearalenone and / or trichothecenes are produced, thereby contaminating the infected material.

[0003] In more than 17,000 samples, almost three quarters were found to contain at least one mycotoxin (Streit et al., 2013. J Sci Food Agric. 93(12): 2892-2899). One of the most prominent groups of mycotoxins is fumonisins. Fumonisins are produced, inter alia, by Fusarium spp. and Alternaria spp. fungi and, upon ingestion, impair sphingolipid metabolism, thereby causing a number of adverse effects. In particular, fumonisins are associated with immunosuppression, inflammation, carcinogenicity, embryonic malformations, etc. In animal husbandry, animals ingesting fumonisins not only suffer from health hazards associated with fumonisin mycotoxicosis, but also show decreased performance, thereby causing considerable economic losses.

[0004] Based on structural similarity, the fumonisin group includes fumonisin Al (CAS No. 117415-48-2), A2 (CAS No. 117415-47-1), Bl (CAS No. 116355-83-0), B2 (CAS No. 116355-84-1), B3 (CAS No. 136379-59-4), B4 (CAS No. 136379-60-7), Cl (PubChem CID 42608361), C2 (PubChem CID 42608362), C3 (PubChem CID 42608363), C4 (PubChem CID 42608364), and HFC1; and Alternaria alternata lycopersici toxin (AAL-T) Al (CAS No. 79367-52-5), A2 (CAS No. 79367-51-4), Bl (CAS No. 149849-90-1), and B2 (CAS No. 149849-91-2); and partially hydrolyzed derivatives thereof. In the partially hydrolyzed fumonisins, one of the two propanetriol acid residues has been removed from the non-hydrolyzed molecule. The following formula exemplarily illustrates fumonisin Al, A2, Bl, B2, B3, B4, Cl, C2, C3, and C4, where for fumonisin Al, R1=OH, R2=OH, R3=CH2CO, R4=CH3; for fumonisin A2, R1=H, R2=OH, R3=CH2CO, R4=CH3; for fumonisin Bl, R1=OH, R2=OH, R3=H, R4=CH3; for fumonisin B2, R1=H, R2=OH, R3=H, R4=CH3; for fumonisin B3, R1=OH, R2=H, R3=H, R4=CH3; for fumonisin B4, R1=H, R2=H, R3=H, R4=CH3; for fumonisin Cl, R1=OH, R2=OH, R3=H, R4=H; for fumonisin C2, R1=H, R2=OH, R3=H, R4=H; for fumonisin C3, R1=OH, R2=H, R3=H, R4=H; and for fumonisin C4, R1=H, R2=H, R3=H, R4=H. Within the fumonisin group, fumonisin Bl (FB1) is the most common mycotoxin. FB1 has been described as hepatotoxic and nephrotoxic, and has been linked to at least equine leukoencephalomalacia and porcine pulmonary edema syndrome.

[0005]

[0006] By applying "good agricultural practice", in particular by avoiding moisture as far as possible to the disadvantage of fungal growth, or by applying fungicides to the food / feed material, it is possible to attempt to contain the extent of fungal contamination. Furthermore, contaminated material should be removed and discarded before processing. However, due to the high prevalence and low median toxic dose, further measures have to be considered.

[0007] In WO 2006 / 053357 A2, the microbial conversion of fumonisins into non-toxic products is described. In WO 2010 / 031101 A1, polypeptides for the enzymatic degradation of fumonisins are described. In WO 2016 / 134387 A1, fumonisin esterase variants with improved temperature stability are described. However, in order to achieve even more efficient and faster removal of fumonisins molecules, there is still a need for fumonisin cleaving enzymes with improved kinetics.

[0008] In view of the prior art as described above, it is an object of the present application to provide devices and methods for modifying fumonisins, in particular for detoxifying fumonisins.

[0009] This object is achieved by providing a method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1; the method comprising: substituting the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; substituting the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and / or substituting the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine. When one or more of these substitutions are performed, it is possible to generate a fumonisin esterase with improved enzyme kinetics, i.e. improved enzyme activity. Such improved fumonisin esterase allows for a faster and more efficient removal of fumonisins, e.g. from nutritional compositions.

[0010] A fumonisin esterase as referred to herein is an esterase capable of modifying one or more fumonisins, in particular such esterase capable of modifying at least one of fumonisins Al, A2, Bl, B2, B3, B4, Cl, C2, C3, C4, AAL-TAl, AAL-TA2, AAL-TBl, AAL-TB2. Upon fumonisin modification by the fumonisin esterase, the tricarballylic acid residue is cleaved from the reactant fumonisin.

[0011] The term “amino acid” is to be interpreted as known to the person skilled in the art. Preferably, the term refers to proteinogenic amino acids. As known in the art, amino acids can be referred to using a three-letter code or a one-letter code. Therein, “Ala” and “A” refer to alanine, “Arg” and “R” refer to arginine, “Asn” and “N” refer to asparagine, “Asp” and “D” refer to aspartate (also known as aspartic acid), “Cys” and “C” refer to cysteine, “Gln” and “Q” refer to glutamine, “Glu” and “E” refer to glutamate (also known as glutamic acid), “Gly” and “G” refer to glycine, “His” and “H” refer to histidine, “Ile” and “I” refer to isoleucine, “Leu” and “L” refer to leucine, “Lys” and “K” refer to lysine, “Met” and “M” refer to methionine, “Phe” and “F” refer to phenylalanine, “Pro” and “P” refer to proline, “Ser” and “S” refer to serine, “Thr” and “T” refer to threonine, “Trp” and “W” refer to tryptophan, “Tyr” and “Y” refer to tyrosine, “Val” and “V” refer to valine.

[0012] When referring to a certain amino acid at a certain position in a polypeptide, the amino acid is typically described by indicating the amino acid by its one-letter code letter, followed by indicating the position, i.e. the number of the amino acid in the amino acid chain of the polypeptide. For example, T251 relative to the amino acid sequence of SEQ ID NO: 1 indicates a threonine residue at position 25 in a polypeptide having the amino acid sequence of SEQ ID NO: 1. Similarly, to indicate a substitution of a certain amino acid at a certain position with another amino acid, the amino acid to be substituted (i.e. the “original” amino acid) is first indicated by its one-letter code letter, followed by its position in the amino acid chain of the polypeptide, followed by the one-letter code letter of the amino acid to substitute the original amino acid. For example, T251 relative to the amino acid sequence of SEQ ID NO: 1 indicates that the threonine at position 25 in a polypeptide having the amino acid sequence of SEQ ID NO: 1 is to be replaced (i.e. substituted) with an isoleucine. Similarly, for example the term “149F” indicates that the amino acid at position 149 is to be replaced with a phenylalanine.

[0013] For completeness only and as known in the art, "aliphatic amino acids" are amino acids selected from the group consisting of alanine, glycine, isoleucine, leucine, methionine, proline, and valine; "polar amino acids" are amino acids selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, and aspartic acid; "charged amino acids" are amino acids selected from the group consisting of lysine, arginine, histidine, glutamic acid, and aspartic acid.

[0014] The term "polypeptide" is to be interpreted in the manner normally used in the art and encompasses, for example, polypeptides, proteins, peptides, enzymes.

[0015] The term "sequence identity" is used to describe the degree of relatedness between two or more nucleic acid sequences (e.g., DNA or RNA polynucleotides) contained in a polynucleotide or two or more amino acid sequences contained in a polypeptide. Sequence identity can be determined by routine methods known to the skilled person. Herein, the preferred method for determining sequence identity between two amino acid sequences is using the Clustal Omega alignment tool EMBL-EBI (https: / / www.ebi.ac.uk / Tools / msa / clustalo / ; Sievers et al., 2011. Mol. Syst. Biol. 7: 539) with default settings. Alternatively, the Needleman-Wunsch algorithm for global sequence alignment can be used, e.g., as provided by the National Center for Biotechnology Information using the algorithm ("Needleman-Wunsch Global Align Protein Sequences") with default settings (Gap Costs: Existence: 11 Extension 1). A polypeptide comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of a polypeptide of interest can be a polypeptide comprising an amino acid sequence having 70% or more % sequence identity, e.g., 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or even 100% sequence identity to the amino acid sequence of the polypeptide of interest. For example, a polypeptide comprising an amino acid sequence having 100% sequence identity to SEQ ID NO: 1 is encompassed by the group of "a polypeptide comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1". It is common practice in the art to construct fusion polypeptides that link one or more peptide tags to the N- and / or C-terminus of a polypeptide, e.g., in order to facilitate easier purification or to increase solubility, without substantially altering the enzymatic properties of the tagged polypeptide. In general, the term "fusion polypeptide" or "fusion protein" describes two or more polypeptide chains that are linked to each other, typically through a peptide bond. Examples of commonly used tags are poly(His) tags (e.g., hexa-histidine tags), maltose binding protein (MBP) tags, Strep tags, Strep II tags, etc. Thus, the present application encompasses enzymatically active fusion polypeptides, including polypeptides comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO. 1.Such fusion polypeptides may, for example, contain a hexahistidine tag fused to the N-terminus of a polypeptide comprising an amino acid sequence having 91 % sequence identity to the amino acid sequence of SEQ ID NO: 1, or, for example, a maltose binding protein tag fused to the C-terminus of a polypeptide comprising an amino acid sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO: 1 via a linker peptide (e.g. GG; GPG; EA or EA repeat; GSG etc.). In other words, said larger polypeptide comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 as part of a larger polypeptide is also encompassed by the present application, as long as the enzymatic activity of said amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 is not essentially eliminated. For the sake of clarity only, the enzymatically active polypeptide according to the present application is capable of and thus suitable for modifying, in particular cleaving and / or detoxifying at least one fumonisin, preferably fumonisin B1.

[0016] In some embodiments, a method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity) of a fumonisin esterase is provided, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1; the method comprising: substituting the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; substituting the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and / or substituting the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine; further comprising the step of substituting at least one further amino acid relative to the amino acid sequence of SEQ ID NO: 1 at a position selected from the group consisting of 10, 33, 66, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490. By introducing said further amino acid substitution at one or more of these positions, the fumonisin esterase can be further improved, in particular not only with respect to improving the enzyme kinetic properties, but also with respect to improving the temperature stability.In preferred embodiments, the at least one further amino acid substitution is selected from 10Q, 33E, 66A, 66C, 66F, 66L, 66M, 66T, 66W, 66E, 66H, 66I, 66K, 66P, 66R, 66Y, 66G, 66Q, 66S, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371V, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N, and 490P, relative to the amino acid sequence of SEQ ID NO: 1.

[0017] In one aspect, the present application relates to a method for improving the enzyme kinetics (e.g., reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax), and / or increased specific activity) of a thermostable fumonisin esterase, the method comprising: (i) providing the thermostable fumonisin esterase, wherein the thermostable fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the thermostable fumonisin esterase comprises an aspartic acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1, and at least one additional mutation at a position relative to the amino acid sequence of SEQ ID NO: 1 selected from the group consisting of 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487, and 490, preferably wherein the at least one additional mutation is selected from the group consisting of 10Q, 33E, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N, and 490P relative to the amino acid sequence of SEQ ID NO: 1; and (ii) substituting the amino acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1 with an amino acid other than aspartic acid.

[0018] In particular, "amino acids other than aspartate" are alanine, arginine, asparagine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine.

[0019] A thermostable fumonisin esterase is described in WO 2016 / 134387 Al and comprises an aspartate at position 66 relative to the amino acid sequence of SEQ ID NO: 1 and at least one additional mutation at a position selected from 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490 relative to the amino acid sequence of SEQ ID NO: 1. Preferably, the at least one additional mutation of the thermostable fumonisin esterase is selected from 10Q, 33E, 107E, 140P, 144M, 149F, 151 R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371 M, 372F, 377V, 389L, 391 V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N and 490P relative to the amino acid sequence of SEQ ID NO: 1.

[0020] In another aspect, the present application relates to a fumonisin esterase obtainable by the method for improving enzyme kinetics according to the present application. Advantageously, such enzyme allows for a faster and more efficient modification, in particular detoxification or bioconversion, of fumonisins.

[0021] In some embodiments, the present application relates to a fumonisin esterase comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the fumonisin esterase comprises: (i) an amino acid substitution of the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and / or (ii) an amino acid substitution of the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and / or (iii) an amino acid substitution of the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine. In some embodiments, the fumonisin esterase according to the present application comprises only one of the mutations (i), (ii) and (iii). In some embodiments, the fumonisin esterase according to the present application comprises mutations (i) and (ii); (i) and (iii); (ii) and (iii); or (i), (ii) and (iii).

[0022] In some embodiments, the fumonisin esterase according to the application further comprises at least one additional amino acid substitution at a position selected from the group consisting of 10, 33, 66, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490 relative to the amino acid sequence of SEQ ID NO: 1. Thus, a fumonisin esterase is provided which shows improved enzyme kinetics as well as improved stability, in particular temperature stability, compared to the non-mutated fumonisin esterase, in particular compared to the fumonisin esterase of SEQ ID NO: 1. In a preferred embodiment, the fumonisin esterase according to the application comprises at least one additional amino acid substitution relative to the amino acid sequence of SEQ ID NO: 1 selected from the group consisting of 10Q, 33E, 66A, 66C, 66F, 66L, 66M, 66T, 66W, 66E, 66H, 66I, 66K, 66P, 66R, 66Y, 66G, 66Q, 66S, 107E, 140P, 144M, 149F, 151 R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371 V, 371 M, 372F, 377V, 389L, 391 V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N and 490P.

[0023] Similarly, the present application relates to a heat-stable fumonisin esterase comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the heat-stable fumonisin esterase comprises at least one mutation at a position selected from 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490 relative to the amino acid sequence of SEQ ID NO: 1, preferably wherein the at least one mutation is selected from 10Q, 33E, 107E, 140P, 144M, 149F, 151 R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371 M, 372F, 377V, 389L, 391 V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N and 490P relative to the amino acid sequence of SEQ ID NO: 1 ; and wherein the heat-stable fumonisin esterase comprises an amino acid other than aspartic acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1.

[0024] In certain embodiments, the fumonisin esterase according to the application comprises or consists of an amino acid sequence which is identical to the amino acid sequence of SEQ ID NO: 1, with the difference that the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1 ; and / or with the difference that the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1 ; and / or with the difference that the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1. In certain embodiments, the fumonisin esterase according to the application comprises or consists of an amino acid sequence which is identical to the amino acid sequence of SEQ ID NO: 1, with the difference that the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1 is replaced by an aliphatic amino acid, more preferably by an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably by an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably by isoleucine; and / or with the difference that the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1 is replaced by a polar amino acid, more preferably by an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably by an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably by glutamine; and / or with the difference that the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1 is replaced by a charged amino acid, more preferably by an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably by an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably by arginine. In certain embodiments, the fumonisin esterase according to the application comprises or consists of an amino acid sequence which is identical to the amino acid sequence of SEQ ID NO: 1, with the difference that the amino acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1 is an amino acid other than aspartic acid; and with the difference that any one or more of the amino acids at positions selected from the group consisting of 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490 relative to the amino acid sequence of SEQ ID NO: 1 is an amino acid other than aspartic acid.In certain embodiments, a fumonisin esterase according to the application comprises or consists of an amino acid sequence that is identical to the amino acid sequence of SEQ ID NO: 1, except that (relative to the amino acid sequence of SEQ ID NO: 1) the amino acid at position 66 is an amino acid other than aspartic acid; and except that (relative to the amino acid sequence of SEQ ID NO: 1) at least one mutation selected from the group consisting of 10Q, 33E, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N, and 490P.In certain embodiments, a fumonisin esterase according to the application comprises or consists of an amino acid sequence identical to the amino acid sequence of SEQ ID NO: 1, except for (relative to the amino acid sequence of SEQ ID NO: 1) at least one mutation selected from the group consisting of 10Q, 33E, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N, and 490P. In certain embodiments, a fumonisin esterase according to the application comprises or consists of the amino acid sequence of any one of SEQ ID NO. 2 to SEQ ID NO. 71.

[0025] In another aspect, the present application relates to a composition (e.g., an additive for food and / or feed; a feed or food) for modifying (e.g., detoxifying) fumonisins, the composition comprising at least one fumonisin esterase according to the application.

[0026] The composition according to the present application can be, for example, a food additive; a foodstuff additive; a forage additive; a feed additive; a nutritional supplement; intermediates thereof; and / or mixtures thereof. Such compositions can comprise additional components, such as prebiotics and / or probiotics. For the sake of clarity only, a forage or feed can for example comprise or consist of corn, hay, straw litter, soybean or products obtained therefrom. Also, a forage or feed can comprise or consist of an expanded feed product, such as a pellet. Additives for food, foodstuff, forage or feed are often used to improve or increase properties of the foodstuff, forage or feed. For example, such additives can be added to improve organoleptic properties, for example to improve the taste, smell, appearance, colour of the food, foodstuff, forage or feed. Also, additives can be added to improve palatability, nutrient utilization, or to add probiotic microorganisms to the food, foodstuff, forage or feed, or to add or enhance prebiotic activity of the food, foodstuff, forage or feed. Also, such additives can be added to counteract potential undesired effects of the food, foodstuff, forage or feed, such as to remove or reduce one or more undesired components comprised in the food, foodstuff, forage or feed.

[0027] In some embodiments, the compositions of the present application comprise at least one carrier. The carrier can be any suitable carrier. The composition can comprise one, two, three, or even more carriers. The carrier can be a dietary supplement, a nutritional formulation, and / or a pharmaceutical, such as a vitamin, a mineral, an amino acid, an essential fatty acid, a fiber, a trace element, an antioxidant, a plant extract, a Chinese herb extract, and / or an essential oil. The carrier can also be a carrier for enzymes. Carriers for enzymes can be both inorganic and organic sources. Potential inorganic materials for immobilized enzymes are silicon dioxide (sol-gel silica, fumed silica, colloidal silica nanoparticles, and silica gel) and oxides such as titanium dioxide, aluminum oxide, and zirconium oxide. In addition, clay materials such as bentonite, halloysite, kaolinite, montmorillonite, sepiolite, and calcium apatite can also be carriers. Additionally, carbon-based materials such as activated carbon can be carriers. Organic enzyme carriers can be biopolymers (e.g., carbohydrates, proteins, maltodextrin, trehalose, inulin, collagen, cellulose, keratin, carrageenan, chitin, deacetylated chitin, and alginate) or synthetic polymers (e.g., polyaniline, polyamide, polystyrene, polyurethane, polypropylene, polyvinyl alcohol, and ion exchange resins). Liquid carriers can be, for example, buffer substances and / or polyols, such as polyalkylene oxide, polyvinyl alcohol, polyethylene-maleic anhydride copolymer, polystyrene-malic anhydride copolymer, dextran, cellulose, deacetylated chitin hydrolysate, starch, glycogen, sorbitol, agarose and its derivatives, guar gum, amylopectin, inulin, xanthan gum, carrageenan, pectin, alginic acid hydrolysate, biopolymer, sorbitol, glycerol, cellobiose, and monopropylene glycol. The carrier can additionally or alternatively be an edible component, preferably a non-toxic component and / or a component that provides texture. In particular embodiments, the carrier is selected from bentonite, silica, maltodextrin, and a carbohydrate; preferably, the carrier is bentonite and / or maltodextrin.

[0028] In another aspect, the present application relates to a method for modifying (e.g., detoxifying) a fumonisin in a composition (e.g., a nutritional composition), the method comprising contacting the composition with a fumonisin esterase according to the present application. In some embodiments of the method for modifying a fumonisin in a composition, the fumonisin esterase is included in a composition (e.g., an additive for food and / or feed; a feed or food) for modifying (e.g., detoxifying) a fumonisin according to the present application. In particular embodiments of the present application, the method for modifying (e.g., detoxifying) a fumonisin in a composition (e.g., in a nutritional composition) involves treating a composition comprising a fumonisin, but does not involve treating the human or animal body. Such embodiments can be achieved because the fumonisin esterase according to the present application is able to well modify a fumonisin comprised in a composition in the absence of a human or animal body. In other words, in particular embodiments, the present application relates to a non-medical method for modifying (e.g., detoxifying) a fumonisin in a composition (e.g., in a nutritional composition), the non-medical method comprising contacting the composition with a fumonisin esterase according to the present application.

[0029] A nutritional composition as referred to herein is a composition comprising one or more components having nutritional value. Often such components provide energy to a consumer of the nutritional composition. The nutritional composition can be based in whole or at least in part on herbs or plants, e.g. commonly used animal feed compositions.

[0030] In yet another aspect, the present application relates to a method for modifying (e.g., detoxifying or biotransforming) a fumonisin (i.e., at least one fumonisin) in a water body (e.g., fresh water, brackish water, or salt water; preferably fresh water or salt water) comprising (at least one) fumonisin, said method comprising contacting said water body with a fumonisin esterase, preferably a fumonisin esterase according to the present application. In some embodiments of the method for modifying a fumonisin in a water body, the fumonisin esterase is comprised in a composition (e.g., an additive for food and / or feed; a feed or food) for use in modifying (e.g., detoxifying) a fumonisin according to the present application. In a particular embodiment of the present application, the method for modifying (e.g., detoxifying) a fumonisin (i.e., at least one fumonisin) in a water body (e.g., fresh water, brackish water, or salt water; preferably fresh water or salt water) comprising (at least one) fumonisin involves treating a water body comprising a fumonisin, but does not involve treating the human or animal body. In other words, in a particular embodiment, the present application relates to a non-medical method for modifying (e.g., detoxifying or biotransforming) a fumonisin (i.e., at least one fumonisin) in a water body (e.g., fresh water, brackish water, or salt water; preferably fresh water or salt water) comprising (at least one) fumonisin, said non-medical method comprising contacting said water body with a fumonisin esterase, preferably a fumonisin esterase according to the present application.

[0031] In some embodiments, the method for modifying a fumonisin in a water body involves a method for modifying (e.g., detoxifying) a fumonisin in aquaculture, e.g., for farming fish, e.g., carp, salmon, tilapia, tuna, catfish, trout, rainbow trout; crustaceans, e.g., crabs, lobsters, crayfish, prawns, shrimps, krill; mollusks, e.g., oysters, mussels, clams, squid, cuttlefish, and octopus; echinoderms, e.g., sea cucumbers, sea urchins; jellyfish; algae; and / or seaweed.

[0032] In another aspect, the present application relates to a method for modifying (e.g., detoxifying) fumonisins in gastric juice, said method comprising contacting the gastric juice with a fumonisin esterase, preferably a fumonisin esterase according to the present application. In some embodiments, the gastric juice is an animal gastric juice, in particular a gastric juice of a ruminant, a pig or a poultry. In some embodiments of the method for modifying fumonisins in gastric juice, the fumonisin esterase is comprised in a composition (e.g., an additive for food and / or feed; a feed or food) for use in the modification (e.g., detoxification) of fumonisins according to the present application. In a particular embodiment of the present application, the method for modifying (e.g., detoxifying) fumonisins in gastric juice involves the treatment of gastric juice, but not the treatment of the human or animal body. In other words, in a particular embodiment, the present application relates to a non-medical method for modifying (e.g., detoxifying) fumonisins in gastric juice, said non-medical method comprising contacting the gastric juice with a fumonisin esterase, preferably a fumonisin esterase according to the present application.

[0033] It will be understood by the skilled person that in the method for modifying (e.g., detoxifying) fumonisins according to the present application, a reaction mixture is formed when the fumonisin esterase is contacted with the fumonisins. The reaction mixture can further be contacted with water. The water can be from the surrounding or ambient environment of the contact of the fumonisin esterase with the fumonisins (e.g., in a composition, such as a nutritional composition; in an aquatic body or aquaculture; in gastric juice), and / or from moisture or water present in the environment, and / or the water can be added by the person performing the method of the present application, and / or the water can originate from any other source, such as from saliva (e.g., after ingestion of the reaction mixture or a composition comprising the reaction mixture).

[0034] In yet another aspect, the present application relates to the use of a fumonisin esterase according to the present application and / or a composition (e.g., an additive for food and / or feed; a feed or food) comprising a fumonisin esterase according to the present application for modifying (e.g., detoxifying) fumonisins. In a particular embodiment of the present application, the use of a fumonisin esterase according to the present application and / or a composition (e.g., an additive for food and / or feed; a feed or food) comprising a fumonisin esterase according to the present application for modifying (e.g., detoxifying) fumonisins involves the use of the fumonisin esterase for treating a composition comprising fumonisins, but not the treatment of the human or animal body. In other words, in a particular embodiment, the present application relates to the non-medical use of a fumonisin esterase according to the present application and / or a composition (e.g., an additive for food and / or feed; a feed or food) comprising a fumonisin esterase according to the present application for modifying (e.g., detoxifying) fumonisins.

[0035] In yet another aspect, the present application relates to the use of a fumonisin esterase according to the present application and / or a composition comprising a fumonisin esterase according to the present application (e.g. an additive for food and / or feed; a feed or food) for the manufacture of an additive for feed and / or food, a feed and / or food composition, or a pharmaceutical composition.

[0036] In another aspect, the present application relates to the use of a fumonisin esterase according to the present application and / or a composition comprising a fumonisin esterase according to the present application (e.g. an additive for food and / or feed; a feed or food) for the manufacture of biogas; bioethanol, DDGS; sugar, preferably sugar from sugar cane or sugar beet; corn oil, corn germ, corn germ meal, corn fiber, corn gluten, starch, in particular corn starch; and / or silage.

[0037] In another aspect, the present application relates to a host cell comprising at least one fumonisin esterase according to the present application and / or comprising at least one polynucleotide encoding at least one fumonisin esterase according to the present application.

[0038] The term "host cell" refers to any cell capable of producing a recombinant protein. In particular, "host cell" refers to a prokaryotic and / or eukaryotic cell, preferably a Pichia pastoris, Escherichia coli, Bacillus sp. (e.g. Bacillus subtilis or Bacillus amyloliquefaciens), Streptomyces sp., Hansenula sp., Trichoderma sp., Lactobacillus sp., Aspergillus sp., a plant cell and / or spore of Bacillus, Trichoderma or Aspergillus. Notably, the strains that are usually associated with "Pichia pastoris" are sometimes referred to as Komagataella pastoris, Komagataella phaffii or Komagataella pseudopastoris, depending on the respective systematic classification at the time of the reference.

[0039] In another aspect, the present invention relates to a fumonisin esterase for treating, ameliorating and / or preventing or prophylaxis of symptoms caused by mycotoxin poisoning, in particular fumonisin mycotoxin poisoning, wherein the fumonisin esterase is a fumonisin esterase according to the present invention. In some embodiments, the fumonisin esterase is provided to a subject, such as an animal, in need of mycotoxin poisoning prevention, in particular fumonisin mycotoxin poisoning prevention. A subject may be considered in need of fumonisin mycotoxin poisoning prevention when there is a risk of ingesting fumonisins exceeding non-toxic concentrations. For example, a subject may be considered in need of fumonisin mycotoxin poisoning prevention when the subject is about to ingest and / or has ingested a nutritional composition comprising one or more detectable levels of fumonisins. Due to the improved characteristics of the fumonisin esterase according to the present invention, this enzyme is advantageously suitable for treating and / or improving symptoms of mycotoxin poisoning, in particular fumonisin mycotoxin poisoning, by reducing the subject's exposure to fumonisins and thereby reducing the symptoms associated with fumonisin exposure. Furthermore, the fumonisin esterase according to the present invention can be used to prevent symptoms of mycotoxicosis, in particular symptoms of fumonisin mycotoxicosis. For example, a fumonisin esterase according to the present invention can be administered to a subject known or suspected of having ingested fumonisin before the onset of symptoms, thereby preventing the occurrence of such symptoms caused by mycotoxicosis, in particular symptoms of fumonisin mycotoxicosis.

[0040] The present invention is further characterized by the following items:

[0041] Item 1. A method for improving the enzyme kinetics of a fumonisin esterase (e.g., decreased Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity), wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0042] The method comprises: replacing the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from isoleucine, leucine, valine, most preferably with isoleucine;

[0043] substitution of the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine;

[0044] and / or substitution of the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine.

[0045] Item 2. A method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0046] the method comprising substitution of the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine.

[0047] Item 3. A method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0048] the method comprising substitution of the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine.

[0049] Item 4. A method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0050] the method comprising: substituting the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine.

[0051] Item 5. A method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0052] the method comprising: substituting the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and

[0053] substituting the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine.

[0054] Item 6. A method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax) and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0055] The method comprises: substituting the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and

[0056] substituting the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and

[0057] Item 7. A method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax), and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0058] The method comprises: substituting the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and

[0059] substituting the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and

[0060] substituting the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and

[0061] Item 8. A method for improving the enzyme kinetics (e.g. reduced Michaelis constant (Km), increased turnover number (kcat), increased catalytic efficiency, increased maximum reaction rate (Vmax), and / or increased specific activity) of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1,

[0062] the method comprising: substituting the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and

[0063] substituting the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine.

[0064] Item 9. The method according to any one of the preceding items, further comprising: substituting the amino acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1 with an amino acid that is not aspartic acid; preferably with an amino acid that is not aspartic acid, valine; more preferably with an amino acid selected from the group consisting of alanine, cysteine, phenylalanine, leucine, methionine, threonine, tryptophan, glutamic acid, histidine, isoleucine, lysine, proline, arginine, tyrosine, glycine, glutamine, serine, and valine; most preferably with an amino acid selected from the group consisting of alanine, phenylalanine, leucine, methionine, asparagine, threonine, tryptophan, and histidine.

[0065] Item 10. The method according to item 9, wherein the amino acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1 is substituted with an amino acid selected from the group consisting of phenylalanine, methionine, tryptophan, histidine, and asparagine.

[0066] Item 11. The method according to any one of items 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, wherein the substitution is at least one additional amino acid substitution relative to the amino acid sequence of SEQ ID NO: 1 at a position selected from the group consisting of 10, 33, 66, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490, preferably wherein the at least one additional amino acid substitution relative to the amino acid sequence of SEQ ID NO: 1 is selected from the group consisting of 10Q, 33E, 66A, 66C, 66F, 66L, 66M, 66T, 66W, 66E, 66H, 66I, 66K, 66P, 66R, 66Y, 66G, 66Q, 66S, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371V, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N and 490P.

[0067] Item 12: A fumonisin esterase obtainable by the method according to any one of the preceding items, in particular item 1-11.

[0068] Item 13. A method for improving the enzyme kinetics of a thermostable fumonisin esterase, the method comprising:

[0069] (i) providing said thermostable fumonisin esterase, wherein said thermostable fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein said thermostable fumonisin esterase comprises an aspartic acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1, and wherein said thermostable fumonisin comprises at least one additional mutation at a position selected from the group consisting of 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487, and 490 relative to the amino acid sequence of SEQ ID NO: 1, preferably wherein said at least one additional mutation is selected from the group consisting of 10Q, 33E, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N, and 490P relative to the amino acid sequence of SEQ ID NO: 1; and

[0070] (ii) substituting the amino acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1 with an amino acid other than aspartic acid.

[0071] Clause 14. A method for improving the enzyme kinetics of a thermostable fumonisin esterase, the method comprising:

[0072] (i) providing said heat-stable fumonisin esterase, wherein said heat-stable fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein said heat-stable fumonisin esterase comprises an aspartic acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1 and at least one additional mutation at a position selected from the group consisting of 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490 relative to the amino acid sequence of SEQ ID NO: 1, preferably wherein said at least one additional mutation is selected from the group consisting of 10Q, 33E, 107E, 140P, 144M, 149F, 151 R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371 M, 372F, 377V, 389L, 391 V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N and 490P relative to the amino acid sequence of SEQ ID NO: 1 ; and

[0073] (ii) comprising a step of introducing an amino acid other than aspartic acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1.

[0074] Item 15. The method according to item 13 or 14, wherein the amino acid substitution at position 66 relative to the amino acid sequence of SEQ ID NO: 1 is with an amino acid other than aspartic acid, other than valine, preferably with an amino acid selected from the group consisting of alanine, cysteine, phenylalanine, leucine, methionine, asparagine, threonine, tryptophan, glutamic acid, histidine, isoleucine, lysine, proline, arginine, tyrosine, glycine, glutamine, serine and valine; more preferably selected from the group consisting of alanine, phenylalanine, leucine, methionine, asparagine, threonine, tryptophan and histidine.

[0075] Item 16: A fumonisin esterase obtainable by the method according to any one of the preceding items.

[0076] Item 17: A fumonisin esterase obtainable by the method according to any one of items 13-15.

[0077] Item 18: A fumonisin esterase comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the fumonisin esterase comprises:

[0078] (i) an amino acid substitution at the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and / or

[0079] (ii) an amino acid substitution at the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and / or

[0080] (iii) an amino acid substitution at the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine.

[0081] Item 19: A fumonisin esterase comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the fumonisin esterase comprises:

[0082] (i) an amino acid substitution to the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and

[0083] (ii) an amino acid substitution to the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine.

[0084] Item 20: A fumonisin esterase comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the fumonisin esterase comprises:

[0085] (i) an amino acid substitution to the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and

[0086] (ii) an amino acid substitution to the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine.

[0087] Item 21 : A fumonisin esterase comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the fumonisin esterase comprises:

[0088] (i) an amino acid substitution at the amino acid located at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and

[0089] (ii) an amino acid substitution at the amino acid located at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine.

[0090] Item 22: A fumonisin esterase comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the fumonisin esterase comprises:

[0091] (i) an amino acid substitution at the amino acid located at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from the group consisting of alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from the group consisting of isoleucine, leucine, valine, most preferably with isoleucine; and

[0092] (ii) an amino acid substitution at the amino acid located at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from the group consisting of tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and

[0093] (iii) an amino acid substitution at the amino acid located at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from the group consisting of lysine, arginine, histidine, most preferably with arginine.

[0094] Item 23: The fumonisin esterase according to any of the preceding items, in particular according to any of items 16-22, wherein the fumonisin esterase further comprises at least one additional amino acid substitution at a position relative to the amino acid sequence of SEQ ID NO: 1 selected from the group consisting of 10, 33, 66, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487, and 490.

[0095] Item 24: The fumonisin esterase according to any of the preceding items, in particular according to any of items 16-22, wherein the fumonisin esterase further comprises at least one additional amino acid substitution relative to the amino acid sequence of SEQ ID NO: 1 selected from the group consisting of 10Q, 33E, 66A, 66C, 66F, 66L, 66M, 66T, 66W, 66E, 66H, 66I, 66K, 66P, 66R, 66Y, 66G, 66Q, 66S, 107E, 140P, 144M, 149F, 151 R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371 V, 371 M, 372F, 377V, 389L, 391 V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N, and 490P.

[0096] Item 25: A fumonisin esterase, in particular a thermostable fumonisin esterase, comprising or consisting of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the fumonisin esterase, in particular the thermostable fumonisin esterase, comprises at least one mutation at a position selected from 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487, and 490, relative to the amino acid sequence of SEQ ID NO: 1, preferably wherein the at least one mutation is selected from 10Q, 33E, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, 465G, 469K, 473A, 478D, 487N, and 490P, relative to the amino acid sequence of SEQ ID NO: 1; and wherein the fumonisin esterase, in particular the thermostable fumonisin esterase, comprises an amino acid other than aspartic acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1.

[0097] Item 26: A fumonisin esterase comprising or consisting of the amino acid sequence according to any one of SEQ ID NO. 2 to SEQ ID NO. 71.

[0098] Item 27: A composition (e.g., an additive for food and / or feed; a feed or food) for modifying (e.g., detoxifying) fumonisins, the composition comprising a fumonisin esterase according to any one of the preceding items.

[0099] Item 28: A composition (e.g., an additive for food and / or feed; a feed or food) for modifying (e.g., detoxifying) fumonisins, the composition comprising at least one fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26.

[0100] Item 29: The composition according to any one of the preceding items, wherein the composition further comprises at least one carrier, preferably selected from bentonite, silicon dioxide, maltodextrin and a carbohydrate, more preferably selected from maltodextrin and bentonite.

[0101] Item 30: A method for modifying (e.g., detoxifying) fumonisins in a composition (e.g., in a nutritional composition), the method comprising contacting the composition with a fumonisin esterase according to any one of the preceding items.

[0102] Item 31: A method for modifying (e.g., detoxifying) fumonisins in a composition (e.g., in a nutritional composition), the method comprising contacting the composition with a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26.

[0103] Item 32: A method for modifying (e.g., detoxifying) fumonisins in a water body (e.g., in fresh water, brackish water or salt water; preferably in fresh water or salt water), the method comprising contacting the water body with a fumonisin esterase.

[0104] Item 33: A method for modifying (e.g., detoxifying) fumonisins in a water body (e.g., in fresh water, brackish water or salt water; preferably in fresh water or salt water), the method comprising contacting the water body with a fumonisin esterase according to any one of the preceding items.

[0105] Item 34: A method for modifying (e.g., detoxifying) fumonisins in a water body (e.g., in fresh water, brackish water or salt water; preferably in fresh water or salt water), the method comprising contacting the water body with a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26.

[0106] Item 35: A method for modifying (e.g., detoxifying) a fumonisin in an aquaculture (e.g., in an aquaculture for farming fish, such as carp, salmon, tilapia, tuna, catfish, trout, rainbow trout; crustaceans, such as crabs, lobsters, crayfish, prawns, shrimps, krill; mollusks, such as oysters, mussels, clams, squid, cuttlefish, and octopus; echinoderms, such as sea cucumbers, sea urchins; jellyfish; algae; and / or seaweed), the method comprising contacting the aquaculture with a fumonisin esterase.

[0107] Item 36: A method for modifying (e.g., detoxifying) a fumonisin in an aquaculture (e.g., in an aquaculture for farming fish, such as carp, salmon, tilapia, tuna, catfish, trout, rainbow trout; crustaceans, such as crabs, lobsters, crayfish, prawns, shrimps, krill; mollusks, such as oysters, mussels, clams, squid, cuttlefish, and octopus; echinoderms, such as sea cucumbers, sea urchins; jellyfish; algae; and / or seaweed), the method comprising contacting the aquaculture with a fumonisin esterase according to any of the preceding items.

[0108] Item 37: A method for modifying (e.g., detoxifying) a fumonisin in an aquaculture (e.g., in an aquaculture for farming fish, such as carp, salmon, tilapia, tuna, catfish, trout, rainbow trout; crustaceans, such as crabs, lobsters, crayfish, prawns, shrimps, krill; mollusks, such as oysters, mussels, clams, squid, cuttlefish, and octopus; echinoderms, such as sea cucumbers, sea urchins; jellyfish; algae; and / or seaweed), the method comprising contacting the aquaculture with a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26.

[0109] Item 38: A method for modifying (e.g., detoxifying) a fumonisin in a gastric juice (e.g., a gastric juice of a ruminant, a pig, or a poultry), the method comprising contacting the gastric juice with a fumonisin esterase.

[0110] Item 39: A method for modifying (e.g., detoxifying) a fumonisin in a gastric juice (e.g., a gastric juice of a ruminant, a pig, or a poultry), the method comprising contacting the gastric juice with a fumonisin esterase according to any of the preceding items.

[0111] Item 40: A method for modifying (e.g., detoxifying) a fumonisin in a gastric juice (e.g., a gastric juice of a ruminant, a pig, or a poultry), the method comprising contacting the gastric juice with a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26.

[0112] Item 41 : The method for modifying (e.g., detoxifying) a fumonisin according to any of the preceding items, in particular any of items 30-41, wherein the fumonisin esterase is comprised in a composition according to any of items 27-29.

[0113] Item 42: The method according to any of items 30-41, wherein the method comprises forming a reaction mixture by contacting the fumonisin esterase with the fumonisin, and incubating the reaction mixture for at least 1 minute, e.g., at least 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, or 15 minutes, etc., e.g., at least 30 minutes, 60 minutes, 90 minutes, or 120 minutes, or even longer, e.g., at least 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, or even longer.

[0114] Item 43: The method according to item 42, wherein the method comprises incubating the reaction mixture at a temperature of at least 5°C and at most 90°C, preferably at a temperature of at least 5°C and at most 50°C, more preferably at a temperature of at least 5°C and at most 40°C, more preferably at a temperature of at least 10°C and at most 40°C, even more preferably at a temperature of at least 20°C and at most 40°C.

[0115] Item 44: The method according to any of items 30-42, further comprising contacting the reaction mixture with water.

[0116] Item 45: Use of a fumonisin esterase according to any of the preceding items (e.g., a fumonisin esterase according to any of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26) and / or a composition according to any of items 27-29 for modifying (e.g., detoxifying) a fumonisin.

[0117] Item 46: Use of a fumonisin esterase according to any of the preceding items (e.g., a fumonisin esterase according to any of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26) for the manufacture of an additive for feed and / or food, a feed and / or food composition, or a pharmaceutical composition.

[0118] Item 47: Use of a composition according to any one of items 27-29 for the manufacture of an additive for feed and / or food, a feed and / or food composition, or a pharmaceutical composition.

[0119] Item 48: Use of a fumonisin esterase according to any one of the preceding items (e.g., a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26) and / or a composition according to any one of items 27-29 for the manufacture of an additive for feed and / or food, a feed and / or food composition, or a pharmaceutical composition.

[0120] Item 49: A method for the manufacture of an additive for feed and / or food, a feed and / or food composition, or a pharmaceutical composition, the method comprising contacting a fumonisin esterase according to any one of the preceding items (e.g., a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26) with at least one additional component of an additive for feed and / or food, a feed and / or food composition, or a pharmaceutical composition.

[0121] Item 50: Use of a fumonisin esterase according to any one of the preceding items (e.g., a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26) and / or a composition according to any one of items 27-29 for the manufacture of biogas; bioethanol, DDGS; sugar, preferably sugar from sugar cane or sugar beet; corn oil, corn germ, corn germ meal, corn fiber, corn gluten meal, starch, in particular corn starch; and / or silage.

[0122] Item 51: A method for the manufacture of biogas; bioethanol, DDGS; sugar, preferably sugar from sugar cane or sugar beet; corn oil, corn germ, corn germ meal, corn fiber, corn gluten meal, starch, in particular corn starch; and / or silage, the method comprising contacting a fumonisin esterase according to any one of the preceding items (e.g., a fumonisin esterase according to any one of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26) and / or a composition according to any one of items 27-29 with at least one additional component of biogas; bioethanol, DDGS (dried distillers grains with solubles); sugar, preferably sugar from sugar cane or sugar beet; corn oil, corn germ, corn germ meal, corn fiber, corn gluten meal, starch, in particular corn starch; and / or silage.

[0123] Item 52: A host cell comprising at least one fumonisin esterase according to any of the preceding items (e.g., a fumonisin esterase according to any of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26); and / or at least one polynucleotide encoding the at least one fumonisin esterase.

[0124] Item 53: A fumonisin esterase for use in the treatment, amelioration and / or prevention or prophylaxis of symptoms caused by mycotoxin intoxication, in particular fumonisin mycotoxin intoxication, wherein the fumonisin esterase is a fumonisin esterase according to any of the preceding items (e.g., a fumonisin esterase according to any of items 12, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26); and / or wherein the fumonisin esterase is comprised in a composition according to any of items 27-29.

[0125] Item 54: The method, fumonisin esterase, composition, use and / or fumonisin esterase for use according to any of the preceding items, wherein the fumonisin is one or more selected from the group consisting of fumonisin B1, B2, B3 and B4, preferably wherein the fumonisin is fumonisin B1. EMBODIMENT

[0126] In the following, the present application is further described by non-limiting embodiments. The present application as disclosed herein is not limited to the specific embodiments, methodologies, examples, protocols, etc. described herein, but only by the claims.

[0127] EMBODIMENT 1

[0128] The fumonisin esterases described herein were recombinantly produced in Pichia pastoris essentially as described in WO 2016 / 134387 Al. The skilled person in the art of recombinant protein production will be aware of suitable analogous, similar or alternative methods for producing fumonisin esterases. The fumonisin esterases were designed for secretion into the culture supernatant and quantified photometrically (NanoDrop spectrophotometer), by Bradford assay and / or by BCA assay. Enzyme preparations were diluted with 20 mM Tris-HCl, pH 8.0 to a concentration of 1 mg / mL. These 1 mg / mL dilutions were further diluted to 10 ng / mL or 20 ng / mL using 1x FE buffer (20 mM Tris-HCl, pH 8.0, 0.1 mg / mL bovine serum albumin). Fumonisin stock solutions were prepared by dissolving crystalline fumonisin B1 (FB1) in 1x FE buffer to a concentration of 1 mM FB1.

[0129] The assay was started by mixing 50 μΐ^of 10 ng / mL or 20 ng / mL enzyme solution with 450 μΐ^of fumonisin solution. The latter was prepared from the fumonisin stock solution to reach one of the following final FB1 concentrations in the assay reaction: 5 μΜ, 10 μΜ, 15 μΜ, 20 μΜ, 25 μΜ, 50 μΜ, 75 μΜ, 100 μΜ of FB1. The reactions were performed in 96 deep well plates at 30 °C with 600 rpm shaking. At 15 min, 30 min, 45 min, 60 min and 120 min, 80 μΐ^samples were withdrawn from the reaction mixture and incubated at 99 °C for 5 min to stop the reaction.

[0130] For LC-MS / MS analysis, samples were diluted with HPLC eluent as follows: 1 :30 for 5-20 μΜ FB1, 1 :50 for 25 μΜ and 50 μΜ FB1 and 1 : 100 for 75 μΜ and 100 μΜ FB1. The analysis was performed on an Agilent 1290 series UHPLC system coupled with a 5500 QTrap mass spectrometer. The column temperature was set to 30 °C and the flow rate was set to 1.0 mL / min. The mobile phase consisted of methanol / water / acetic acid (5 / 94 / 1, v / v / v). The injection volume was 1 μΐ^and the total run time was 0.4 min. Separation was performed on an Agilent ZORBAX Eclipse Plus C18 RRHD column (50 x 2.1 mm, 1.8 μιη). SRM parameters are shown in Table 1.

[0131] Table 1. Selected reaction monitoring transitions in negative ion mode. Declustering potential (DP), collision energy (CE), collision cell exit potential (CXP). Inlet potential (EP) was -10.0 V for all analytes.

[0132]

[0133] HPLC-eluent: Ultrapure water containing 0.0385% formic acid (pH about 2.85, corresponding to 192.5 μΐ^of formic acid in 500 mL water) was adjusted to pH 3.0 by adding 6.1 mM ammonium formate (192.5 mg of ammonium formate in 500 mL water) in ultrapure water. This solution was mixed with acetonitrile in a ratio of 75 (HPLC buffer pH 3.0):25 (acetonitrile) (v / v). The HPLC eluent was stored at room temperature.

[0134] Data analysis was performed using MultiQuant 3.0.3 software (Sciex) after LC-MS / MS measurements. Results were exported to Microsoft Excel for statistical analysis. Michaelis-Menten curves were generated by using SigmaPlot software. The improved kinetic parameters of the exemplary fumonisin esterase variants relative to the fumonisin esterase of SEQ ID NO: 1 are shown in Tables 2 and 3 below.

[0135] Table 2: Relative kinetic parameters of exemplary fumonisin esterases. Michaelis constant (Km) in mM; turnover number (kcat) in seconds -1 catalytic efficiency in mM -1 seconds -1 determined.

[0136]

[0137]

[0138] Table 3: Relative kinetic parameters of exemplary fumonisin esterases. Specific activity determined in U / mg, where 1 unit (U) is defined as the amount of enzyme that releases 1 pmol of tricarballylic acid (TCA) per minute from 100 mM fumonisin B1 (FB1) in 20 mM Tris-Cl buffer pH 8.0 containing 0.1 mg / mL bovine serum albumin (BSA) at 30 °C.

[0139]

[0140]

[0141] Example 2

[0142] To determine temperature stability, stability tests were performed in Teorell-Stenhagen buffer, pH 6.5 (Teorell and Stenhagen. 1938. Biochem. Ztschrft. 299:416-419) containing 0.1 mg / L bovine serum albumin. Fumonisin esterase was incubated at 75 °C for 5 minutes at a concentration of 10 pg / mL in 70 pL of buffer. An aliquot of the incubation mixture was stored on ice without incubation at 75 °C as an untreated control to determine initial activity. After incubation at 75 °C, the enzyme solution was stored on ice until residual activity was determined compared to initial activity. The FB1 assay described in Example 1 was used to determine initial activity and residual activity. Residual activity was determined as a % of untreated initial activity. Table 4 below shows improvement in temperature stability of exemplary fumonisin esterase variants relative to fumonisin esterase of SEQ ID NO: 1. For example, where the initial activity of fumonisin esterase A was 100 U / mg and the residual activity after incubation was 50 U / mg, the residual activity was found to be 50%. Where the residual activity of fumonisin esterase B was 80%, the relative residual activity of fumonisin esterase B relative to fumonisin esterase A was 80*100 / 50 = 160%.

[0143] Table 4: Relative residual activity of exemplary fumonisin esterases relative to fumonisin esterase of SEQ ID NO: 1. Residual activity was determined as a % of untreated initial activity.

[0144]

[0145] As a further assay for determining temperature stability, thermal shift analysis (also known as thermal fluorescence assay) was performed using SYPRO Orange as a fluorescent indicator. While fumonisin esterase of SEQ ID NO: 1 showed a fluorescence peak at 45 °C, indicating temperature-induced unfolding of the polypeptide, fumonisin esterases of any one of SEQ ID NO: 9 to SEQ ID NO: 71 showed a fluorescence peak at 70-75 °C, thereby suggesting that unfolding occurs at significantly higher temperatures than fumonisin esterase of SEQ ID NO: 1.

[0146] Example 3

[0147] To investigate the suitability of fumonisin esterases in gastric juice, exemplary fumonisin esterases were tested at a final concentration of 2.0 ng / mL, 1.5 ng / mL, 1.0 ng / mL or 0.5 ng / mL diluted in gastric simulated buffer (GSB: 118.5 mM NaCl, 8.55 mM acetic acid, 14.9 mM sodium acetate, pH 5.0, 0.1 mg / mL bovine serum albumin). The assay was performed in a 37 °C water bath. The assay was started by adding FB1 to a final concentration of down to 5 mM in a total assay volume of 500 pL. After 15 min, 30 min, 45 min, 60 min and 120 min, 80 pL samples were withdrawn and incubated at 99 °C for 5 min. Specific activity was determined as described in Example 1. All fumonisin esterases were found to be active in gastric juice, see Table 5, although in acetic acid conditions and at lower substrate concentrations.

[0148] Table 5: Specific activity of exemplary fumonisin esterases in gastric juice.

[0149]

[0150] Example 4

[0151] To investigate whether fumonisin esterases are suitable for application in water, in particular in aquaculture, a 56-day trial with 180 salmon (rainbow trout) was performed. In the trial the following three groups were compared: a control without fumonisins and without fumonisin esterases (control), a control group contaminated with fumonisins (FUM), and a trial group receiving fumonisins and 15 U / kg feed of the improved fumonisin esterase according to the application (any one of SEQ ID NO. 2 to SEQ ID NO. 71) (esterase). In the FUM and esterase groups, a contamination of about 10 ppm of fumonisins was applied, which comprised a mixture of fumonisins B1, B2 and B3.

[0152] The fish arrived as eyed eggs and were kept for a period of approximately 9 months before the trial. On the first day of the trial, the fish were individually weighed and allocated to 15 tanks based on stratified randomization with the aim of balancing the average body weight per tank. The fish were kept in 15 sub- square recirculating aquaculture system (RAS) tanks of 500 L each, with twelve fish allocated to each tank. The tanks were divided into three trial groups: three tanks for the control group, six tanks for the FUM group, and six tanks for the esterase group. The tanks were the experimental units. Fumonisin degradation was followed by analyzing fumonisins (FB1, FB2, FB3) and their metabolites, hydrolyzed fumonisins (HFB1, HFB2, HFB3) and partially hydrolyzed fumonisins (pHFB1 a+b, pHFB2a+b, pHFB3a+b) in the content of the proximal and distal parts of the gastrointestinal tract (GIT). In addition, the levels of dihydro sphingosine (Sa) and sphingosine (So) in the plasma and their ratio (Sa / So) were analyzed as an indicator of exposure to toxic levels of fumonisins (Meredith et al., 1998. J Food Prot. 61 (8): 1034-8).

[0153] For analysis of samples from GIT, 100 mg of lyophilized and homogenized sample was weighed into a 5 mL Eppendorf tube. 3 mL of extraction solvent (acetonitrile / water / formic acid, 74 / 25 / 1, v / v / v) was added, vortexed, and the tube was shaken on a horizontal shaker for 30 minutes at room temperature (fast shaking) before a centrifugation step (1880 rcf for 5 minutes). The supernatant was collected into a 15 mL tube and the residue was re-extracted with 3 mL of extraction solvent. After resuspension of the residue by vortexing, the sample was again shaken on a horizontal shaker for 20 minutes at room temperature (fast shaking) before a centrifugation step (1880 rcf for 5 minutes). The supernatant from the first extraction step was transferred to a 15 mL tube by pouring, and for the final extraction step, 2 mL of extraction solvent was added to the residue. After vortexing and shaking for 10 minutes on a horizontal shaker at room temperature, a centrifugation was performed at 3200 rcf for 5 minutes, and the supernatant was combined with the supernatants from the previous two extraction steps. The combined extract was vortexed and centrifuged at 3200 rcf for 10 minutes. The final step of sample preparation was to transfer 500 μL of the supernatant to the HPLC and dilute with 500 μL of extraction solvent. For plotting of the calibration curve, standards of fumonisins (FB1, 2, 3), HFB and pHFB were diluted in dilution solvent (acetonitrile / water / formic acid, 50 / 49 / 1, v / v / v) to 600 ng / mL, 300 ng / mL, 150 ng / mL, 30 ng / mL, 15 ng / mL, 3 ng / mL, 1.5 ng / mL and 0.3 ng / mL. The analysis was performed on an Agilent 1290 series UHPLC system coupled to a 5500 QTrap mass spectrometer. The column temperature was set to 30 °C and the flow rate was set to 0.8 mL / min. The mobile phase A consisted of methanol / water / acetic acid (40 / 59.8 / 0.2; v / v / v) and the mobile phase B consisted of methanol / acetic acid (99.8 / 0.2; v / v). The gradient started at 100% A for 0.5 minutes, then continued with a linear increase to 73% B at 5.9 minutes and 100% B at 6.0 minutes, then to 100% B at 7.9 minutes, and a sharp decrease to 0% B between 7.9 and 8.0 minutes. The total run time for each sample was 10.5 minutes. The injection volume was 1 μL. The separation was performed on a Phenomenex Gemini 5 μ C18 110A column (150 x 4.6 mm).

[0154] Mass spectrometry detection was performed with negative electrospray ionization, in multiple reaction monitoring mode and according to the parameters of Table 6.

[0155] Table 6. Parameters mass transfer. DP = de-clustering potential; EP = entrance potential; CE = collision energy; CXP = chamber exit potential.

[0156]

[0157]

[0158] Data analysis for LC-MS / MS: Peak integration and concentration calculation were performed using Multiquant (version 3.0.3). For concentration calculation, a 1 / x weighted linear calibration function was used. The measured concentration in the extracted sample was multiplied by the dilution factor to receive the result for the GIT sample.

[0159] For the analysis of plasma samples, 600 μΐ^of acetonitrile / water (50 / 50, v / v) was added to 50 μΐ^of plasma in a 2 mL tube. After homogenization by vortexing, the samples were shaken for 30 minutes at room temperature using a vortexer equipped with an Eppendorf tube adapter at level 5-6, followed by a centrifugation step (2700 rcf for 5 minutes). The supernatant was collected into a new Eppendorf tube (decanting) and the pellet was re-extracted with 300 μΐ^of 80% (v / v) methanol for 30 seconds while vortexing. After centrifugation (19 000 rcf for 5 minutes), the supernatant was combined and dried using a SpeedVac at 30°C heating. When dry, the extract was re-dissolved in 300 μΐ^of 80% (v / v) methanol for 30 minutes at room temperature using a vortexer at level 5-6. Sample preparation ended with a centrifugation step (19 000 rcf for 10 minutes) and 200 μΐ^of supernatant was transferred to a HPLC vial with a glass insert. The analysis was performed on an Agilent 1290 series UHPLC system coupled to a 5500 QTrap mass spectrometer. The column temperature was set to 30°C and the flow rate was set to 0.5 mL / min. The mobile phase A consisted of methanol / water / acetic acid (40 / 59.8 / 0.2; v / v / v) and the mobile phase B consisted of methanol / acetic acid (99.8 / 0.2; v / v). The gradient started with 65% B for 1.7 minutes, followed by a linear increase to 100% B at 1.71 minutes, then to 100% B at 2.5 minutes, and a sharp decrease to 65% B between 2.5 and 2.51 minutes. The injection volume was 2 μΐ^. Separation was performed on a Phenomenex Kinetex C18 column (150 x 2.1 mm, 2.6 μιη). Quantitation was performed based on a calibration using external standards of dihydro-sphingosine and sphingosine at a concentration range of 0.1 ppb to 300 ppb. SRM parameters are shown in Table 7.

[0160] Table 7. Transitions of selected reaction monitoring in positive ion mode. DP = de-clustering potential; EP = entrance potential; CE = collision energy; CXP = chamber exit potential.

[0161]

[0162]

[0163] With any of the fumonisin esterases, the concentration of fumonisins was significantly (p<0.05) reduced compared to the contaminated group (FUM). In addition, the levels of partially and completely hydrolyzed fumonisins (HFB1, HFB2, HFB3, pHFB1a+b, pHFB2a+b, pHFB3a+b) were significantly higher in both the proximal and distal part of the GIT of the esterase groups compared to the FUM group, see Table 8 and Table 9. Also, the Sa / So ratio exhibited by the fish in the esterase groups was significantly lower than in the FUM group, see Table 10.

[0164] Table 8: Fumonisins and their metabolites in the proximal GI tract of rainbow trout (group means in pg / g lyophilized GIT content) (n=6 for FUM and esterase groups and n=3 for control group). p values refer to comparison between FUM and esterase groups.

[0165] Parameter Control FUM Esterase p-value FB1 0.06 16.99 4.66 <0.001 FB2 0.11 4.12 1.40 0.002 FB3 0.03 6.70 2.10 0.002 HFB1 0.17 0.16 5.91 0.002 HFB2 0.32 0.32 0.52 0.001 HFB3 0.25 0.25 0.88 0.001 pHFB1a 0.03 0.23 0.43 0.001 pHFB1b 0.02 0.31 1.05 0.002 pHFB2a 0.02 0.04 0.10 0.004 pHFB2b 0.02 0.09 0.39 <0.001 pHFB3a 0.04 0.08 0.15 0.004 pHFB3b 0.02 0.15 0.43 <0.001

[0166] Table 9: Fumonisins and their metabolites in the distal GI tract of rainbow trout (group means in pg / g lyophilized GIT content) (n=6 for FUM and esterase groups and n=3 for control group). p values refer to comparison between FUM and esterase groups.

[0167]

[0168]

[0169] Table 10: Plasma dihydro- to sphingosine ratio (Sa / So) of rainbow trout (n=6 for FUM and esterase and n=3 for control group).

[0170] Group Sa / So Control 0.044 FUM 0.079 Esterase 0.053 p-value <0.001

Claims

1. A method for improving the enzyme kinetics of a fumonisin esterase, wherein the fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence of SEQ ID NO: 1, The method comprises: Substituting the amino acid at position 25 relative to the amino acid sequence of SEQ ID NO: 1, preferably with an aliphatic amino acid, more preferably with an amino acid selected from alanine, glycine, isoleucine, leucine, proline, valine, even more preferably with an amino acid selected from isoleucine, leucine, valine, most preferably with isoleucine; Replacing the amino acid at position 45 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a polar amino acid, more preferably with an amino acid selected from tyrosine, threonine, glutamine, glycine, serine, cysteine, asparagine, lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from asparagine, aspartic acid, glutamine, glutamic acid, most preferably with glutamine; and / or replacing the amino acid at position 259 relative to the amino acid sequence of SEQ ID NO: 1, preferably with a charged amino acid, more preferably with an amino acid selected from lysine, arginine, histidine, glutamic acid, aspartic acid, even more preferably with an amino acid selected from lysine, arginine, histidine, most preferably with arginine.

2. The method of claim 1 , wherein the substitution is at least one additional amino acid selected from the group consisting of: 10, 33, 66, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 366, 7, 371, 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490, preferably wherein a sequence selected from the group consisting of the following relative to said SEQ ID at least one additional amino acid substitution of the amino acid sequence of NO: 1: 10Q, 33E, 66A, 66C, 66F, 66L, 66M, 66T, 66W, 66E, 66H, 66I, 66K, 66P, 66R, 66Y, 66G, 66Q, 66S, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 32 9F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 371V, 371M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430 A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, ​​465G, 469K, 473A, 478D, 487N and 490P.

3. A method for improving the enzyme kinetics of a thermostable fumonisin esterase, the method comprising: (i) providing the thermostable fumonisin esterase, wherein the thermostable fumonisin esterase comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, wherein the thermostable fumonisin esterase comprises an aspartic acid at position 66 relative to the amino acid sequence of SEQ ID NO: 1, and at least one additional mutation at a position selected from the group consisting of: 10, 33, 107, 140, 144, 149, 151, 157, 199, 266, 267, 270, 272, 275, 280, 284, 286, 293, 302, 312, 329, 332, 360, 363, 364, 365, 367, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, ​​383, 384, 386, 387, 388, 389, 390, 401, 403, 404, 405, 406, 407, 408, 410, 411, 412, 413, 414, 415 , 372, 377, 389, 391, 394, 418, 419, 424, 427, 429, 430, 436, 440, 443, 447, 453, 455, 456, 457, 462, 463, 464, 465, 469, 473, 478, 487 and 490, preferably wherein the at least one additional mutation is selected from the group consisting of relative to said SEQ ID 10Q, 33E, 107E, 140P, 144M, 149F, 151R, 157Y, 199I, 266S, 267P, 270F, 272H, 275E, 275A, 280D, 280P, 284T, 284P, 286P, 286R, 293E, 302I, 312F, 329F, 332E, 360V, 363T, 364H, 364L, 365I, 367H, 3 71M, 372F, 377V, 389L, 391V, 394P, 418A, 419V, 424A, 424K, 427V, 429P, 430A, 436A, 436S, 440G, 440S, 443T, 447A, 453R, 455S, 456Q, 457T, 462Y, 463D, 464I, 465H, 465S, ​​465G, 469K, 473A, 478D, 487N, and 490P; and (ii) substituting an amino acid other than aspartic acid for the amino acid at position 66 relative to the amino acid sequence of SEQ ID NO:

1.

4. A fumonisin esterase obtainable by the method according to any one of the preceding claims.

5. A composition for modifying fumonisin, comprising at least one fumonisin esterase according to claim 4.

6. The composition according to claim 5, further comprising at least one carrier, preferably selected from bentonite, silicon dioxide, maltodextrin and carbohydrates.

7. A method for modifying fumonisins in a composition, the method comprising contacting the composition with the fumonisin esterase according to claim 4.

8. A method for modifying fumonisins in a body of water, the method comprising contacting the body of water with a fumonisin esterase.

9. A method for modifying fumonisins in gastric juice, the method comprising contacting the gastric juice with a fumonisin esterase.

10. The method according to claim 8 or 9, wherein the fumonisin esterase is the fumonisin esterase according to claim 4 and / or wherein the fumonisin esterase is comprised in the composition according to claim 5 or 6.

11. Use of the fumonisin esterase according to claim 4 and / or the composition according to claim 5 or 6 for modifying, for example, detoxifying, fumonisin.

12. Use of the fumonisin esterase according to claim 4 and / or the composition according to claim 5 or 6 for the manufacture of an additive for feed and / or food, a feed and / or food composition, or a pharmaceutical composition.

13. Use of the fumonisin esterase according to claim 4 and / or the composition according to claim 5 or 6 for producing biogas; bioethanol, DDGS; sugar, preferably from sugar cane or sugar beet; corn oil, corn germ, corn germ meal, corn fiber, corn gluten, starch, in particular corn starch; and / or silage.

14. A host cell comprising at least one fumonisin esterase according to claim 4 and / or at least one polynucleotide encoding the at least one fumonisin esterase.

15. A fumonisin esterase for treating, ameliorating and / or preventing symptoms caused by mycotoxin poisoning, in particular fumonisin mycotoxin poisoning, wherein the fumonisin esterase is the fumonisin esterase according to claim 4.

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