Herbicide resistant plants

Abstract

The present invention relates to plants and parts thereof that have been modified to comprise a BIO3-BIO1 (BioDA) enzyme which confers at least partial resistance to compounds which inhibit the biotin synthesis pathway, such as herbicides, wherein the enzymes comprises modifications which confer resistance. The invention further relates to such modified BIO3- BIO1 enzymes, having modifications which improve resistance to such compounds, as well as polynucleotides and proteins encoding such enzymes. The invention also relates to methods of growing and propagating such plants, improving plant growth and controlling unwanted vegetation using such plants and parts thereof.

Description

[0001] HERBICIDE RESISTANT PLANTS

[0002] FIELD OF THE INVENTION

[0003] The invention relates to modified BIO3-BIO1 enzymes, having modifications which confer at least partial resistance to compounds which inhibit the biotin synthesis pathway, such as herbicides, as well as polynucleotides and proteins encoding such enzymes. The invention also relates to methods of growing and propagating such plants, improving plant growth and controlling unwanted vegetation using such plants and parts thereof.

[0004] BACKGROUND

[0005] The present invention relates to the production of plants that are resistant to herbicides that inhibit the biotin synthesis pathway, specifically to herbicides which inhibit the BIO3-BIO1 enzyme in plants.

[0006] Biotin, also known as vitamin B7, is an essential co-factor for enzymes involved in cellular processes including metabolism of fats, proteins or carbohydrates. Plants and most fungi / bacteria are able to synthesise biotin in contrast to animals which derive biotin from dietary sources or gut bacteria.

[0007] In bacteria, biotin is synthesised from pimeloyl-CoA and Alanine via the activity of four enzymes (BioF, BioA, BioD and BioB) which are located in an operon. BioF catalyses the production of 7-keto-8-Aminopelargonic Acid (KAPA) from pimeloyl-CoA and Alanine. BioA, also known as 7,8-diaminopelargonic acid aminotransferase (EC. 2.6.1.62) then carries out the next step, converting KAPA to 7,8 Diaminopelargonic Acid (DAPA). BioD, also known as dethiobiotin synthase (EC 6.3.3.3), subsequently converts DAPA to Dethiobiotin which is in turn converted to Biotin via the activity of BioB. (Entcheva et al, Applied Microbiology and Biotechnology volume 61 , 21-31 (2003).

[0008] In plants, the pathway is similar, but the BioA and BioD enzyme activities are found in a single bifunctional protein known as BIO3-BIO1 , BioDA, or bifunctional dethiobiotin synthetase. The BioD activity is found in the BIO3 sequence and BioA activity is provided by BIO1. Loss of function mutations in the BIO3-BIO1 gene of plants leads to an embryo lethal effect which can be rescued by exogenous application of biotin (Meinke et al, Plant Physiol. 2008 Jan; 146(1): 60-73.) Given that the biotin pathway is essential to the survival of plants, it has been identified as a herbicidal target. Recently, effective herbicidal compounds which inhibit one or more of the enzymes of this pathway, especially BIO3-BIO1 , have been developed. Upon contact with plants, the herbicides cause cell death and eventually death of the plant. Industrially, herbicides are used in agriculture to remove unwanted vegetation such as weeds from cultivated crops. However non-specific herbicides which target essential pathways such as those that target the biotin synthesis pathway, will affect both crops and the unwanted vegetation, making them difficult to use without destroying the valuable crop. Therefore in order to effectively use these herbicides, it would be desirable to protect the crop plants so that they have resistance to the herbicidal compounds. Plants that have resistance to the herbicides can then be contacted with the herbicide and will not be affected, whilst non- resistant unwanted vegetation is affected and controlled. Means to protect crop plants from other herbicides in the past have involved providing the plant with a mutated form of the enzyme which is targeted by the herbicide, thereby imparting resistance to the plant.

[0009] However, there has not yet been a link shown between modification of the BIO3-BIO1 genes, by mutagenesis, and increased resistance to herbicidal compounds which target the biotin pathway in plants.

[0010] SUMMARY OF INVENTION

[0011] In a first aspect there is provided, a modified BIO3-BIO1 enzyme or a functional fragment thereof comprising at least two modifications at amino acid positions selected from one of the following combinations: a) amino acid position 348 and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto; b) amino acid position 391 and one or more of amino acid positions 388, 421 , 348, 506, 507, 508, 517 and / or 520 of SEQ ID NO: 1 or positions corresponding thereto; c) amino acid position 508 and one or more of amino acid positions 700, 348, 388, 391 , 392, 506, 507, 511 , 513, 517, 520, 609 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto; or d) amino acid position 609 and one or more of amino acid positions 348, 512, 507, 508, 511 , 520, 700 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. For example, in one aspect there is provided a modified BIO3-BIO1 enzyme or a functional fragment thereof comprising at least two modifications at amino acid positions selected from amino acid position 348 and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 391, 392, 506, 509, 511, 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto.

[0012] For example, in one aspect there is provided a modified BIO3-BIO1 enzyme or a functional fragment thereof comprising at least two modifications at amino acid positions selected from amino acid position 391 and one or more of amino acid positions 388, 421, 348, 506, 507, 508, 517 and / or 520 of SEQ ID NO: 1 or positions corresponding thereto.

[0013] For example, in one aspect there is provided a modified BIO3-BIO1 enzyme or a functional fragment thereof comprising at least two modifications at amino acid positions selected from amino acid position 508 and one or more of amino acid positions 700, 348, 388, 391, 392, 506, 507, 511 , 513, 517, 520, 609 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto.

[0014] For example, in one aspect there is provided a modified BIO3-BIO1 enzyme or a functional fragment thereof comprising at least two modifications at amino acid positions selected from amino acid position 508 and one or more of amino acid positions 348, 512, 507, 508, 511, 520, 700 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto.

[0015] In a second aspect there is provided an isolated nucleic acid comprising a polynucleotide encoding at least one modified BIO3-BIO1 enzyme or functional fragment thereof of the first aspect .

[0016] In a third aspect there is provided an expression construct comprising a polynucleotide encoding at least one modified BIO3-BIO1 enzyme or functional fragment thereof of the first aspect and / or a polynucleotide of the second aspect, operably linked to one or more expression elements

[0017] In a fourth aspect there is provided a recombinant vector comprising the isolated nucleic acid of the second aspect or the expression construct of the third aspect. In a fifth aspect there is provided a modified plant, or part thereof, comprising at least one modified BIO3-BIO1 enzyme or functional fragment thereof according to the first aspect, an isolated nucleic acid according to the second aspect, an expression construct according to the third aspect or a recombinant vector according to the fourth aspect.

[0018] In a sixth aspect there is provided a method of producing a modified plant or part thereof as described herein, the method comprising: modifying the plant or part thereof to comprise a modified BIO3-BIO1 enzyme according to the first aspect which provides the plant with increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

[0019] In a seventh aspect there is provided method of controlling undesired vegetation in the vicinity of a modified plant or part thereof as described herein, or at the locus for growth of a modified plant as described herein, the method comprising applying an effective amount of at least one compound which inhibits the biotin synthesis pathway to the undesired vegetation and the plant, or applying an effective amount of at least one compound which inhibits the biotin synthesis pathway to the locus and planting a seed at said locus, wherein the seed is capable of producing the modified plant as described herein.

[0020] In an eighth aspect there is provided use of a compound which inhibits the biotin synthesis pathway in combination with a modified plant or part thereof as described herein.

[0021] In some embodiments, the modified BIO3-BIO1 enzyme or the functional fragment thereof comprises: a. amino acid position 348, or a position corresponding thereto, is substituted with an A, D, S, T, M, V, C, or N amino acid residue; b. amino acid position 350, or a position corresponding thereto, is substituted with an H amino acid residue; c. amino acid position 388, or a position corresponding thereto, is substituted with a T amino acid residue; d. amino acid position 391 , or a position corresponding thereto, is substituted with a L or M amino acid residue; e. amino acid position 392, or a position corresponding thereto, is substituted with a S amino acid residue; f. amino acid position 421, or a position corresponding thereto, is substituted with a L amino acid residue; g. amino acid position 506, or a position corresponding thereto, is substituted with an A amino acid residue; h. amino acid position 507, or a position corresponding thereto, is substituted with a S or K amino acid residue; i. amino acid position 508, or a position corresponding thereto, is substituted with a T or L amino acid residue; j. amino acid position 509, or a position corresponding thereto, is substituted with a V amino acid residue; k. amino acid position 511, or a position corresponding thereto, is substituted with an E, W or M amino acid residue; l. amino acid position 512 is substituted with a C amino acid residue; m. amino acid position 513, or a position corresponding thereto, is deleted (Del); n. amino acid position 517, or a position corresponding thereto, is substituted with an H, F or T amino acid residue; o. amino acid position 520, or a position corresponding thereto, is substituted with a W or N amino acid residue; p. amino acid position 608, or a position corresponding thereto, is substituted with an A amino acid residue; q. amino acid position 609, or a position corresponding thereto, is substituted with a Y, T, M or V amino acid residue; r. amino acid position 700, or a position corresponding thereto, is substituted with a S or A amino acid residue; and / or s. amino acid position 704, or a position corresponding thereto, is substituted with an H amino acid residue.

[0022] In some embodiments, the modified BIO3-BIO1 enzyme or functional fragment thereof comprises modifications at amino acid positions selected from one of the following combinations: Positions 348, 388, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto;

[0023] Positions 348, 388 and 517 of SEQ ID NO: 1 or positions corresponding thereto;

[0024] Positions 348, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto;

[0025] Positions 388, 391 and 421 of SEQ ID NO: 1 or positions corresponding thereto;

[0026] Positions 348 and 700 of SEQ ID NO: 1 or positions corresponding thereto;

[0027] Positions 348, 507, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto;

[0028] Positions 348, 509, 517, 608 and 700 of SEQ ID NO: 1 or positions corresponding thereto;

[0029] Positions 348, 350, 506 and 507 of SEQ ID NO: 1 or positions corresponding thereto;

[0030] Positions 348 and 508 of SEQ ID NO: 1 or positions corresponding thereto;

[0031] Positions 348, 388 and 508 of SEQ ID NO: 1 or positions corresponding thereto;

[0032] Positions 348 and 506 of SEQ ID NO: 1 or positions corresponding thereto;

[0033] Positions 348, 508 and 513 of SEQ ID NO: 1 or positions corresponding thereto;

[0034] Positions 348, 507 and 609 of SEQ ID NO: 1 or positions corresponding thereto;

[0035] Positions 348, 350, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto;

[0036] Positions 348, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto;

[0037] Positions 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto;

[0038] Positions 512 and 609 of SEQ ID NO: 1 or positions corresponding thereto;

[0039] Positions 348, 507 and 508 of SEQ ID NO: 1 or positions corresponding thereto;

[0040] Positions 348, 506, 508 and 517 of SEQ ID NO: 1 or positions corresponding thereto;

[0041] Positions 348, 511 , 512, 609 and 704 of, SEQ ID NO: 1 or positions corresponding thereto;

[0042] Positions 348, 350, 506 and 507 of SEQ ID NO: 1 or positions corresponding thereto;

[0043] Positions 348, 388, 508, 511 and 520 of SEQ ID NO: 1 or positions corresponding thereto;

[0044] Positions 391 and 507 of SEQ ID NO: 1 or positions corresponding thereto;

[0045] Positions 391 and 517 of SEQ ID NO: 1 or positions corresponding thereto;

[0046] Positions 391 , 506, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto;

[0047] Positions 348, 506 and 512 of SEQ ID NO: 1 or positions corresponding thereto; Positions 348, 506, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto;

[0048] Positions 348, 508 and 511 of SEQ ID NO: 1 or positions corresponding thereto;

[0049] Positions 348 and 512 of SEQ ID NO: 1 or positions corresponding thereto;

[0050] Positions 348 and 391 of SEQ ID NO: 1 or positions corresponding thereto;

[0051] Positions 348 and 507 of SEQ ID NO: 1 or positions corresponding thereto;

[0052] Positions 348, 350 and 512 of SEQ ID NO: 1 or positions corresponding thereto;

[0053] Positions 348 and 509 of SEQ ID NO: 1 or positions corresponding thereto;

[0054] Positions 520 and 609 of SEQ ID NO: 1 or positions corresponding thereto;

[0055] Positions 348, 391 and 506 of SEQ ID NO: 1 or positions corresponding thereto;

[0056] Positions 348, 517 and 520 of SEQ ID NO: 1 or positions corresponding thereto;

[0057] Positions 391 and 508 of SEQ ID NO: 1 or positions corresponding thereto;

[0058] Positions 348 and 520 of SEQ ID NO: 1 or positions corresponding thereto;

[0059] Positions 348, 507, 508, 700 and 704 of SEQ ID NO: 1 or positions corresponding thereto;

[0060] 348, 507, 508, 700 and 609 of SEQ ID NO: 1 or positions corresponding thereto;

[0061] Positions 348, 507, 508, 700 and 392 of SEQ ID NO: 1 or positions corresponding thereto; or

[0062] Positions 507, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0063] In some embodiments, the modified BIO3-BIO1 enzyme or functional fragment thereof comprises modifications selected from any one of the following combinations:

[0064] F348C, C388T, A507S and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0065] F348A, C388T and Q517H of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0066] C388T, W391 L and P421 L of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0067] F348A, A507S and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; F348A, A507S, P508T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0068] F348A, P508T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0069] F348C and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0070] F348C, P508T and G513Del of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0071] F348C, S509V, Q517F, G608A and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0072] F348M, A507S and A609T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0073] F348M, A507S, P508T and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0074] F348M and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0075] F348N and P508L of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0076] F348S, C388T and P508T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0077] F348S and Q506A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0078] F348T, Q350H, Q506A and A507S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0079] F348T and Q506A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0080] F348V and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0081] P508T and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0082] T512C and A609M of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; F348M, Q350H, A507S and Y520W of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0083] F348S, A507S and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0084] F348T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0085] F348T, P508L and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0086] F348D, Q506A, P508L and Q517T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0087] F348N, Y511E, T512C, A609Y and S704H of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0088] F348D, C388T, P508T, Y511W and Y520W of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0089] F348D and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0090] W391L and A507S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0091] W391M and Q517F of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0092] W391L, Q506A, A507S and Y520W of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0093] F348S, Q506A and T512C of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0094] F348D, Q506A, A507S and Y520W of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0095] F348T, P508T and Y511M of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0096] F348S and T512C of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0097] F348M and W391L of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; F348T and A507K of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0098] F348A, Q350H and T512C of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0099] F348S and S509V of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0100] F348N and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0101] Y520W and A609V of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0102] W391 L and Q517T of SEQ ID NO: of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0103] F348M, W391 L and Q506A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0104] F348M, Q517F and Y520W of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0105] W391 L and P508T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0106] F348A and Y520N of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0107] F348A, A507S and P508T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0108] A507S, P508T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0109] F348T and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;

[0110] F348A, A507S, P508T, G700A and S704H of SEQ I D NO: 1 or the same modifications at positions corresponding thereto;

[0111] F348A, A507S, P508T, G700A and W392S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; or

[0112] F348A, A507S, P508T, G700A and A609V of SEQ I D NO: 1 or the same modifications at positions corresponding thereto. In some embodiments, the modified BIO3-BIO1 enzyme or functional fragment thereof comprises an amino acid sequence having at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 98%, at least 99% or 100% identity to any of SEQ ID NO: 40 to 663.

[0113] In some embodiments, the modified BIO3-BIO1 enzyme consists of a sequence having at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 98% or at least 99% identity to any of SEQ ID NO: 40 to 663.

[0114] In some embodiments, the modified BIO3-BIO1 enzyme or functional fragment thereof, comprises an increased resistance to a compound that inhibits a BIO3-BIO1 enzyme relative to a control BIO3-BIO1 enzyme and / or has decreased percentage inhibition in response to a compound that inhibits a BIO3-BIO1 enzyme relative to a control BIO3-BIO1 enzyme.

[0115] In some embodiments, the modified BIO3-BIO1 enzyme or functional fragment thereof is modified to comprise a heterologous targeting peptide, preferably a heterologous mitochondrial targeting peptide.

[0116] In some embodiments, the modified BIO3-BIO1 enzyme provides the plant or part thereof with increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

[0117] In some embodiments, the modified BIO3-BIO1 enzyme is derived from a plant, from an algae, from an oomycete, or from a fungus. In some embodiments, the modified BIO3-BIO1 enzyme is derived from a plant. In some embodiments, the modified BIO3-BIO1 enzyme is derived from any of the following plant species: Setaria italica, Arabidopsis thaliana, Helianthus annuus, Quercus robur, Phoenix dactylifera, Physcomitrium patens, Taxus chinensis, Adiantum nelumboides, Zea mays, Ostreococcus tauri, Hordeum vulgare, Brassica napus, Gossypium hirsutum, Oryza sativa, Glycine max, and Triticum aestivum. In some embodiments, the modified BIO3-BIO1 enzyme is derived from a BIO3-BIO1 enzyme heterologous to the plant or part thereof. That is to say that the modified BIO3-BIO1 enzyme is derived from an organism such as a plant, an algae, an oomycete, or a fungus that is different to the plant into which the modified BIO3-BIO1 enzyme is introduced.

[0118] In some embodiments, the functional fragment comprises at least DAPA synthase activity; optionally wherein the functional fragment comprises the BIO1 portion of the BIO3-BIO1 enzyme; further optionally wherein the functional fragment comprises amino acid residues 322 to 833 of SEQ ID NO: 1 or residues corresponding thereto.

[0119] In some embodiments, the functional fragment comprises at least dethiobiotin synthase activity; optionally wherein the functional fragment comprises the BIO3 portion of the BIO3- BIO1 enzyme; further optionally wherein the functional fragment comprises amino acid residues 24 to 287 of SEQ ID NO: 1 or residues corresponding thereto.

[0120] In some embodiments, modifying the plant or part thereof comprises transforming the plant or part thereof with an isolated nucleic acid as described herein, an expression construct as described herein, a vector as described herein, or gene editing the endogenous BIO3-BIO1 enzyme encoding gene of the plant or plant part to encode a modified BIO3-BIO1 enzyme or functional fragment thereof as described herein, the expression of which provides increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

[0121] In some embodiments, the compound which inhibits the biotin synthesis pathway inhibits one or more of the enzymes in the biotin synthesis pathway or related pathways. In some embodiments, the compound inhibits one or more of pimeloyl CoA synthetase, KAPA synthase, DAPA synthase, DTB synthetase, and biotin synthase.

[0122] In some embodiments, the compound is a herbicide. In some embodiments, the compound is a herbicide selected from: a herbicidal cinnoline compound such as ethyl 5-acetyl-4-oxo-1- (2,2,3,3-tetrafluoro-1 ,4-benzodioxin-6-yl)cinnoline-3-carboxylate (Compound A); a herbicidal compound such as [5-carboxy-1-(1-hydroxyethyl)pentyl]ammonium;chloride (compound B); a herbicidal pyridone compound such as 2-(3,4-dichlorophenyl)-5-(2,4-difluorophenyl)-1-ethyl- 6-methyl-4-oxo-pyridine-3-carboxylic acid (Compound C) or 5-[2-chloro-5- (trifluoromethoxy)phenyl]-2-(3,4-dichlorophenyl)-1-ethyl-6-methyl-4-oxo-pyridine-3-carboxylic acid (Compound D); a herbicidal compound such as [(1 R)-2-(6-carboxypyridin-1-ium-2-yl)oxy- 1-methyl-ethyl]ammonium;dichloride (Compound E); a herbicidal pyrrolidine-2-one such as 2- [1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]-N-(2-methyl-1 ,2,4-triazol-3-yl)acetamide (Compound F), 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]acetic acid (Compound G), 2-[5-oxo-1-[(2,3,5-trifluorophenyl)methyl]pyrrolidin-2-yl]acetic acid (Compound H), 2-(4- fluorophenoxy)ethyl 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]acetate

[0123] (Compound I), 2-methoxyethyl 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]acetate (Compound J), 2-[5-oxo-1-[(2,3,4-trifluorophenyl)methyl]pyrrolidin-2-yl]acetic acid (Compound K); a N-benzyl azole herbicidal compound such as 2-(3,4-dichlorophenyl)-1-ethyl-4-oxo-6-[[3- (trifluoromethyl)pyrazol-1-yl]methyl]pyridine-3-carboxylic acid (Compound L); and a herbicidal quinolone compound such as 2-(3,4-dichlorophenyl)-1-ethyl-6-fluoro-4-oxo-quinoline-3- carboxylic acid (Compound M).

[0124] In some embodiments, increased resistance to the compound comprises an increase of at least 2-fold, 3-fold, 4-fold, 5-fold 6-fold, 7-fold, 8-fold, 9-fold, 10-fold in resistance to the compound compared to a reference plant or part thereof.

[0125] The invention will now be further described under the following headed sections, features under any given section may apply to any embodiment or aspect in any combination.

[0126] DETAILED DESCRIPTION

[0127] Modified plants

[0128] The invention primarily relates to plants that have been modified to comprise a modified BIO3- BIO1 enzyme which provides the plants with increased resistance to a compound which inhibits the biotin synthesis pathway. Suitable compounds which inhibit the biotin synthesis pathway are defined hereinbelow.

[0129] As used herein, reference to a “BIO3-BIO1 enzyme” and the amino acid sequences thereof also refers to and is intended to encompass isolated polynucleotides encoding such an enzyme. BIO3-BIO1 refers to an enzyme that catalyses the conversion of 7-keto-8-Aminopelargonic Acid (KAPA) into Dethiobiotin, the final intermediate in the biotin pathway before the formation of biotin. BIO3-BIO1 is identified by the enzyme number EC 2.6.1.62. BIO3-BIO1 enzyme may refer to any protein that is capable of carrying out the conversion of KAPA into Dethiobiotin. Examples of suitable BIO3-BIO1 enzymes are provided herein in SEQ ID NOs 1 to 21 . BIOS- BIOI may also be referred to as ‘BioDA’ or bifunctional dethiobiotin synthetase, these terms are used interchangeably herein. The BIO1 part of BIO3-BIO1 is a DAPA synthase and the BIO3 part of BIO3-BIO1 is a dethiobiotin synthase.

[0130] Suitably the plant has been modified to increase expression of a modified BIO3-BIO1 enzyme, suitably within the plant or a part thereof. Suitably therefore, the modified plant comprises increased expression of a modified BIO3-BIO1 enzyme. Suitably relative to a reference plant. For example, expression may be increased in comparison to expression of a wild-type BIOS- BIOI enzyme in a reference plant.

[0131] By ‘reference’ plant it is meant a plant that does not include a modified BIO3-BIO1 enzyme of the invention. Reference plants includes wild-type plants (that include a wild-type BIO3-BIO1 enzyme); or modified plants that include modifications other than the modified BIO3-BIO1 enzymes of the invention. Suitably, the term reference plant may be used interchangeably with the term control or control plant.

[0132] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150% greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0133] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 150%, from 10% to 150%, 20% to 150%, 30% to 150%, 40% to 150%, 50% to 150%, 60% to 150%, 70% to 150%, 80% to 150%, 90% to 150%, 100% to 150%, 110% to 150%, 120% to 150%, 130% to 150% or 140% to 150% greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0134] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 100%, from 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100% greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant. Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 90%, from 10% to 90%, 20% to 90%, 30% to 90%, 40% to 90%, 50% to 90%, 60% to 90%, 70% to 90%, 80% to 90% greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0135] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 80%, from 10% to 80%, 20% to 80%, 30% to 80%, 40% to 80%, 50% to 80%, 60% to 80%, 70% to 80%, greater than the expression than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0136] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 70%, from 10% to 70%, 20% to 70%, 30% to 70%, 40% to 70%, 50% to 70%, 60% to 70% greater than the expression than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0137] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 60%, from 10% to 60%, 20% to 60%, 30% to 60%, 40% to 60%, 50% to 60%, greater than the expression thereof than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0138] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 50%, from 10% to 50%, 20% to 50%, 30% to 50%, 40% to 50% greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0139] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 40%, from 10% to 40%, 20% to 40%, 30% to 40%, greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0140] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 30%, from 10% to 30%, 20% to 30%, greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0141] Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 20%, from 10% to 20% greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant. Suitably the expression of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 10% greater than the expression of a reference BIO3-BIO1 enzyme in a reference plant.

[0142] Suitably increases in expression of an enzyme may be determined by measuring an increase in expression of the gene encoding the enzyme, such as by known molecular biology techniques including RT-PCR, qPCR, RNA-seq and the like. Alternatively, expression of the enzyme may be measured directly by other known molecular biology techniques including western blots, or fluorescence based imaging techniques.

[0143] Suitably the plant has been modified to increase activity of a BIO3-BIO1 enzyme, suitably within the plant or a part thereof. Suitably the plant may have been modified to comprise a modified BIO3-BIO1 enzyme as described herein having increased activity when compared to a reference BIO3-BIO1 enzyme. Suitably a reference enzyme is an enzyme which is unmodified, suitably a wildtype enzyme, and which may be considered a control or reference enzyme herein, suitably which may already exist within the plant or a part thereof. Suitably the plant may have been modified to comprise a modified BIO3-BIO1 enzyme having increased activity when compared to a reference BIO3-BIO1 enzyme. Suitably, a reference BIO3-BIO1 enzyme may also be a modified BIO3-BIO1 enzyme that comprises one or more modifications other than those of the invention.

[0144] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level which provides the plant with increased resistance to a compound that inhibits the biotin synthesis pathway relative to an reference plant. In particular, the activity of the modified BIO3-BIO1 enzyme is increased to a level which provides the plant with increased resistance to a compound that inhibits a BIO3-BIO1 enzyme. Suitably, the modified BIO3-BIO1 enzymes described herein may have increased DAPA synthase activity. For example, the modifications may provide for increased resistance to DAPA synthase inhibitors. Suitably, the modified BIO3-BIO1 enzymes described herein may have increased dethiobiotin synthase activity. For example, the modifications may provide for increased resistance to dethiobiotin synthase inhibitors.

[0145] As used herein the terms “resistance” and “tolerance” are used interchangeably to refer to a decrease in the negative effects of compounds on an enzyme or plant in comparison to a reference enzyme or plant. For example, resistance or tolerance refer to an enzyme, such as a modified BIO3-BIO1 enzyme described herein, that is less affected by exposure to compounds that are inhibitory to a reference enzyme (such as a wild-type enzyme). Resistance or tolerance may be imparted by any mechanism such as an increase in activity which means the compound has less effect on the modified enzyme and a plant including the enzyme.

[0146] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0147] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 150%, from 10% to 150%, 20% to 150%, 30% to 150%, 40% to 150%, 50% to 150%, 60% to 150%, 70% to 150%, 80% to 150%, 90% to 150%, 100% to 150%, 110% to 150%, 120% to 150%, 130% to 150% or 140% to 150% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0148] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 100%, from 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0149] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 90%, from 10% to 90%, 20% to 90%, 30% to 90%, 40% to 90%, 50% to 90%, 60% to 90%, 70% to 90%, 80% to 90% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0150] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 80%, from 10% to 80%, 20% to 80%, 30% to 80%, 40% to 80%, 50% to 80%, 60% to 80%, 70% to 80%, greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0151] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 70%, from 10% to 70%, 20% to 70%, 30% to 70%, 40% to 70%, 50% to 70%, 60% to 70% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0152] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 60%, from 10% to 60%, 20% to 60%, 30% to 60%, 40% to 60%, 50% to 60%, greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant. Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 50%, from 10% to 50%, 20% to 50%, 30% to 50%, 40% to 50% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0153] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 40%, from 10% to 40%, 20% to 40%, 30% to 40%, greater than the activity of a reference BIO3- BIO1 enzyme in a reference plant.

[0154] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 30%, from 10% to 30%, 20% to 30%, greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0155] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 20%, from 10% to 20% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0156] Suitably the activity of the modified BIO3-BIO1 enzyme is increased to a level from 5% to 10% greater than the activity of a reference BIO3-BIO1 enzyme in a reference plant.

[0157] Suitably an increase in the activity of an enzyme may be measured by an enzyme assay, which measures the consumption of a substrate or production of a product over time, suitably in an in vitro environment. Such assays may be spectrophotometric, fluorometric, calorimetric, chemiluminescent, light scattering or microscale thermophoresis. In one embodiment, a fluorometric assay, in which the fluorescent product produced by the reaction of DAPA with o- phthalaldehyde and p-mercaptoethanol is measured.

[0158] Suitably the plant may have been modified to increase the expression and / or the activity of a BIO3-BIO1 enzyme, suitably within the plant or a part thereof. Suitably the increase in expression and the increase in activity are as defined above.

[0159] As noted above, the plant or part thereof may be modified to comprise a BIO3-BIO1 enzyme which provides the plant or part thereof with increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant. For example, a BIO3-BIO1 enzyme including the modifications described herein may have increased resistance to compounds which inhibit DAPA synthase (e.g. the BIO1 part of a BIO3-BIO1 enzyme). For example, a BIO3-BIO1 enzyme including the modifications described herein may have increased resistance to compounds which inhibit dethiobiotin synthase (e.g. the BIO3 part of a BIO3- BIO1 enzyme).

[0160] The BIO3-BIO1 enzyme is found in plants, algae, fungi, or oomycetes. Therefore, suitably the modified BIO3-BIO1 enzyme may be an endogenous or a heterologous enzyme to the plant. Suitably the modified BIO3-BIO1 enzyme may be derived from a plant, from an algae, or a from a fungus. In one embodiment, the modified BIOI3-BIO1 enzyme is derived from a plant. In one embodiment, the modified BIO3-BIO1 enzyme is introduced into a heterologous plant species. In another embodiment, the modified BIO3-BIO1 enzyme is introduced into a plant of the same species or to a crossable plant species. In one embodiment, the modified BIOI3- BIO1 enzyme is derived from an algae. In one embodiment, the modified BIOI3-BIO1 enzyme is derived from a fungus.

[0161] Suitably the modified BIO3-BIO1 enzyme may be derived from any plant species. Suitably the modified BIO3-BIO1 enzyme may be derived from any of the following plant species: Arabidopsis thaliana, Zea mays, Quercus robur, Triticum aestivum, Glycine max, Setaria italica, Oryza sativa, Heliosperma pusilium, Taxus chinensis, Carpinus fangiana, Cinnamomum micranthum, Apostasia shenzhenica, Asparagus officinalis, Phoenix dactylifera, Zostera marina, Amborella trichopoda, Adiantum nelumboides Echinochloa crus- galli, Zingiber officinale, Thlaspi arvense, Vitis vinifera, Helianthus annuus, Brassica oleracea, Hordeum vulgare, Brassica napus, Selaginella moellendorffii, Gossypium hirsutum and Amaranthus palmeri. In one embodiment, the modified BIO3-BIO1 enzyme may be derived from any of the following plant species: Setaria italica, Arabidopsis thaliana, Helianthus annuus, Quercus robur, Phoenix dactylifera, Physcomitrium patens, Taxus chinensis, Adiantum nelumboides, Zea mays, Hordeum vulgare, Brassica napus, Gossypium hirsutum, Oryza sativa, Triticum aestivum, Selaginella moellendorffii and Glycine max. In one embodiment, the modified BIO3-BIO1 enzyme is derived from Arabidopsis thaliana, or Zea mays. In other embodiments, the modified BIO3-BIO1 enzyme is the endogenous BIO3-BIO1 enzyme from the plant of interest.

[0162] Suitably the modified BIO3-BIO1 enzyme may be derived from any algal species. Suitably the modified BIO3-BIO1 enzyme may be derived from any of the following species of algae: Nannochloropsis gaditana, Pedinophyceae sp., Trebouxia sp., Ostreococcus tauri, and Micromonas pusilla.\r\ one embodiment, the modified BIO3-BIO1 enzyme may be derived from any of the following species of algae: Nannochloropsis gaditana, and Ostreococcus tauri. In one embodiment, the modified BIO3-BIO1 enzyme is derived from Nannochloropsis gaditana. Suitably the modified BIO3-BIO1 enzyme may be derived from any fungal species. Suitably the modified BIO3-BIO1 enzyme may be derived from any of the following species of fungi: Aspergillus candidus, Blastocladiella emersonii, Paraphysoderma sedebokerense, Talaromyces proteolyticus, Pseudomassariella vexata, Microthyrium microscopicum, Lophium mytilinum, Monilinia fructicola, Cryomyces minteri, Coniosporium apollinis, Polytolypa hystricis, Xylona heveae, Calocera cornea, Rhinocladiella mackenziei, Coniosporium apollinis, Schizosaccharomyces japonicus, Yarrowia lipolytica, Aspergillus niger, Gibberella zeae, and Aspergillus nidulans. In one embodiment, the modified BIO3-BIO1 enzyme may be derived from any of the following species of fungi: Schizosaccharomyces japonicus and Gibberella zeae.

[0163] Suitably the modified BIO3-BIO1 enzyme may be derived from any oomycete species. Suitably the modified BIO3-BIO1 enzyme may be derived from any of the following species of oomycete: Thraustotheca clavata, Albugo laibachii, Achlya hypogyna, Phytophthora spp. such as Phytophthora cactorum, Phytophthora rubi, Phytophthora capsici, and Phytophthora sojae. In one embodiment the modified BIO3-BIO1 enzyme may be derived from any of the following species of oomycete: Thraustotheca clavata,

[0164] Suitably the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 30% identity to an amino acid sequence selected from: SEQ ID NO: 1 to 21 , (or SEQ ID NO: 22 and 39), or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0165] Suitably the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to an amino acid sequence selected from: SEQ ID NO: 1 to 21 , (or SEQ ID NO: 22 and 39), or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0166] Suitably the modified BIO3-BIO1 enzyme may comprise an amino acid sequence having at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to an amino acid sequence selected from: SEQ ID NO: 1 to 21 , (or SEQ ID NO: 22 and 39), or a functional fragment thereof, and comprises two or more of the modifications described herein. In one embodiment, the modified BIO3-BIO1 enzyme is derived from Arabidopsis thaliana and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:1 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0167] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Zea mays and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:2 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0168] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Nannochloropsis gaditana and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:3 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0169] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Setaria italica and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:4 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0170] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Oryza sativa and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:5 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0171] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Ostreococcus tauri and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:6 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0172] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Schizosaccharomyces japonicus and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:7 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0173] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Selaginella moellendorffii and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:8 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0174] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Helianthus annuus and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:9 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0175] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Taxus chinensis and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 10 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0176] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Physcomitrium patens and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 11 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0177] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Adiantum nelumboides and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 12 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0178] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Phoenix dactylifera and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 13 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0179] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Quercus robur and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:14 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0180] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Thraustotheca clavate and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 15 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0181] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Gibberella zeae and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:16 or a functional fragment thereof, and comprises two or more of the modifications described herein. In one embodiment, the modified BIO3-BIO1 enzyme is derived from Hordeum vulgare and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 17 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0182] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Brassica napus and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 18 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0183] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Gossypium hirsutum and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO: 19 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0184] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Glycine max and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NQ:20 or a functional fragment thereof, and comprises two or more of the modifications described herein.

[0185] In one embodiment, the modified BIO3-BIO1 enzyme is derived from Triticum aestivum and comprises an amino acid sequence having at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity according to SEQ ID NO:21 or a functional fragment thereof, and comprises two or more of the modifications described herein. In some embodiments, the modified BIO3-BIO1 enzyme comprises two or more modifications, as described herein. Suitably in such embodiments, the plant has been modified to comprise a BIO3-BIO1 enzyme having two or more modifications.

[0186] In one embodiment, the plant or part thereof may be modified to comprise a modified BIO3- BIO1 enzyme, wherein the enzyme comprises two or more modifications which provide the plant or part thereof with increased resistance to a compound which inhibits the biotin synthesis pathway relative to an reference plant. For example, a BIO3-BIO1 enzyme including the modifications described herein may have increased resistance to compounds which inhibit DAPA synthase (e.g. the Biol part of a BIO3-BIO1 enzyme).

[0187] Suitably the two or more modifications provide the enzyme, and therefore the plant, with increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

[0188] Suitably in some embodiments, the BIO3-BIO1 enzyme may be modified and may also be overexpressed in the plant or part thereof. Suitable increases in expression and overexpression are described above. Suitable modifications are described below. Suitably the two or more modifications and / or the increased expression provide the plant with increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

[0189] Suitably the two or more modifications are mutations. Suitably any of the modifications herein may be referred to interchangeably as ‘mutations’, and therefore the modified plant or modified BIO3-BIO1 enzyme maybe referred to as a ‘mutant’ plant or ‘mutant’ BIO3-BIO1 enzyme. Suitably the mutations are selected from deletions, insertions, substitutions etc. In one embodiment the mutations are amino acid substitutions. Suitable modifications to the BIO3- BIO1 enzymes are defined in the relevant sections hereinbelow. As explained therein, in one embodiment, the BIO3-BIO1 enzyme is modified, in one embodiment the BIO3-BIO1 enzyme comprises two or more amino acid substitutions. In another embodiment, the native mitochondrial targeting peptide of the modified BIO3-BIO1 enzyme is replaced with a heterologous mitochondrial targeting peptide, optionally in addition to the two or more amino acid modifications.

[0190] The plants of the present invention include both non-transgenic plants and transgenic plants.

[0191] Suitably, the term ‘modified plant’ includes both non-transgenic plants and transgenic plants. By "non-transgenic plant" is intended to mean a plant lacking recombinant DNA in its genome, but containing a mutant nucleic acid molecule in the plant cell genome which has been mutated using mutagenic techniques, such as chemical mutagenesis, gene editing or by those methods provided herein. Non-transgenic plants may encompass those plants having mutant or modified sequences as a result of natural processes. In specific embodiments, the non- transgenic plant comprises a modified BIO3-BIO1 enzyme that has been altered through gene editing to comprise at least two or more of the modifications disclosed herein. Such gene editing modifications will increase the resistance of the plant to the herbicide of interest.

[0192] As used herein, a “non-natural mutation” or “non-natural modification” refers to a mutation that is generated through human intervention and differs from a mutation found in the same gene that have occurred in nature (e.g., occurred naturally). A “non-natural mutation” or “non- natural modification” as used herein does not include a mutation generated in a gene of a plant species through human intervention if the same mutation is also naturally occurring mutation in that gene and in that plant species.

[0193] By "transgenic plant" is intended to mean a plant comprising recombinant DNA in its genome. As used herein, "recombinant," when referring to nucleic acid or polypeptide, indicates that such material has been altered as a result of human application of a recombinant technique, such as by polynucleotide restriction and ligation, by polynucleotide overlap- extension, or by genomic insertion or transformation. In one embodiment, recombinant in relation to nucleic acids or polypeptides refers to nucleic acids or polypeptides that are produced or altered outside of a host cell into which they are intended to be transformed (such as a plant cell or plant as described herein). Therefore ‘transgenic plants’ as referred to herein refer to a plant comprising at least one recombinant DNA that is inserted into the genome not employing an SDN-1 or SDN-2 technique.. A gene sequence open reading frame is recombinant if that nucleotide sequence has been removed from its natural context and cloned into any type of artificial nucleic acid vector. The term recombinant also can refer to an organism having a recombinant material, e.g., a plant that comprises a recombinant nucleic acid can be considered a recombinant plant. Such a transgenic plant can be produced by introducing recombinant DNA into the genome of the plant. When such recombinant DNA is incorporated into the genome of the transgenic plant, progeny of the plant can also comprise the recombinant DNA. A progeny plant that comprises at least a portion of the recombinant DNA of at least one progenitor transgenic plant is also a transgenic plant.

[0194] As used herein, "heterologous" in reference to a polypeptide or polynucleotide sequence is a sequence that originates, for example, from a cell or an organism from a foreign species. Alternatively, if the sequence originates from the same species, it is derived from a cell organism having a different genetic background; or if from the same genetic background, it is substantially modified from its native form in composition and / or genomic locus by deliberate human intervention. As such, heterologous sequences are in a configuration not found in nature.

[0195] The term “spontaneous mutant” refers to mutants or variants that arise from the parent strain without the intentional use of mutagens i.e. they are considered as not genetically modified (non-GMO). Spontaneous mutants in respect of plants may also be known as sports, breaks, or chimeras.

[0196] Suitably the plant or part thereof of the invention is transgenic. In other embodiments, the plant or plant part thereof of the invention is non-transgenic and comprises a gene edit that increases the plant’s or plant part’s tolerance to a herbicide of interest.

[0197] In one embodiment, therefore, the plant or part thereof comprises a recombinant polynucleotide encoding a modified BIO3-BIO1 enzyme. Suitably the recombinant polynucleotide may be operable to express the modified BIO3-BIO1 enzyme at increased levels compared to a reference plant. Suitably wherein the increased expression of said polynucleotide provides or confers to the plant or part thereof an increased resistance to a compound which inhibits the biotin synthesis pathway as defined above.

[0198] In another embodiment, the plant or part thereof comprises a polynucleotide encoding a modified BIO3-BIO1 enzyme. Suitably the polynucleotide encoding the modified BIO3-BIO1 enzyme may comprise two or more modifications as described herein. Suitably therefore the polynucleotide may be operable to express a modified BIO3-BIO1 enzyme having two or more modifications as described herein. Suitably wherein the expression of said polynucleotide provides or confers to the plant or part thereof increased resistance to a compound which inhibits the biotin synthesis pathway. Suitably the or each modification in the BIO3-BIO1 enzyme provides increased resistance to a compound which inhibits the biotin synthesis pathway. Suitable such modifications are defined hereinbelow. In specific embodiments, the modified BIO3-BIO1 enzyme is encoded by a recombinant polynucleotide stably integrated into the plant's genome. Alternatively, the modified BIO3-BIO1 enzyme is encoded by a polynucleotide, suitably a gene, comprising a non-transgenic modification, such as an edit, within the genome of the plant. Suitably in such embodiments, the polynucleotide encoding a modified BIO3-BIO1 enzyme may also be operable to express the modified BIO3-BIO1 enzyme at increased levels compared to a reference plant. In one embodiment, the plant has been transformed with said recombinant polynucleotide. Suitable means of transformation are described hereinbelow.

[0199] The transformed parts of plants, transformed plant cells or a transformed plant protoplasts as described herein may be regenerated to produce a modified plant as described herein.

[0200] When adequate numbers of transformed cells or protoplasts containing a recombinant modified BIO3-BIO1 enzyme are obtained, the cells can be cultured, then regenerated into whole plants. “Regeneration” refers to the process of growing a plant from a plant cell (for example, plant protoplast or explant). Such regeneration techniques rely on manipulation of certain phytohormones in a tissue culture growth medium, typically relying on a biocide and / or herbicide marker that has been introduced together with the desired nucleotide sequences. Choice of methodology for the regeneration step is not critical. See, for example, Ammirato et al., Handbook of Plant Cell Culture — Crop Species. Macmillan Publ. Co. (1984); Shimamoto et al., Nature 338:274-276 (1989); Fromm, UCLA Symposium on Molecular Strategies for Crop Improvement, Apr. 16-22, 1990. Keystone, Colo. (1990); Vasil et al., Bio / Technology 8:429-434 (1990); Vasil et al., Bio / Technology 10:667-674 (1992); Hayashimoto, Plant Physiol. 93:857-863 (1990); and Datta et al., Bio-technology 8:736-740 (1990). Such regeneration techniques are described generally in Klee et al., Ann. Rev. Plant Phys. 38:467- 486 (1987).

[0201] Modified BIO3-BIO1 enzyme

[0202] As described above, the BIO3-BIO1 enzyme may be modified with two or more modifications as described herein. Suitably the two or more modifications provide the enzyme, and therefore the plant, with increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant. For example, a compound that inhibits a wild-type, unmodified or control BIO3-BIO1 enzyme.

[0203] The invention provides a modified BIO3-BIO1 enzyme. In some aspects and embodiments of the invention, the BIO3-BIO1 enzyme may comprise two or more modifications, suitably two or more mutations.

[0204] Mutations may include deletions or substitutions or combinations thereof. For example, the mutations may be conservative or non-conservative amino acid substitutions. “Conservative amino acid substitutions” refer to the interchangeability of residues having similar side chains, and thus typically involves substitution of an amino acid in a polypeptide with amino acids within the same or similar defined class of amino acids. By way of example, an amino acid with an aliphatic side chain may be substituted with another aliphatic amino acid, e.g., alanine, valine, leucine, and isoleucine; an amino acid with hydroxyl side chain may be substituted with another amino acid with a hydroxyl side chain, e.g., serine and threonine; an amino acids having aromatic side chains may be substituted with another amino acid having an aromatic side chain, e.g., phenylalanine, tyrosine, tryptophan, and histidine; an amino acid with a basic side chain may be substituted with another amino acid with a basic side chain, e.g., lysine and arginine; an amino acid with an acidic side chain may be substituted with another amino acid with an acidic side chain, e.g., aspartic acid or glutamic acid; and a hydrophobic or hydrophilic amino acid may be substituted with another hydrophobic or hydrophilic amino acid, respectively. Exemplary conservative substitutions are provided below:

[0205] RESIDUE Possible Conservative Substitutions

[0206] A, L, V, I Other aliphatic (A, L, V, I )

[0207] Other non-polar (A, L, V, I, G, M)

[0208] G, M Other non-polar (A, L, V, I, G, M)

[0209] D, E Other acidic (D, E)

[0210] K, R Other basic (K, R)

[0211] N, Q, S, T Other polar

[0212] H, Y, W, F Other aromatic (H, Y, W, F) C None

[0213] P None

[0214] “Non-conservative substitution” refers to substitution of an amino acid in a polypeptide with an amino acid with significantly differing side chain properties. Non-conservative substitutions may use amino acids between, rather than within, the defined groups and may affect (a) the structure of the peptide backbone in the area of the substitution (e.g., proline for glycine) (b) the charge or hydrophobicity, or (c) the bulk of the side chain. By way of example, an exemplary non-conservative substitution can be an acidic amino acid substituted with a basic or aliphatic amino acid; an aromatic amino acid substituted with a small amino acid; and a hydrophilic amino acid substituted with a hydrophobic amino acid.

[0215] “Deletion” refers to modification of a polypeptide by removal of one or more amino acids in comparison to a wild-type or reference polypeptide. Deletions can comprise removal of 1 or more amino acids, 2 or more amino acids, or 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids of the polypeptide while retaining enzymatic activity. Deletions can comprise a continuous segment or can be discontinuous.

[0216] In one embodiment, the modified BIO3-BIO1 enzyme comprises a modification at two or more of amino acid positions 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto.

[0217] Suitably, the corresponding positions thereto may be those in any homologous sequence to that of SEQ ID NO:1 , such as those in SEQ ID NOs 2-21 , for example as shown in Table 1.

[0218] In one embodiment, the modified BIO3-BIO1 enzyme may comprise two or more modifications selected from: F348, Q350, C388, W391 , W392, P421 , Q506, A507, P508, S509, Y511 , T512, G513, Q517, Y520, G608, A609, G700, and / or S704 of SEQ ID NO:1 (WT Arabidopsis thaliana sequence), or said modifications at corresponding residues thereto. Suitably, the corresponding residues thereto may be those in any homologous sequence to that of SEQ ID NO:1 , such as those in SEQ ID NOs 2-21 , for example as shown in Table 1.

[0219] Suitably the modified BIO3-BIO1 enzyme may comprise two or more modifications at positions selected from any of those described in Table 1 below:

[0220] Table 1 : corresponding positions for modification in homologous BIO3-BIO1 enzymes

[0221] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 , wherein at least two residues selected from F348, Q350, C388, W391 , W392, P421 , Q506, A507, P508, S509, Y511 , T512, G513, Q517, Y520, G608, A609, G700, and / or S704 thereof, or corresponding residues thereto, include a modification. Suitably wherein at least two residues selected from F348, Q350, C388, W391 , W392, P421 , Q506, A507, P508, S509, Y511 , T512, G513, Q517, Y520, G608, A609, G700, and / or S704 of SEQ ID NO: 1 , or corresponding residues thereto, include a modification.

[0222] In some embodiments, the two or more modifications include a substitution of amino acid residue F348 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of phenylalanine 348 of SEQ ID NO: 1 or a residue corresponding thereto with an alanine, aspartic acid, serine, threonine, methionine, valine, cysteine or asparagine. For example, the two or more modifications include the substitution F348A, F348D, F348S, F348T, F348M, F348V, F348C, or F348N with reference to SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0223] In some embodiments, the two or more modifications include a substitution of amino acid residue Q350 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of glutamine 350 of SEQ ID NO: 1 or a residue corresponding thereto with a histidine. For example, the two or more modifications include the substitution Q350H with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0224] In some embodiments, the two or more modifications include a substitution of amino acid residue C388 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of cysteine 388 of SEQ ID NO: 1 or a residue corresponding thereto with a threonine. For example, the two or more modifications include the substitution C388T with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0225] In some embodiments, the two or more modifications include a substitution of amino acid residue W391 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of tryptophan 391 of SEQ ID NO: 1 or a residue corresponding thereto with a leucine or methionine. For example, the two or more modifications include the substitution W391 L or W391M with reference to SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0226] In some embodiments, the two or more modifications include a substitution of amino acid residue W392 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of tryptophan 392 of SEQ ID NO: 1 or a residue corresponding thereto with a serine. For example, the two or more modifications include the substitution W392S with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0227] In some embodiments, the two or more modifications include a substitution of amino acid residue P421 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of proline 421 of SEQ ID NO: 1 or a residue corresponding thereto with a leucine. For example, the two or more modifications include the substitution P421 L with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0228] In some embodiments, the two or more modifications include a substitution of amino acid residue Q506 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of glutamine 506 of SEQ ID NO: 1 or a residue corresponding thereto with an alanine. For example, the two or more modifications include the substitution Q506A with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0229] In some embodiments, the two or more modifications include a substitution of amino acid residue A507 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of alanine 507 of SEQ ID NO: 1 or a residue corresponding thereto with a serine or lysine. For example, the two or more modifications include the substitution A507S or A507K with reference to SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0230] In some embodiments, the two or more modifications include a substitution of amino acid residue P508 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of proline 508 of SEQ ID NO: 1 or a residue corresponding thereto with a threonine or leucine. For example, the two or more modifications include the substitution P508T or P508L with reference to SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0231] In some embodiments, the two or more modifications include a substitution of amino acid residue S509 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of serine S509 of SEQ I D NO: 1 or a residue corresponding thereto with a valine. For example, the two or more modifications include the substitution S509V with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0232] In some embodiments, the two or more modifications include a substitution of amino acid residue Y511 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of tyrosine Y511 of SEQ ID NO: 1 or a residue corresponding thereto with a glutamic acid, tryptophan or methionine. For example, the two or more modifications include the substitution Y511 E, Y511W or Y511M with reference to SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0233] In some embodiments, the two or more modifications include a substitution of amino acid residue T512 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of threonine 512 of SEQ ID NO: 1 or a residue corresponding thereto with a cysteine. For example, the two or more modifications include the substitution T512C with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0234] In some embodiments, the two or more modifications include a deletion of amino acid residue G513 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a deletion of glycine G513 of SEQ ID NO: 1 or a residue corresponding thereto. For example, the two or more modifications include the deletion of G513 (G513Del) with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0235] In some embodiments, the two or more modifications include a substitution of amino acid residue Q517 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of glutamine 517 of SEQ ID NO: 1 or a residue corresponding thereto with a histidine, phenylalanine or threonine. For example, the two or more modifications include the substitution Q517H, Q517F, or Q517T with reference to SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0236] In some embodiments, the two or more modifications include a substitution of amino acid residue Y520 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of tyrosine 520 of SEQ ID NO: 1 or a residue corresponding thereto with a histidine, tryptophan or asparagine. For example, the two or more modifications include the substitution Y520W or Y520N with reference to SEQ I D NO: 1 or the same modifications at residues corresponding thereto. In some embodiments, the two or more modifications include a substitution of amino acid residue G608 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of glycine 608 of SEQ ID NO: 1 or a residue corresponding thereto with an alanine. For example, the two or more modifications include the substitution G608A with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0237] In some embodiments, the two or more modifications include a substitution of amino acid residue A609 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of alanine 609 of SEQ I D NO: 1 or a residue corresponding thereto with a tyrosine, threonine, methionine or valine. For example, the two or more modifications include the substitution A609Y, A609T, A609M or A609V with reference to SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0238] In some embodiments, the two or more modifications include a substitution of amino acid residue G700 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of glycine 700 of SEQ ID NO: 1 or a residue corresponding thereto with a serine or alanine. For example, the two or more modifications include the substitution G700S or G700A with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0239] In some embodiments, the two or more modifications include a substitution of amino acid residue A704 of SEQ ID NO: 1 or a residue corresponding thereto. Suitably, the two or more modifications include a substitution of alanine 704 of SEQ I D NO: 1 or a residue corresponding thereto with a histidine. For example, the two or more modifications include the substitution A704H with reference to SEQ ID NO: 1 or the same modification at residues corresponding thereto.

[0240] A modified BIO3-BIO1 enzyme of the invention may comprise an amino acid sequence according to any of SEQ ID NO: 1 to 21 (or SEQ ID NOs: 22 to 39), wherein at least two positions selected from 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and / or 704 as defined in relation to SEQ ID NO:1 , or corresponding positions thereto in any of SEQ ID NOs 2-21 , include a modification. Suitably wherein at least two positions selected from 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and / or 704 as defined in relation to SEQ ID NO:1 , or corresponding positions thereto in any of SEQ ID NOs 2-21 , include a modification. Positions 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F333, Q335, C372, W375, W376, P405, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G595, A596, G687, and T691 of SEQ ID NO: 2 (which relates to Zea mays BIO3-BIO1). As such, the modified BIO3- BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 2 wherein at least two of residues F333, Q335, C372, W375, W376, P405, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G595, A596, G687, and T691 thereof include a modification.

[0241] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 2, wherein at least two of residues F333, Q335, C372, W375, W376, P405, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G595, A596, G687, and T691 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 2 wherein at least two of residues F333, Q335, C372, W375, W376, P405, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G595, A596, G687, and T691 include a modification as described herein.

[0242] Positions 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to positions F291 , Q293, C339, W342, W343, P371 , A467, A468, P469, T470, F472, T474, G475, H477, Y480, G600, A601 , G692, and T696 of SEQ ID NO: 3 (which relates to Nannochloropsis gaditana BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 3 wherein at least two of residues F333, Q335, C372, W375, W376, P405, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G595, A596, G687, and T691 include a include a modification.

[0243] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 3, wherein at least two of residues F291 , Q293, C339, W342, W343, P371 , A467, A468, P469, T470, F472, T474, G475, H477, Y480, G600, A601 , G692, and T696 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 3 wherein at least two of residues F291 , Q293, C339, W342, W343, P371 , A467, A468, P469, T470, F472, T474, G475, H477, Y480, G600, A601 , G692, and T696 include a modification as described herein.

[0244] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F334, Q336, C375, W378, W379, P408, Q497, A498, P499, S500, Y502, T503, S504, Q508, Y511 , G601 , A602, G693, and T697 of SEQ ID NO: 4 (which relates to Setaria italica BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 4 wherein at least two of residues F334, Q336, C375, W378, W379, P408, Q497, A498, P499, S500, Y502, T503, S504, Q508, Y511 , G601 , A602, G693, and T697 thereof include a modification.

[0245] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 4, wherein at least two of residues F334, Q336, C375, W378, W379, P408, Q497, A498, P499, S500, Y502, T503, S504, Q508, Y511 , G601 , A602, G693, and T697 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ I D NO: 4 wherein at least two of residues F334, Q336, C375, W378, W379, P408, Q497, A498, P499, S500, Y502, T503, S504, Q508, Y511 , G601 , A602, G693, and T697 thereof include a modification as described herein.

[0246] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F332, Q334, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 of SEQ ID NO: 5 (which relates to Oryza sativa BIO3-BIO1). As such, the modified BIO3- BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 5 wherein at least two of residues F332, Q334, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 thereof include a modification.

[0247] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 5, wherein at least two of residues F332, Q334, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ I D NO: 5 wherein at least two of residues F332, Q334, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 thereof include a modification as described herein.

[0248] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F332, Q334, A374, W377, W378, P407, Q503, S504, P505, S506, F508, T509, G510, T514, Y517, G621 , A622, G713, and T717 of SEQ ID NO: 6 (which relates to Ostreococcus tauri BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 6 wherein at least two of residues F332, Q334, A374, W377, W378, P407, Q503, S504, P505, S506, F508, T509, G510, T514, Y517, G621 , A622, G713, and T717 thereof include a modification.

[0249] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 6, wherein at least two of residues F332, Q334, A374, W377, W378, P407, Q503, S504, P505, S506, F508, T509, G510, T514, Y517, G621 , A622, G713, and T717 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 6 wherein residues F332, Q334, A374, W377, W378, P407, Q503, S504, P505, S506, F508, T509, G510, T514, Y517, G621 , A622, G713, and T717 thereof include a modification as described herein.

[0250] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F263, Q265, S324, W327, W328, P356, C433, P434, P435, N436, Y438, N439, - (G513 has no corresponding residue), V443, Y445, G525, A526, G633, and T637 of SEQ ID NO: 7 (which relates to Schizosaccharomyces japonicus BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 7 wherein at least two of residues F263, Q265, S324, W327, W328, P356, C433, P434, P435, N436, Y438, N439, V443, Y445, G525, A526, G633, T637 thereof include a modification. The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 7, wherein at least two of residues F263, Q265, S324, W327, W328, P356, C433, P434, P435, N436, Y438, N439, V443, Y445, G525, A526, G633, and T637 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 7 wherein residues F263, Q265, S324, W327, W328, P356, C433, P434, P435, N436, Y438, N439, V443, Y445, G525, A526, G633, T637 thereof include a modification as described herein.

[0251] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F358, Q360, C397, W400, W401 , P430, Q514, A515, P516, S517, Y519, T520, S521 , Q525, Y528, A615, A616, G707, and T711 of SEQ ID NO: 8 (which relates to Selaginella moellendorffi BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 8 wherein at least two of residues F358, Q360, C397, W400, W401 , P430, Q514, A515, P516, S517, Y519, T520, S521 , Q525, Y528, A615, A616, G707, and T711 thereof include a modification.

[0252] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 8, wherein at least two of residues F358, Q360, C397, W400, W401 , P430, Q514, A515, P516, S517, Y519, T520, S521 , Q525, Y528, A615, A616, G707, and T711 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 8 wherein at least two of residues F358, Q360, C397, W400, W401 , P430, Q514, A515, P516, S517, Y519, T520, S521 , Q525, Y528, A615, A616, G707, and T711 thereof include a modification as described herein.

[0253] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F344, Q346, C384, W387, W388, P417, Q506, A507, P508, S509, Y511 , T512, G513, Q517, Y520, G610, A611 , G702, and S706 of SEQ ID NO: 9 (which relates to Helianthus annuus BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 9 wherein at least two residues F344, Q346, C384, W387, W388, P417, Q506, A507, P508, S509, Y511 , T512, G513, Q517, Y520, G610, A611 , G702, and S706 thereof include a modification.

[0254] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 9, wherein at least two of residues F344, Q346, C384, W387, W388, P417, Q506, A507, P508, S509, Y511 , T512, G513, Q517, Y520, G610, A611 , G702, and S706 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 9 wherein at least two of residues F344, Q346, C384, W387, W388, P417, Q506, A507, P508, S509, Y511 , T512, G513, Q517, Y520, G610, A611 , G702, and S706 thereof include a modification as described herein.

[0255] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F374, Q376, C418, W421 , W422, P451 , A533, F534, F535, I536, M538, T539, M540, E545, Y548, A589, S590, G681 , and S685 of SEQ ID NO: 10 (which relates to Taxus chinensis BIO3-BIO1). As such, the modified BIO3- BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 10 wherein at least two of residues F374, Q376, C418, W421 , W422, P451 , A533, F534, F535, I536, M538, T539, M540, E545, Y548, A589, S590, G681 , and S685 thereof include a modification.

[0256] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 10, wherein at least two of residues F374, Q376, C418, W421 , W422, P451 , A533, F534, F535, I536, M538, T539, M540, E545, Y548, A589, S590, G681 , and S685 include at least one modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 10 wherein at least two of residues F374, Q376, C418, W421 , W422, P451 , A533, F534, F535, I536, M538, T539, M540, E545, Y548, A589, S590, G681 , and S685 thereof include a modification as described herein. Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F443, Q445, C486, W489, W490, P519, Q605, A606, P607, S608, Y610, T611 , G612, Q616, H619, G708, A709, G800, T804 of SEQ ID NO: 11 (which relates to Physcomitrium patens BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 11 wherein at least two of residues F443, Q445, C486, W489, W490, P519, Q605, A606, P607, S608, Y610, T611 , G612, Q616, H619, G708, A709, G800, T804 thereof include a modification.

[0257] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 11 , wherein at least two of residues F443, Q445, C486, W489, W490, P519, Q605, A606, P607, S608, Y610, T611 , G612, Q616, H619, G708, A709, G800, T804 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 11 wherein at least two of residues F443, Q445, C486, W489, W490, P519, Q605, A606, P607, S608, Y610, T611 , G612, Q616, H619, G708, A709, G800, T804 thereof include a modification as described herein.

[0258] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F324, Q326, S373, W376, W377, P405, C515, E516, G517, N518, F520, N521 , - (G513 has no corresponding residue), V524, Y527, G622, A623, G744, and T748 of SEQ ID NO: 12 (which relates to Adiantum nelumboides BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 12 wherein at least two of residues F324, Q326, S373, W376, W377, P405, C515, E516, G517, N518, F520, N521 , V524, Y527, G622, A623, G744, and T748 thereof include a modification.

[0259] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 12, wherein at least two of residues F324, Q326, S373, W376, W377, P405, C515, E516, G517, N518, F520, N521 , V524, Y527, G622, A623, G744, and T748 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 12 wherein at least two of residues F324, Q326, S373, W376, W377, P405, C515, E516, G517, N518, F520, N521 , V524, Y527, G622, A623, G744, and T748 thereof include a modification as described herein.

[0260] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F366, Q368, C407, W410, W411 , P440, Q529, A530, P531 , S532, Y534, T535, G536, Q540, Y543, A633, A634, G725, and S729 of SEQ ID NO: 13 (which relates to Phoenix dactylifera BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 13 wherein at least two of residues F366, Q368, C407, W410, W411 , P440, Q529, A530, P531 , S532, Y534, T535, G536, Q540, Y543, A633, A634, G725, and S729 thereof include a modification.

[0261] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 13, wherein at least two of residues F366, Q368, C407, W410, W411 , P440, Q529, A530, P531 , S532, Y534, T535, G536, Q540, Y543, A633, A634, G725, and S729 include at least one modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 13 wherein at least two of residues F366, Q368, C407, W410, W411 , P440, Q529, A530, P531 , S532, Y534, T535, G536, Q540, Y543, A633, A634, G725, and S729 thereof include a modification as described herein.

[0262] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F360, Q362, C400, W403, W404, P433, Q522, A523, P524, S525, Y527, T528, G529, Q533, Y536, G626, A627, G718, and S722 of SEQ ID NO: 14 (which relates to Quercus robur BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 14 wherein at least two of residues F360, Q362, C400, W403, W404, P433, Q522, A523, P524, S525, Y527, T528, G529, Q533, Y536, G626, A627, G718, and S722 thereof include a modification. The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 14, wherein at least two of residues F360, Q362, C400, W403, W404, P433, Q522, A523, P524, S525, Y527, T528, G529, Q533, Y536, G626, A627, G718, and S722 include at least one modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 14 wherein at least two of residues F360, Q362, C400, W403, W404, P433, Q522, A523, P524, S525, Y527, T528, G529, Q533, Y536, G626, A627, G718, and S722 thereof include a modification as described herein.

[0263] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F304, Q306, C341 , W344, W345, P373, A456, E457, P458, S459, Y461 , N462, - (G513 has no corresponding residue), H466, Y469, G552, A553, G644, T648 of SEQ ID NO: 15 (which relates to Thraustotheca clavata BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 15 wherein at least two of residues F304, Q306, C341 , W344, W345, P373, A456, E457, P458, S459, Y461 , N462, H466, Y469, G552, A553, G644, T648 thereof include a modification.

[0264] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 15, wherein at least two of residues F304, Q306, C341 , W344, W345, P373, A456, E457, P458, S459, Y461 , N462, H466, Y469, G552, A553, G644, T648 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 15 wherein at least two of residues F304, Q306, C341 , W344, W345, P373, A456, E457, P458, S459, Y461 , N462, H466, Y469, G552, A553, G644, T648 thereof include a modification as described herein.

[0265] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F270, Q272, S310, W313, W314, A342, A421 , E422, P423, C424, Y426, N427, -(G513 has no corresponding residue), I430, Y433, G516, A517, G626, and T630 of SEQ ID NO: 16 (which relates to Gibberella zeae BIO3- BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ I D NO: 16 wherein at least two of residues F270, Q272, S310, W313, W314, A342, A421 , E422, P423, C424, Y426, N427, I430, Y433, G516, A517, G626, and T630 thereof include a modification.

[0266] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 16, wherein at least two of residues F270, Q272, S310, W313, W314, A342, A421 , E422, P423, C424, Y426, N427, I430, Y433, G516, A517, G626, and T630 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 16 wherein at least two of residues F270, Q272, S310, W313, W314, A342, A421 , E422, P423, C424, Y426, N427, I430, Y433, G516, A517, G626, and T630 thereof include a modification as described herein.

[0267] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F330, Q332, C371 , W374, W375, P404, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G598, A599, G690, and T694 of SEQ ID NO: 17 (which relates to Hordeum vulgare BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 17 wherein at least two of residues F330, Q332, C371 , W374, W375, P404, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G598, A599, G690, and T694 thereof include a modification.

[0268] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 17, wherein at least two of residues F330, Q332, C371 , W374, W375, P404, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G598, A599, G690, and T694 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 17 wherein at least two of residues F330, Q332, C371 , W374, W375, P404, Q494, A495, P496, S497, Y499, T500, S501 , Q505, Y508, G598, A599, G690, and T694 thereof include a modification as described herein. Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F348, Q350, C388, W391 , W392, P421 , Q510, A511 , P512, S513, Y515, T516, G517, Q521, Y524, G613, A614, G705, and S709 of SEQ ID NO: 18 (which relates to Brassica napus BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 18 wherein at least two of residues F348, Q350, C388, W391 , W392, P421 , Q510, A511 , P512, S513, Y515, T516, G517, Q521 , Y524, G613, A614, G705, and S709thereof include a modification.

[0269] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 18, wherein at least two of residues F348, Q350, C388, W391 , W392, P421 , Q510, A511 , P512, S513, Y515, T516, G517, Q521 , Y524, G613, A614, G705, and S709 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 18 wherein at least two of residues F348, Q350, C388, W391 , W392, P421 , Q510, A511 , P512, S513, Y515, T516, G517, Q521 , Y524, G613, A614, G705, and S709 thereof include a modification as described herein.

[0270] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F356, Q358, C396, W399, W400, P429, Q518, A519, P520, S521 , F523, T524, G525, Q529, Y532, G623, A624, G715, and S719 of SEQ ID NO: 19 (which relates to Gossypium hirsutum BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 19 wherein at least two of residues F356, Q358, C396, W399, W400, P429, Q518, A519, P520, S521 , F523, T524, G525, Q529, Y532, G623, A624, G715, and S719 thereof include a modification.

[0271] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 19, wherein at least two of residues F356, Q358, C396, W399, W400, P429, Q518, A519, P520, S521 , F523, T524, G525, Q529, Y532, G623, A624, G715, and S719 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 19 wherein at least two of residues F356, Q358, C396, W399, W400, P429, Q518, A519, P520, S521 , F523, T524, G525, Q529, Y532, G623, A624, G715, and S719 thereof include a modification as described herein.

[0272] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F331 , Q333, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, G500, Q504, Y507, G594, A595, G686, and S690 of SEQ ID NO: 20 (which relates to Glycine max BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 20 wherein at least two of residues F331 , Q333, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, G500, Q504, Y507, G594, A595, G686, and S690 thereof include a modification.

[0273] The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 20, wherein at least two of residues F331 , Q333, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, G500, Q504, Y507, G594, A595, G686, and S690 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 20 wherein at least two of residues F331 , Q333, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, G500, Q504, Y507, G594, A595, G686, and S690 thereof include a modification as described herein.

[0274] Positions, 348, 350, 388, 391 , 392, 421 , 506, 507, 508, 509, 511 , 512, 513, 517, 520, 608, 609, 700, and 704 of SEQ ID NO: 1 correspond to residues F330, Q332, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 of SEQ ID NO: 21 (which relates to Triticum aestivum BIO3-BIO1). As such, the modified BIO3-BIO1 enzyme may comprise an amino acid sequence according to SEQ ID NO: 21 wherein at least two of residues F330, Q332, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 thereof include a modification. The modified BIO3-BIO1 enzyme may comprise an amino acid sequence that has at least 70% sequence identity to an amino acid sequence according to SEQ ID NO: 21 , wherein at least two of residues F330, Q332, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 include a modification. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 21 wherein at least two of residues F330, Q332, C371 , W374, W375, P404, Q493, A494, P495, S496, Y498, T499, S500, Q504, Y507, G597, A598, G689, and T693 thereof include a modification as described herein.

[0275] Position 348 modifications and combinations thereof

[0276] In one embodiment, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at one or more of amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein position 348 is modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 thereof, or positions corresponding thereto, are modified.

[0277] Suitably the modification at position 348 is a substitution selected from F348A, F348D, F348S, F348T, F348M, F348V, F348C, or F348N with reference to SEQ ID NO 1 or the same modifications at a position corresponding thereto.

[0278] Suitably, the modification at position 350 is the substitution Q350H with reference to SEQ ID NO 1 or the same modification at a position corresponding thereto. Suitably, the modification at position 388 is the substitution C388T with reference to SEQ ID NO 1 or the same modification at a position corresponding thereto. Suitably the modification at position 391 is a substitution selected from the substitution W391 L or W391 M with reference to SEQ ID NO: 1 or the same modifications at a position corresponding thereto. Suitably, the modification at position 392 is the substitution W392S with reference to SEQ ID NO 1 or the same modifications at positions corresponding thereto. Suitably, the modification at position 506 is the substitution Q506A with reference to SEQ ID NO 1 or the same modification at positions corresponding thereto. Suitably the modification at position 507 is a substitution selected from the substitution A507S or A507K with reference to SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably the modification at position 508 is a substitution selected from the substitution P508T or P508L with reference to SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably, the modification at position 509 is the substitution S509V with reference to SEQ ID NO 1 or the same modification at positions corresponding thereto. Suitably the modification at position 511 is a substitution selected from the substitution Y511 E, Y511W or Y511M with reference to SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably, the modification at position 512 is the substitution T512C with reference to SEQ ID NO 1 or the same modification at positions corresponding thereto. Suitably, the modification at position 513 is the deletion of G513 (G513Del) with reference to SEQ ID NO 1 or the same modification at positions corresponding thereto. Suitably the modification at position 517 is a substitution selected from the substitution Q517H, Q517F, or Q517M with reference to SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably the modification at position 520 is a substitution selected from the substitution Y520W or Y520N with reference to SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably the modification at position 608 is the substitution G608A with reference to SEQ ID NO: 1 or the same modification at positions corresponding thereto. Suitably the modification at position 609 is a substitution selected from the substitution A609Y, A609T, A609M or A609V with reference to SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably the modification at position 700 is a substitution selected from the substitution G700S or G700A with reference to SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably the modification at position 704 is the substitution A704H with reference to SEQ ID NO: 1 or the same modification at positions corresponding thereto.

[0279] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 388 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 388 or positions corresponding thereto are modified and one or more of amino acid positions 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified. Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 507 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 507 or positions corresponding thereto are modified and one or more of amino acid positions 388, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 thereof or positions corresponding thereto are modified.

[0280] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 517 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 517 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0281] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 700 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 700 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 thereof or positions corresponding thereto are modified. Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 508 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein position positions 348 and 508 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0282] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 350 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 350 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0283] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 391 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 391 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified. Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 392 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 392 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0284] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 506 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 506 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 509, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0285] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 509 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 509 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 511 , 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified. Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position

[0286] 511 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 511 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 509, 512, 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0287] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position

[0288] 512 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 512 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 509, 511 , 513, 520, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0289] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position

[0290] 513 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 513 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 509, 511 , 512, 520, 608, 609, and / or 704 or positions corresponding thereto are modified. Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 520 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 520 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 509, 511 , 512, 513, 608, 609, and / or 704 or positions corresponding thereto are modified.

[0291] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position

[0292] 608 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 608 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 509, 511 , 512, 513, 520, 609, and / or 704 or positions corresponding thereto are modified.

[0293] Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position

[0294] 609 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 609 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 509, 511 , 512, 513, 520, 608, and / or 704 or positions corresponding thereto are modified. Suitably, the modified BIO3-BIO1 enzyme comprises a modification at amino acid position 348 of SEQ ID NO: 1 or a position corresponding thereto and a modification at amino acid position 704 of SEQ ID NO: 1 or a position corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme further includes one or more additional modifications at amino acid positions 388, 507, 517, 700, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, and / or 609 of SEQ ID NO: 1 or positions corresponding thereto. Other variants of such an enzyme may comprise an amino acid sequence of at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to an amino acid sequence according to SEQ ID NO: 1 wherein positions 348 and 704 or positions corresponding thereto are modified and one or more of amino acid positions 388, 507, 517, 700, 508, 350, 392, 391 , 506, 509, 511 , 512, 513, 520, 608, and / or 609 or positions corresponding thereto are modified.

[0295] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 388, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 388, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0296] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348C, C388T, A507S and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348C, C388T, A507S and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto.

[0297] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 40. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 40.

[0298] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 372, 495 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 372, 495 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0299] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333C, C372T, A495S and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333C, C372T, A495S and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0300] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 92. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 92.

[0301] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 375, 498 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 375, 498 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0302] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334C, C375T, A498S and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334C, C375T, A498S and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0303] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 144. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 144.

[0304] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 371 , 494 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 371 , 494 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0305] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332C, C371T, A494S and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332C, C371T, A494S and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0306] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 196. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 196.

[0307] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 384, 507 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 384, 507 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0308] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344C, C384T, A507S and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344C, C384T, A507S and G702A of SEQ ID NO: 9 or the same modifications at positions corresponding thereto.

[0309] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 248. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 248.

[0310] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 371 , 494 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 371 , 494 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0311] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331C, C371T, A494S and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331C, C371T, A494S and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0312] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 300. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 300.

[0313] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 388 and 517 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 388 and 517 of SEQ ID NO: 1 or positions corresponding thereto.

[0314] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348A, C388T and Q517H of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348A, C388T and Q517H of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0315] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 41. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 41. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 372 and 505 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 372 and 505 of SEQ ID NO: 2 or positions corresponding thereto.

[0316] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333A, C372T and Q505H of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333A, C372T and Q505H of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0317] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 93. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 93.

[0318] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 375 and 508 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 375 and 508 of SEQ ID NO: 4 or positions corresponding thereto.

[0319] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334A, C375T and Q508H of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334A, C375T and Q508H of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0320] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 145. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 145. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 371 and 504 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 371 and 504 of SEQ ID NO: 5 or positions corresponding thereto.

[0321] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332A, C371T and Q504H of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332A, C371T and Q504H of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0322] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 197. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 197.

[0323] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 384 and 517 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 384 and 517 of SEQ ID NO: 9 or positions corresponding thereto.

[0324] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344A, C384T and Q517H of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344A, C384T and Q517H of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0325] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 249. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 249. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 371 and 504 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 371 and 504 of SEQ ID NO: 20 or positions corresponding thereto.

[0326] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331A, C371T and Q504H of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331 A, C371T and Q504H of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0327] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 301. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 301.

[0328] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0329] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348A, A507S and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348A, A507S and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0330] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 42. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 42.

[0331] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 495 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 495 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0332] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333A, A495S and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333A, A495S and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0333] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 94. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 94.

[0334] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 498 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 498 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0335] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334A, A498S and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334A, A498S and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 146. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 146.

[0336] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 494 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 494 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0337] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332A, A494S and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332A, A494S and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0338] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 198. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 198.

[0339] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 507 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 507 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0340] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344A, A507S and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344A, A507S and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 250 Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 250.

[0341] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 494 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 494 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0342] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331A, A494S and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331 A, A494S and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0343] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 302. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 302.

[0344] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348S, A507S and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348S, A507S and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0345] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 53. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 53. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 495 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 495 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0346] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333S, A495S and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333S, A495S and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0347] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 105. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 105.

[0348] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 498 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 498 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0349] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334S, A498S and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334S, A498S and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0350] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 157. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 157. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 494 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 494 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0351] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332S, A494S and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332S, A494S and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0352] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 209. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 209.

[0353] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 507 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 507 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0354] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344S, A507S and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344S, A507S and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0355] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 261. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 261. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 494 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 494 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0356] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331S, A494S and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ I D NO: 20 and comprises the modifications F331 S, A494S and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0357] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 313. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 313.

[0358] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 507, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 507, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0359] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348A, A507S, P508T and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348A, A507S, P508T and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0360] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 43. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 43.

[0361] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 495, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 495, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0362] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333A, A495S, P496T and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333A, A495S, P496T and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0363] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 95. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 95.

[0364] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 498, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 498, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0365] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334A, A498S, P499T and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334A, A498S, P499T and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 147. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 147.

[0366] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 494, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 494, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0367] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332A, A494S, P495T and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332A, A494S, P495T and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0368] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 199. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 199.

[0369] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 507, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 507, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0370] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344A, A507S, P508T and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344A, A507S, P508T and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 251. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 251.

[0371] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 494, 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 494, 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0372] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331A, A494S, P495T and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331A, A494S, P495T and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0373] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 303. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 303.

[0374] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348M, A507S, P508T and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348M, A507S, P508T and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0375] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 49. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 49. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 495, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 495, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0376] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333M, A495S, P496T and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333M, A495S, P496T and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0377] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 101. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 101.

[0378] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 498, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 498, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0379] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334M, A498S, P499T and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334M, A498S, P499T and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0380] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 153. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 153. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 494, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 494, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0381] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332M, A494S, P495T and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332M, A494S, P495T and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0382] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 205. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 205.

[0383] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 507, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 507, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0384] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344M, A507S, P508T and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344M, A507S, P508T and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0385] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 257. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 257. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 494, 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 494, 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0386] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331 M, A494S, P495T and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331M, A494S, P495T and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0387] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 309. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 309.

[0388] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0389] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348A, P508T and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348A, P508T and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0390] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 44. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 44.

[0391] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0392] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333A, P496T and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333A, P496T and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0393] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 96. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 96.

[0394] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0395] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334A, P499T and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334A, P499T and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 148. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 148.

[0396] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0397] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332A, P495T and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332A, P495T and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0398] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 200. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 200.

[0399] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0400] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344A, P508T and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344A, P508T and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 252. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 252.

[0401] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0402] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331A, P495T and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331 A, P495T and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0403] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 304. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 304.

[0404] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348T, P508L and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348T, P508L and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0405] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 57. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 57. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 496 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0406] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333T, P496L and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333T, P496L and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0407] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 109. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 109.

[0408] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 499 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0409] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334T, P499L and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334T, P499L and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0410] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 161. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 161. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 495 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0411] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332T, P495L and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332T, P495L and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0412] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 213. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 213.

[0413] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 508 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0414] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344T, P508L and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344T, P508L and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0415] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 265. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 265. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 495 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0416] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331T, P495L and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331T, P495L and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0417] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 317. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 317.

[0418] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348 and 700 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0419] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348C and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348C and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0420] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 45. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 45.

[0421] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0422] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333C and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333C and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0423] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 97. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 97.

[0424] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0425] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334C and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334C and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 149. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 149.

[0426] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0427] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332C and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332C and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0428] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 201. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 201.

[0429] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0430] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344C and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344C and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 253. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 253.

[0431] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0432] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331C and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331C and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0433] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 305. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 305.

[0434] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348 and 700 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 348 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0435] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348M and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348M and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 50 Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 50.

[0436] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0437] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333M and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333M and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0438] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 102. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 102.

[0439] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0440] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334M and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334M and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 154. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 154.

[0441] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0442] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332M and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332M and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0443] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 206. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 206.

[0444] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0445] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344M and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344M and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 258. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 258.

[0446] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0447] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331M and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331M and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0448] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 310. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 310.

[0449] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348T and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348T and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0450] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 56. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 56. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0451] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333T and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333T and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0452] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 108. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 108.

[0453] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0454] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334T and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334T and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0455] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 160. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 160. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0456] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332T and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332T and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0457] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 212. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 212.

[0458] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0459] Suitably, the modified BIO3-BIO1 enzyme comprises the F344T and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3- BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344T and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0460] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 264. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 264. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0461] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331T and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331T and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0462] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 316. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 316.

[0463] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348V and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348V and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0464] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 60 Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 60.

[0465] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333V and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333V and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0466] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 112. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 112.

[0467] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0468] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334V and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334V and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0469] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 164. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 164.

[0470] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332V and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332V and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0471] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 216. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 216.

[0472] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0473] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344V and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344V and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0474] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 268. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 268.

[0475] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331V and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331V and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0476] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 320. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 320.

[0477] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348D and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348D and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0478] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 64. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 64.

[0479] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0480] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333D and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333D and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 116. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 116.

[0481] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0482] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334D and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334D and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0483] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 168. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 168.

[0484] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0485] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332D and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332D and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 220. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 220.

[0486] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0487] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344D and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344D and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0488] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 272. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 272.

[0489] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0490] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331 D and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331 D and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 324. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 324.

[0491] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348N and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348N and G700A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0492] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 73. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 73.

[0493] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0494] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333N and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333N and G687A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0495] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 125. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 125. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0496] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334N and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334N and G693A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0497] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 177. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 177.

[0498] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0499] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332N and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332N and G689A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0500] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 229. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 229. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0501] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344N and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344N and G702A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0502] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 281. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 281.

[0503] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0504] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331 N and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331 N and G686A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0505] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 333. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 333. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348T and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348T and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0506] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 78. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 78.

[0507] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0508] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333T and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333T and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0509] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 130 Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 130.

[0510] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334T and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334T and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0511] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 182. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 182.

[0512] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0513] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332T and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332T and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0514] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 234. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 234.

[0515] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344T and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344T and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0516] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 286. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 286.

[0517] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0518] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331T and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331T and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0519] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 338. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 338.

[0520] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 508 and 513 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 508 and 513 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348C, P508T and G513Del of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348C, P508T and G513Del of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0521] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 46. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 46.

[0522] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 496 and 501 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 496 and 501 of SEQ ID NO: 2 or positions corresponding thereto.

[0523] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333C, P496T and S501 DEL of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333C, P496T and S501 DEL of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0524] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 98. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 98.

[0525] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 499 and 504 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 499 and 504 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334C, P499T and S504DEL of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334C, P499T and S504DEL of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0526] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 150. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 150.

[0527] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 495 and 500 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 495 and 500 of SEQ ID NO: 5 or positions corresponding thereto.

[0528] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332C, P495T and S500DEL of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332C, P495T and S500DEL of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0529] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 202. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 202.

[0530] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 508 and 513 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 508 and 513 of SEQ ID NO: 9 or positions corresponding thereto.

[0531] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344C, P508T and G513DEL of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344C, P508T and G513DEL of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0532] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 254. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 254.

[0533] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 495 and 500 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 495 and 500 of SEQ ID NO: 20 or positions corresponding thereto.

[0534] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331C, P495T and G500DEL of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331C, P495T and G500DEL of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0535] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 306. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 306.

[0536] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 509, 517, 608 and 700 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 509, 517, 608 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

[0537] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348C, S509V, Q517F, G608A and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348C, S509V, Q517F, G608A and G700S of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0538] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 47. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 47.

[0539] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 497, 505, 595 and 687 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 497, 505, 595 and 687 of SEQ ID NO: 2 or positions corresponding thereto.

[0540] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333C, S497V, Q505F, G595A and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333C, S497V, Q505F, G595A and G687S of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0541] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 99. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 99. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 500, 508, 601 and 693 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 500, 508, 601 and 693 of SEQ ID NO: 4 or positions corresponding thereto.

[0542] Suitably, the modified BIO3-BIO1 enzyme comprises the F334C, S500V, Q508F, G601A and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334C, S500V, Q508F, G601A and G693S of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0543] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 151. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 151.

[0544] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 496, 504, 597 and 689 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 496, 504, 597 and 689 of SEQ ID NO: 5 or positions corresponding thereto.

[0545] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332C, S496V, Q504F, G597A and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332C, S496V, Q504F, G597A and G689S of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0546] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 203. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 203.

[0547] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 509, 517, 610 and 702 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 509, 517, 610 and 702 of SEQ ID NO: 9 or positions corresponding thereto.

[0548] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344C, S509V, Q517F, G610A and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344C, S509V, Q517F, G610A and G702S of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0549] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 255. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 255.

[0550] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 496, 504, 594 and 686 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 496, 504, 594 and 686 of SEQ ID NO: 20 or positions corresponding thereto.

[0551] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331C, S496V, Q504F, G594A and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331C, S496V, Q504F, G594A and G686S of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 307. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 307.

[0552] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 507 and 609 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid positions 348, 507 and 609 of SEQ ID NO: 1 or positions corresponding thereto.

[0553] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348M, A507S and A609T of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348M, A507S and A609T of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0554] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 48. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 48.

[0555] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 495 and 596 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 495 and 596 of SEQ ID NO: 2 or positions corresponding thereto.

[0556] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333M, A495S and A596T of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333M, A495S and A596T of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 100 Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 100.

[0557] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 498 and 602 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 498 and 602 of SEQ ID NO: 4 or positions corresponding thereto.

[0558] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334M, A498S and A602T of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334M, A498S and A602T of SEQ ID NO:

[0559] 4 or the same modifications at residues corresponding thereto.

[0560] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 152. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 152.

[0561] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 494 and 598 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 494 and 598 of SEQ ID NO: 5 or positions corresponding thereto.

[0562] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332M, A494S and A598T of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332M, A494S and A598T of SEQ ID NO:

[0563] 5 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 204. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 204.

[0564] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 507 and 611 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 507 and 611 of SEQ ID NO: 9 or positions corresponding thereto.

[0565] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344M, A507S and A611T of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344M, A507S and A611T of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0566] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 256. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 256.

[0567] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 494 and 595 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 494 and 595 of SEQ ID NO: 20 or positions corresponding thereto.

[0568] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331 M, A494S and A595T of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ I D NO: 20 and comprises the modifications F331 M , A494S and A595T of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 308. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 308.

[0569] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 350, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid 348, 350, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto.

[0570] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348M, Q350H, A507S and Y520W of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348M, Q350H, A507S and Y520W of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0571] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 51. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 51.

[0572] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 335, 495 and 508 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 335, 495 and 508 of SEQ ID NO: 2 or positions corresponding thereto.

[0573] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333M, Q335H, A495S and Y508W of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333M, Q335H, A495S and Y508W of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0574] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 103. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 103.

[0575] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 336, 498 and 511 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 336, 498 and 511 of SEQ ID NO: 4 or positions corresponding thereto.

[0576] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334M, Q336H, A498S and Y511W of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334M, Q336H, A498S and Y511W of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0577] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 155. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 155.

[0578] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 334, 494 and 507 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 334, 494 and 507 of SEQ ID NO: 5 or positions corresponding thereto.

[0579] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332M, Q334H, A494S and Y507W of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332M, Q334H, A494S and Y507W of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0580] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 207. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 207.

[0581] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 346, 507 and 520 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 346, 507 and 520 of SEQ ID NO: 9 or positions corresponding thereto.

[0582] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344M, Q346H, A507S and Y520W of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344M, Q346H, A507S and Y520W of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0583] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 259. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 259.

[0584] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 333, 494 and 507 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 333, 494 and 507 of SEQ ID NO: 20 or positions corresponding thereto.

[0585] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331M, Q333H, A494S and Y507W of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331M, Q333H, A494S and Y507W of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0586] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 311. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 311.

[0587] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348 and 508 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid 348 and 508 of SEQ ID NO: 1 or positions corresponding thereto.

[0588] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348N and P508L of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348N and P508L of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0589] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 52. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 52.

[0590] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 496 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 496 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333N and P496L of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333N and P496L of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0591] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 104. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 104.

[0592] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 499 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 499 of SEQ ID NO: 4 or positions corresponding thereto.

[0593] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334N and P499L of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334N and P499L of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0594] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 156. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 156.

[0595] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 495 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 495 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332N and P495L of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332N and P495L of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0596] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 208. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 208.

[0597] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 508 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 508 of SEQ ID NO: 9 or positions corresponding thereto.

[0598] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344N and P508L of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344N and P508L of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0599] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 260. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 260.

[0600] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 495 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 495 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331 N and P495L of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331 N and P495L of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0601] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 312. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 312.

[0602] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348, 388 and 508 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid 348, 388 and 508 of SEQ ID NO: 1 or positions corresponding thereto.

[0603] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348S, C388T and P508T of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348S, C388T and P508T of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0604] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 54. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 54.

[0605] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333, 372 and 496 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333, 372 and 496 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333S, C372T and P496T of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333S, C372T and P496T of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0606] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 106. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 106.

[0607] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334, 375 and 499 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334, 375 and 499 of SEQ ID NO: 4 or positions corresponding thereto.

[0608] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334S, C375T and P499T of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334S, C375T and P499T of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0609] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 158. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 158.

[0610] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332, 371 and 495 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332, 371 and 495 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332S, C371T and P495T of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332S, C371T and P495T of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0611] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 210. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 210.

[0612] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344, 384 and 508 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344, 384 and 508 of SEQ ID NO: 9 or positions corresponding thereto.

[0613] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344S, C384T and P508T of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344S, C384T and P508T of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0614] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 262. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 262.

[0615] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 , 371 and 495 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 , 371 and 495 of SEQ ID NO: 20 or positions corresponding thereto.

[0616] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331S, C371T and P495T of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331S, C371T and P495T of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0617] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 314. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 314.

[0618] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 348 and 506 of SEQ ID NO: 1 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises modifications at amino acid 348 and 506 of SEQ ID NO: 1 or positions corresponding thereto.

[0619] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348S and Q506A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348S and Q506A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0620] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 55. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 55.

[0621] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 494 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 494 of SEQ ID NO: 2 or positions corresponding thereto.

[0622] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333S and Q494A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333S and Q494A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0623] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 107. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 107.

[0624] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 334 and 497 of SEQ ID NO: 4 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises modifications at amino acid positions 334 and 497 of SEQ ID NO: 4 or positions corresponding thereto.

[0625] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F334S and Q497A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 4 and comprises the modifications F334S and Q497A of SEQ ID NO: 4 or the same modifications at residues corresponding thereto.

[0626] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 159. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 159.

[0627] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 332 and 493 of SEQ ID NO: 5 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises modifications at amino acid positions 332 and 493 of SEQ ID NO: 5 or positions corresponding thereto.

[0628] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F332S and Q493A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 5 and comprises the modifications F332S and Q493A of SEQ ID NO: 5 or the same modifications at residues corresponding thereto.

[0629] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 211. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 211.

[0630] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 344 and 506 of SEQ ID NO: 9 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises modifications at amino acid positions 344 and 506 of SEQ ID NO: 9 or positions corresponding thereto.

[0631] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F344S and Q506A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 9 and comprises the modifications F344S and Q506A of SEQ ID NO: 9 or the same modifications at residues corresponding thereto.

[0632] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 263. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 263. In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 331 and 493 of SEQ ID NO: 20 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises modifications at amino acid positions 331 and 493 of SEQ ID NO: 20 or positions corresponding thereto.

[0633] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F331S and Q493A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 20 and comprises the modifications F331S and Q493A of SEQ ID NO: 20 or the same modifications at residues corresponding thereto.

[0634] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 315. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 315.

[0635] Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F348T and Q506A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 1 - 39 and comprises the modifications F348T and Q506A of SEQ ID NO: 1 or the same modifications at residues corresponding thereto.

[0636] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 59. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence comprising or consisting of SEQ ID NO: 59.

[0637] In one embodiment, the modified BIO3-BIO1 enzyme comprises modifications at amino acid positions 333 and 494 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises modifications at amino acid positions 333 and 494 of SEQ ID NO: 2 or positions corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises the modifications F333T and Q494A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto. Suitably, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2 and comprises the modifications F333T and Q494A of SEQ ID NO: 2 or the same modifications at residues corresponding thereto.

[0638] For example, the modified BIO3-BIO1 enzyme comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%...

Claims

CLAIMS1. A method of controlling undesired vegetation in the vicinity of a modified plant or part thereof, or at the locus for growth of a modified plant, the method comprising applying an effective amount of at least one compound which inhibits the biotin synthesis pathway to the undesired vegetation and the plant, or applying an effective amount of at least one compound which inhibits the biotin synthesis pathway to the locus and planting a seed at said locus, wherein the seed is capable of producing the modified plant, wherein the modified plant comprises at least one herbicide resistant modified BIO3- BIO1 enzyme or functional fragment thereof; at least one isolated nucleic acid comprising a polynucleotide encoding at least one herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof; an expression construct comprising a polynucleotide encoding at least one modified herbicide resistant BIO3-BIO1 enzyme or functional fragment thereof operably linked to one or more expression elements or; a recombinant vector comprising the isolated nucleic acid or the expression construct and wherein the herbicide resistant modified BIO3-BIO1 enzyme or a functional fragment thereof comprises at least two modifications at amino acid positions selected from one of the following combinations: a) amino acid position 348 and one or more of amino acid positions 700, 388, 507, 517, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto; b) amino acid position 391 and one or more of amino acid positions 388, 421 , 348, 506, 507, 508, 517 and / or 520 of SEQ ID NO: 1 or positions corresponding thereto; c) amino acid position 508 and one or more of amino acid positions 700, 348, 388, 391 , 392, 506, 507, 511 , 513, 517, 520, 609 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto; d) amino acid position 609 and one or more of amino acid positions 348, 512, 507, 508, 511 , 520, 700 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto.

2. A herbicide resistant modified BIO3-BIO1 enzyme or a functional fragment thereof comprising at least two modifications at amino acid positions selected from one of the following combinations: e) amino acid position 348 and one or more of amino acid positions 700, 388, 507, 517, 508, 350, 391 , 392, 506, 509, 511 , 512, 513, 520, 608, 609, and / or 704 of SEQ ID NO: 1 or positions corresponding thereto; f) amino acid position 391 and one or more of amino acid positions 388, 421 , 348, 506, 507, 508, 517 and / or 520 of SEQ ID NO: 1 or positions corresponding thereto; g) amino acid position 508 and one or more of amino acid positions 700, 348, 388, 391, 392, 506, 507, 511 , 513, 517, 520, 609 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto; h) amino acid position 609 and one or more of amino acid positions 348, 512, 507, 508, 511 , 520, 700 and / or 704 of SEQ ID NO: 1 or positions corresponding thereto.

3. The herbicide resistant modified BIO3-BIO1 enzyme or the functional fragment thereof of claim 2, wherein: a. amino acid position 348, or a position corresponding thereto, is substituted with an A, D, S, T, M, V, C, or N amino acid residue; b. amino acid position 350, or a position corresponding thereto, is substituted with an H amino acid residue; c. amino acid position 388, or a position corresponding thereto, is substituted with a T amino acid residue; d. amino acid position 391 , or a position corresponding thereto, is substituted with a L or M amino acid residue; e. amino acid position 392, or a position corresponding thereto, is substituted with a S amino acid residue; f. amino acid position 421 , or a position corresponding thereto, is substituted with a L amino acid residue;g. amino acid position 506, or a position corresponding thereto, is substituted with an A amino acid residue; h. amino acid position 507, or a position corresponding thereto, is substituted with a S or K amino acid residue; i. amino acid position 508, or a position corresponding thereto, is substituted with a T or L amino acid residue; j. amino acid position 509, or a position corresponding thereto, is substituted with a V amino acid residue; k. amino acid position 511 , or a position corresponding thereto, is substituted with an E, W or M amino acid residue; l. amino acid position 512, or a position corresponding thereto, is substituted with a C amino acid residue; m. amino acid position 513, or a position corresponding thereto, is deleted (Del); n. amino acid position 517, or a position corresponding thereto, is substituted with an H, F or T amino acid residue; o. amino acid position 520, or a position corresponding thereto, is substituted with a W or N amino acid residue; p. amino acid position 608, or a position corresponding thereto, is substituted with an A amino acid residue; q. amino acid position 609, or a position corresponding thereto, is substituted with a Y, T, M or V amino acid residue; r. amino acid position 700, or a position corresponding thereto, is substituted with a S or A amino acid residue; and / or s. amino acid position 704, or a position corresponding thereto, is substituted with an H amino acid residue.

4. The herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof of claim 2 or 3, comprising modifications at amino acid positions selected from one of the following combinations: a. Positions 348 and 700 of SEQ ID NO: 1 or positions corresponding thereto; b. Positions 348, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto;c. Positions 348, 507, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto; d. Positions 348, 509, 517, 608 and 700 of SEQ ID NO: 1 or positions corresponding thereto; e. Positions 348, 388, 507 and 700 of SEQ ID NO: 1 or positions corresponding thereto; f. Positions 348, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto; g. 348, 507, 508, 700 and 609 of SEQ ID NO: 1 or positions corresponding thereto; h. Positions 348, 507, 508, 700 and 392 of SEQ ID NO: 1 or positions corresponding thereto; i. Positions 348, 388 and 517 of SEQ ID NO: 1 or positions corresponding thereto; j. Positions 388, 391 and 421 of SEQ ID NO: 1 or positions corresponding thereto; k. Positions 348, 350, 506 and 507 of SEQ ID NO: 1 or positions corresponding thereto; l. Positions 348 and 508 of SEQ ID NO: 1 or positions corresponding thereto; m. Positions 348, 388 and 508 of SEQ ID NO: 1 or positions corresponding thereto; n. Positions 348 and 506 of SEQ ID NO: 1 or positions corresponding thereto; o. Positions 348, 508 and 513 of SEQ ID NO: 1 or positions corresponding thereto; p. Positions 348, 507 and 609 of SEQ ID NO: 1 or positions corresponding thereto; q. Positions 348, 350, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto; r. Positions 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto; s. Positions 512 and 609 of SEQ ID NO: 1 or positions corresponding thereto;t. Positions 348, 507 and 508 of SEQ ID NO: 1 or positions corresponding thereto; u. Positions 348, 506, 508 and 517 of SEQ ID NO: 1 or positions corresponding thereto; v. Positions 348, 511 , 512, 609 and 704 of, SEQ ID NO: 1 or positions corresponding thereto; w. Positions 348, 350, 506 and 507 of SEQ ID NO: 1 or positions corresponding thereto; x. Positions 348, 388, 508, 511 and 520 of SEQ ID NO: 1 or positions corresponding thereto; y. Positions 391 and 507 of SEQ ID NO: 1 or positions corresponding thereto; z. Positions 391 and 517 of SEQ ID NO: 1 or positions corresponding thereto; aa. Positions 391 , 506, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto; bb. Positions 348, 506 and 512 of SEQ ID NO: 1 or positions corresponding thereto; cc. Positions 348, 506, 507 and 520 of SEQ ID NO: 1 or positions corresponding thereto; dd. Positions 348, 508 and 511 of SEQ ID NO: 1 or positions corresponding thereto; ee. Positions 348 and 512 of SEQ ID NO: 1 or positions corresponding thereto; ff. Positions 348 and 391 of SEQ ID NO: 1 or positions corresponding thereto; gg. Positions 348 and 507 of SEQ ID NO: 1 or positions corresponding thereto; hh. Positions 348, 350 and 512 of SEQ ID NO: 1 or positions corresponding thereto; ii. Positions 348 and 509 of SEQ ID NO: 1 or positions corresponding thereto; jj. Positions 520 and 609 of SEQ ID NO: 1 or positions corresponding thereto; kk. Positions 348, 391 and 506 of SEQ ID NO: 1 or positions corresponding thereto;II. Positions 348, 517 and 520 of SEQ ID NO: 1 or positions corresponding thereto;mm. Positions 391 and 508 of SEQ ID NO: 1 or positions corresponding thereto; nn. Positions 348 and 520 of SEQ ID NO: 1 or positions corresponding thereto; or oo. Positions 507, 508 and 700 of SEQ ID NO: 1 or positions corresponding thereto.

5. The herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof of claim 2 or 3, comprising modifications selected from any one of the following combinations: a. F348A, A507S and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; b. F348A, A507S, P508T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; c. F348A, P508T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; d. F348C and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; e. F348C, S509V, Q517F, G608A and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; f. F348C, C388T, A507S and G700A of SEQ I D NO: 1 or the same modifications at positions corresponding thereto; g. F348M, A507S, P508T and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; h. F348M and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; i. F348V and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; j. F348S, A507S and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; k. F348T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;l. F348T, P508L and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; m. F348D and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; n. F348N and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; o. F348T and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; p. F348A, A507S, P508T, G700A and S704H of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; q. F348A, A507S, P508T, G700A and W392S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; r. F348A, A507S, P508T, G700A and A609V of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; s. F348A, C388T and Q517H of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; t. C388T, W391L and P421L of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; u. F348C, P508T and G513Del of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; v. F348M, A507S and A609T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; w. F348N and P508L of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; x. F348S, C388T and P508T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; y. F348S and Q506A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; z. F348T, Q350H, Q506A and A507S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; aa. F348T and Q506A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;bb. P508T and G700S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; cc. T512C and A609M of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; dd. F348M, Q350H, A507S and Y520Wof SEQ ID NO: 1 or the same modifications at positions corresponding thereto; ee. F348D, Q506A, P508L and Q517T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; ff. F348N, Y511 E, T512C, A609Y and S704H of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; gg. F348D, C388T, P508T, Y511W and Y520W of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; hh. W391 L and A507S of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; ii. W391 M and Q517F of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; jj. W391 L, Q506A, A507S and Y520Wof SEQ ID NO: 1 or the same modifications at positions corresponding thereto; kk. F348S, Q506A and T512C of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;II. F348D, Q506A, A507S and Y520Wof SEQ ID NO: 1 or the same modifications at positions corresponding thereto; mm. F348T, P508T and Y511M of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; nn. F348S and T512C of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; oo. F348M and W391 L of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; pp. F348T and A507K of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; qq. F348A, Q350H and T512C of SEQ ID NO: 1 or the same modifications at positions corresponding thereto;rr. F348S and S509V of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; ss. Y520W and A609V of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; tt. W391 L and Q517T of SEQ I D NO: of SEQ I D NO: 1 or the same modifications at positions corresponding thereto; uu. F348M, W391 L and Q506A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; vv. F348M, Q517F and Y520W of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; ww. W391 L and P508T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; xx. F348A and Y520N of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; yy. F348A, A507S and P508T of SEQ ID NO: 1 or the same modifications at positions corresponding thereto; or zz. A507S, P508T and G700A of SEQ ID NO: 1 or the same modifications at positions corresponding thereto.

6. The herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof of any of claims 2 to 5, wherein the herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof comprises an amino acid sequence having at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 98%, at least 99% or 100% identity to any of SEQ ID NO: 40 to 663.

7. The herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof of any of claims 2 to 6, wherein the herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof, comprises an increased resistance to a compound that inhibits a BIO3-BIO1 enzyme relative to a control BIO3-BIO1 enzyme and / or has decreased percentage inhibition in response to a compound that inhibits a BIO3-BIO1 enzyme relative to a control BIO3-BIO1 enzyme.

8. The herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof of any of claims 2 to 7, wherein the herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof is modified to comprise a heterologous targeting peptide, preferably a heterologous mitochondrial targeting peptide.

9. The herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof of any of claims 2 to 8, wherein the functional fragment comprises at least DAPA synthase activity; optionally wherein the functional fragment comprises the BIO1 portion of the BIO3-BIO1 enzyme; further optionally wherein the functional fragment comprises amino acid residues 322 to 833 of SEQ ID NO: 1 or residues corresponding thereto.

10. An isolated nucleic acid comprising a polynucleotide encoding at least one herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any of claims 2 to 9.

11. An expression construct comprising a polynucleotide encoding at least one herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof of any of claims 2 to 9 and / or a polynucleotide of claim 10, operably linked to one or more expression elements.

12. A recombinant vector comprising the isolated nucleic acid of claim 10 or the expression construct of claim 11.

13. The method of controlling undesired vegetation of claim 1 , wherein the at least one herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof comprises at least one herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any one of claims 2 to 9.

14. A modified plant, or part thereof, comprising at least one herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any one of claims 2 to 9, an isolated nucleic acid according to claim 10, an expression construct according to claim 11 , or a recombinant vector according to claim 12.

15. The modified plant, or part thereof, according to claim 14, wherein the herbicide resistant modified BIO3-BIO1 enzyme provides the plant or part thereof with increasedresistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

16. The modified plant, or part thereof, according to claim 14 or claim 15, wherein the herbicide resistant modified BIO3-BIO1 enzyme is derived from a plant, from an algae, from an oomycete, or a from a fungus, preferably wherein the herbicide resistant modified BIO3-BIO1 enzyme is derived from a plant, more preferably the herbicide resistant modified BIO3-BIO1 enzyme is derived from any of the following plant species: Setaria italica, Arabidopsis thaliana, Helianthus annuus, Quercus robur, Phoenix dactylifera, Physcomitrium patens, Taxus chinensis, Adiantum nelumboides, Zea mays, Ostreococcus tauri, Hordeum vulgare, Brassica napus, Gossypium hirsutum, Oryza sativa, Glycine max, and Triticum aestivum.

17. The modified plant, or part thereof, according to any of claims 14 to 16, wherein the herbicide resistant modified BIO3-BIO1 enzyme is derived from a BIO3-BIO1 enzyme heterologous to the plant or part thereof.

18. The modified plant, or part thereof, according to any of claims 14 to 17, wherein the herbicide resistant modified BIO3-BIO1 enzyme comprises or consists of a sequence having at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 98% or at least 99% identity to any of SEQ ID NO: 40 to 663.

19. The method of controlling undesired vegetation of claim 1 , wherein the modified plant comprises a modified plant according to any one of claims 14 to 18.

20. A method of producing a modified plant or part thereof according to any of claims 14- 18, the method comprising: modifying the plant or part thereof to comprise a herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any one of claims 2 to 9 which provides the plant with increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

21. The method according to claim 20, wherein modifying the plant or part thereof comprises transforming the plant or part thereof with an isolated nucleic acid according to claim 10, an expression construct according to claim 11 , or a vector according toclaim 12, or gene editing the endogenous Bio3-Bio1 enzyme encoding gene of the plant or part thereof to encode a herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any one of claims 2 to 9, the expression of which provides increased resistance to a compound which inhibits the biotin synthesis pathway relative to a reference plant.

22. Use of a compound which inhibits the biotin synthesis pathway in combination with a modified plant or part thereof according to any of claims 14 to 18.

23. A method of controlling undesired vegetation according to any one of claims 1 , 13, or 19, a herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any of claims 7 to 9, a modified plant, or part thereof according to any of claims 14-18, a method of producing a modified plant or part thereof according to claims 20 or 21 , and a use according to claim 22, wherein the compound which inhibits the biotin synthesis pathway inhibits one or more of the enzymes in the biotin synthesis pathway or related pathways, preferably wherein the compound inhibits one or more of pimeloyl CoA synthetase, KAPA synthase, DAPA synthase, DTB synthetase, and biotin synthase.

24. A method of controlling undesired vegetation according to any one of claims 1 , 13, 19, and 23, a herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any of claims 7 to 9 and 23, a modified plant, or part thereof according to any of claims 14-18 and 23, a method of producing a modified plant or part thereof according to any one of claims 20, 21 , and 23 and a use according to claim 22 or 23, wherein the compound is a herbicide, preferably wherein the compound is a herbicide selected from: a herbicidal cinnoline compound such as ethyl 5-acetyl-4-oxo-1-(2,2,3,3-tetrafluoro-1 ,4-benzodioxin-6-yl)cinnoline-3-carboxylate (Compound A); a herbicidal compound such as [5-carboxy-1-(1-hydroxyethyl)pentyl]ammonium;chloride (compound B); a herbicidal pyridone compound such as 2-(3,4-dichlorophenyl)-5-(2,4- difluorophenyl)-1-ethyl-6-methyl-4-oxo-pyridine-3-carboxylic acid (Compound C) or 5- [2-chloro-5-(trifluoromethoxy)phenyl]-2-(3,4-dichlorophenyl)-1-ethyl-6-methyl-4-oxo- pyridine-3-carboxylic acid (Compound D); a herbicidal compound such as [(1 R)-2-(6- carboxypyridin-1-ium-2-yl)oxy-1-methyl-ethyl]ammonium;dichloride (Compound E); a herbicidal pyrrolidine-2-one such as 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]-N-(2-methyl-1 ,2,4-triazol-3-yl)acetamide (Compound F), 2-[1-[(2,3- difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]acetic acid (Compound G), 2-[5-oxo-1- [(2,3,5-trifluorophenyl)methyl]pyrrolidin-2-yl]acetic acid (Compound H), 2-(4-fluorophenoxy)ethyl 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]acetate (Compound I), 2-methoxyethyl 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2- yl]acetate (Compound J), 2-[5-oxo-1-[(2,3,4-trifluorophenyl)methyl]pyrrolidin-2- yl]acetic acid (Compound K); a N-benzyl azole herbicidal compound such as 2-(3,4- dichlorophenyl)-1-ethyl-4-oxo-6-[[3-(trifluoromethyl)pyrazol-1-yl]methyl]pyridine-3- carboxylic acid (Compound L); and a herbicidal quinolone compound such as 2-(3,4- dichlorophenyl)-1-ethyl-6-fluoro-4-oxo-quinoline-3-carboxylic acid (Compound M).

25. A method of controlling undesired vegetation according to any one of claims 1 , 13, 19, 23 and 24, a herbicide resistant modified BIO3-BIO1 enzyme or functional fragment thereof according to any of claims 7 to 9, 23 and 24, a modified plant, or part thereof according to any of claims 14-18, 23 and 24, a method of producing a modified plant or part thereof according to any one of claims 20, 21 , 23 and 24 and a use according to any one of claims 22to 24,, wherein increased resistance to the compound comprises an increase of at least 2-fold, 3-fold, 4-fold, 5-fold 6-fold, 7-fold, 8-fold, 9- fold, 10-fold in resistance to the compound compared to a reference plant or part thereof.

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