Methods for identifying herbicidal compounds

Abstract

The present invention relates to an in vitro method for determining whether a candidate compound is a diaminononanoic acid aminotransferase (DAPA-AT) inhibitor, said method comprising (i) determining the candidate compound in an assay mixture comprising (a) the DAPA-AT, (b) an amine donor, (c) pyridoxal 5 '-phosphate (PLP) and (d) 7-keto-8-aminononanoic acid (KAPA), and (ii) detecting the inhibition of the candidate compound on the DAPA-AT; characterized in that the amine donor comprises an aliphatic diamine or an aliphatic polyamine.

Description

[0001] Biotin (also known as vitamin B7) is an essential cofactor for enzymes involved in cellular processes, including the metabolism of fats, proteins, or carbohydrates. In contrast to animals, which obtain biotin from dietary sources or intestinal bacteria, plants and most fungi / bacteria are able to synthesize biotin. Therefore, inhibiting enzymes along the biotin synthesis pathway in plants offers attractive herbicidal targets.

[0002] In bacteria, biotin is synthesized from pimeloyl-CoA and alanine through the activities of four enzymes (BioF, BioA, BioD, and BioB), encoded by genes located in an operon. BioF (also known as 7-keto-8-aminononanoic acid (KAPA) synthase (EC 2.3.1.47)) catalyzes the conversion of pimeloyl-CoA and L-alanine to KAPA. BioA (also known as 7,8-diaminononanoic acid aminotransferase (DAPA-AT) or DAPA synthase (EC.2.6.1.62)) then proceeds to the next step, converting KAPA to 7,8-diaminononanoic acid (DAPA). Subsequently, BioD (also known as dethiobiotin synthase (DTBS) (EC 6.3.3.3)) converts DAPA to dethiobiotin (DTB), which is in turn converted to biotin through the activity of BioB (also known as biotin synthase (EC 2.8.1.6)).

[0003] In plants, DAPA-AT is represented by the BioA ortholog BIO1, and DTBS is represented by the BioD ortholog BIO3. The genes encoding BIO1 and BIO3 are present at a single genetic locus in the plant genome, and in principle, through alternative splicing events, a single transcript and a chimeric BIO3-BIO1 transcript can be generated, potentially allowing the generation of: a bicistronic transcript capable of producing separate BIO3 and BIO1 proteins; or a monocistronic transcript producing the bifunctional fusion protein BIO3-BIO1 (also known as BioDA (dethiobiotin synthetase / 7,8-diaminononanoic acid aminotransferase)). However, it should be understood that the BIO3-BIO1 fusion protein is the major, if not the only, protein produced by the BIO3-BIO1 locus in plants. Therefore, BIO3-BIO1 is a single, bifunctional enzyme that catalyzes two independent steps in biotin biosynthesis. Therefore, inhibition of the BIO3-BIO1 enzyme, in particular the BIO1 catalytic domain, represents an attractive herbicidal target.The present invention therefore relates to methods for identifying DAPA-AT inhibitors.

[0004] DAPA-AT is a PLP (pyridoxal 5'-phosphate) dependent enzyme, and several biochemical assays for examining DAPA-AT activity have been developed and reported. Many of these assays were developed using the bacterial BioA enzyme, but Cobessi et al. (2012) The Plant Cell, Vol. 24: 1608-1625 describe the development of an assay involving the Arabidopsis BIO3-BIO1 enzyme. However, a common feature of all of these assays reported to date is that they all employ S-adenosyl-L-methionine (SAM) as the amino group donor in the assay, which is converted to S-adenosyl-4-methylthio-2-oxobutyrate (Scheme 1). Option 1: DAPA-AT (DAPA synthase) activity with SAM as amine donor.

[0005]

[0006] The present invention is based on the development of a new assay that utilizes alternative amine donors.

[0007] Thus, according to the present invention, there is provided an in vitro method for determining whether a candidate compound is an inhibitor of diaminononanoic acid aminotransferase (DAPA-AT), the method comprising (i) assaying the candidate compound in an assay mixture comprising (a) the DAPA-AT, (b) an amine donor, (c) pyridoxal 5'-phosphate (PLP) and (d) 7-keto-8-aminononanoic acid (KAPA), and (ii) detecting inhibition of the DAPA-AT by the candidate compound; characterised in that the amine donor comprises an aliphatic diamine or an aliphatic polyamine or a carbamate derivative thereof.

[0008] Aliphatic diamines are low molecular weight aliphatic compounds containing two amino (NH2) groups, such as cadaverine or putrescine. Aliphatic polyamines are low molecular weight aliphatic compounds containing more than two amino (NH2) groups, such as spermidine or spermine. Therefore, in a preferred embodiment of the present invention, the amine donor is a compound having formula (I):

[0009]

[0010] wherein X is NH or CH2;

[0011] R 1 is hydrogen or -[CH2]pNH2;

[0012] m and n are independently selected from 0, 1, 2, 3 or 4; and

[0013] p = 1, 2, 3 or 4,

[0014] or its carbamate derivatives, wherein R 1 It is -C(O)OH.

[0015] In another embodiment of the present invention, the amine donor is selected from the group consisting of: cadaverine (X = CH2, R 1 =H, m=2, n=2), putrescine (X=CH2, R 1 =H, m=2, n=1), spermidine (X=NH, R 1 =H, m=3, n=4) and spermine (X=NH, R 1 =-[CH2] p NH2, m=3, n=4, p=3). In a particularly preferred embodiment of the present invention, the amine donor is spermidine.

[0016] It has been found that the use of aliphatic diamine or polyamine donors (rather than SAM), especially spermidine, can provide significantly improved assays, especially for assays utilizing plant BIO1 or BIO3-BIO1 enzymes (see Figure 1 Without being bound by any particular theory, it is believed that when spermidine is utilized in an assay, it is converted to 4-(3-aminopropylamino)butanal (or 3-(4-aminobutylamino)propanal) (Scheme 2).

[0017] Option 2: DAPA-AT (DAPA synthase) activity with spermidine as amine donor.

[0018]

[0019] Therefore, providing new functional assays would be particularly useful for methods of identifying DAPA-AT (especially plant BIO1 or BIO3-BIO1) inhibitors, which could aid in the discovery and development of herbicidal compounds.

[0020] The candidate compound can be any compound that is a putative DAPA-AT inhibitor. Examples of known DAPA-AT inhibitors that can be used to develop this method include: herbicidal cinnoline compounds, such as 5-acetyl-4-oxo-1-(2,2,3,3-tetrafluoro-1,4-benzodioxin-6-yl)cinnoline-3-carboxylic acid ethyl ester (Compound A); herbicidal compounds, such as [5-carboxy-1-(1-hydroxyethyl)pentyl]ammonium chloride (Compound B); herbicidal pyridone compounds, such as 2-(3,4-dichlorophenyl)-5-(2,4-difluorophenyl)-1-ethyl-6-methyl-4-oxo-pyridine-3-carboxylate; 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); herbicidal compounds such as [(1R)-2-(6-carboxypyridin-1-ium-2-yl)oxy-1-methyl-ethyl]ammonium; dichloride (Compound E); herbicidal pyrrolidin-2-ones such as 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]-N-(2-methyl-1,2,4-triazole -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-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]acetic acid 2-(4-fluorophenoxy)ethyl ester (Compound I), 2-[1-[(2,3-difluorophenyl)methyl]-5-oxo-pyrrolidin-2-yl]acetic acid -yl] acetic acid 2-methoxyethyl ester (Compound J), 2-[5-oxo-1-[(2,3,4-trifluorophenyl)methyl]pyrrolidin-2-yl]acetic acid (Compound K); N-benzylazole herbicidal compounds, such as 2-(3,4-dichlorophenyl)-1-ethyl-4-oxo-6-[[3-(trifluoromethyl)pyrazol-1-yl]methyl]pyridine-3-carboxylic acid (Compound L); and herbicidal quinolone compounds, such as 2-(3,4-dichlorophenyl)-1-ethyl-6-fluoro-4-oxo-quinoline-3-carboxylic acid (Compound M). The results obtained using these compounds to develop the method of the present invention are shown in Table 1; and a typical IC50 graph is shown in Table 1. Figure 2 Shown in.

[0021] The assay mixture comprises (a) DAPA-AT, (b) an amine donor, (c) 5'-pyridoxal phosphate (PLP), and (d) 7-keto-8-aminononanoic acid (KAPA). The term diaminononanoic acid aminotransferase (DAPA-AT), represented by component (a), refers to an enzyme that catalyzes the conversion of 7-keto-8-aminononanoic acid (KAPA) to 7,8-diaminononanoic acid (DAPA). Examples of DAPA-AT can include, for example, bacterial BioA enzymes or functional derivatives or fragments thereof. More preferably, DAPA-AT is a plant BIO1 enzyme, which can be present as a bifunctional BIO3-BIO1 enzyme or a functional derivative or fragment of BIO1 or BIO3-BIO1 enzyme. BIO3-BIO1 refers to an enzyme that can catalyze the conversion of KAPA to desthiobiotin. The exact nature of DAPA-AT is not relevant to the present invention, and many suitable DAPA-AT enzyme sequences are reported in the literature. Thus, the methods of the present invention can utilize various plant BIO3-BIO1 enzymes, including, for example, enzymes from monocot species (e.g., millet (Setaria italica) - see Figure 3 ) and the mature BIO3-BIO1 enzymes from dicot species (e.g. Arabidopsis thaliana - see Figure 1 ) of the mature BIO3-BIO1 enzyme.

[0022] Thus, in a preferred embodiment of the present invention, the DAPA-AT is a functional derivative of a plant BIO1 or a fragment thereof. In a more preferred embodiment, the DAPA-AT is provided within a bifunctional BIO3-BIO1 enzyme or a functional derivative or fragment thereof. In a preferred embodiment, the DAPA-AT is a BIO3-BIO1 enzyme selected from the group consisting of SEQ ID NOs 1 to 37, or a functional derivative or fragment thereof. In one embodiment of the present invention, the DAPA-AT is mature Arabidopsis BIO3-BIO1 (SEQ ID NO. 21) or Setaria BIO3-BIO1 (SEQ ID NO. 25), or a functional derivative or fragment thereof. In an embodiment of the present invention, the DAPA-AT is a plant BIO3-BIO1 enzyme that is not from the genus Arabidopsis.

[0023] Functional derivatives are those in which some amino acids in wild-type DAPA-AT are substituted and / or deleted but which still retain DAPA-AT activity. Typically, such functional derivatives will retain 85%, more preferably 90%, even more preferably 95%, even more preferably 99% sequence identity at the amino acid level.

[0024] By fragment is meant a truncated version of the DAPA-AT enzyme in which DAPA-AT activity is retained. For example, the wild-type BIO3-BIO1 enzyme includes an N-terminal mitochondrial targeting peptide (e.g., for Arabidopsis BIO3-BIO1 of SEQ ID NO. 1, this peptide contains 22 amino acids), and it will be understood that this N-terminal mitochondrial targeting peptide can be removed, allowing the truncated mature DAPA-AT to be used in the methods of the present invention. Examples of BIO3-BIO1 enzymes with the putative N-terminal mitochondrial targeting peptide removed are provided as SEQ ID NOs. 21 to 37.

[0025] It is contemplated that DAPA-AT may be provided for use in the assay in a number of art-recognized ways. For example, it may be provided in the form of a crude extract, but more preferably, DAPA-AT will be produced by recombinant means and used in the assay in a suitably purified form.

[0026] The amine donor can be obtained from any suitable source. For example, it can be obtained from a suitable biological source and contained in, for example, a crude extract or clarified biological lysate. Alternatively, and preferably, the amine donor can be chemically synthesized and thus provided in the assay mixture in a suitably pure form. It will be understood that component (b) may contain more than one amine donor.

[0027] Since DAPA-AT is a PLP-dependent enzyme, the assay mixture further includes (c) PLP. PLP may already be present in association with DAPA-AT and thus provided in the assay mixture in this manner. Alternatively, exogenous PLP, which is readily available from commercial sources, may be provided in the assay mixture.

[0028] The substrate 7-keto-8-aminononanoic acid (KAPA) (d) is also readily available from commercial sources.

[0029] Those skilled in the art will appreciate that additional assay components will be included in the assay mixture. These may include, for example, buffers such as 3-[4-(2-hydroxyethyl)piperazin-1-yl]propane-1-sulfonic acid (EPPS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), N-[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]glycine (Tricine) or 2-amino-2-(hydroxymethyl)propane-1,3-diol (Tris), salts such as sodium chloride, potassium chloride or magnesium chloride, detergents such as polyoxyethylene (20) sorbitan monolaurate (Polysorbate 4-Hydroxyethyl)piperazine-1-yl]propane-1-sulfonic acid (EPPS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), N-[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]glycine (Tricine) or 2-amino-2-(hydroxymethyl)propane-1,3-diol (Tris), 20), 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol (Triton X-100) or 3-{dimethyl[3-(3α,7α,12α-trihydroxy-5β-cholan-24-amido)propyl]ammonio}propane-1-sulfonate (CHAPS), reducing agents such as (2S,3S)-1,4-bis(mercapto)butane-2,3-diol (dithiothreitol), chelating agents such as N,N′-(ethane-1,2-diyl)bis[N-(carboxymethyl)glycine] (EDTA) or other proteins such as bovine serum albumin (BSA). The exact nature of these conventional components is not relevant to the present invention.

[0030] Conveniently, the method of the present invention will be carried out in at least two distinct stages. In the first stage, the candidate compound is introduced into an enzyme mixture comprising (a) DAPA-AT. The enzyme mixture is then optionally incubated for a period of time to allow the candidate compound to bind to DAPA-AT. In the second stage, an assay mixture is provided by adding a substrate mixture comprising an amine donor (b) and KAPA (d) to the enzyme mixture. PLP (c) will typically be included in the enzyme mixture, but may additionally / alternatively be provided in the substrate mixture. The assay will typically be carried out at room temperature for 60 minutes.

[0031] The "detection inhibition" portion of the method can be performed using several art-recognized methods readily apparent to those skilled in the art. For example, alternative methods include directly detecting DAPA formation or KAPA loss in these functional assays, or by coupling the assay to desthiobiotin synthetase (DTBS) and detecting the formation of the product DTB. In the case of utilizing BIO3-BIO1 in the assay, (e) ATP and (f) CO2 can be further included in the assay mixture and the formation of DAPA and / or DTB detected. These components can be monitored using, for example, a suitable LC-MS program. Other assays are also contemplated, including radiolabeled assays and spectrophotometric assays. However, in a preferred embodiment of the present invention, the formation of DAPA is monitored in the assay. In a more preferred embodiment, DAPA is conveniently converted into a fluorescent derivative, such as DAPA:o-phthalaldehyde.

[0032]

[0033] The use of fluorescent derivatives is particularly useful in the methods of the invention as this allows the methods of the invention to be adapted for high throughput assays using, for example, multiwell plates. Thus, in a particular embodiment of the invention, the assay is terminated using a termination mixture comprising o-phthalaldehyde and 2-mercaptoethanol.

[0034] Inhibition can be determined by comparing the results obtained from this assay with the results obtained from an assay performed in the absence of the candidate compound, under otherwise identical conditions. % Inhibition can then be calculated using the following calculation: % Inhibition = 100 x ((((MAX-MIN)-(X-MIN)) / (MAX-MIN))), where MIN and MAX are the averages of the minimum and maximum response controls, respectively, and X is the sample of interest, to characterize whether the candidate compound is an inhibitor of DAPA-AT.

[0035] The present invention further provides the use of an amine donor in an in vitro diaminononanoic acid aminotransferase (DAPA-AT) assay, the amine donor comprising a compound having formula (I):

[0036]

[0037] wherein X is NH or CH2;

[0038] R 1 is hydrogen or -[CH2] p NH2;

[0039] m and n are independently selected from 0, 1, 2, 3 or 4; and

[0040] p = 1, 2, 3, or 4;

[0041] or its carbamate derivatives, wherein R 1 It is -C(O)OH.

[0042] Examples

[0043] The following non-limiting examples provide specific methods representative of the present invention.

[0044] Example 1 - Expression of Arabidopsis thaliana BIO3-BIO1 (BioDA) in Escherichia coli

[0045] Twist Bioscience synthesized a pET-24 plasmid containing a DNA sequence (SEQ ID NO: 38) encoding BIO3-BIO1 derived from Arabidopsis thaliana, which was codon-optimized for expression in Escherichia coli, with the predicted N-terminal mitochondrial transit peptide removed and an N-terminal 6xHis-tag and thrombin cleavage site, under the control of a T7 promoter. The plasmid was expressed in Escherichia coli BL21 (DE3) bioA under the screening conditions of 50 μg / mL kanamycin and 25 μg / mL chloramphenicol. Overnight cultures grown at 37°C were inoculated in 8X 500mL AIM in a shake flask at a ratio of 1:100. The culture was grown at 37°C, 200rpm for 3.5 hours and then overnight at 20°C. The cells were harvested by centrifugation at 10,000g for 30 minutes. The cells were stored at -80°C until extraction. 30 g of cell pellet was thawed in 120 mL of IMAC buffer A (20 mM HEPES pH 7.5, 25 mM imidazole, 500 mM sodium chloride, 0.15 mM pyridoxal phosphate, 0.5 mM TCEP, 1X complete protease inhibitor tablet (Roche)). The resuspension was passed through a cell disruptor at 18,000 PSI and centrifuged at 50,000 g for 25 minutes at 4°C. The His-tagged BIO3-BIO1 proteins were purified from the extract supernatant using a HisTrap FF column on a Cytiva chromatography system (Cytiva). The proteins were eluted in IMAC buffer B (20 mM HEPES pH 7.5, 500 mM imidazole, 500 mM sodium chloride, 0.15 mM pyridoxal phosphate, 0.5 mM TCEP) and subsequently eluted using a HiLoad 26 / 60 Superdex200 column in SEC buffer (25 mM HEPES pH 7.5, 150 mM sodium chloride, 0.15 mM pyridoxal phosphate, 0.5 mM TCEP). Protein concentration was measured and calculated using the absorbance at 280 nm. The extract was diluted 1 to 4, and 10 uL was analyzed by SDS PAGE.

[0046] Example 2 - DAPA-AT assay.

[0047] To measure DAPA-AT enzyme activity in the presence of herbicides (candidate compounds), 0.5 μL of herbicide was incubated with 20 μL of enzyme mix (100 mM EPPS pH 8.6, 0.1 mM pyridoxal phosphate, 0.1 mg / mL BSA, 25 nM BIO3-BIO1 enzyme) in a well of a 384-well black plate at 20°C for 10 minutes, followed by the addition of 20 μL of substrate mix (final concentration 100 mM EPPS pH 8.6, 0.1 mM pyridoxal phosphate, 0.1 mg / mL BSA, 10 μM spermidine, 1 μM KAPA, 100 mM NaHCO3) and incubated at 20°C for an additional 60 minutes. The reaction was terminated by adding 50 μL of a derivatization mixture consisting of 185 mM borate NaOH, pH 9.4, 3 mM o-phthalaldehyde, 6 mM β-mercaptoethanol, 22.5% (v / v) ethanol, and 3% (v / v) methanol. The fluorescent product generated by the reaction of DAPA with o-phthalaldehyde and β-mercaptoethanol was measured at 470 nm after excitation at 410 nm. The percent inhibition was calculated using the following equation: % Inhibition = 100 x ((((MAX-MIN)-(X-MIN)) / (MAX-MIN))), where MIN and MAX are the average of the minimum and maximum response controls, respectively, and X is the test sample. Compounds were typically added directly to the assay plate starting at a maximum final concentration of 100 μM and serially diluted 3x at 10 different concentrations. Example results are shown in Table 1 below. Using the equation Y = Bottom + (Top - Bottom) / (1 + ((X 希尔斜率 ) / (IC 50 The Hill slope))) The percent inhibition values ​​were fitted to the dose-response to calculate the compound concentration that gave 50% inhibition (IC) 50 ).

[0048] Table 1. % inhibition of mature Arabidopsis BIO3-BIO1 (SEQ ID NO: 21) by various inhibitor compounds.

[0049]

[0050] Example 3 - Measurement of DAPA-AT and desthiobiotin synthetase enzyme activities in the presence of different amine donors.

[0051] To measure the enzymatic activity of DAPA-AT and dethiobiotin synthetase in the presence of different amine donors, 10 μL of candidate amine donors were placed in the wells of a 96-well plate (final concentration of 3.3 mM). 10 μL of substrate mixture (final concentration of 100 mM EPPS pH 8.6, 0.1 mM pyridoxal phosphate, 1 mM ATP, 5 mM MgCl2, 20 μM KAPA, 100 mM NaHCO3) was added to each well. The assay was started by adding 10 μL of BIO3-BIO1 enzyme (final concentration of 1 μM) and incubated at 20°C for up to 90 minutes. The reaction was stopped by adding 100 μL of 1:1 acetonitrile / methanol, and dethiobiotin (DTB) was quantified by LC-MS using an Orbitrap mass spectrometer in positive ionization and full scan mode using a heated electrospray ionization source. The analytes were separated by reverse phase chromatography using a UPLC 2.1 mm × 50 mm C18 column. 5 μL of sample was injected at a flow rate of 0.7 ml / min onto a column equilibrated with 95% solution A (0.2% formic acid) and 5% solution B (acetonitrile). DTB was eluted with a 0.7 ml gradient from 5% to 95% B and compared with a commercial standard of DTB. The results obtained are summarized in Figure 1 middle. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 Comparison of Arabidopsis BIO3-BIO1 activities with various amine donors.

[0053] Figure 2 Typical IC50 plot.

[0054] Figure 3 Setaria BIO3-BIO1 enzyme activity at different amine donor (spermidine) concentrations.

[0055] sequence BIODA / BIO3-BIO1 protein sequence - full length (putative mitochondrial targeting sequence is underlined)

[0056] Arabidopsis thaliana BioDA protein (SEQ ID NO: 1)

[0057] MIPVTATLIRHRLRHLRHRIRF KSTSVSPFHLPLNHPTYLIWSANTSL

[0058] GKTLVSTGIAASFLLQQPSSSATKLLYLKPIQTGFPSDSDSRFVFSKLD

[0059] SLSLRRQIPISISNSVLHSSLPAAKSLGLNVEVSESGMCSLNFRDEKTV

[0060] TGAPELLCKTLYAWEAAISPHLAAERENATVEDSVVLQMIEKCLKE

[0061] EMECGVKSEXDLLCLVETAGGVASPGPSGTLQCDLYRPFRLPGILV

[0062] GDGRLGGISGTIAAYESLKLRGYDIAAVVFEDHGLVNEVPLTSYLRN

[0063] KVPVLVLPPVPKDPSDDLIEWFVESDGVFKALKETMVLANLERLERL

[0064] NGMAKLAGEVFWWPFTQHKLVHQETTVTVIDSRCGENFSIYKASDNS

[0065] SLSQQFDACASWWTQGPDPTFQAELAREMGYTAARFGHVMFPENV

[0066] YEPALKCAELLLDGVGKGWASRVYFSDNGSTAIEIALKMAFRKFCV

[0067] DHNFCEATEEEKHIVVKVIALRGSYHGDTLGAMEAQAPSPYTGFLQ

[0068] QPWYTGRGLFLDPPTVFLSNGSWNISLPESFSEIAPEYGTFTSRDEIFD

[0069] KSRDASTLARIYSAYLSKHLQEHSGVRQSAHVGALIIEPVIHGAGGM

[0070] HMVDPLFQRVLVNECRNRKIPVIFDEVFTGFWRLGVETTTELLGCK

[0071] PDIACFAKLLTGGMVPLAVTLATDAVFDSFSGDSKLKALLHGHSYSA

[0072] HAMGCATAAKAIQWFKDPETNHNITSQGKTLRELWDEELVQQISSH

[0073] SAVQRVVVIGTLFALELKADASNSGYASLYAKSLLIMLREDGIFTRPL

[0074] GNVIYLMCGPCTSPEICRRLLTKLYKRLGEFNRT

[0075] BioDA protein sequence of Zea mays (SEQ ID NO: 2) MVRRLFLHHARRRLHSGTASSIPLST PIFAIFGANTGVGKTLVSAGLATALLSSPSPAVSSVAYLKPLQTGYPVDSDASFVFSRTPALLRAFPPCRATRLIAS CRTFF QSPAVEAKAESLHESQEKVVTYGAGAAEQTKVLACCTAYAWREPVSPHLAAEREGMAAGDDEVKGCVEQWLLEEGIGEGGEVWKILETAGGVASPSASGTLQCDLYRPFRLPAILVGDGRLGGISSTLSAYETLLLRGYDVSAVILEDRELSNDKFLLSYLRNRVHVLILPQIPEDPLDDLTDWFSESSSIFIFLKDELQSFHSRRIERLNSMQRKSKALLWWPFTQHNLVPQDSVTVIDSRYGENFSVYKIKDKMLIPQFDACASWWTQGPDSNLQIELARDMGYAAARYGHVMFPENVHEPALHSAEVLLGGVGKGWASRVYYSDNGSTAIEIALKMAFRKFSLDHGIMASSENSTRNERNIQLKVLALNGSYHGDTLGAMEAQAPSAYTSFLQQPWYSGRGLFLDPPTVYIKSEVYNISLPQSMQHDPQTYGFFSQAEVFCKTRDKTFAADLYSSYIKQKLSEFSLSSSSEHLAALIIEPVIQGAGGMLMIDPLFQRVLVSECRSRKIPVIFDEVFTGFWRLGVESASELLGCLPDIACYAKLMTGGIIPLAATLATEEVFESFESDSKLTALLHGHSYTAHAMGCTAALKAMQWYRGPSTNSNLDTDHMKLKELWDGTLVKQLSSLPNVKRVVSLGTLCAIELKAEGADAGYASLYASSLVQQLRKEDDIYIRPLGNVIYLMCGPCTPRDSCTRQLLKVHRRLCALN

[0076] Nannochloropsis gaditana BioDA protein (SEQ ID NO:3) MCSLTIQVFGANTDVGKTVISAGLLRAASNRPLRRRTTYIKPLQTGKIHDGSFVQRYTSADVSCHTLYSWETPLSPHVAATKEGKTISSEELLGMLRTCLQQAQESHTVGGDGGGVRQREGSNVGDLVVVETAGGVLSPGPSPLQTQADLYRPLRLPVLLVGDGRLGGISSTLAAYESLYCRGYEVHAIAVLEVPHYIENAAAVSEYITGTLGHAETPVLSLPHLPEVSSVSFSTGDVDKDVLLGGLDQWYHQHEHVFHHLLDTLIDRERVRIQRLQKMTTQAAQVFWYPFTQHSGLSGTDLHVVDSAYGDNYTLISKSSEDGDAKERVPCSRTLIDACASWWTQGLGHGRVDVALAVGAAAGRFGHVIFPGNVHEPALRLAELMLAGPGQGWASRVFFSDNGSTAMEIAVKMALKKNSVDLKYEKGCSKEEFRVEQPNDKISATPLTVVTQRDCYHGDTLGCMDAAAPTIFNTGQHPWYQPRTLSLTVPTFGYVSGHLQVEIPPEITRAAGQSALQLPDSGQVLKFPGGLATLLDVHAREGSDAAVIYRKAIRAQFIAFERGEEGILSSNTVIPLRGEAGQGHRCRKLGAVLLEPVLMGAAGMVMVDPLFQRILTLEGRAYGLPVVFDEIASGLYRLGCASAAELLGVAPDIACYGKTLTGGYLPMALTLTTEAVFEAFHGTNKSDALLHGHSYTANPLGCAAAVHALKAYAVIHETNIRTSQECCQRKEDFFPNKNKCSSHHNICRYNEDQVAALSNLSGVRRAVALGTVLAVELTEVESSNRKGYSSTASTHIVNDLREAGIYARPLGNVVYLIASQVSSGGTFECLVDTLDRVLRRRGDTLGKFTHDMTTSGVVI

[0077] Taxus chinensis BioDA protein (SEQ ID NO:4) MASLALTRRRFNPVSRLSL HKCRSTFSRSGQAGEDRTPKQKLHIPSGFEVDLSHPTYVIWGSNTGVGKTLVSTGLAASLLNASQNLLYVKPVQTGFPQDSDSRYVSHKVKDILKQRKPSNNVHISHQVLKASPRAWELLPGSKGFGEVASYERAIQGMRNVCDYEGQTVQGLYKGLADLKLVCKTIYGWYEAVSPHLAAAGEKCSVDDCEVVENLQNSLREFNVDENNTNRWSIIETAGGVASPGPSGTLQCDLYRPFRLPSILVGDGRLGGISGTISAYETLCIRGYDICAVILADTGLSNEKCLSTYLHDRVPVFVLPKIPDDTNNTLLEWFSESYKVFSEIKDTLQTSYFKRIYRLHEMPMKARQILWWPFTQHSLVTEQSISLIDSRFGENFGVYKGKNHKDVKESIVQQFDACASWWTQGPSAILQTELAREVGYTTGRFGHVMYPENVYEPALHCAELLLEGVGKGWATRVFLSDNGSTAIEISLKMAFRKFAFDHGFLPEHFNEDLRKICPQFKVEIGTISYTTCAFFIEMTMEEFSEESYTGRGLFLDPPTVFLHNEEWQLHLPDALMYSDLKYPGLVIHASGGMHMIDPLFQRDLVKECRKRRIPVIFDEVFTGFWRLGAESAAEILGCAPDIACYAKLLTGGIVPLAATLATESIFEAFEGKSKLFALLHGHSYSGHAIGCAAAIKAIQWFKDAEMNLNICTDGSRLQELWDSKLVAKISALSSVKRVIALGTICALELQADESNTGYGSLLASSLVQKLRDDGVYMRPLGNVIYLMCGPATPPAMCSTLLQKLHKRLDEC

[0078] BioDA protein of Physcomitrium patens (SEQ ID NO:5) MHLLLLLPLRRRCTNP IAPRIAHQSRFLVSTAGACSPLPRHLLSGIWGRCLSSHPSPIEDADEARCVAENVADALDVSYPTYMVWGSNTGVGKTLVSAGLCSAILRLSKCQGNSGAKDLLYLKPVQTGFPSDSDARYVYQEVSRVGRLHPEAISEGLFLCNHTLQVSPAVQISARAGASVPVQPQSAWNAGGLSSSLNGTGMKDFCSYESQLLQGVKSRNIPADGDVYGRAQLRNTLSRFTCKTMWAWYEPISPHLAAAREGSTVPDTMVVEAIQRSLQALSSSRKTSGHREDAGDCAEKWAILETAGGVASPGPSGSLQCDMYRSLRFSGLLVGDGRLGGISTTIAAYESLRSRGYDIPAIVIVDSGLSNEEPVLQYLRHKIPVVVLPPLPEDPNKRLEEWFRLAEGTFLELMNVLENSHSKRLRRLEEMPKMAGEILWWPFTQHDLVAQDSITLIDSRSGENFSVYKSGTPSTGKTIATQFDACASWWTQGPDSILQPEIAREVAYTAGRYGHVMFPENVHEPALRCAEILLAGAGRGWAERVFFSDNGSTAIEVAIKMAFRKYIVDHGLSDPSTIERRGGQLKVVALKGSYHGDTLGAMEAQAPSAYTGFKQQPWHVGRGFFLEPPTVFQRQGRWHLQLPRVFHDSDSSDSASCWKSRDEIFDICRDSTPLANLYIRSINQQLAAAVREENNVPAALIIEPVMHGAGGMDMIDPLYQRMLVRECRRRGIPTIFDEVLSGCWRFGVESAVDLLGCSPDIACYSKLLSGGLVPLAATLATQAVFEAFRGISKLDALLHGHSYTAHAVGCASAVVSLQYFSDPSKNSNLLPGGRRLRELWKPELVSLISSHQSVERVVSVGTIFALELRANAAESGYASMLSKELVENLRQEGIYTRPLGNVVYLMCGPLTQPKTCTTMLEKLLALLG

[0079] Adiantum nelumboides BioDA protein (SEQ ID NO:6) MAPPLSHLYPHLRIHQIFGANTDVGKTIFTTALALASAALPIGSKNGKIERDGQGSLNFQENGEAVHYIKPISTGPLRDSDSLHIARYLPSVNAHTLFQFNDPVSPHVAAARYKAEDKNKQDLDVSDEAVIHALSNKISTFTQAAIRPGAAYIETAGGVHSPAPSGSSISQLLRPLRMPTILIGDSNLGGISTTKSAYDSLLMAGYDIEALLLFSDAGRGWGNAEYLTKWGEEIGLPVWALTGPSSSTVWGLPPAREGSDEEDARRMQEYYAGLVAGRGDAAENDESSLGAAPIIEHLRRRHEKRLENIGSMAKRTRDTCWWPFTQHMLAKSDEDVNVIDSAYGDFFTVLNQSSENASKHGNTSLLRPLLDGSASWWTQCLGHADPGLTRAAARAAGRYGHVLFPMCANEPSLQLSETLVGKNSSDVIGTSPGEGWASRVFFSDDGSTGMEVALKMAISSASSRYAPKAESQLAKSKVQQGRSAGNQSGRPPREWMVLGLKGSYHGDTIGAMDACEGNIFNEKVEWYRGRGSWFEPPTVSIKDGQAMVDLPLQDADWQHLSGKVESKKNFNNVSDIYNVSDRMDNDELKNIYSKLIYGWLDHLTRMEGNRYGALILEPLIMGAGGMICVDPLFQRCFVDVIRANPDLFSQVDPPLRGTSHIRESDVPGSGDWKGLPIIFDEVFTGLYRLGYSTPAQAIGVSPDISVLAKILTGGMVPMSVTLASDSIFSTFSKSDQKIDALLHGHSYTAHPIGCEIANEALRRIETMQVENFWGDAKADWESRTSSLQRDNSMKNVPSPWSLWSREAVEAFSRSANVESSMALGTVMILHLSDSAGEKGYSSSAAVDILQHLRTTQDTGFNKHVSFNIHARPLGNVVYMMCSLNTPVNVLRETEAALKHVLKV

[0080] Setaria italica BioDA protein (SEQ ID NO:7)

[0081] MLPRLLLRSRHRRRY SSAAATVPLSTPTFAVFGANTGVGKTLVSAGL

[0082] VAALLNSRSPSVSAVSYLKPLQTGYPADSDARFVFARAPALLRASSSP

[0083] RATRLVASCRTLFPSPAVGPEAEPLHQSQEKVVTYGGDGAEEETKV

[0084] LACRTVYAWREPVSPHLAAEREGMAAGDDEVRGCVDQWLMEEGV

[0085] GEGGEVWKVLETAGGVASPSASGTLQCDLYRPFRLPAILVGDGRLG

[0086] GISSTLSAYETLLLRGYDVSTVILEDRDLTNDTFLLSYLRNRVHVLAL

[0087] PQIPEDPSDDLTDWFSESSSVFNLLKDSLQSFHSKRVERLNGMQKKS

[0088] KDLLWWPFTQHNLVPQDSVTVIDSRCGENFSAYKIKDNTMMLVPQF

[0089] DACASWWTQGPDSDLQIELARDMGYAAARYGHVMFPENVHEPALR

[0090] AAELLLGGVGKGWASRVYYSDNGSTAVEIALKMAFRKFSLDHGILA

[0091] DSEKSIRNERNIQLKVLALNGSYHGDTLGAMEAQAPSAYTSFIQQPW

[0092] YSGRGLFLDPPTVCIKSETCNVSLPQSMQHYPRTSGDTCFPSLAEVFC

[0093] KTRDTSSAADLYLSYIKQQLSEFSQSSNSDHLAALIIEPVIQGAGGML

[0094] MIDPLFQRVLVRECRSRKIPVIFDEVFTGFWRLGVESASELLGCLPDI

[0095] ACYAKLMTGGIVPLAATLATEEVFESFKSDSKLTALLHGHSYTAHA

[0096] MGCMAALKAIQWYRDPSTNSNLDLDHMKLKELWDGALVKQLSSLP

[0097] NVKRVVSLGTLCAIELKAEGSDAGYASLYASSLVQQLRREDDIYIRPL

[0098] GNVIYLMCGPCTPRDSCSRQLHKVLRRLCDLN

[0099] Date palm (Phoenix dactylifera) BioDA protein (SEQ ID NO:8) MPSCPLLPLLPRRRPPPSPLL LLLLSRSLHSSPPELDLSHPTLVVWGSNTGVGKTIVSAGLAASVLSPSSPNTATFLYLKPVQTGFPRDSDSRFVYRKVSDLFRHRLGGFSAQLLASNHALKASVPAAREVLGARTERPGEEGEEASTGLCCYEETRVGTGESTTGDFKLVCKTLYGWKEAISPHLAVEREGMAVEDSSLREVLGRLLNSSMGGGGDRRKENVWRVIETAGGVASPGPSGSIQCDLYRPFRLPAVLVGDGHLGGISTTLSAYESLTLRGYDVIAIILADHGLSNEIPLLSNLRHRVPVLVLPPVPEDPLNDLVDWFCESSKAFSSLQDLVQSTHSKRIQRLHDMPRKAGNILWWPFTQHKLVPEEIVTVIDSRCGENFAVHKVRNDREMIVRQFDACASWWTQGPDFALQIELARDMGYSAARYGHVMFPENVYEPALHCAELLLEGVGKGWASRTYYSDNGSTAIEIALKMAFRKFLLDNEMLMDSGNGISSERCIELRVLALNGSYHGDTLGAMEAQAPSSYTGFLQQPWYSGRGLFLDPPTIFIHKEIWNLSVPESFESDQLKPEDMRFSSSDALFCPSRDSSAIAGIYSAYISNQLLQFSKLSCSTQIGALIIEPVIQAAGGMHMIDPLFQRILVRECRNRRIPVVFDEVFTGFWRLGKESAAELLGCVPDVACFGKLMSGGIIPLAVTLTTEAVFEAFKGDSKLMALLHGHSYSAHAMGCTAAAKSIQWFRDPFTNPNVEFKEKKLKELWEMKLVLQISSLPAVQRVITLGTLCAVELKAEGSDAGYASLYASLLVQQLREDGLYVRPLGNVIYLMCGPCTSSHFCSQQLQQVYQRLHEFSQHQLRDSQSFQREG

[0100] Ostreococcus tauri ATCC 22665 BioDA protein (SEQ ID NO:9) MSAPIA RRASSVARGRTRWLTSTSIERSREWFVRSLGSASDGAASGNGRALELSIASFAIWGANTGVGKTLASAGLARAATRRGTATTFLKPVQTGYPEDSDAGFVARQTKGNETAGEHAAVASGSTEGGGEGFAAGVTRAHTEFAWRRAAGPHLTAAEEGRPVSDEEIVRATRNYLREFAESAREGNARGFAVIETAGGVASPGPSGTLQCEILRALRLPAILIGDGRLGGISTTTSAYESLHARGYDVVGVLMADDGYANHEAVAKILPSGTKVFALPPLPPQGKADAWLEKSAGVFSEAFETAMNWHEKRIQALQKLPGEALEKIWWPFTQHATVEPDTVTVIDGRYGEDFEIYDQEKGRDGAIDLRFDGAASWWTQGVSAELQQELNSAAANAAGRYGHVMFPENVHQPSMDATNALLEGPGKGWGSRVFYSDNGSTATEVGLKMAFRKYYVDAGLVSKDGHQRAKDLINDGSCHLPQLRIIALDGSYHGDTLGAMDMQSPSVFTGPMQTPWYQPRGLFLQPPSLAIRNGEWTVALPPHGLLNDGEDRLFHDGKDLESNTTWKSKADAFDIGKRLDSPLANEYRQAVDKALERAEQDAKAGHMGPVGALMMEPVLHGAGGMVLIDPLFQNILMKVCRERKIPIVLDEVFAGIWRLGTEGAWEIIGEKPDISCYAKLLTGGLMPMSATVATADVFDVFYGPGKAQGLLHGHSYTAYPIGCAVASKALSLYKDPAKNPNLIVDASSKGDTPTAYLRELWDESKLKEISNKSSVRRVFGQGCVLAVELEVEGEGGYSSNAAKEVVLRLRPHSVQARPLGNVVYLMCAPTTPKATCDRLLNVLAEELSE

[0101] Helianthus annuus BioDA protein (SEQ ID NO:10) MFHPPLLFTLSRHFLRRHHH RHLSTLSHTLSHPIYTIWSANTGLGKTLVSAGIAAASVASPPSKLVYIKPIQTGYPLDSDAAFVYRKIPQILHRRNPNLTLTASNQTLNVSIPAAKVKIGDNFSDSRNGFGEFGFCEEMKVGEGVGEGSELICKTIYAWNEAISPHLASEREGALVGDEEVLVLLKKCLEIGVGSEGQGSVLSLVETAGGVASPGSSGSLQCDLYRPFRLPSILVGDGKLGGISGTISAYESLTIRGYDVVAIVLEDHGLENEVPLLSYLRNRVPVLVLPSIPKDPLDNLMEWYDESQSVFDSLKGIMLSSYEKQMYRLHEMPKKAQEIFWWPFTQHKLVPQEKVTVIDSRCGENFAVHKVKDHDYITQMFDACASWWTQGPNAALQIKLAREMGYTAARYGHVMFPENVYEPALQCAELLLQGVGKGWASRVFFSDNGSTAIEIALKMAFRKFLADNKLLMDLPHYNTDESNIELKVVALNGSYHGDTLGAMDAQAPSPYTGFLQQPWYTGRGVFLDPPFVSMCDGVWKIAVPKKMHYEDVILEDTSFNSRDQVFNRSRDDSKLASIYSTYISQELLPSSGSTKCSYAGALIIEPVIQGAGGMLMVDPLFQRILVKECKNRKIPVIFDEVFTGYWRLGVESAAELLFCQPDIACFAKLMTGGMIPLAATLATNDVFESFVGDSKLNALLHGHSYSAHAVGCATACNSIKWFKDSQTNPNLVPGQNLLRELWDAELVRKISFHPAVQRVVSLGTLFALELKVEDNDAGYASLSGTSLLLKLREDGIYMRPLGNVIYMMCGPCTSHRVCLQMLEKLYMRLDEFKQEKIGTARDYDHDHRLPIA

[0102] BioDA protein (SEQ ID NO:11) of Quercus robur MYFLISLHRRGHRHLRHVKDLSTTAHSQLHVPLNHPTFLVWASNTSLGKTLVSTGLAASSLLNPTPKPRKFLYLKPIQTGFPSESDSSSVFHKLSRLSLLRRHNLPHFSLFSSNHILKASFPAANSALGQEISETPHSGMRDLGYYEEAKPEGDVGGGAASQLVCKTLYAWREAVSPHLAAERESGVVEDAAVLELLQKCLRFELEGGGEKGKETEIFCVVETAGGVASPGPSGSLQCDLYRPFRLPGILVGDGRLGGISGTISAYESLKLRGYDIAAVVFEDHGLVNEMPLLSYLRNRVPVLVLPSVPKDLSNDLMEWFDESRGVFDSLKEEMLSAYSKRMMRLQDMPKKAGEIFWWPFTQHQLVPEEAVTVIDSRHGENFAVFKAQNNEFITQQFDACASWWTQGPDTTLQTELARDMGYAAARFGHVMFPENVYEPALECAELLLEGVGKGWASRTFFSDNGSTAIEIALKMAFRKFSYDHGILSDGRKDNTASQRVKLMVLALQGSYHGDTLGAMEAQAPSSYTGFLQQPWYTGRGLFLDPPTVYMCNGVWKLSLPEGLHLEIPKLENKGFSSRDEIFHKIRDKSDLARIYSSYISEQLSQYSGSGGFYHIGALIMEPVILGAGGMHMIDPLFQRVLVNECRSKKIPVIFDEVFTGFWRLGRETAAELLGCVPDIACFAKLMTGGIIPLAATLATNAIFNSFIGESKLKALLHGHSYSGHAMGCTAAVKSIKWFKDPQTNLNITDEGRSLRELWDEDVVQQISSHPAVERVVALGTLFALELRAEGSNAGYASQYASSLLQNLRQDGIYTRPLGNVIYLMCGPCTSPEICSQLLVKLYKRLEDFDQFHQIREENSESYGG

[0103] Thraustotheca clavata BioDA protein (SEQ ID NO:12) MLTLSMRMRPRT LMKMSFSTRAGLGQLGCLTSLDQSVPNAVPTSLHAPIVQVFGSNTDVGKTVIAAGLCRAALLSSKNATVGYIKPLQTGGDAMMDAEFLNRHAGNADNRLTSHTLFSWETAVSPHLAASLENKAVDDATVVAKLSKTLKQIQKEEKGSALTVVETAGGVCSPSASGKFQCDVYRPLRLPTVLVGDGCLGGISATMSALDSLLIRGYDVSCIVLIEQDNLSNGEAISERASELGIPIFSLDKLPPQPEPLDLWYENSTETFRSVHDTIASTHKARVARLQELPSIAKKALWWPFTQHKSNKGVLVIDSAYGDTFATYNQKNQAIEPMYDACASWWTQGVGHGNAKMALSLATAAGRYGHVMFPENAHEPAIALSEMLLKSVGKNWAGKVFFSDDGSTAVEIGIKMALRKYAVDHGLGMSFEGINDVLVLAQEGCYHGDTLGTMNIAEPSVYNKTQHPWYKPKALFVHPPSVKVASNGQMQVTWQNVNPTLVTHLPTLDDLFDISRDTSPLTQQYTEYVESLLDSVPSTTVVGALIVEPILIGAGGMVFVDPLFQRTMVRVCQQRKIPVIFDEVFSGFWRLGAESARDLLQVNPDIACYAKLLTGGAVPMSTTLATNEVFDAFYTDSKADALLHGHSFTANPVGCAAAVTALDMYKTIEKKNSTRVHWNVDQVAALAARPGIARAFQLGTVLVIELESTSRGYTDTTGQDVLVRLRNHGIYARSLGNVLYIMASPLSTPATCIKIFNTVVDNLFA

[0104] Schizosaccharomyces japonicus osaccharomyces japonicus)BioDA(SEQ ID NO:13)(Gibberella zeae)BioDA(SEQ ID NO:14) MAPVPALLWRSLRTHQVYGANTDVGKTIFSTVLCNAASKKGDRTWFLKPVSTGAVDEADGCHIQRFASSTNHETLFQYDIPCSPHIAAKVSGKPIPSDEAVLAKIHDSVSRYASEGPGWLFLETAGGVHSPGPSGTPQADLYTPLRAPVILVGDSKLGGISQTISAYESLRMRGHDIESILLFQDMKYENYQYLKDYFSKQGGIPVDTVPEPPSRLPDIQQDTEQMREYYASQTSNVSHVLDHLDKRGKQRISRLESLSEKASKSIWYPFTQQKLVTADTISAIDSAHGDYFQVLNKTYENLLQPAFDGSASWWSQGLGHANSRLTLAAAYAAGRYGHVMFAEAIHEPALALAEMMLEGAQNSRFSRVFYSDNGSTGCEVAVKMALRAARLRYGWGPNDNLQILGLKGSYHGDTIGAMDCAEPCVYNEKIEWYEGKGYWFDYPTIQCVEGKWVVSAPNGMGQDQKLESISEVFNLESRLKTEQYRGYEEYIEKTLKKLQDQGRKFGALMMEPIILGAGGMIFVDPLFQRALVDVVRRSPHLFGTKDALPAQGPQWSGLPVIFDEVFTGLYRLGRFTAASFLGTEADISVNAKLLTGGLVPLCTTMASESIFDAFKSDDKSDALLHGHSYTAHAVGCQVAVESVSEMQSMEKNGEWEWAEKDWAEDETQAWSVWSRDFVTDMSHNPQVLGVWALGSVLAISLRDDDGVGYKSLAAKKIQSHLRQGTGSWNAHSRVLGNVFYVMASQKTSRQSIRELQGLLVDALSK

[0105] Barley (Hordeum vulgare) BIO3 - BIO1 (SEQ ID NO:15) MLRRLLLRHARRLHYSSSFSSALPLSSPTFAVFGANTGVGKTLVSAGLAASLLRSHAASPSAVLYLKPLQTGFPDDSDAGFLYRRVPALLRPSHAAHLAASIDTLSPAPSVETQSPSREEAAFSYDGEGEAVRKLLACRTLYAWREAVSPHLAAEREGMAVEDAHVRSLLAACLVEGENLGKGEGDVWKLVETAGGVASPGPSGTLQCDLYRQFRLPTILVGDGRLGGISCTLSAYETLLLRGYDVSAVILEDHGLSNNKFLLSYLRNRVPVLVLPPIPEDPSDDLTDWFSESSSVFGLLEDALRSFHLTRIQRLNSMQRKSKDLLWWPFTQHNLVPVDCVTVIDSRCGESFSAYKVKDNKMMMIPQFDACASWWTQGPDSKLQIELARDMGYAAARYGHVMFPENAHEPALRCAELLLGGIGKDWASRVYYSDNGSTAIEIALKMAFRKFSLDRGILLDSDKSIITNGRNIQLKVLALKGSYHGDTLGAMEAQAPSAYTSFLQQPWYSGRGLFLDPPTVFIKNGTCTLSLPQAIQNCHLSPGDKCFPSLAEVFCKTRDSSSAADLYSTYISQQLSEYSLSSNTEHIAALIIEPVIQGAGGMHMIDPLFQRVLVHECRDRKIPVIFDEVFTGFWRLGVESASELLGCLPDVACYAKLMTGGLVPLAATVTTEAVFEAFKSDSKLTALLHGHSYTAHAMGCSAAVKAIQWFRDPSTNSNLDFDCMKLKELWDGTLVNQLSSLPNVKRIVSLGTLCAIELQAEGSDAGYASLYARSLIQQLREEDDIYVRPLGNVIYLMCGPCTTRDVCTEQLSKVYRRISHFNPVHRG

[0106] Brassica napus BIO3 - BIO1 (SEQ ID NO:16) MSPVTATLLRHRLRHLRHHHI RLNSTAVPSHFNLPLNHPTYLIWSANTSLGKTLVSTGIASSFLLHQPSSSPPAAHSAKLLYLKPIQTGFPSDSDSRFVFSKLDSLSLRRRIPLSVSNSVLRSSLPVAESMRRNIKVSESGMCDLNFREEKTVTGAPELLCKTLYAWEAAISPHLAAEREHATVEDSVVLKMVEQEMECGSKANVLCLVETAGGVASPGPSGTLQCDLYRPFRLPGILVGDGRLGGISGTIAAYESLKLRGYDVAAVVFEDHGLVNEVPLTSYLRNKVPVLVLPPVPKDPSDDLIEWFVESDGVFKALKEVMVSVYLERVERLNGMAKQAGEVFWWPFTQHKLVPEDNVTIIDSRCGENFSVFKASDNNSITQQFDACASWWTQGPDPAFQAELAREMGYTAARFGHVMFPENVYEPALKCAELLLDGVGKGWASRVYFSDNGSTAIEIGLKMAFRKFCVDHETLFGLSEDAEEKKHVDVKVLALRGSYHGDTLGAMEAQAPSPYTGFLQQPWYTGRGLFLEPPTVFLSNGTWNLSLPESFSQTASEECGTFTTRDEIFDKSRDTSVLATVYSTYVSEQLQEYSGISQSAHVGALIIEPVIHGAGGMHMVDPLFQRVLVNECRNRKIPVIFDEVFTGFWRLGVETTADLLGCKPDIACFAKLLTGGMIPLAVTLATDAVFDSFSGDSKLKALLHGHSYSAHAMGCATATKAIEWFKDPETNHNIDSQRGTLRELWDEELVQHISCHCAVERVVVLGTLFALELKVDASNSGYASLYAKSLLEMLREDGIFMRPLGNVVYLMCGPCTSPEICRRLLTKLHKRLGEFNRA Upland cotton (Gossypium hirsutum) BIO3 - BIO1 (SEQ ID NO:17) MLLRRHHHHRLFRLRLLLLHHHHHQFQSFSTLTIPLSHPTYIIWSSNTSLGKTLVSTGLSSSFLLSPSSSSKKFLYLKPLQTGFPSDSDSRFLFQKLSSLSLSRNLPLFSSHSVLLSSLPAAKSFKPNEFSLNESREMCELGFYEEKKVLEAGRVAPELVSETIYAWEGALSPHFAAEREGGAVEDSELVKRLETRLKEGLLEGGVERGKLDGFCVVETAGGVASPGPSGTLQCDLYRPLRFPGVLVGDGRLGGISGTISAYESLKLRGYDVVAIVIEDHGLINEIPLKSYLRSRVPVFVLPPIPQDPSNDLMEWFDESCNVFNSLKDIMLEAYLERMRRLNEMPRKAGDVFWWPFTQHKLVPPSTVTVIDSRCGENFSVYKVQNKEYITQQFDACASWWTQGPDATLQTELARDMGYAAARFGHVMFPENVYEPALECAELLLDGVGKGWATRVYFSDNGSTAIEIALKMAFRKFSSDHGILPELLKNNPTESCTELMVLALKGSYHGDTLGAMEAQAPSSFTGFLQQPWYTGRGLFLDPPTVFMHNGKWTVSLPEVFHSATLKPEDTTFRSRDEIFLKSRDESNLAGLYSLYVSQQLSQYSTLSETNKQIGALIMEPIIQGAGGMHMVDPLFQRMLVNECRHCRIPVIFDEVFTGFWRLGVESAAELLGCVPDIACFAKLMTGGIIPLAATMATDAIFDSFTGDSKLKALLHGHSYSAHAMGCTAAAKSIKWFKDPNTNLNITSEKSQKMLLRELWDAELVQQFSSHPSISRVVTLGTLFALELQADESDAGYASLYARSLVQMLREDCIYTRPLGNVIYVMCGPCTSPKMCTLQLLKLYTKLEEFTLVKTKMNII

[0107] Rice (Oryza sativa) BIO3 - BIO1 (SEQ ID NO:18)

[0108] MLRLLRHARRH STSSSSSAAAAAVPLTSPAFAVFGANTGVGKTLVSA

[0109] GLVASLLASPSPSPSTVAYLKPLQTGFPDDSDARFVFDRAPALLRRLR

[0110] LAGGGASTRLVASNHTLFPSPAVDPLPERQDTVVNYGGEEGVEEKA

[0111] LVCRTVYAWREPVSPHLAAEREGMPVEDEEVRWLVDRWLAEEDGG

[0112] GEVWKVLETAGGVASPGPSGTLQCDLYRSSRLPAVLVGDGRLGGIS

[0113] STLSAYETLLLRGYDVGSVILEDRGLSNDRFLLSYLRKRVPVHVLPPI

[0114] PEDPKDDLTDWFSESSSAFSSLKDSLQSFHSRRVQRLNSMQRKSKYL

[0115] LWWPFTQHDLVPVDSVTVIDSRFGENFSAYKVKDKTIVPQFDACAS

[0116] WWTQGPDSNLQIELARDMGYAAARYGHVMFPENVHEPALRCAELL

[0117] LGGVGKDWASRVYFSDNGSTAIEIALKMAFRKYACDHGIIVDSEKDI

[0118] RSEGSVHFKVLALNGSYHGDTLGAMEAQAPSAYTSFLQQPWYSGRG

[0119] LFLDPPTVYIKNKSANLSLPPSIMHDQLSSCDTCFSSLTEVFCKTRDTS

[0120] SAANVYVSYISQQLSQYAMSNNSEHIAALIIEPVIQGAGGMHLIDPLF

[0121] QRLLVKECKNRKIPVIFDEVFTGFWRLGVESASELLGCFPDISCYAK

[0122] LMTGGIVPLAATLATEPIFEAFRSDSKLTALLHGHSYTAHPMGCTAA

[0123] VKAIQWYKDPSTNSNIDLDRMKLKELWDSALVNHLSSLPNVKRVVS<000%307>

[0124] LGTLCAIELKAEGSDAGYASLYASSLIRQLREEDNIYARPLGNVIYLM

[0125] CGPCTTQDSCTRQLAKVHRRLQKLN

[0126] Soybean (Glycine max) BIO3 - BIO1 (SEQ ID NO:19) MFRFPSVLLSRRLHRREL It should be noted that there seems to be an error in the tag in line 3, it might be " " instead of "<000%307>". If this is not a typo, it needs to be corrected according to the actual situation for accurate translation.SSSTTSHPPSLPLSHPIYLIWGSNTGVGKTLVSAGIAAAFLLSSPTPSQFHYLKPLQTGFPSDSDSRFVFNKLRQLSLIRNPHIPLSASHRVINVSPAVTASNPLAGEEGVGPPPELLCKTLYAWEEAVSPHLAAEREGLVVKDSAVLETLGECLEDVAECGAGKERSEVLCVVETAGGVASPGPSGSLQCDLYRPFRIPAVLVGDGRLGGISGTISAYESLKLRGYDVVAVVFEDHGLLNEGPLMSYMRNKVPVLVLPPIPKDPLNDLMEWFESSHYIFSNLKEIMLSAYFERIKKLHDMPREARDIIWWPFTQHKLVPDGGVTVIDSRCGENFSIFKVQKTEVIAPQFDACASWWTQGPDAIMQTELAREMGYAAARFGHVMFPENVHEPALNCAELLLQGVGKGWASRTYFSDNGSTAIEIALKMAFRKFSVDHGLIVHCHEDTTNERSTELMVLALQRSYHGDTLGAMEAQAPSSYTGFLQQPWYTGRGLFLDPPSVFMHNSTWNVSIPEGYHLESLKGKSITFASPDEIFHKGRDKSELATVYSSYISKVLSGFRGSNNVGALIMEPVIQGAGGMHMVDPLFQRVLVNECRSRKIPVIFDEVFTGFWRLGVETAVELIHCVPDIACFGKLLTGGIIPLAVTLATNAVFDSFIGDSKLKALLHGHSYSAHAMGCAAAVKSIQWFKDPCSNPNITSEGRLLRELWDDKMVHRISSHPAIQRVVALGTLCALELKAEGNNAGYGSLYARPLLEKLREDGVYMRPLGNVIYLLCGPCTSPEVCNQLLVKLLRRLEEFDVGKN

[0127] Common wheat (Triticum aestivum) BIO3 - BIO1 (SEQ ID NO:20) MLRRLLLRQARRRHCSSS FSSALPLSSPTFAVFGANTGVGKTLVSAGLAASLLRSHAASPSAVLYLKPLQTGFPADSDAGFLYRRVPALVRPSHAAHLVASIDTLSPAPSVETPSPSREEAAFSYDGEGKAVRKLLACRTLYAWREAVSPHLAAEREGMTVEDAHVRSLLAACLAEGETLGKGEGDVWKLVETAGGVASPGPSGTLQCDLYRHFRLPIILVGDGRLGGISGTLSAYETLLLRGYDVSAVILEDCGLSNDKFLLSYLRNRVPVLVLPPIPEDPSDDLTDWFSESSSVFGLLEDALRSFHLKRIQRLNSMQRKSKDLLWWPFTQHNLVPVDCVTVIDSRCGESFSAYKVKDNKLMMIPQFDACASWWTQGPDSKLQIELARDMGYAAARYGHVMFPENAHEPALRCAELLLGGIGKDWASRVYYSDNGSTAIEIALKMAFRKFSLDRGILLDSDKSITNERNIQLKVLALKGSYHGDTLGAMEAQAPSAYTSFLQQPWYSGRGLFLDPPTVFIRNGTCALSLPQSIQNCHLSPGDKCFPSFAEVFCKSRDSSAAADLYSTYISQQLSEYSMSSNTEHIAALIIEPVIQGAGGMHMIDPLFQRVLVHECRDRKIPVIFDEVFTGFWRLGVESASELLGCLPDVACYAKLMTGGIVPLAATVTTEAVFEAFKSDSKLTALLHGHSYTAHAMGCSAAVKAIQWFRDPSTNSNLDFDCMKLKELWDGTLVNQLSSLPNVKRVVSLGTLCAIELQAEGSDAGYASLYARSLIQQLREEDDIYVRPLGNVIYLMCGPCTTRDVCTEQLSKVYRRISHFNPIH

[0128] BIODA / BIO3-BIO1 protein sequence - MTP removal

[0129] Arabidopsis thaliana BioDAΔMTP protein (SEQ ID NO:21)

[0130] MKSTSVSPFHLPLNHPTYLIWSANTSLGKTLVSTGIAASFLLQQPSSS

[0131] ATKLLYLKPIQTGFPSDSDSRFVFSKLDSLSLRRQIPISISNSVLHSSLP

[0132] AAKSLGLNVEVSESGMCSLNFRDEKTVTGAPELLCKTLYAWEAAISP

[0133] HLAAERENATVEDSVVLQMIEKCLKEEMECGVKSEKSDLLCLVETA

[0134] GGVASPGPSGTLQCDLYRPFRLPGILVGDGRLGGISGTIAAYESLKLR

[0135] GYDIAAVVFEDHGLVNEVPLTSYLRNKVPVLVLPPVPKDPSDDLIEW

[0136] FVESDGVFKALKETMVLANLERLERLNGMAKLAGEVFWWPFTQHK

[0137] LVHQETVTVIDSRCGENFSIYKASDNSSLSQQFDACASWWTQGPDPT

[0138] FQAELAREMGYTAARFGHVMFPENVYEPALKCAELLLDGVGKGWA

[0139] SRVYFSDNGSTAIEIALKMAFRKFCVDHNFCEATEEEKHIVVKVIALR

[0140] GSYHGDTLGAMEAQAPSPYTGFLQQPWYTGRGLFLDPPTVFLSNGS

[0141] WNISLPESFSEIAPEYGTFTSRDEIFDKSRDASTLARIYSAYLSKHLQE

[0142] HSGVRQSAHVGALIIEPVIHGAGGMHMVDPLFQRVLVNECRNRKIP

[0143] VIFDEVFTGFWRLGVETTTELLGCKPDIACFAKLLTGGMVPLAVTL

[0144] ATDAVFDSFSGDSKLKALLHGHSYSAHAMGCATAAKAIQWFKDPET

[0145] NHNITSQGKTLRELWDEELVQQISSHSAVQRVVVIGTLFALELKADA

[0146] SNSGYASLYAKSLLIMLREDGIFTRPLGNVIYLMCGPCTSPEICRRLL

[0147] TKLYKRLGEFNRT

[0148] Maize BioDAΔMTP protein sequence (SEQ ID NO:22)

[0149] MQSPAVEAKAESLHESQEKVVTYGAGAAEQTKVLACCTAYAWREP

[0150] VSPHLAAEREGMAAGDDEVKGCVEQWLLEEGIGEGGEVWKILETA

[0151] GGVASPSASGTLQCDLYRPFRLPAILVGDGRLGGISSTLSAYETLLLR

[0152] GYDVSAVILEDRELSNDKFLLSYLRNRVHVLILPQIPEDPLDDLTDWF

[0153] SESSSIFIFLKDELQSFHSRRIERLNSMQRKSKALLWWPFTQHNLVPQ

[0154] DSVTVIDSRYGENFSVYKIKDKMLIPQFDACASWWTQGPDSNLQIEL

[0155] ARDMGYAAARYGHVMFPENVHEPALHSAEVLLGGVGKGWASRVY

[0156] YSDNGSTAIEIALKMAFRKFSLDHGIMASSENSTRNERNIQLKVLALN

[0157] GSYHGDTLGAMEAQAPSAYTSFLQQPWYSGRGLFLDPPTVYIKSEV

[0158] YNISLPQSMQHDPQTYGFFSQAEVFCKTRDKTFAADLYSSYIKQKLS

[0159] EFSLSSSSEHLAALIIEPVIQGAGGMLMIDPLFQRVLVSECRSRKIPVI

[0160] FDEVFTGFWRLGVESASELLGCLPDIACYAKLMTGGIIPLAATLATE

[0161] EVFESFESDSKLTALLHGHSYTAHAMGCTAALKAMQWYRGPSTNSN

[0162] LDTDHMKLKELWDGTLVKQLSSLPNVKRVVSLGTLCAIELKAEGAD

[0163] AGYASLYASSLVQQLRKEDDIYIRPLGNVIYLMCGPCTPRDSCTRQL

[0164] LKVHRRLCALN

[0165] Taxus chinensis BioDAΔMTP (SEQ ID NO:23)

[0166] MSRSGQAGEDRTPKQKLHIPSGFEVDLSHPTYVIWGSNTGVGKTLVS

[0167] TGLAASLLNASQNLLYVKPVQTGFPQDSDSRYVSHKVKDILKQRKPS

[0168] NNVHISHQVLKASPRAWELLPGSKGFGEVASYERAIQGMRNVCDYE

[0169] GQTVQGLYKGLADLKLVCKTIYGWYEAVSPHLAAAGEKCSVDDCE

[0170] VVENLQNSLREFNVDENNTNRWSIIETAGGVASPGPSGTLQCDLYRP

[0171] FRLPSILVGDGRLGGISGTISAYETLCIRGYDICAVILADTGLSNEKCL

[0172] STYLHDRVPVFVLPKIPDDTNNTLLEWFSESYKVFSEIKDTLQTSYFK

[0173] RIYRLHEMPMKARQILWWPFTQHSLVTEQSISLIDSRFGENFGVYKG

[0174] KNHKDVKESIVQQFDACASWWTQGPSAILQTELAREVGYTTGRFGH

[0175] VMYPENVYEPALHCAELLLEGVGKGWATRVFLSDNGSTAIEISLKM

[0176] AFRKFAFDHGFLPEHFNEDLRKICPQFKVEIGTISYTTCAFFIEMTME

[0177] EFSEESYTGRGLFLDPPTVFLHNEEWQLHLPDALMYSDLKYPGLVIH

[0178] ASGGMHMIDPLFQRDLVKECRKRRIPVIFDEVFTGFWRLGAESAAEI

[0179] LGCAPDIACYAKLLTGGIVPLAATLATESIFEAFEGKSKLFALLHGHS

[0180] YSGHAIGCAAAIKAIQWFKDAEMNLNICTDGSRLQELWDSKLVAKIS

[0181] ALSSVKRVIALGTICALELQADESNTGYGSLLASSLVQKLRDDGVYM

[0182] RPLGNVIYLMCGPATPPAMCSTLLQKLHKRLDEC

[0183] Physcomitrella patens BioDAΔMTP (SEQ ID NO:24)

[0184] MSSHPSPIEDADEARCVAENVADALDVSYPTYMVWGSNTGVGKTLV

[0185] SAGLCSAILRLSKCQGNSGAKDLLYLKPVQTGFPSDSDARYVYQEVS

[0186] RVGRLHPEAISEGLFLCNHTLQVSPAVQISARAGASVPVQPQSAWNA

[0187] GGLSSSLNGTGMKDFCSYESQLLQGVKSRNIPADGDVYGRAQLRNT

[0188] LSRFTCKTMWAWYEPISPHLAAAREGSTVPDTMVVEAIQRSLQALSS

[0189] SRKTSGHREDAGDCAEKWAILETAGGVASPGPSGSLQCDMYRSLRF

[0190] SGLLVGDGRLGGISTTIAAYESLRSRGYDIPAIVIVDSGLSNEEPVLQY

[0191] LRHKIPVVVLPPLPEDPNKRLEEWFRLAEGTFLELMNVLENSHSKRL

[0192] RRLEEMPKMAGEILWWPFTQHDLVAQDSITLIDSRSGENFSVYKSGT

[0193] PSTGKTIATQFDACASWWTQGPDSILQPEIAREVAYTAGRYGHVMF

[0194] PENVHEPALRCAEILLAGAGRGWAERVFFSDNGSTAIEVAIKMAFRK

[0195] YIVDHGLSDPSTIERRGGQLKVVALKGSYHGDTLGAMEAQAPSAYT

[0196] GFKQQPWHVGRGFFLEPPTVFQRQGRWHLQLPRVFHDSDSSDSASC

[0197] WKSRDEIFDICRDSTPLANLYIRSINQQLAAAVREENNVPAALIIEPV

[0198] MHGAGGMDMIDPLYQRMLVRECRRRGIPTIFDEVLSGCWRFGVES

[0199] AVDLLGCSPDIACYSKLLSGGLVPLAATLATQAVFEAFRGISKLDALL

[0200] HGHSYTAHAVGCASAVVSLQYFSDPSKNSNLLPGGRRLRELWKPEL

[0201] VSLISSHQSVERVVSVGTIFALELRANAAESGYASMLSKELVENLRQE

[0202] GIYTRPLGNVVYLMCGPLTQPKTCTTMLEKLLALLG

[0203] Xiaomi BioDAΔMTP (SEQ ID NO:25)

[0204] MSSAAATVPLSTPTFAVFGANTGVGKTLVSAGLVAALLNSRSPSVSA

[0205] VSYLKPLQTGYPADSDARFVFARAPALLRASSSPRATRLVASCRTLFP

[0206] SPAVGPEAEPLHQSQEKVVTYGGDGAEEETKVLACRTVYAWREPVS

[0207] PHLAAEREGMAAGDDEVRGCVDQWLMEEGVGEGGEVWKVLETAG

[0208] GVASPSASGTLQCDLYRPFRLPAILVGDGRLGGISSTLSAYETLLLRG

[0209] YDVSTVILEDRDLTNDTFLLSYLRNRVHVLALPQIPEDPSDDLTDWFS

[0210] ESSSVFNLLKDSLQSFHSKRVERLNGMQKKSKDLLWWPFTQHNLVP

[0211] QDSVTVIDSRCGENFSAYKIKDNTMMLVPQFDACASWWTQGPDSDL

[0212] QIELARDMGYAAARYGHVMFPENVHEPALRAAELLLGGVGKGWAS

[0213] RVYYSDNGSTAVEIALKMAFRKFSLDHGILADSEKSIRNERNIQLKVL

[0214] ALNGSYHGDTLGAMEAQAPSAYTSFIQQPWYSGRGLFLDPPTVCIKS

[0215] ETCNVSLPQSMQHYPRTSGDTCFPSLAEVFCKTRDTSSAADLYLSYIK

[0216] QQLSEFSQSSNSDHLAALIIEPVIQGAGGMLMIDPLFQRVLVRECRSR

[0217] KIPVIFDEVFTGFWRLGVESASELLGCLPDIACYAKLMTGGIVPLAAT

[0218] LATEEVFESFKSDSKLTALLHGHSYTAHAMGCMAALKAIQWYRDPS

[0219] TNSNLDLDHMKLKELWDGALVKQLSSLPNVKRVVSLGTLCAIELKA

[0220] EGSDAGYASLYASSLVQQLRREDDIYIRPLGNVIYLMCGPCTPRDSCS

[0221] RQLHKVLRRLCDLN

[0222] Phoenix dactylifera BioDAΔMTP (SEQ ID NO:26)

[0223] MPPELDLSHPTLVVWGSNTGVGKTIVSAGLAASVLSPSSPNTATFLY

[0224] LKPVQTGFPRDSDSRFVYRKVSDLFRHRLGGFSAQLLASNHALKASV

[0225] PAAREVLGARTERPGEEGEEASTGLCCYEETRVGTGESTTGDFKLV

[0226] CKTLYGWKEAISPHLAVEREGMAVEDSSLREVLGRLLNSSMGGGGD

[0227] RRKENVWRVIETAGGVASPGPSGSIQCDLYRPFRLPAVLVGDGHLG

[0228] GISTTLSAYESLTLRGYDVIAIILADHGLSNEIPLLSNLRHRVPVLVLP

[0229] PVPEDPLNDLVDWFCESSKAFSSLQDLVQSTHSKRIQRLHDMPRKAG

[0230] NILWWPFTQHKLVPEEIVTVIDSRCGENFAVHKVRNDREMIVRQFD

[0231] ACASWWTQGPDFALQIELARDMGYSAARYGHVMFPENVYEPALHC

[0232] AELLLEGVGKGWASRTYYSDNGSTAIEIALKMAFRKFLLDNEMLMD

[0233] SGNGISSERCIELRVLALNGSYHGDTLGAMEAQAPSSYTGFLQQPWY

[0234] SGRGLFLDPPTIFIHKEIWNLSVPESFESDQLKPEDMRFSSSDALFCPS

[0235] RDSSAIAGIYSAYISNQLLQFSKLSCSTQIGALIIEPVIQAAGGMHMID

[0236] PLFQRILVRECRNRRIPVVFDEVFTGFWRLGKESAAELLGCVPDVAC

[0237] FGKLMSGGIIPLAVTLTTEAVFEAFKGDSKLMALLHGHSYSAHAMG

[0238] CTAAAKSIQWFRDPFTNPNVEFKEKKLKELWEMKLVLQISSLPAVQ

[0239] RVITLGTLCAVELKAEGSDAGYASLYASLLVQQLREDGLYVRPLGN

[0240] VIYLMCGPCTSSHFCSQQLQQVYQRLHEFSQHQLRDSQSFQREG Chlorella minutissima ATCC 22665 BioDAΔMTP (SEQ ID NO:27) MLGSASDGAASGNGRALELSIASFAIWGANTGVGKTLASAGLARAATRRGTATTFLKPVQTGYPEDSDAGFVARQTKGNETAGEHAAVASGSTEGGGEGFAAGVTRAHTEFAWRRAAGPHLTAAEEGRPVSDEEIVRATRNYLREFAESAREGNARGFAVIETAGGVASPGPSGTLQCEILRALRLPAILIGDGRLGGISTTTSAYESLHARGYDVVGVLMADDGYANHEAVAKILPSGTKVFALPPLPPQGKADAWLEKSAGVFSEAFETAMNWHEKRIQALQKLPGEALEKIWWPFTQHATVEPDTVTVIDGRYGEDFEIYDQEKGRDGAIDLRFDGAASWWTQGVSAELQQELNSAAANAAGRYGHVMFPENVHQPSMDATNALLEGPGKGWGSRVFYSDNGSTATEVGLKMAFRKYYVDAGLVSKDGHQRAKDLINDGSCHLPQLRIIALDGSYHGDTLGAMDMQSPSVFTGPMQTPWYQPRGLFLQPPSLAIRNGEWTVALPPHGLLNDGEDRLFHDGKDLESNTTWKSKADAFDIGKRLDSPLANEYRQAVDKALERAEQDAKAGHMGPVGALMMEPVLHGAGGMVLIDPLFQNILMKVCRERKIPIVLDEVFAGIWRLGTEGAWEIIGEKPDISCYAKLLTGGLMPMSATVATADVFDVFYGPGKAQGLLHGHSYTAYPIGCAVASKALSLYKDPAKNPNLIVDASSKGDTPTAYLRELWDESKLKEISNKSSVRRVFGQGCVLAVELEVEGEGGYSSNAAKEVVLRLRPHSVQARPLGNVVYLMCAPTTPKATCDRLLNVLAEELSE

[0241] Helianthus annuus BioDAΔMTP (SEQ ID NO:28)

[0242] MLSTLSHTLSHPIYTIWSANTGLGKTLVSAGIAAASVASPPSKLVYIK

[0243] PIQTGYPLDSDAAFVYRKIPQILHRRNNPNLTLTASNQTLNVSIPAAKV

[0244] KIGDNFSDSRNGFGEFGFCEEMKVGEGVGEGSELICKTIYAWNEAIS

[0245] PHLASEREGALVGDEEVLVLLKKCLEIGVGSEGQGSVLSLVETAGG

[0246] VASPGSSGSLQCDLYRPFRLPSILVGDGKLGGISGTISAYESLTIRGYD

[0247] VVAIVLEDHGLENEVPLLSYLRNRVPVLVLPSIPKDPLDNLMEWYDE

[0248] SQSVFDLSKGIMLSSYEKQMYRLHEMPKKAQEIFWWPFTQHKLVPQ

[0249] EKVTVIDSRCGENFAVHKVKDHDYITQMFDACASWWTQGPNAALQI

[0250] KLAREMGYTAARYGHVMFPENVYEPALQCAELLLQGVGKGWASR

[0251] VFFSDNGSTAIEIALKMAFRKFLADNKLLMDLPHYNTDESNIELKVV

[0252] ALNGSYHGDTLGAMDAQAPSPYTGFLQQPWYTGRGVFLDPPFVSM

[0253] CDGVWKIAVPKKMHYEDVILEDTSFNSRDQVFNRSRDSKLASIYST

[0254] YISQELLPSSGSTKCSYAGALIIEPVIQGAGGMLMVDPLFQRILVKEC

[0255] KNRKIPVIFDEVFTGYWRLGVESAAELLFCQPDIACFAKLMTGGMIP

[0256] LAATLATNDVFESFVGDSKLNALLHGHSYSAHAVGCATACNSIKWF

[0257] KDSQTNPNLVPGQNLLRELWDAELVRKISFHPAVQRVVSLGTLFALE

[0258] LKVEDNDAGYASLSGTSLLLKLREDGIYMRPLGNVIYMMCGPCTSH

[0259] RVCLQMLEKLYMRLDEFKQEKIGTARDYDHDHRLPIA

[0260] Quercus robur BioDAΔMTP protein (SEQ ID NO:29)

[0261] MSTTAHSQLHVPLNHPTFLVWASNTSLGKTLVSTGLAASSLLNPTPK

[0262] PRKFLYLKPIQTGFPSESDSSSVFHKLSRLSLLRRHNLPHFSLFSSNHI

[0263] LKASFPAANSALGQEISETPHSGMRDLGYYEEAKPEGDVGGGAASQ

[0264] LVCKTLYAWREAVSPHLAAERESGVVEDAAVLELLQKCLRFELEGG

[0265] GEKGKETEIFCVVETAGGVASPGPSGSLQCDLYRPFRLPGILVGDGR

[0266] LGGISGTISAYESLKLRGYDIAAVVFEDHGLVNEMPLLSYLRNRVPV

[0267] LVLPSVPKDLSNDLMEWFDESRGVFDSLKEEMLSAYSKRMMRLQD

[0268] MPKKAGEIFWWPFTQHQLVPEEAVTVIDSRHGENFAVFKAQNNEFI

[0269] TQQFDACASWWTQGPDTTLQTELARDMGYAAARFGHVMFPENVY

[0270] EPALECAELLLEGVGKGWASRTFFSDNGSTAIEIALKMAFRKFSYDH

[0271] GILSDGRKDNTASQRVKLMVLALQGSYHGDTLGAMEAQAPSSYTGF

[0272] LQQPWYTGRGLFLDPPTVYMCNGVWKLSLPEGLHLEIPKLENKGFS

[0273] SRDEIFHKIRDKSDLARIYSSYISEQLSQYSGSGGFYHIGALIMEPVIL

[0274] GAGGMHMIDPLFQRVLVNECRSKKIPVIFDEVFTGFWRLGRETAAE

[0275] LLGCVPDIACFAKLMTGGIIPLAATLATNAIFNSFIGESKLKALLHGH

[0276] SYSGHAMGCTAAVKSIKWFKDPQTNLNITDEGRSLRELWDEDVVQQ

[0277] ISSHPAVERVVALGTLFALELRAEGSNAGYASQYASSLLQNLRQDGI

[0278] YTRPLGNVIYLMCGPCTSPEICSQLLVKLYKRLEDFDQFHQIREENSE

[0279] SYGG

[0280] Thielavia bacillispora BioDAΔMTP (SEQ ID NO:30)

[0281] MSLDQSVPNAVPTSLHAPIVQVFGSNTDVGKTVIAAGLCRAALLSSK

[0282] NATVGYIKPLQTGGDAMMDAEFLNRHAGNADNRLTSHTLFSWETA

[0283] VSPHLAASLENKAVDDATVVAKLSKTLKQIQKEEKGSALTVVETAG

[0284] GVCSPSASGKFQCDVYRPLRLPTVLVGDGCLGGISATMSALDSLLIR

[0285] GYDVSCIVLIEQDNLSNGEAISERASELGIPIFSLDKLPPQPEPLDLWY

[0286] ENSTETFRSVHDTIASTHKARVARLQELPSIAKKALWWPFTQHKSNK

[0287] GVLVIDSAYGDTFATYNQKNQAIEPMYDACASWWTQGVGHGNAKM

[0288] ALSLATAAGRYGHVMFPENAHEPAIALSEMLLKSVGKNWAGKVFFS

[0289] DDGSTAVEIGIKMALRKYAVDHGLGMSFEGINDVLVLAQEGCYHGD

[0290] TLGTMNIAEPSVYNKTQHPWYKPKALFVHPPSVKVASNGQMQVTW

[0291] QNVNPTLVTHLPTLDDLFDISRDTSPLTQQYTEYVESLLDSVPSTTVV

[0292] GALIVEPILIGAGGMVFVDPLFQRTMVRVCQQRKIPVIFDEVFSGFW

[0293] RLGAESARDLLQVNPDIACYAKLLTGGAVPMSTTLATNEVFDAFYT

[0294] DSKADALLHGHSFTANPVGCAAAVTALDMYKTIEKKNSTRVHWNV

[0295] DQVAALAARPGIARAFQLGTVLVIELESTSRGYTDTTGQDVLVRLRN

[0296] HGIYARSLGNVLYIMASPLSTPATCIKIFNTVVDNLFA

[0297] Gibberella zeae BioDAΔMTP (SEQ ID NO:31)

[0298] MCNAASKKGDRTWFLKPVSTGAVDEADGCHIQRFASSTNHETLFQY

[0299] DIPCSPHIAAKVSGKPIPSDEAVLAKIHDSVSRYASEGPGWLFLETAG

[0300] GVHSPGPSGTPQADLYTPLRAPVILVGDSKLGGISQTISAYESLRMRG

[0301] HDIESILLFQDMKYENYQYLKDYFSKQGGIPVDTVPEPPSRLPDIQQD

[0302] TEQMREYYASQTSNVSHVLDHLDKRGKQRISRLESLSEKASKSIWYP

[0303] FTQQKLVTADTISAIDSAHGDYFQVLNKTYENLLQPAFDGSASWWS

[0304] QGLGHANSRLTLAAAYAAGRYGHVMFAEAIHEPALALAEMMLEGA

[0305] QNSRFSRVFYSDNGSTGCEVAVKMALRAARLRYGWGPNDNLQILGL

[0306] KGSYHGDTIGAMDCAEPCVYNEKIEWYEGKGYWFDYPTIQCVEGK

[0307] WVVSAPNGMGQDQKLESISEVFNLESRLKTEQYRGYEEYIEKTLKK

[0308] LQDQGRKFGALMMEPIILGAGGMIFVDPLFQRALVDVVRRSPHLFG

[0309] TKDALPAQGPQWSGLPVIFDEVFTGLYRLGRFTAASFLGTEADISVN

[0310] AKLLTGGLVPLCTTMASESIFDAFKSDDKSDALLHGHSYTAHAVGC

[0311] QVAVESVSEMQSMEKNGEWEWAEKDWAEDETQAWSVWSRDFVTD

[0312] MSHNPQVLGVWALGSVLAISLRDDDGVGYKSLAAKKIQSHLRQGTG

[0313] SWNAHSRVLGNVFYVMASQKTSRQSIRELQGLLVDALSK

[0314] Barley BIO3 - BIO1ΔMTP (SEQ ID NO:32)

[0315] MSSSFSSALPLSSPTFAVFGANTGVGKTLVSAGLAASLLRSHAASPSA

[0316] VLYLKPLQTGFPDDSDAGFLYRRVPALLRPSHAAHLAASIDTLSPAPS

[0317] VETQSPSREEAAFSYDGEGEAVRKLLACRTLYAWREAVSPHLAAER [[ID=I8]]

[0318] EGMAVEDAHVRSLLAACLVEGENLGKGEGDVWKLVETAGGVASPG

[0319] PSGTLQCDLYRQFRLPTILVGDGRLGGISCTLSAYETLLLRGYDVSA

[0320] VILEDHGLSNNKFLLSYLRNRVPVLVLPPIPEDPSDDLTDWFSESSSVF

[0321] GLLEDALRSFHLTRIQRLNSMQRKSKDLLWWPFTQHNLVPVDCVTV

[0322] IDSRCGESFSAYKVKDNKMMMIPQFDACASWWTQGPDSKLQIELAR

[0323] DMGYAAARYGHVMFPENAHEPALRCAELLLGGIGKDWASRVYYSD

[0324] NGSTAIEIALKMAFRKFSLDRGILLDSDKSIITNGRNIQLKVLALKGS

[0325] YHGDTLGAMEAQAPSAYTSFLQQPWYSGRGLFLDPPTVFIKNGTCT

[0326] LSLPQAIQNCHLSPGDKCFPSLAEVFCKTRDSSSAADLYSTYISQQLS

[0327] EYSLSSNTEHIAALIIEPVIQGAGGMHMIDPLFQRVLVHECRDRKIPVI

[0328] FDEVFTGFWRLGVESASELLGCLPDVACYAKLMTGGLVPLAATVTT

[0329] EAVFEAFKSDSKLTALLHGHSYTAHAMGCSAAVKAIQWFRDPSTNS

[0330] NLDFDCMKLKELWDGTLVNQLSSLPNVKRIVSLGTLCAIELQAEGSD

[0331] AGYASLYARSLIQQLREEDDIYVRPLGNVIYLMCGPCTTRDVCTEQL

[0332] SKVYRRISHFNPVHRG

[0333] Brassica napus BIO3 - BIO1ΔMTP (SEQ ID NO:33)

[0334] MSTAVPSHFNLPLNHPTYLIWSANTSLGKTLVSTGIASSFLLHQPSSSP

[0335] PAAHSAKLLYLKPIQTGFPSDSDSRFVFSKLDSLSLRRRIPLSVSNSVL

[0336] RSSLPVAESMRRNIKVSESGMCDLNFREEKTVTGAPELLCKTLYAW

[0337] EAAISPHLAAEREHATVEDSVVLKMVEQEMECGSKANVLCLVETAG

[0338] GVASPGPSGTLQCDLYRPFRLPGILVGDGRLGGISGTIAAYESLKLRG

[0339] YDVAAVVFEDHGLVNEVPLTSYLRNKVPVLVLPPVPKDPSDDLIEWF

[0340] VESDGVFKALKEVMVSVYLERVERLNGMAKQAGEVFWWPFTQHK

[0341] LVPEDNVTIIDSRCGENFSVFKASDNNSITQQFDACASWWTQGPDPA

[0342] FQAELAREMGYTAARFGHVMFPENVYEPALKCAELLLDGVGKGWA

[0343] SRVYFSDNGSTAIEIGLKMAFRKFCVDHETLFGLSEDAEEKKHVDVK

[0344] VLALRGSYHGDTLGAMEAQAPSPYTGFLQQPWYTGRGLFLEPPTVF

[0345] LSNGTWNLSLPESFSQTASEECGTFTTRDEIFDKSRDTSVLATVYSTY

[0346] VSEQLQEYSGISQSAHVGALIIEPVIHGAGGMHMVDPLFQRVLVNEC

[0347] RNRKIPVIFDEVFTGFWRLGVETTADLLGCKPDIACFAKLLTGGMIP

[0348] LAVTLATDAVFDSFSGDSKLKALLHGHSYSAHAMGCATATKAIEWF

[0349] KDPETNHNIDSQRGTLRELWDEELVQHISCHCAVERVVVLGTLFAL

[0350] ELKVDASNSGYASLYAKSLLEMLREDGIFMRPLGNVVYLMCGPCTS

[0351] PEICRRLLTKLHKRLGEFNRA[[ID=​​​​​​​PLQTGFPSDSDSRFLFQKLSSLSLSRNLPLFSSHSVLLSSLPAAKSFKP

[0355] NEFSLNESREMCELGFYEEKKVLEAGRVAPELVSETIYAWEGALSPH

[0356] FAAEREGGAVEDSELVKRLETRLKEGLLEGGVERGKLDGFCVVETA

[0357] GGVASPGPSGTLQCDLYRPLRFPGVLVGDGRLGGISGTISAYESLKL

[0358] RGYDVVAIVIEDHGLINEIPLKSYLRSRVPVFVLPPIPQDPSNDLMEW

[0359] FDESCNVFNSLKDIMLEAYLERMRRLNEMPRKAGDVFWWPFTQHK

[0360] LVPPSTVTVIDSRCGENFSVYKVQNKEYITQQFDACASWWTQGPDA

[0361] TLQTELARDMGYAAARFGHVMFPENVYEPALECAELLLDGVGKGW

[0362] ATRVYFSDNGSTAIEIALKMAFRKFSSDHGILPELLKNNPTESCTELM

[0363] VLALKGSYHGDTLGAMEAQAPSSFTGFLQQPWYTGRGLFLDPPTVF

[0364] MHNGKWTVSLPEVFHSATLKPEDTTFRSRDEIFLKSRDESNLAGLYS

[0365] LYVSQQLSQYSTLSETNKQIGALIMEPIIQGAGGMHMVDPLFQRML

[0366] VNECRHCRIPVIFDEVFTGFWRLGVESAAELLGCVPDIACFAKLMTG

[0367] GIIPLAATMATDAIFDSFTGDSKLKALLHGHSYSAHAMGCTAAAKSI

[0368] KWFKDPNTNLNITSEKSQKMLLRELWDAELVQQFSSHPSISRVVTLG

[0369] TLFALELQADESDAGYASLYARSLVQMLREDCIYTRPLGNVIYVMC [[ID=,7]]

[0370] GPCTSPKMCTLQLLKLYTKLEEFTLVKTKMNII

[0371] Oryza BIO3 - BIOY1ΔMTP (SEQ ID NO:35)

[0372] MSTSSSSSAAAAAVPLTSPAFAVFGANTGVGKTLVSAGLVASLLASPS

[0373] PSPSTVAYLKPLQTGFPDDSDARFVFDRAPALLRRLRLAGGGASTRL

[0374] VASNHTLFPSPAVDPLPERQDTVVNYGGEEGVEEKALVCRTVYAWR

[0375] EPVSPHLAAEREGMPVEDEEVRWLVDRWLAEEDGGGEVWKVLETA

[0376] GGVASPGPSGTLQCDLYRSSRLPAVLVGDGRLGGISSTLSAYETLLL

[0377] RGYDVGSVILEDRGLSNDRFLLSYLRKRVPVHVLPPIPEDPKDDLTD

[0378] WFSESSSAFSSLKDSLQSFHSRRVQRLNSMQRKSKYLLWWPFTQHD

[0379] LVPVDSVTVIDSRFGENFSAYKVKDKTIVPQFDACASWWTQGPDSNL

[0380] QIELARDMGYAAARYGHVMFPENVHEPALRCAELLLGGVGKDWAS

[0381] RVYFSDNGSTAIEIALKMAFRKYACDHGIIVDSEKDIRSEGSVHFKVL It should be noted that "稻BIO3 - BIO1ΔMTP" is translated as "Oryza BIO3 - BIOY1ΔMTP" here. It's possible that there is an error in the original text or a specific context - related naming convention that needs further clarification. If this is a biological or technical term with a more specific standard translation, it may need to be adjusted accordingly.

[0382] ALNGSYHGDTLGAMEAQAPSAYTSFLQQPWYSGRGLFLDPPTVYIK

[0383] NKSANLSLPPSIMHDQLSSCDTCFSSLTEVFCKTRDTSSAANVYVSYIS

[0384] QQLSQYAMSNNSEHIAALIIEPVIQGAGGMHLIDPLFQRLLVKECKN

[0385] RKIPVIFDEVFTGFWRLGVESASELLGCFPDISCYAKLMTGGIVPLAA

[0386] TLATEPIFEAFRSDSKLTALLHGHSYTAHPMGCTAAVKAIQWYKDPS

[0387] TNSNIDLDRMKLKELWDSALVNHLSSLPNVKRVVSLGTLCAIELKAE

[0388] GSDAGYASLYASSLIRQLREEDNIYARPLGNVIYLMCGPCTTQDSCT

[0389] RQLAKVHRRLQKLN

[0390] Soybean BIO3 - BIO1ΔMTP (SEQ ID NO:36)

[0391] MSSSTTSHPPSLPLSHPIYLIWGSNTGVGKTLVSAGIAAAFLLSSPTPS

[0392] QFHYLKPLQTGFPSDSDSRFVFNKLRQLSLIRNPHIPLSASHRVINVSP

[0393] AVTASNPLAGEEGVGPPPELLCKTLYAWEEAVSPHLAAEREGLVVK

[0394] DSAVLETLGECLEDVAECGAGKERSEVLCVVETAGGVASPGPSGSL

[0395] QCDLYRPFRIPAVLVGDGRLGGISGTISAYESLKLRGYDVVAVVFED

[0396] HGLLNEGPLMSYMRNKVPVLVLPPIPKDPLNDLMEWFESSHYIFSNL

[0397] KEIMLSAYFERIKKLHDMPREARDIIWWPFTQHKLVPDGGVTVIDSR

[0398] CGENFSIFKVQKTEVIAPQFDACASWWTQGPDAIMQTELAREMGYA

[0399] AARFGHVMFPENVHEPALNCAELLLQGVGKGWASRTYFSDNGSTAI

[0400] EIALKMAFRKFSVDHGLIVHCHEDTTNERSTELMVLALQRSYHGDT

[0401] LGAMEAQAPSSYTGFLQQPWYTGRGLFLDPPSVFMHNSTWNVSIPE

[0402] GYHLESLKGKSITFASPDEIFHKGRDKSELATVYSSYISKVLSGFRGS

[0403] NNVGALIMEPVIQGAGGMHMVDPLFQRVLVNECRSRKIPVIFDEVFT

[0404] GFWRLGVETAVELIHCVPDIACFGKLLTGGIIPLAVTLATNAVFDSFI

[0405] GDSKLKALLHGHSYSAHAMGCAAAVKSIQWFKDPCSNPNITSEGRL

[0406] LRELWDDKMVHRISSHPAIQRVVALGTLCALELKAEGNNAGYGSLY

[0407] ARPLLEKLREDGVYMRPLGNVIYLLCGPCTSPEVCNQLLVKLLRRL

[0408] EEFDVGKN

[0409] Common wheat BIO3 - BIO1ΔMTP (SEQ ID NO:37)

[0410] MLSSPTFAVFGANTGVGKTLVSAGLAASLLRSHAASPSAVLYLKPLQ

[0411] TGFPADSDAGFLYRRVPALVRPSHAAHLVASIDTLSPAPSVETPSPSR

[0412] EEAAFSYDGEGKAVRKLLACRTLYAWREAVSPHLAAEREGMTVED

[0413] AHVRSLLAACLAEGETLGKGEGDVWKLVETAGGVASPGPSGTLQC

[0414] DLYRHFRLPIILVGDGRLGGISGTLSAYETLLLRGYDVSAVILEDCGL

[0415] SNDKFLLSYLRNRVPVLVLPPIPEDPSDDLTDWFSESSSVFGLLEDAL

[0416] RSFHLKRIQRLNSMQRKSKDLLWWPFTQHNLVPVDCVTVIDSRCGE

[0417] SFSAYKVKDNKLMMIPQFDACASWWTQGPDSKLQIELARDMGYAA

[0418] ARYGHVMFPENAHEPALRCAELLLGGIGKDWASRVYYSDNGSTAIEI

[0419] ALKMAFRKFSLDRGILLDSDKSITNERNIQLKVLALKGSYHGDTLGA

[0420] MEAQAPSAYTSFLQQPWYSGRGLFLDPPTVFIRNGTCALSLPQSIQN

[0421] CHLSPGDKCFPSFAEVFCKSRDSSAAADLYSTYISQQLSEYSMSSNTE

[0422] HIAALIIEPVIQGAGGMHMIDPLFQRVLVHECRDRKIPVIFDEVFTGF

[0423] WRLGVESASELLGCLPDVACYAKLMTGGIVPLAATVTTEAVFEAFK

[0424] SDSKLTALLHGHSYTAHAMGCSAAVKAIQWFRDPSTNSNLDFDCMK

[0425] LKELWDGTLVNQLSSLPNVKRVVSLGTLCAIELQAEGSDAGYASLY

[0426] ARSLIQQLREEDDIYVRPLGNVIYLMCGPCTTRDVCTEQLSKVYRRIS

[0427] HFNPIH

[0428]

Claims

1. An in vitro method for determining whether a candidate compound is a diaminononanoic acid aminotransferase (DAPA-AT) inhibitor, the method comprising (i) assaying the candidate compound in an assay mixture comprising (a) the DAPA-AT, (b) an amine donor, (c) pyridoxal 5'-phosphate (PLP), and (d) 7-keto-8-aminononanoic acid (KAPA), and (ii) detecting inhibition of the DAPA-AT by the candidate compound; The amine donor comprises an aliphatic diamine or an aliphatic polyamine or a carbamate derivative thereof. 2 . The method according to claim 1 , wherein the DAPA-AT is plant BIO1 or a functional derivative or fragment thereof.

3. The method of claim 1 or claim 2, wherein the DAPA-AT is provided as a bifunctional BIO3-BIO1 enzyme or a functional derivative or fragment thereof.

4. The method according to any one of the preceding claims, wherein the DAPA-AT is Arabidopsis BIO3-BIO1 or a functional derivative or fragment thereof.

5. The method of claim 3, wherein the DAPA-AT is SEQ ID NO: 21 or a functional derivative or fragment thereof.

6. The method of any one of the preceding claims, wherein the DAPA-AT is recombinantly produced.

7. The method according to any one of the preceding claims, wherein the amine donor is a compound of formula (I): wherein X is NH or CH2; R 1 is hydrogen or -[CH2]pNH2; m and n are independently selected from 0, 1, 2, 3 or 4; and p = 1, 2, 3 or 4, or its carbamate derivatives, wherein R 1 It is -C(O)OH.

8. The method according to any one of the preceding claims, wherein the amine donor is selected from the group consisting of cadaverine, putrescine, spermidine and spermine.

9. The method of claim 8, wherein the amine donor is spermidine.

10. The method of any preceding claim, wherein detecting inhibition comprises monitoring the formation of diaminononanoic acid (DAPA).

11. The method of any one of the preceding claims, wherein the method utilizes an assay mixture wherein the candidate compound is first incubated with the DAPA-AT (a) and PLP (c) in an enzyme mixture, and then the enzyme mixture is combined with a substrate mixture comprising the amine donor (b) and the 7-keto-8-aminononanoic acid (KAPA) (d).

12. The method according to any one of the preceding claims, wherein the formation of DAPA is monitored by converting the DAPA into a fluorescent derivative.

13. The method according to claim 12, wherein the fluorescent derivative is DAPA: o-phthalaldehyde.

14. The method of claim 3, wherein the assay mixture further comprises (e) ATP and (f) CO2, and detecting inhibition comprises monitoring the formation of DAPA and / or desthiobiotin (DTB).

15. Use of an amine donor in an in vitro diaminononanoic acid aminotransferase (DAPA-AT) assay, the amine donor comprising a compound having formula (I): wherein X is NH or CH2; R 1 Selected from hydrogen, -C(O)OH and -[CH2] p The group composed of NH2; m and n are independently selected from 0, 1, 2, 3 or 4; and p = 1, 2, 3, or 4; or its carbamate derivatives, wherein R 1 It is -C(O)OH.

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