CN113817696B - Amine oxidase ASAO, preparation method and application - Google Patents

Amine oxidase ASAO, preparation method and application Download PDF

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CN113817696B
CN113817696B CN202111133638.9A CN202111133638A CN113817696B CN 113817696 B CN113817696 B CN 113817696B CN 202111133638 A CN202111133638 A CN 202111133638A CN 113817696 B CN113817696 B CN 113817696B
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asao
amine oxidase
gly
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thr
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CN113817696A (en
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李中媛
夏忠强
张同存
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Tianjin University of Science and Technology
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
    • C12N9/0014—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on the CH-NH2 group of donors (1.4)
    • C12N9/0022—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3)
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    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
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    • C12Y—ENZYMES
    • C12Y104/00—Oxidoreductases acting on the CH-NH2 group of donors (1.4)
    • C12Y104/03—Oxidoreductases acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3)
    • C12Y104/03006—Amine oxidase (copper-containing)(1.4.3.6)

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Abstract

The invention relates to an amine oxidase ASAO, the amino acid sequence of which is SEQ ID NO.1. The degradation rate of the amine oxidase ASAO to HFB1 is 100 percent; the amine oxidase ASAO has excellent property, and can be applied to industries such as agriculture, feed, food and the like, so that the damage of fumonisin FB1 to animal and human health is reduced.

Description

Amine oxidase ASAO, preparation method and application
Technical Field
The invention belongs to the technical field of agricultural biology, and particularly relates to an amine oxidase ASAO, a preparation method and application thereof.
Background
Fumonisins (FB) is a structurally similar secondary metabolite consisting of different polyols and tricarballylic acids produced by fungi of the genus fusarium and the like under specific conditions. Grain crops are a necessity for human survival. However, some mycotoxins are particularly prone to contaminating food crops, and are not well removed when these food crops are processed into some produce, and can enter the human body along with the food chain, causing some harm to the human body. Fumonisins are mainly found in animal feed raw materials and in their preparations, can cause acute poisoning of certain domestic animals, and have potential carcinogenicity to most poultry, and at the same time have the possibility of human infection, and some researches indicate that fumonisins are closely related to the onset of human esophageal cancer. Based on many researches, the fumonisins have the characteristics of wide distribution, large harm and the like, so that the fumonisins are necessary to be effectively prevented and controlled.
The currently known degradation methods of fumonisins mainly comprise physical, chemical and biological degradation. Since biodegradation has more advantages over physical and chemical degradation, the research associated therewith remains relatively few and the biodegradable methods available for commercial use are more rare. The biological degradation is to clone genes related to fumonisin detoxification enzyme separated from microorganisms, and construct genetic engineering bacteria by using genetic engineering technology, so that the expression related proteins of the genetic engineering bacteria degrade toxins, and the biological degradation has a good effect.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides an amine oxidase ASAO, a preparation method and application thereof, wherein the amine oxidase ASAO can degrade amino groups in fumonisins to relieve toxicity.
The technical scheme adopted for solving the technical problems is as follows:
an amine oxidase ASAO having the amino acid sequence of SEQ ID No.1.
The preparation method of the amine oxidase ASAO comprises the following steps:
(1) Recombinant strains are obtained by adopting a molecular biological means to recombine genes containing the coded amine oxidase ASAO into a vector and then transforming host cells;
(2) Culturing the recombinant strain and inducing the strain to express amine oxidase ASAO;
(3) Isolating and purifying the amine oxidase ASAO;
(4) Protein expression is verified to be correct through protein immunoblotting western blot.
And, the host cell in the step (1) is an E.coli cell, a Saccharomyces cerevisiae cell, a Pichia pastoris cell or a polytype of a yeast cell.
Use of an amine oxidase ASAO as described above for degrading fumonisins.
An amine oxidase ASAO gene encoding an amine oxidase ASAO as described above.
Furthermore, the nucleotide sequence of the gene is SEQ ID NO.2.
A recombinant vector comprising an amine oxidase ASAO gene as described above.
Recombinant vector pET28a (+) -ASAO comprising an amine oxidase ASAO gene as described above.
A recombinant strain comprising an amine oxidase ASAO gene as described above.
Moreover, the recombinant strain is E.coli.
The invention has the advantages and positive effects that:
1. the amine oxidase ASAO of the invention can degrade and catalyze the amino group of the fumonisin FB1 (HFB 1) to hydrolyze the fumonisin, and eliminate the toxicity of the fumonisin. Since amino groups are one of the key functional groups that make fumonisins toxic, removal of amino groups is a key point for detoxication of fumonisins. The amine oxidase ASAO has the activity of degrading and hydrolyzing fumonisin HFB1, can be applied to industries such as agriculture, feed, food and the like, and reduces the harm of the fumonisin to animal and human health.
Drawings
FIG. 1 is a SDS-PAGE purification of an amine oxidase ASAO of the present invention; wherein M: protein marker;1:300mM imidazole eluted protein;
FIG. 2 is a graph showing the western blot results of the amine oxidase ASAO of the present invention, wherein M: protein marker;1, amine oxidase ASAO;
FIG. 3 is a schematic diagram showing the degradation effect of amine oxidase ASAO in the present invention, wherein FIG. A is a graph showing the liquid phase result of standard hydrolysis of fumonisins, and FIG. B is a graph showing the liquid phase result of degradation of hydrolysis of fumonisins by amine oxidase ASAO;
Detailed Description
The following describes the embodiments of the present invention in detail, but the present embodiments are illustrative and not limitative, and are not intended to limit the scope of the present invention.
The raw materials used in the invention are conventional commercial products unless specified; the methods used in the present invention are conventional in the art unless otherwise specified.
An amine oxidase ASAO having the amino acid sequence of SEQ ID No.1.
The preparation method of the amine oxidase ASAO comprises the following steps:
(1) Recombinant strains are obtained by adopting a molecular biological means to recombine genes containing the coded amine oxidase ASAO into a vector and then transforming host cells;
(2) Culturing the recombinant strain and inducing expression of amine oxidase ASAO;
(3) Separating and purifying amine oxidase ASAO;
(4) And verifying correct protein expression by western blot.
Preferably, the host cell in the above step is an E.coli cell, a Saccharomyces cerevisiae cell or a polytype yeast cell, preferably E.coli BL21 (DE 3).
Use of an amine oxidase ASAO as described above for degrading fumonisins.
An amine oxidase ASAO gene encoding an amine oxidase ASAO as described above.
Preferably, the nucleotide sequence of the gene is SEQ ID NO.2.
A recombinant vector comprising an amine oxidase ASAO gene as described above.
Recombinant vector pET28a (+) -ASAO comprising an amine oxidase ASAO gene as described above.
A recombinant strain comprising an amine oxidase ASAO gene as described above.
Preferably, the recombinant strain is E.coli.
Specifically, the amino acid sequence of the amine oxidase ASAO is shown in SEQ ID NO. 1:
SEQ ID NO.1：
MDAFDKAPRVYDVVVVGAGLSGLQAAHSICAAGFSVCVLEATNRVGGKTLSVRSCEKGINDLGAAWINDTNQSEMFKLHQRYSLDGEIQYTHGDDIIQSTDGLICKIPYGQTPLSTGLVEDLVRILREKSAPIDLHNPASSPAAKEVDNETFQDFCIRRTGCEDAVHIADSISAALLGVDSIEVSALYMLYYFKSGSGIDNLLSDEKNGGQYLRTRQGTQTISQKMAEELDPETLFLGTPVTSIDQSRPDGICLVQTLDNATFHCRRVIVSVPTTLYQSISFSPPLPQTKNSLSHDTVMGYYSKMIFVFNEPWWRNAGFTGIFDSADGPVSFTRDTSVPRDDQWSITCFIVGGRGRTWSKLSKADRHTQVWEQFCRYFGEFVEKIPEPANVLEMEWSKQPSFLGAPCPVMTPGTMTTVGTELATPFGRVHFVGTETSIVWRGYMEGAIRSGQRGAAEVLAALNGDLCSEA
wherein the enzyme gene encodes 470 amino acids without a signal peptide, and thus the theoretical molecular weight of the mature amine oxidase ASAO is 51.55kDa.
The present invention provides a polypeptide encoding the amine oxidase ASAO described above. The genome sequence of the gene is shown as SEQ ID NO. 2:
SEQ ID NO.2：
ATGGATGCCTTCGATAAGGCTCCAAGGGTCTACGACGTTGTTGTTGTCGGTGCCGGATTGAGTGGTCTTCAAGCTGCTCACTCTATTTGTGCCGCCGGATTCTCTGTTTGCGTCCTTGAGGCCACCAATAGAGTCGGAGGAAAGACCCTTTCTGTTAGGTCTTGCGAGAAGGGTATCAACGATCTTGGTGCTGCTTGGATCAATGACACTAACCAGAGTGAGATGTTCAAATTGCATCAAAGATATAGTCTTGACGGAGAGATTCAGTACACTCACGGAGACGACATTATTCAGTCTACCGACGGACTTATCTGCAAGATCCCATATGGACAGACCCCTTTGTCTACCGGACTTGTCGAGGACTTGGTTAGAATCTTGAGGGAAAAGAGTGCCCCAATCGACCTTCACAACCCAGCTTCTAGTCCAGCTGCCAAAGAGGTTGATAACGAAACCTTCCAAGATTTCTGCATTAGAAGGACCGGTTGTGAGGATGCCGTTCACATCGCCGATTCTATCAGTGCTGCCCTTTTGGGTGTCGATAGTATCGAGGTCTCTGCCCTTTACATGTTGTATTACTTCAAATCTGGTTCTGGAATCGATAATTTGCTTAGTGACGAGAAAAACGGAGGACAGTATTTGAGGACTAGGCAAGGAACTCAAACCATCAGTCAGAAGATGGCCGAGGAGTTGGACCCAGAAACCCTTTTCTTGGGAACTCCAGTTACCAGTATCGATCAGAGTAGGCCAGATGGTATCTGCTTGGTTCAAACCTTGGACAACGCCACCTTCCATTGTAGAAGGGTTATCGTCAGTGTCCCAACCACCTTGTACCAATCTATCAGTTTCTCTCCTCCATTGCCTCAGACCAAGAACTCTTTGTCTCATGATACTGTTATGGGATATTATAGTAAAATGATCTTCGTTTTCAACGAGCCTTGGTGGAGGAATGCCGGTTTCACCGGAATCTTCGACTCTGCTGATGGACCAGTCAGTTTCACCAGAGACACCAGTGTCCCTAGGGATGACCAGTGGTCTATCACTTGTTTCATCGTTGGTGGTAGGGGTAGGACTTGGAGTAAATTGTCTAAAGCTGATAGACATACCCAAGTTTGGGAGCAGTTTTGTAGATACTTCGGTGAGTTCGTCGAGAAGATCCCAGAGCCAGCCAATGTCTTGGAGATGGAGTGGTCTAAGCAGCCTTCTTTCTTGGGTGCTCCATGCCCAGTTATGACTCCCGGAACTATGACCACCGTCGGAACTGAATTGGCTACCCCATTTGGTAGGGTCCACTTCGTTGGAACCGAGACCTCTATTGTCTGGAGGGGATACATGGAGGGTGCCATTAGGTCTGGACAAAGGGGTGCCGCTGAAGTCTTGGCCGCCTTGAACGGTGATTTGTGCTCTGAGGCC
the invention separates and clones amine oxidase ASAO by PCR method, and the DNA complete sequence analysis result shows that the open reading frame sequence (ORF) of amine oxidase ASAO gene is 1410bp in full length.
The invention also provides a recombinant vector comprising the amine oxidase ASAO, and the preferred recombinant vector is named pET28a (+) -ASAO. The amine oxidase ASAO gene of the present invention is inserted between proper restriction enzyme sites of expression vector to make its nucleotide sequence operably linked with expression regulating sequence. As a most preferred embodiment of the present invention, it is preferable that the detoxification enzyme gene of the present invention is inserted between EcoRI and NotI restriction sites on the plasmid pET28a (+) such that the nucleotide sequence is located downstream of and under the control of the T7 promoter, to obtain the recombinant E.coli expression plasmid pET28a (+) -ASAO. More specifically, the relevant preparation and detection are as follows:
test materials and reagents:
1. strains and vectors: the invention provides a novel amine oxidase ASAO. The E.coli expression vector pET28a (+) and strain BL21 (DE 3) were stored for laboratory use.
2. Enzymes and other biochemical reagents: endonucleases were purchased from TransGen Biotech company and ligases were purchased from TransGen Biotech company. The protein intetech His tag antibody was purchased from beijing jejie science and technology, the protein marker was purchased from Thermo Scientific, the chromatographic grade methanol was purchased from Fisher Chemical, and the other were domestic reagents (all available from general biochemical reagent company).
3. Culture medium:
coli LB medium (10 g/L peptone, 5g/L yeast extract, 10g/L sodium chloride, pH 7.0).
Description: the following molecular biology experimental methods, which are not specifically described, are carried out with reference to the specific methods listed in the "guidelines for molecular cloning experiments" (third edition) J.Sam Brookfield, or according to the kit and product specifications.
1. Cloning of amine oxidase ASAO
The gene fragment of the amine oxidase ASAO is obtained by utilizing an artificial chemical synthesis method, and an endonuclease site EcoRI is introduced at the 5 'end, and an endonuclease site NotI is introduced at the 3' end.
2. Preparation of recombinant amine oxidase ASAO
The expression vector pET28a (+) is subjected to double digestion (EcoRI+NotI), the gene encoding the amine oxidase ASAO is subjected to double digestion (EcoRI+NotI), the cut gene fragment encoding the mature amine oxidase is connected with the expression vector pET28a (+) through T4 DNA ligase, and the recombinant plasmid pET28a (+) -ASAO containing the amine oxidase gene ASAO is obtained to transform the escherichia coli BL21 (DE 3), so that the recombinant escherichia coli strain BL21/ASAO is obtained.
The recombinant strain BL21/ASAO was activated overnight and inoculated into 100mL of LB medium, after shaking culture at 37℃and 220rpm for about 2.5 hours, 0.2mM IPTG was added, and induction was performed at 25℃and 220rpm, and after about 24 hours, intracellular and extracellular amine oxidase activities were measured. The activity of amine oxidase was detected in cells, and 80mM, 100mM and 300mM of imidazole were collected by nickel column purification. SDS-PAGE results show that recombinant amine oxidase is expressed. And verifying correct protein expression by further western blot. As shown in FIG. 1, lane 1 is the result after purification. FIG. 2 shows the result of the western blot verification.
3. Determination of degradation Properties of recombinant amine oxidase
The high performance liquid chromatography is used for detecting the degradability of the amine oxidase, and the specific method is as follows:
(1) HFB1 standard stock: preparing standard solution with concentration of 100 mug/mL, and preserving at-20 ℃.
(2) Preparation of the samples: taking 45 mu L of purified amine oxidase enzyme solution, adding 5 mu L of HFB1 standard stock solution to make the final concentration of HFB1 be 10 mu g/mL, placing in a 37 ℃ water bath for 10min, and then placing in an incubator for inactivating enzyme at 100 ℃ for 10min.
(3) Sample derivatization: and adding 50 mu L of acetonitrile water (V: V=1:1) into 50 mu L of a sample to be detected, mixing 250 mu L of OPA derivative liquid uniformly for 30s, sampling within 2min of derivative, and filtering to obtain the sample to be detected. The enzymatic activity of the amine oxidase ASAO was determined by comparison with the peak pattern of the standard of HFB 1.
Although embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the disclosure of the embodiments.
Sequence listing
<110> university of Tianjin science and technology
<120> an amine oxidase ASAO, preparation method and application
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 470
<212> PRT
<213> amino acid sequence of amine oxidase ASAO (Unknown)
<400> 1
Met Asp Ala Phe Asp Lys Ala Pro Arg Val Tyr Asp Val Val Val Val
1 5 10 15
Gly Ala Gly Leu Ser Gly Leu Gln Ala Ala His Ser Ile Cys Ala Ala
20 25 30
Gly Phe Ser Val Cys Val Leu Glu Ala Thr Asn Arg Val Gly Gly Lys
35 40 45
Thr Leu Ser Val Arg Ser Cys Glu Lys Gly Ile Asn Asp Leu Gly Ala
50 55 60
Ala Trp Ile Asn Asp Thr Asn Gln Ser Glu Met Phe Lys Leu His Gln
65 70 75 80
Arg Tyr Ser Leu Asp Gly Glu Ile Gln Tyr Thr His Gly Asp Asp Ile
85 90 95
Ile Gln Ser Thr Asp Gly Leu Ile Cys Lys Ile Pro Tyr Gly Gln Thr
100 105 110
Pro Leu Ser Thr Gly Leu Val Glu Asp Leu Val Arg Ile Leu Arg Glu
115 120 125
Lys Ser Ala Pro Ile Asp Leu His Asn Pro Ala Ser Ser Pro Ala Ala
130 135 140
Lys Glu Val Asp Asn Glu Thr Phe Gln Asp Phe Cys Ile Arg Arg Thr
145 150 155 160
Gly Cys Glu Asp Ala Val His Ile Ala Asp Ser Ile Ser Ala Ala Leu
165 170 175
Leu Gly Val Asp Ser Ile Glu Val Ser Ala Leu Tyr Met Leu Tyr Tyr
180 185 190
Phe Lys Ser Gly Ser Gly Ile Asp Asn Leu Leu Ser Asp Glu Lys Asn
195 200 205
Gly Gly Gln Tyr Leu Arg Thr Arg Gln Gly Thr Gln Thr Ile Ser Gln
210 215 220
Lys Met Ala Glu Glu Leu Asp Pro Glu Thr Leu Phe Leu Gly Thr Pro
225 230 235 240
Val Thr Ser Ile Asp Gln Ser Arg Pro Asp Gly Ile Cys Leu Val Gln
245 250 255
Thr Leu Asp Asn Ala Thr Phe His Cys Arg Arg Val Ile Val Ser Val
260 265 270
Pro Thr Thr Leu Tyr Gln Ser Ile Ser Phe Ser Pro Pro Leu Pro Gln
275 280 285
Thr Lys Asn Ser Leu Ser His Asp Thr Val Met Gly Tyr Tyr Ser Lys
290 295 300
Met Ile Phe Val Phe Asn Glu Pro Trp Trp Arg Asn Ala Gly Phe Thr
305 310 315 320
Gly Ile Phe Asp Ser Ala Asp Gly Pro Val Ser Phe Thr Arg Asp Thr
325 330 335
Ser Val Pro Arg Asp Asp Gln Trp Ser Ile Thr Cys Phe Ile Val Gly
340 345 350
Gly Arg Gly Arg Thr Trp Ser Lys Leu Ser Lys Ala Asp Arg His Thr
355 360 365
Gln Val Trp Glu Gln Phe Cys Arg Tyr Phe Gly Glu Phe Val Glu Lys
370 375 380
Ile Pro Glu Pro Ala Asn Val Leu Glu Met Glu Trp Ser Lys Gln Pro
385 390 395 400
Ser Phe Leu Gly Ala Pro Cys Pro Val Met Thr Pro Gly Thr Met Thr
405 410 415
Thr Val Gly Thr Glu Leu Ala Thr Pro Phe Gly Arg Val His Phe Val
420 425 430
Gly Thr Glu Thr Ser Ile Val Trp Arg Gly Tyr Met Glu Gly Ala Ile
435 440 445
Arg Ser Gly Gln Arg Gly Ala Ala Glu Val Leu Ala Ala Leu Asn Gly
450 455 460
Asp Leu Cys Ser Glu Ala
465 470
<210> 2
<211> 1410
<212> DNA/RNA
<213> nucleotide sequence of amine oxidase ASAO Gene (Unknown)
<400> 2
atggatgcgt ttgataaagc gccgcgcgtg tatgatgtgg tggtggtggg cgcgggcctg 60
agcggcctgc aggcggcgca tagcatttgc gcggcgggct ttagcgtgtg cgtgctggaa 120
gcgaccaacc gcgtgggcgg caaaaccctg agcgtgcgca gctgcgaaaa aggcattaac 180
gatctgggcg cggcgtggat taacgatacc aaccagagcg aaatgtttaa actgcatcag 240
cgctatagcc tggatggcga aattcagtat acccatggcg atgatattat tcagagcacc 300
gatggcctga tttgcaaaat tccgtatggc cagaccccgc tgagcaccgg cctggtggaa 360
gatctggtgc gcattctgcg cgaaaaaagc gcgccgattg atctgcataa cccggcgagc 420
agcccggcgg cgaaagaagt ggataacgaa acctttcagg atttttgcat tcgccgcacc 480
ggctgcgaag atgcggtgca tattgcggat agcattagcg cggcgctgct gggcgtggat 540
agcattgaag tgagcgcgct gtatatgctg tattatttta aaagcggcag cggcattgat 600
aacctgctga gcgatgaaaa aaacggcggc cagtatctgc gcacccgcca gggcacccag 660
accattagcc agaaaatggc ggaagaactg gatccggaaa ccctgtttct gggcaccccg 720
gtgaccagca ttgatcagag ccgcccggat ggcatttgcc tggtgcagac cctggataac 780
gcgacctttc attgccgccg cgtgattgtg agcgtgccga ccaccctgta tcagagcatt 840
agctttagcc cgccgctgcc gcagaccaaa aacagcctga gccatgatac cgtgatgggc 900
tattatagca aaatgatttt tgtgtttaac gaaccgtggt ggcgcaacgc gggctttacc 960
ggcatttttg atagcgcgga tggcccggtg agctttaccc gcgataccag cgtgccgcgc 1020
gatgatcagt ggagcattac ctgctttatt gtgggcggcc gcggccgcac ctggagcaaa 1080
ctgagcaaag cggatcgcca tacccaggtg tgggaacagt tttgccgcta ttttggcgaa 1140
tttgtggaaa aaattccgga accggcgaac gtgctggaaa tggaatggag caaacagccg 1200
agctttctgg gcgcgccgtg cccggtgatg accccgggca ccatgaccac cgtgggcacc 1260
gaactggcga ccccgtttgg ccgcgtgcat tttgtgggca ccgaaaccag cattgtgtgg 1320
cgcggctata tggaaggcgc gattcgcagc ggccagcgcg gcgcggcgga agtgctggcg 1380
gcgctgaacg gcgatctgtg cagcgaagcg 1410

Claims (1)

1. Use of an amine oxidase ASAO having the amino acid sequence of SEQ ID No.1 for degradation of fumonisin B1 not for therapeutic purposes.
CN202111133638.9A 2021-09-27 2021-09-27 Amine oxidase ASAO, preparation method and application Active CN113817696B (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020047656A1 (en) * 2018-09-05 2020-03-12 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture And Agri-Food Recombinant expression of fumonisin amine oxidase
CN112301010A (en) * 2020-09-22 2021-02-02 天津科技大学 Amine oxidase ACAO, preparation method and application

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020047656A1 (en) * 2018-09-05 2020-03-12 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture And Agri-Food Recombinant expression of fumonisin amine oxidase
CN112301010A (en) * 2020-09-22 2021-02-02 天津科技大学 Amine oxidase ACAO, preparation method and application

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
amine oxidase [Aspergillus sclerotiicarbonarius CBS 121057];GenBank;《Genbank》;20180612;CDS、ORIGIN部分 *
GenBank.amine oxidase [Aspergillus sclerotiicarbonarius CBS 121057].《Genbank》.2018,CDS、ORIGIN部分. *

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