CN112301010B - Amine oxidase ACAO, preparation method and application - Google Patents
Amine oxidase ACAO, preparation method and application Download PDFInfo
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- CN112301010B CN112301010B CN202010999974.0A CN202010999974A CN112301010B CN 112301010 B CN112301010 B CN 112301010B CN 202010999974 A CN202010999974 A CN 202010999974A CN 112301010 B CN112301010 B CN 112301010B
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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)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- 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
Abstract
The invention relates to amine oxidase ACAO, and the amino acid sequence of the amine oxidase ACAO is SEQ ID NO. 1. The degradation rate of the amine oxidase ACAO to HFB1 is 100%; the amine oxidase ACAO has excellent property, can be applied to agriculture, feed, food and other industries, and reduces the harm of fumonisins FB1 to the health of animals and human beings.
Description
Technical Field
The invention belongs to the technical field of agricultural biology, and particularly relates to an amine oxidase ACAO, a preparation method and application thereof.
Background
Fumonisins (also called fumonisins) are mycotoxins, which were discovered in 1989 to be water-soluble metabolites produced by fungi such as Fusarium proliferatum, Fusarium moniliforme, and Fusarium verticillioides, and are structurally similar diester compounds composed of different types of polyhydric alcohols and tricarballylic acid. The fumonisins are mainly present in animal feed raw materials, can cause acute poisoning of some livestock, has potential carcinogenicity to most poultry, and has the possibility of human infection, so that how to maximally degrade the fumonisins becomes another research hotspot after aflatoxin. The popularity of our country is wide, and the feed in some areas is easily affected by environmental factors to generate mycotoxins such as fumonisins and the like.
The currently known fumonisins degradation methods mainly comprise physical degradation, chemical degradation and biological degradation, the physical methods have the defects that some toxins are not completely degraded, the chemical methods easily cause the quality reduction of food, and the physical methods and the chemical methods have some defects. The biodegradation is to separate and clone genes related to fumonisin detoxification enzyme from microorganisms, construct genetic engineering bacteria by utilizing a genetic engineering technology, express related proteins and degrade toxins, and has a good effect.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides amine oxidase ACAO, a preparation method and application thereof.
The technical scheme adopted by the invention for solving the technical problems is as follows:
an amine oxidase ACAO, wherein the amino acid sequence of the amine oxidase ACAO is SEQ ID NO. 1.
The preparation method of the amine oxidase ACAO comprises the following steps:
(1) adopting a molecular biology method to recombine a gene containing the amine oxidase ACAO to a vector and then transforming a host cell to obtain a recombinant strain;
(2) culturing the recombinant strain, and inducing amine oxidase ACAO expression;
(3) separating and purifying the obtained amine oxidase ACAO;
(4) the correct expression of the protein was verified by western blot.
Moreover, the host cell in the step (1) is a large intestine cell, a beer yeast cell or a Torulopsis torulosa cell.
The application of the amine oxidase ACAO in degrading fumonisins.
An amine oxidase ACAO gene encoding the amine oxidase ACAO as described above.
Moreover, the nucleotide sequence of the gene is SEQ ID NO. 2.
A recombinant vector comprising the amine oxidase ACAO gene as described above.
The recombinant vector pET28a (+) -ACAO comprising the amine oxidase ACAO gene as described above.
A recombinant strain comprising the amine oxidase ACAO gene as described above.
Moreover, the recombinant strain is Escherichia coli.
The invention has the advantages and positive effects that:
1. the amine oxidase ACAO can degrade and catalyze the amino group of fumonisins FB1(HFB1) to hydrolyze, and eliminate the activity of fumonisins. Since amino group is one of the key functional groups that make fumonisins toxic, removal of amino group is the key point for detoxification of fumonisins. The amine oxidase ACAO has the activity of degrading and hydrolyzing fumonisins HFB1, can be applied to industries such as agriculture, feed, food and the like, and reduces the harm of fumonisins to the health of animals and human beings.
Drawings
FIG. 1 is a SDS-PAGE purification of the amine oxidase ACAO of the present invention; wherein, M: protein marker; 1: protein of 80mM imidazole, 2: protein eluted with 100mM imidazole, 3: 300mM imidazole-eluted protein;
FIG. 2 is a western blot result chart of the amine oxidase ACAO of the present invention, wherein M: protein marker; amine oxidase ACAO;
FIG. 3 is a schematic diagram showing the degradation effect of amine oxidase ACAO in the present invention, wherein FIG. A is a liquid phase result of hydrolyzing fumonisins by a standard substance, and FIG. B is a liquid phase result of hydrolyzing fumonisins by degrading by amine oxidase ACAO;
Detailed Description
The following detailed description of the embodiments of the present invention is provided for the purpose of illustration and not limitation, and should not be construed as limiting the scope of the invention.
The raw materials used in the invention are conventional commercial products unless otherwise specified; the methods used in the present invention are conventional in the art unless otherwise specified.
An amine oxidase ACAO, wherein the amino acid sequence of the amine oxidase ACAO is SEQ ID NO. 1.
The preparation method of the amine oxidase ACAO comprises the following steps:
(1) adopting a molecular biology method to recombine a gene containing the amine oxidase ACAO to a vector and then transforming a host cell to obtain a recombinant strain;
(2) culturing the recombinant strain, and inducing amine oxidase ACAO expression;
(3) separating and purifying the obtained amine oxidase ACAO;
(4) the correct expression of the protein was verified by western blot.
Preferably, in the first step, the host cell is a large intestine cell, a beer yeast cell or a polytypic yeast cell, preferably escherichia coli BL21(DE 3).
The application of the amine oxidase ACAO in degrading fumonisins.
An amine oxidase ACAO gene encoding the amine oxidase ACAO as described above.
Preferably, the nucleotide sequence of the gene is SEQ ID NO. 2.
A recombinant vector comprising the amine oxidase ACAO gene as described above.
The recombinant vector pET28a (+) -ACAO comprising the amine oxidase ACAO gene as described above.
A recombinant strain comprising the amine oxidase ACAO gene as described above.
Preferably, the recombinant strain is escherichia coli.
Specifically, the amino acid sequence of the amine oxidase ACAO is shown in SEQ ID NO. 1:
SEQ ID NO.1：
MSITNSFPTPAAPKEVDVIVVGAGLSGLRAALDVQAAGLSCIVVEAVDRVGGKTLSVP SKLSGPGVNDIGAAWINDTSQSEMYGLLRKYGLHGEIQRAEGMSLSLTAEGVITHPYGTLPL DEHDQVLVASVLQTIQGLVDQIDLKNPAAGSIGKELDSMTASEYCSKTFQSDLVVGIVDTVT QSLLGVEAASISMLSLVHYIKAATGVDAALSDGKDGGQYLRVREGTQSFSRKMAEALEPGT LFLSTPVTSIEQSSQTCICTVRTSSPTNSTFHAKKVILSIPTPLYHKISFDPPLPEDKTRLASDNI LGYYSKMIYVFDQPWWRTAGLSGVMETDLGPILFSRDTSIPDDDQWSITCFIVGKRGREWS LLSSSERRRTAWDQFRTTFESASGLSGKLDVPEPINVLEIEWTKQEFFGGAPCPVSPPGLLSS VDASAAQCPFKDVHFVGTETAFVWRGYMEGAVRSGIRGAKEVIDLLKG
wherein, the enzyme gene codes 481 amino acids without signal peptide, so the theoretical molecular weight of mature amine oxidase ACAO is 51.69 kDa.
The invention provides a method for coding the above amine oxidase ACAO. The genome sequence of the gene is shown in SEQ ID NO. 2:
SEQ ID NO.2：
AACCCCAGCTGCCCCAAAGGAGGTTGACGTTATCGTTGTCGGAGCCGGATTGAGTGGTTTGAGA GCTGCCCTTGACGTCCAAGCTGCTGGTCTTAGTTGCATCGTTGTCGAAGCTGTCGATAGGGTCGGAGG TAAAACCTTGTCTGTCCCATCTAAGTTGTCTGGTCCCGGAGTCAACGACATTGGTGCTGCTTGGATCA ACGATACCTCTCAGTCTGAGATGTATGGATTGTTGAGGAAGTATGGACTTCACGGAGAGATCCAGAGG GCCGAAGGAATGAGTCTTTCTTTGACCGCCGAGGGTGTCATCACTCATCCATACGGAACCTTGCCACT TGACGAACACGATCAAGTTTTGGTCGCCTCTGTTCTTCAGACCATCCAAGGATTGGTCGATCAGATCG ATTTGAAGAACCCAGCCGCTGGAAGTATCGGTAAGGAGCTTGACTCTATGACTGCCAGTGAATATTGT TCTAAGACTTTCCAGTCTGACTTGGTCGTCGGAATCGTTGATACCGTCACTCAGTCTTTGCTTGGTGTC GAGGCTGCCTCTATCTCTATGCTTTCTCTTGTCCACTATATCAAAGCCGCTACCGGTGTTGATGCCGCTT TGAGTGACGGTAAAGACGGTGGTCAATATTTGAGAGTTAGGGAGGGAACTCAGAGTTTCAGTAGGAA GATGGCTGAGGCTCTTGAACCCGGAACCCTTTTCTTGTCTACCCCAGTCACCTCTATCGAGCAGAGTT CTCAGACTTGCATCTGCACTGTTAGAACCAGTAGTCCAACTAATTCTACTTTTCACGCTAAAAAAGTTA TTCTTTCTATCCCAACCCCATTGTATCACAAGATCAGTTTCGACCCTCCTCTTCCAGAGGACAAGACTA GATTGGCCTCTGACAACATTTTGGGTTATTACTCTAAGATGATTTACGTCTTCGACCAACCTTGGTGGA GAACCGCTGGTTTGTCTGGAGTCATGGAGACCGACCTTGGACCTATCCTTTTCAGTAGGGACACCAGT ATCCCAGACGACGACCAGTGGTCTATCACTTGTTTTATCGTCGGAAAAAGGGGTAGGGAGTGGAGTTT GTTGTCTAGTAGTGAGAGGAGGAGAACCGCTTGGGATCAGTTTAGGACCACCTTCGAATCTGCCAGT GGTTTGAGTGGAAAGTTGGACGTCCCAGAGCCTATCAACGTCTTGGAGATCGAGTGGACCAAGCAAG AATTTTTCGGAGGTGCTCCATGCCCAGTCTCTCCACCCGGATTGCTTAGTTCTGTTGATGCCAGTGCCG CCCAATGCCCTTTCAAGGATGTCCACTTCGTCGGAACTGAGACCGCTTTCGTCTGGAGGGGATACATG GAGGGAGCCGTTAGGTCTGGTATTAGAGGAGCCAAGGAGGTTATCGATTTGCTTAA
the invention separates and clones amine oxidase ACAO by a PCR method, and the analysis result of the DNA complete sequence shows that the total length of an open reading frame sequence (ORF) of the amine oxidase ACAO gene is 1416 bp.
The invention also provides a recombinant vector containing the amine oxidase ACAO, and a preferred recombinant vector is named as pET28 a-ACAO. The amine oxidase ACAO gene of the present invention is inserted between appropriate restriction sites of an expression vector so that the nucleotide sequence thereof is operably linked to an expression regulatory sequence. As a most preferred embodiment of the present invention, it is preferred that the detoxification enzyme gene of the present invention is inserted between restriction sites EcoR I and Not I on the plasmid pET28a so that the nucleotide sequence is located downstream of and under the control of the T7 promoter to give a recombinant large intestine expression plasmid pET28 a-ACAO.
More specifically, the preparation and detection are as follows:
test materials and reagents:
1. bacterial strains and vectors: the invention provides a novel amine oxidase ACAO. The Escherichia coli expression vector pET28a (+) and the strain BL21(DE3) are stored in the laboratory.
2. Enzymes and other biochemical reagents: the endonuclease was purchased from Fermentas, and the ligase was purchased from Thermo Scientific. Proteitech His tag antibodies were obtained from the Scientific and technical company of Western America, Beijing, protein marker was obtained from Thermo Scientific, chromatographic grade methanol was obtained from Fisher Chemical, and others were made in the home (all available from general Biochemical Co.).
3. Culture medium:
escherichia coli Medium LB (10g/L peptone, 5g/L yeast extract, 10g/L sodium chloride, pH 7.0).
Description of the drawings: the following molecular biology experiments, which are not specifically described, are performed by referring to the specific methods listed in molecular cloning, a laboratory manual (third edition) J. SammBruke, or according to the kit and product instructions.
Cloning of mono-amine oxidase ACAO
The gene segment of amine oxidase ACAO is obtained by artificial chemical synthesis method, and endonuclease site EcoR I is introduced at 5 'end and endonuclease site Not I is introduced at 3' end.
Preparation of di-and recombinant amine oxidase ACAO
The expression vector pET28a is subjected to double enzyme digestion (EcoR I + Not I), meanwhile, the gene coding the amine oxidase ACAO is subjected to double enzyme digestion (EcoR I + Not I), the gene fragment coding mature amine oxidase is cut out and connected with the expression vector pET28a, the recombinant plasmid pET28a-ACAO containing the amine oxidase gene ACAO is obtained, escherichia coli BL21(DE3) is transformed, and the recombinant escherichia coli strain BL21/ACAO is obtained.
BL21(DE3) strain containing plasmid was inoculated in 100mL LB medium, cultured with shaking at 37 ℃ and 220rpm for about 2 hours, then 0.5mM IPTG was added, and the mixture was induced at 25 ℃ and 220rpm, and after about 24 hours, the activity of intracellular and extracellular amine oxidases was measured. The activity of amine oxidase was detected intracellularly, and 80mM, 100mM, and 300mM of imidazole were collected by nickel column purification, respectively. SDS-PAGE results show that the recombinant amine oxidase is expressed. The correct expression of the protein was further verified by western blot. As shown in FIG. 1, 3 lanes show the results after purification. FIG. 2 shows the results of western blot validation.
Determination of degradation Properties of recombinant amine oxidase
The high performance liquid chromatography is used for detecting the degradability of amine oxidase, and the specific method comprises the following steps:
(1) HFB1 standard stock solution: prepared into standard solution with the concentration of 100 mu g/mL and stored at the temperature of-20 ℃.
(2) Preparation of a sample: taking 45 μ L of purified amine oxidase enzyme solution, adding 5 μ L of HFB1 standard stock solution to make the final concentration of HFB 110 μ g/mL, 37 deg.C, 220rpm, and standing in dark for 30 min.
(3) Derivatization of the sample: and (3) taking 50 mu L of a sample to be detected, adding 200 mu L of acetonitrile water (V: V ═ 1:1) and 250 mu L of OPA derivative solution, mixing uniformly for 30s, introducing a sample within 2min of derivative, and filtering the sample by using a filter membrane to be detected. The enzymatic activity of the amine oxidase ACAO was determined by comparison with the peak pattern of a standard of HFB 1.
Although the 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 embodiments disclosed.
Sequence listing
<110> Tianjin science and technology university
<120> amine oxidase ACAO, preparation method and application
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 440
<212> PRT
<213> amino acid sequence of amine oxidase ACAO (Unknown)
<400> 1
Met Ser Ile Thr Ala Ser Pro Pro Thr Pro Ala Ala Pro Leu Gly Val
1 5 10 15
Ala Val Ile Val Val Gly Ala Gly Leu Ser Gly Leu Ala Ala Ala Leu
20 25 30
Ala Val Gly Ala Ala Gly Leu Ser Cys Ile Val Val Gly Ala Val Ala
35 40 45
Ala Val Gly Gly Leu Thr Leu Ser Val Pro Ser Leu Leu Ser Gly Pro
50 55 60
Gly Val Ala Ala Ile Gly Ala Ala Thr Ile Ala Ala Thr Ser Gly Ser
65 70 75 80
Gly Met Thr Gly Leu Leu Ala Leu Thr Gly Leu His Gly Gly Ile Gly
85 90 95
Ala Ala Gly Gly Met Ser Leu Ser Leu Thr Ala Gly Gly Val Ile Thr
100 105 110
His Pro Thr Gly Thr Leu Pro Leu Ala Gly His Ala Gly Val Leu Val
115 120 125
Ala Ser Val Leu Gly Thr Ile Gly Gly Leu Val Ala Gly Ile Ala Leu
130 135 140
Leu Ala Pro Ala Ala Gly Ser Ile Gly Leu Gly Leu Ala Ser Met Thr
145 150 155 160
Ala Ser Gly Thr Cys Ser Leu Thr Pro Gly Ser Ala Leu Val Val Gly
165 170 175
Ile Val Ala Thr Val Thr Gly Ser Leu Leu Gly Val Gly Ala Ala Ser
180 185 190
Ile Ser Met Leu Ser Leu Val His Thr Ile Leu Ala Ala Thr Gly Val
195 200 205
Ala Ala Ala Leu Ser Ala Gly Leu Ala Gly Gly Gly Thr Leu Ala Val
210 215 220
Ala Gly Gly Thr Gly Ser Pro Ser Ala Leu Met Ala Gly Ala Leu Gly
225 230 235 240
Pro Gly Thr Leu Pro Leu Ser Thr Pro Val Thr Ser Ile Gly Gly Ser
245 250 255
Ser Gly Thr Cys Ile Cys Thr Val Ala Thr Ser Ser Pro Thr Ala Ser
260 265 270
Thr Pro His Ala Leu Leu Val Ile Leu Ser Ile Pro Thr Pro Leu Thr
275 280 285
His Leu Ile Ser Pro Ala Pro Pro Leu Pro Gly Ala Leu Thr Ala Leu
290 295 300
Ala Ser Ala Ala Ile Leu Gly Thr Thr Ser Leu Met Ile Thr Val Pro
305 310 315 320
Ala Gly Pro Thr Thr Ala Thr Ala Gly Leu Ser Gly Val Met Gly Thr
325 330 335
Ala Leu Gly Pro Ile Leu Pro Ser Ala Ala Thr Ser Ile Pro Ala Ala
340 345 350
Ala Gly Thr Ser Ile Thr Cys Pro Ile Val Gly Leu Ala Gly Ala Gly
355 360 365
Thr Ser Leu Leu Ser Ser Ser Gly Ala Ala Ala Thr Ala Thr Ala Gly
370 375 380
Pro Ala Thr Thr Pro Gly Ser Ala Ser Gly Leu Ser Gly Leu Leu Ala
385 390 395 400
Val Pro Gly Pro Ile Ala Val Leu Gly Ile Gly Thr Thr Leu Gly Gly
405 410 415
Pro Pro Gly Gly Ala Pro Cys Pro Val Ser Pro Pro Gly Leu Leu Ser
420 425 430
Ser Val Ala Ala Ser Ala Ala Gly
435 440
<210> 2
<211> 1416
<212> DNA/RNA
<213> nucleotide sequence of amine oxidase ACAO Gene (Unknown)
<400> 2
aaccccagct gccccaaagg aggttgacgt tatcgttgtc ggagccggat tgagtggttt 60
gagagctgcc cttgacgtcc aagctgctgg tcttagttgc atcgttgtcg aagctgtcga 120
tagggtcgga ggtaaaacct tgtctgtccc atctaagttg tctggtcccg gagtcaacga 180
cattggtgct gcttggatca acgatacctc tcagtctgag atgtatggat tgttgaggaa 240
gtatggactt cacggagaga tccagagggc cgaaggaatg agtctttctt tgaccgccga 300
gggtgtcatc actcatccat acggaacctt gccacttgac gaacacgatc aagttttggt 360
cgcctctgtt cttcagacca tccaaggatt ggtcgatcag atcgatttga agaacccagc 420
cgctggaagt atcggtaagg agcttgactc tatgactgcc agtgaatatt gttctaagac 480
tttccagtct gacttggtcg tcggaatcgt tgataccgtc actcagtctt tgcttggtgt 540
cgaggctgcc tctatctcta tgctttctct tgtccactat atcaaagccg ctaccggtgt 600
tgatgccgct ttgagtgacg gtaaagacgg tggtcaatat ttgagagtta gggagggaac 660
tcagagtttc agtaggaaga tggctgaggc tcttgaaccc ggaacccttt tcttgtctac 720
cccagtcacc tctatcgagc agagttctca gacttgcatc tgcactgtta gaaccagtag 780
tccaactaat tctacttttc acgctaaaaa agttattctt tctatcccaa ccccattgta 840
tcacaagatc agtttcgacc ctcctcttcc agaggacaag actagattgg cctctgacaa 900
cattttgggt tattactcta agatgattta cgtcttcgac caaccttggt ggagaaccgc 960
tggtttgtct ggagtcatgg agaccgacct tggacctatc cttttcagta gggacaccag 1020
tatcccagac gacgaccagt ggtctatcac ttgttttatc gtcggaaaaa ggggtaggga 1080
gtggagtttg ttgtctagta gtgagaggag gagaaccgct tgggatcagt ttaggaccac 1140
cttcgaatct gccagtggtt tgagtggaaa gttggacgtc ccagagccta tcaacgtctt 1200
ggagatcgag tggaccaagc aagaattttt cggaggtgct ccatgcccag tctctccacc 1260
cggattgctt agttctgttg atgccagtgc cgcccaatgc cctttcaagg atgtccactt 1320
cgtcggaact gagaccgctt tcgtctggag gggatacatg gagggagccg ttaggtctgg 1380
tattagagga gccaaggagg ttatcgattt gcttaa 1416
Claims (9)
1. An amine oxidase ACAO characterized in that the amino acid sequence of said amine oxidase ACAO is SEQ ID No. 1.
2. Process for the preparation of the amine oxidase ACAO according to claim 1, characterized in that it comprises the following steps:
transforming a host with a recombinant vector containing a gene encoding the amine oxidase ACAO of claim 1 to obtain a recombinant strain;
culturing recombinant strains to induce the expression of amine oxidase ACAO;
and obtaining the amine oxidase ACAO by separation and purification.
3. The method of claim 2, wherein the host used in the step of performing is escherichia coli or brewer's yeast.
4. Application of amine oxidase ACAO in preparing preparation for degrading fumonisins FB1 is characterized in that the amino acid sequence of the amine oxidase ACAO is SEQ ID NO. 1.
5. A gene encoding the amine oxidase ACAO of claim 1.
6. The gene of claim 5, wherein the nucleotide sequence of the gene is SEQ ID NO. 2.
7. A recombinant vector comprising the gene of claim 5 or 6.
8. A recombinant strain comprising the gene of claim 5 or 6.
9. The recombinant strain of claim 8, wherein the recombinant strain is E.
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| CN108251399A (en) * | 2016-12-29 | 2018-07-06 | 中粮营养健康研究院有限公司 | Fumonisin degrading enzyme, encoding gene, recombinant vector, cell, additive and its application |
| CN109337886A (en) * | 2018-10-09 | 2019-02-15 | 天津科技大学 | Fumonisin degrading enzyme FumDXA and its gene and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108251399A (en) * | 2016-12-29 | 2018-07-06 | 中粮营养健康研究院有限公司 | Fumonisin degrading enzyme, encoding gene, recombinant vector, cell, additive and its application |
| CN109337886A (en) * | 2018-10-09 | 2019-02-15 | 天津科技大学 | Fumonisin degrading enzyme FumDXA and its gene and application |
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