CN107446902A - A kind of zearalenone toxin degradation enzyme ZENdease N2 and its encoding gene and application - Google Patents
A kind of zearalenone toxin degradation enzyme ZENdease N2 and its encoding gene and application Download PDFInfo
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- CN107446902A CN107446902A CN201710506384.8A CN201710506384A CN107446902A CN 107446902 A CN107446902 A CN 107446902A CN 201710506384 A CN201710506384 A CN 201710506384A CN 107446902 A CN107446902 A CN 107446902A
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- Prior art keywords
- zearalenone
- digestive enzyme
- enzyme
- zendease
- amino acid
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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
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
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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
- 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
Abstract
The present invention discloses a kind of zearalenone digestive enzyme, and the zearalenone digestive enzyme is named as ZENdease N2, from eutypa dieback bacterium (Eutypalata), for the protein as shown in (a) or (b)：(a) protein being made up of the amino acid sequence shown in sequence table SED ID NO.1；(b) the obtained protein with degrading zearalenone activity is derived by the substitution of one or more amino acid residues and/or missing and/or insertion as the amino acid sequence shown in sequence table SED ID NO.1.The invention also discloses the expression of zearalenone digestive enzyme and application.Zearalenone digestive enzyme of the present invention has efficient degradation effect to zearalenone.
Description
Technical field
The present invention relates to enzyme engineering field.More particularly, to a kind of zearalenone digestive enzyme and its encoding gene
With application.
Background technology
Zearalenone (Zearalenone, ZEN) is mainly caused by Fusarium (Fusarium species)
Secondary metabolites, it is a kind of fusarium toxin that pollution range is most wide in the world, easily the cereal crops by ZEN pollutions include jade
Rice, barley, wheat, sorghum and rye etc..ZEN chemical name is 6- (- 6 epoxides of 10- hydroxyls-undecenyl) β-thunder lock acid
Lactone, its structure is similar to the structure of female hormone estradiol (estradiol), ZEN can with animal cell membrane it is female swash
Plain acceptor (estrogen receptor, ER) is combined, and causes ER conformational change, and ZEN/ER compounds are further transferred to
In nucleus, combined with estrogen response element, adjust the transcription and translation of target gene, and then influenceed the growth of cell and divide
Split.ZEN is ingested into mammal body, and scorching manmmal vagina, false heat and oestrous cycle can be caused to extend, miscarry, no
Educate and deformity etc. " high female hormone disease " (hyperestrogenism).ZEN is accumulated by food chain in human body, can be led
Sex premature is caused, or even causes the increase of the incidences of disease such as breast cancer, the cancer of the esophagus, grave danger is caused to human health.
At present, domestic and international ZEN detoxicating method mainly has physical removal, chemical treatment etc..To having polluted ZEN fungies poison
Physics, the chemical treatment method of plain cereal mainly have：Flushing and rinsing, heat treatment, ionizing radiation, inorganic adsorbent, chemical reagent
Processing and ozone oxidation etc..But physical chemistry detoxification efficiency is not expected high, and the quality of food is changed, easily
The loss of nutriment is caused, and European Union eliminates mycotoxin not in applied chemistry method in food production.
It is that caused enzyme acts on ZEN in metabolic process using microorganism that microbial degradation method, which eliminates ZEN pollutions, is destroyed
The toxophore of ZEN molecules, it is set to be degraded or be converted into the process of non-toxic products.Biological degradation method degraded ZEN high specificities,
ZEN molecules are converted efficiency high, will not cause environmental pollution, the ZEN pollutions that scientific and effective can be eliminated in cereal crops, have
Fabulous application prospect.
Therefore it provides a kind of enzyme that can effectively degrade ZEN has great importance.
The content of the invention
First purpose of the present invention is to provide a kind of zearalenone digestive enzyme and its encoding gene, and the enzyme can be with
Degrading zearalenone (ZEN).
Second object of the present invention is to provide a kind of method for expressing above-mentioned zearalenone digestive enzyme.
Third object of the present invention is to provide a kind of application of above-mentioned zearalenone digestive enzyme.
To reach above-mentioned purpose, the present invention uses following technical proposals：
The invention provides a kind of zearalenone digestive enzyme, it is named as ZENdease-N2, from grape apical dieback
Germ (Eutypalata), the zearalenone digestive enzyme are the protein as shown in (a) or (b)：
(a) protein being made up of the amino acid sequence shown in sequence table SED ID NO.1；
(b) substitution of one or more amino acid residues is passed through as the amino acid sequence shown in sequence table SED ID NO.1
And/or lack and/or insert and the derivative obtained protein with degrading zearalenone activity.
Wherein, the amino acid sequence shown in the sequence table SED ID NO.1 is made up of 266 amino acid residues.
The encoding gene of above-mentioned zearalenone digestive enzyme falls within protection scope of the present invention in the present invention, described
Encoding gene is such as shown in (a) or (b)：
(a) nucleotide sequence as shown in sequence table SED ID NO.2；
(b) nucleotide sequence of coding amino acid sequence as shown in sequence table SED ID NO.1.
Wherein, by 801 base compositions, it is encoded the nucleotide sequence in the present invention shown in sequence table SED ID NO.2
Sequence is from 5 ' the 1st to the 801st bit bases of end, albumen of the coding with the amino acid sequence shown in sequence table SED ID NO.1
Matter.
It should be noted that expression vector, cell line, engineering bacteria and Host Strains containing the above-mentioned encoding gene of the present invention are equal
Belong to protection scope of the present invention.
It is by containing above-mentioned Gibberella zeae alkene present invention also offers a kind of method for expressing zearalenone digestive enzyme
The recombinant expression carrier of ketone degraded enzyme coding gene imports host cell, and expression obtains zearalenone digestive enzyme.
Wherein, the host includes but is not limited to Escherichia coli, saccharomycete, mammalian cell, insect cell, withered grass bar
Bacterium, bacillus or lactobacillus etc.；Preferably Escherichia coli, more preferably e. coli bl21.
The carrier that sets out for building the recombination bacillus coli and recombinant yeast expression vector can be in above-mentioned host
The expression vector of expression alien gene, such as can be in the pet vector of expression in escherichia coli, and in Pasteur's moral Pichia pastoris
PPIC9K, pPIC9, pPIC3.5K of expression etc. in (Pichia pastoris).
Above-mentioned recombinant expression carrier can be built according to a conventional method.
Present invention also offers application of the zearalenone digestive enzyme in degrading zearalenone.
Beneficial effects of the present invention are as follows：
The zearalenone digestive enzyme that the present invention obtains can degrade to mycotoxin zearalenone in grain,
Effective effect is played to control grain contamination.
Brief description of the drawings
The embodiment of the present invention is described in further detail below in conjunction with the accompanying drawings.
Fig. 1 shows the SDS-PAGE figures of recombinant plasmid PET30a-ZENdease-N2 expression product；
Wherein, swimming lane 1,2 expression products；Swimming lane M, Protein Marker (97KD, 66KD, 45KD, 31KD, 21.5KD,
14.4KD, 6.5KD).
Fig. 2 shows zearalenone digestive enzyme ability measure figure.
Fig. 3 shows degraded situations of the ZENdease-N2 to ZEN molecules under different pH condition.
Fig. 4 shows degraded situations of the ZENdease-N2 to ZEN molecules under condition of different temperatures.
Embodiment
In order to illustrate more clearly of the present invention, the present invention is done further with reference to preferred embodiments and drawings
It is bright.Similar part is indicated with identical reference in accompanying drawing.It will be appreciated by those skilled in the art that institute is specific below
The content of description is illustrative and be not restrictive, and should not be limited the scope of the invention with this.
The zearalenone digestive enzyme ZENdease-N2 of embodiment 1 acquisition and expression
1st, the acquisition of zearalenone degraded enzyme coding gene
ZENdease-N2 full-length genes are obtained by chemical synthesis process, using the gene that this is synthesized as template, in primer 1
Reacted with performing PCR is entered under the guiding of primer 2, expand the sequence of zearalenone degrading enzyme gene.
Primer 1：5'-GAAATTCATATGIts nucleotide sequence of CGGACAAGATCG-3'(such as sequence table SED ID NO.3
Shown, dashed part base is NdeI recognition sites)
Primer 2：5'-CCCGTCGACIts nucleotide sequence of TAGATACCTCCG-3'(such as sequence table SED ID NO.4 institutes
Show, dashed part base is SalI recognition sites)
In PCR reactions, PCR reaction conditions are：94 DEG C, kept for 5 minutes, then circulated by following temperature change program
30 times：94 DEG C are warming up to, is kept for 1 minute, is cooled to 54 DEG C, is kept for 1 minute, is warming up to 68 DEG C, is kept for 2 minutes；Then in 68
DEG C, kept for 10 minutes, be most incubated 10 minutes after 4 DEG C, terminate amplified reaction.About 0.8kb is obtained by agarose electrophoretic analysis
Single band, after expanding PCR primer detection after amplified production DNA gel QIAquick Gel Extraction Kit of the purpose with size is entered
Row recovery, and detect its concentration.PCR primer connection PET30a Vector are reclaimed, convert e. coli bl21, obtain restructuring matter
Grain PET30a-ZENdease-N2 sequencing identifications.The zearalenone degrading enzyme gene DNA sequence dna such as sequence table SED ID
Shown in NO.2, its corresponding amino acid sequence is as shown in sequence table SED IDNO.1.
2nd, the structure of the recombinant expression carrier containing zearalenone digestive enzyme coding gene sequence
Above-mentioned obtained PCR primer both ends have NdeI and SalI restriction endonuclease sites, with (NdeI) and
(SalI) restriction enzyme carries out double digestion reaction, the μ L of digestion system 50 simultaneously to PCR primer and plasmid PET30a：Purpose piece
Section or 20 μ L, 10 × K Buffer of plasmid 5 μ L, NdeI 2 μ L, SalI 2 μ L, ddH2The μ L of O 21, digestion condition are 37 DEG C of reactions
3h.Digestion products are attached after post reclaims, and connection product conversion e. coli bl21, are screened, are chosen through kalamycin resistance
Positive bacterium colony culture is taken, recombinant expression carrier is identified through PCR, digestion identification, sequence verification.The carrier after agarose electrophoresis.Return
The purpose fragment and carrier segments of receipts are 3 quantitatively and in molar ratio:1 ratio carries out Ligation in vitro with T4DNA ligases, connection
The μ L of reaction system 10：μ L, the T4DNA ligases of 5 μ L, PET30a carrier of purpose fragment, 2 μ L 10 × T4DNA connections buffer solution 1
(350U/μL)1μL,ddH2O 1μL.16 DEG C of connections overnight, connection product conversion e. coli bl21, are sieved through kalamycin resistance
Choosing, 37 DEG C of shaken cultivation 6-8h of picking colony, enter performing PCR identification respectively and the digestion of recombinant plasmid is identified.The restructuring table of acquisition
Rear PET30a-ZENdease-N2 is named as up to carrier.
Ultrasonic disruption, supernatant is collected by centrifugation, takes 15 μ L of supernatant to be detected with SDS-PAGE electrophoresis.To PET30a-
ZENdease-N2 is sequenced, and as a result proves that fusion connection enters carrier PET30a DNA sequence dna and sequence table SED ID NO.2
Shown sequence is identical, builds the recombinant expression carrier PET30a-ZENdease- containing zearalenone degrading enzyme gene sequence
N2 is correct.
3rd, expression and purification of the zearalenone digestive enzyme in Escherichia coli
Recombinant expression carrier PET30a-ZENdease-N2 is converted into Escherichia coli, is coated with kalamycin resistance LB flat boards.
Single bacterium on picking flat board flat board falls in 1L LB culture mediums and cultivated, and when bacterial growth to OD values is 1 or so, adds 0.5mL
Concentration is that 0.6mol/L IPTG induced expressions are stayed overnight, and next day receives bacterium purifying protein.The thalline of overnight expression is collected by centrifugation, abandoned
Supernatant, add 30mL and buffer solution is resuspended, ultrasonication bacterium after bacterial precipitation has been hanged, broken cell suspension is transferred at a high speed
In centrifuge tube, supernatant is added in Ni-NTA affinity columns after high speed centrifugation, allows it to flow through affinity media.With 10 times of column volumes
Wash buffer rinses affinity media, washes the foreign protein of non-specific binding off, with 5mL elution buffers by destination protein from parent
Eluted with post.The expression that ZENdease-N2 albumen is carried out with SDS- polyacrylamide gel electrophoresises (SDS-PAGE) is pure
Change analysis.Detected with SDS-PAGE electrophoresis.As a result it is as shown in Figure 1：Swimming lane 1 is expression product, and arrow represents purpose band,
This shows the molecular weight about 29.2KD of the protein of recombinant bacterial strain expression, with pushing away the theoretical molecular that section goes out from amino acid
(29.2KD) is in the same size.
The Activity determination of the zearalenone digestive enzyme of embodiment 2
The μ L of ZENdease-N2 albumen 50 of purifying are taken, are added in the eppendorf pipes that concentration containing ZEN is 10 μ g/mL,
Compared with being not added with the same concentration ZEN solution of ZENdease-N2 albumen, 37 DEG C of reaction 3h, 100 DEG C of heating 10min terminating reactions,
Reaction system is evaporated with Rotary Evaporators, 1mL methanol is added, ultrasonic extraction, after 0.22 μm of membrane filtration, uses high-efficient liquid phase color
Spectrum detection, analyzes whether ZENdease-N2 albumen has degrading activity to ZEN according to the change of ZEN concentration.High-efficient liquid phase color
Spectrum detection ZEN chromatographic condition be：Chromatographic column：C18Post (250mm × 4.6mm, 5 μm, xbridge)；Mobile phase：Ultra-pure water/color
Spectrum level acetonitrile=50/50 (V/V)；Flow rate of mobile phase：1.0mL/min；Chromatogram column temperature is set：25℃；Sample size：10μL；It is glimmering
Photodetector：Excitation wavelength (Ex) 274nm, launch wavelength (Em) 440nm；Acquisition time：13min.Under above-mentioned chromatographic condition,
Control sample has strong absworption peak at retention time RT=11.2min, and reaction group sample detects without ZEN target peaks substantially,
Through absorbing calculated by peak area, existing 90% ZEN molecules are degraded, as shown in Fig. 2 it can be shown that ZENdease-N2 genes gram
The grand digestive enzyme given expression to has the activity of degrading zearalenone.
The reaction condition detection of the zearalenone digestive enzyme of embodiment 3
The present invention explores the optimum reaction condition of ZENdease-N2 degraded ZEN molecules, tests different temperatures, pH value etc.
Under the conditions of ZENdease-N2 to ZEN molecular degradations activity influence.
First, degradation rates of the ZENdease-N2 to ZEN molecules under test different pH condition.Take after purification
ZENdease-N2 digestive enzyme 2mL, it is standby with 100 times of PBS dilution；8 50mL centrifuge tubes are taken, after every adds dilution
Digestive enzyme 5mL, after adjusting pH to place 12h to 2,3,4,5,6,7,8,9,4 DEG C respectively with hydrochloric acid and ammoniacal liquor, draw 500 μ L respectively
Into 8 EP pipes containing 10 μ g ZEN, 37 DEG C of reaction 30min terminating reactions, to be not added with the same concentration of ZENdease-N2 digestive enzymes
ZEN solution compares, with ZENdease-N2 under high performance liquid chromatography detection different pH condition to the degrading activity of ZEN molecules
Influence.As shown in Figure 3, the results showed that, under the conditions of pH=7-8 neutrality and meta-alkalescence, degradeds of the ZENdease-N2 to ZEN
Active highest.
Same method, under conditions of pH=7, test 4 DEG C, 20 DEG C, 37 DEG C, 45 DEG C, 60 DEG C, 70 DEG C of condition of different temperatures
Lower ZENdease-N2 influences on ZEN degrading activity, as shown in Figure 4, the results showed that, ZENdease-N2 degradeds ZEN's is optimal
Reaction temperature is 37 DEG C, and under 60 DEG C of temperature conditionss, ZENdease-N2 still has higher degrading activity to ZEN, explanation
ZENdease-N2 digestive enzymes have preferable heat-resisting quantity.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not pair
The restriction of embodiments of the present invention, for those of ordinary skill in the field, may be used also on the basis of the above description
To make other changes in different forms, all embodiments can not be exhaustive here, it is every to belong to this hair
Row of the obvious changes or variations that bright technical scheme is extended out still in protection scope of the present invention.
SEQUENCE LISTING
<110>Institute of Science and Technology, National Food Bureau
<120>A kind of zearalenone toxin degradation enzyme ZENdease-N2 and its encoding gene and application
<130> JLC17I0312E
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 286
<212> PRT
<213>Zearalenone toxin degradation enzyme ZENdease-N2 amino acid sequences
<400> 1
Met Arg Thr Arg Ser Thr Leu Lys Asp Lys Asn Gly Ile Asn Trp Tyr
1 5 10 15
Tyr Glu Gln Glu Gly Ser Gly Pro His Val Val Leu Ile Pro Asp Gly
20 25 30
Trp Gly Glu Cys Gln Met Met Asp Lys Pro Met Ser Leu Ile Ala Ala
35 40 45
Gln Gly Phe Thr Val Thr Thr Phe Asp Met Pro Gly Phe Ser Arg Ser
50 55 60
Ser Asp Ala Pro Pro Glu Thr Tyr Gln Asp Val Thr Ala Gln Lys Leu
65 70 75 80
Ala Ser Tyr Val Ile Ser Ile Leu Asp Glu Leu His Val Asp Tyr Ala
85 90 95
Thr Phe Trp Gly Cys Ala Ala Gly Gly Ala Thr Val Leu Ala Leu Ala
100 105 110
Ala Asp Tyr Pro Glu Arg Met Arg Asn Gly Leu Pro His Glu Val Pro
115 120 125
Thr Ala Ala Asn Pro Lys Glu Asn Leu Asn Ala Leu Ala Lys Met Glu
130 135 140
Asp Glu Ala Ile Val Lys Ile Met Glu Gly Asp Met Leu Lys His Ile
145 150 155 160
Phe Gly Pro Asp Leu Thr Ala Trp His Ala Leu Gly Glu Glu Ala His
165 170 175
Ala Arg Leu Arg Lys Ala Tyr Pro Arg Trp Ala Arg Gly Tyr Pro Leu
180 185 190
Thr Leu Pro Ser Ser Ala Pro Thr Gly Glu Glu Asp Leu Lys Lys Arg
195 200 205
Pro Leu Asp Trp Thr Val Gly Gly Asp Thr Ala Thr Gln Ser Phe Ile
210 215 220
Asp Asn Ile Ile Thr Ala Ala Lys Ala Gly Ile Pro Ile Gly Thr Ile
225 230 235 240
Pro Gly Met His Phe Pro Tyr Val Ser His Pro Glu Ala Leu Val Lys
245 250 255
His Ile Val Asp Thr Thr Arg Arg Tyr Leu
260 265
<210> 2
<211> 801
<212> DNA
<213>Zearalenone toxin degradation enzyme ZENdease-N2 coding gene sequences
<400> 2
atgcggacaa gatcgactct caaagacaag aatgggatca actggtacta cgaacaagaa 60
gggtctggcc cccacgtggt tcttatcccc gatgggtggg gagagtgcca aatgatggac 120
aagcccatgt ctctaattgc cgcccagggg tttacggtca ccacattcga catgccggga 180
ttctcgaggt cttcagatgc cccgccggag acttaccagg acgtcacagc ccagaagctg 240
gccagctacg tcatcagcat cctagatgag ctgcacgtcg attacgctac gttctggggc 300
tgcgccgccg gcggtgcgac cgtgcttgca ttggcggctg actaccccga gcgcatgcgc 360
aacgggctgc cgcatgaagt tccgacggct gctaatccca aggaaaacct caacgctttg 420
gctaagatgg aagacgaggc tatcgtgaag atcatggaag gggacatgct caagcacatc 480
ttcggtcccg atttgacggc gtggcatgcg ctgggcgagg aggcccacgc caggttgcgg 540
aaagcctatc cccgctgggc ccgcggctac ccccttactc taccgtcgtc tgcgcccact 600
ggagaagagg acttgaagaa gcgaccgctg gattggactg ttggtgggga tacggcgaca 660
cagtcgttca tcgataacat tatcactgct gcgaaggccg gaatcccgat cgggacgatc 720
ccgggcatgc actttccgta tgtctcgcat ccggaggctc tggtgaaaca tattgtggat 780
actactcgga ggtatctatg a 801
<210> 3
<211> 24
<212> DNA
<213>Artificial synthesized primer 1
<400> 3
gaaattcata tgcggacaag atcg 24
<210> 4
<211> 21
<212> DNA
<213>Artificial synthesized primer 2
<400> 4
cccgtcgact agatacctcc g 21
Claims (10)
1. a kind of zearalenone digestive enzyme, it is characterised in that the zearalenone digestive enzyme is such as (a) or (b) institute
The protein shown：
(a) protein being made up of the amino acid sequence shown in sequence table SED ID NO.1；
(b) as the amino acid sequence shown in sequence table SED ID NO.1 by one or more amino acid residues substitution and/or
Lack and/or insert and the derivative obtained protein with degrading zearalenone activity.
2. the encoding gene of zearalenone digestive enzyme described in claim 1.
A kind of 3. encoding gene of zearalenone digestive enzyme as claimed in claim 2, it is characterised in that the Gibberella zeae
The encoding gene of ketenes digestive enzyme is such as shown in (a) or (b)：
(a) nucleotide sequence as shown in sequence table SED ID NO.2；
(b) nucleotide sequence of coding amino acid sequence as shown in sequence table SED ID NO.1.
4. contain the expression vector of the encoding gene of zearalenone digestive enzyme, cell line, engineering described in Claims 2 or 3
Bacterium or Host Strains.
5. application of the zearalenone digestive enzyme in degrading zearalenone described in claim 1.
6. application of the zearalenone degraded enzyme coding gene in degrading zearalenone described in Claims 2 or 3.
A kind of 7. method for expressing the zearalenone digestive enzyme described in claim 1, it is characterised in that：It will contain and have the right to want
The recombinant expression carrier of the zearalenone degraded enzyme coding gene described in 2 or 3 is asked to import host cell, expression obtains corn
Zeranol digestive enzyme.
8. according to the method for claim 7, it is characterised in that：The carrier that sets out for building the recombinant expression carrier is
PET、pPIC9K、pPIC9、pPIC3.5k。
9. according to the method for claim 7, it is characterised in that：The host is that Escherichia coli, saccharomycete, mammal are thin
Born of the same parents, insect cell, hay bacillus, bacillus or lactobacillus.
10. according to the method for claim 9, it is characterised in that：The host is Escherichia coli, it is preferred that is large intestine bar
Bacterium BL21.
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| CN201710506384.8A CN107446902B (en) | 2017-06-28 | 2017-06-28 | Zearalenone toxin degrading enzyme ZENdease-N2 and encoding gene and application thereof |
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Cited By (4)
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| CN108277210A (en) * | 2017-12-18 | 2018-07-13 | 中国农业科学院饲料研究所 | Mould ketenes hydrolase ZEN214 and encoding gene and application |
| CN110029095A (en) * | 2019-04-15 | 2019-07-19 | 南京工业大学 | A kind of zearalenone degrading enzyme and its application |
| CN110592046A (en) * | 2019-09-30 | 2019-12-20 | 湖北大学 | Application of zearalenone degrading enzyme in hydrolysis of zearalenone and derivatives thereof |
| CN112430611A (en) * | 2020-11-30 | 2021-03-02 | 华南理工大学 | Optimized zearalenone degrading enzyme ZHD-P encoding gene, recombinant thallus, surface display system and application |
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| CN102199581A (en) * | 2011-03-31 | 2011-09-28 | 国家粮食局科学研究院 | Zearalenone toxin degradation enzyme and coding gene and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108277210A (en) * | 2017-12-18 | 2018-07-13 | 中国农业科学院饲料研究所 | Mould ketenes hydrolase ZEN214 and encoding gene and application |
| CN108277210B (en) * | 2017-12-18 | 2021-02-23 | 中国农业科学院北京畜牧兽医研究所 | Mycetone hydrolase ZEN214, and coding gene and application thereof |
| CN110029095A (en) * | 2019-04-15 | 2019-07-19 | 南京工业大学 | A kind of zearalenone degrading enzyme and its application |
| CN110029095B (en) * | 2019-04-15 | 2022-06-07 | 南京工业大学 | Zearalenone degrading enzyme and application thereof |
| CN110592046A (en) * | 2019-09-30 | 2019-12-20 | 湖北大学 | Application of zearalenone degrading enzyme in hydrolysis of zearalenone and derivatives thereof |
| CN110592046B (en) * | 2019-09-30 | 2022-03-15 | 湖北大学 | Application of zearalenone degrading enzyme in hydrolysis of zearalenone and derivatives thereof |
| CN112430611A (en) * | 2020-11-30 | 2021-03-02 | 华南理工大学 | Optimized zearalenone degrading enzyme ZHD-P encoding gene, recombinant thallus, surface display system and application |
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| CN107446902B (en) | 2020-03-31 |
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