CN103013944A - Hydrolytic enzyme for diethyl phthalate plasticizer - Google Patents
Hydrolytic enzyme for diethyl phthalate plasticizer Download PDFInfo
- Publication number
- CN103013944A CN103013944A CN2013100084873A CN201310008487A CN103013944A CN 103013944 A CN103013944 A CN 103013944A CN 2013100084873 A CN2013100084873 A CN 2013100084873A CN 201310008487 A CN201310008487 A CN 201310008487A CN 103013944 A CN103013944 A CN 103013944A
- Authority
- CN
- China
- Prior art keywords
- softening agent
- phthalate esters
- diethyl phthalate
- phthalate
- phthalate plasticizer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 102000004190 Enzymes Human genes 0.000 title abstract description 23
- 108090000790 Enzymes Proteins 0.000 title abstract description 23
- FLKPEMZONWLCSK-UHFFFAOYSA-N diethyl phthalate Chemical compound CCOC(=O)C1=CC=CC=C1C(=O)OCC FLKPEMZONWLCSK-UHFFFAOYSA-N 0.000 title abstract 14
- 239000008029 phthalate plasticizer Substances 0.000 title abstract 7
- 230000003301 hydrolyzing effect Effects 0.000 title abstract 5
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 10
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 4
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical class OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 claims description 50
- 239000004902 Softening Agent Substances 0.000 claims description 47
- 108090000604 Hydrolases Proteins 0.000 claims description 21
- 101000925662 Enterobacteria phage PRD1 Endolysin Proteins 0.000 claims description 12
- 239000013612 plasmid Substances 0.000 claims description 12
- 244000005700 microbiome Species 0.000 claims description 10
- 125000003729 nucleotide group Chemical group 0.000 claims description 5
- 239000002773 nucleotide Substances 0.000 claims description 4
- 241000588722 Escherichia Species 0.000 claims 1
- 235000013305 food Nutrition 0.000 abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 7
- 239000002689 soil Substances 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 238000002360 preparation method Methods 0.000 abstract description 2
- 230000002255 enzymatic effect Effects 0.000 abstract 1
- DOIRQSBPFJWKBE-UHFFFAOYSA-N dibutyl phthalate Chemical compound CCCCOC(=O)C1=CC=CC=C1C(=O)OCCCC DOIRQSBPFJWKBE-UHFFFAOYSA-N 0.000 description 14
- KCXZNSGUUQJJTR-UHFFFAOYSA-N Di-n-hexyl phthalate Chemical compound CCCCCCOC(=O)C1=CC=CC=C1C(=O)OCCCCCC KCXZNSGUUQJJTR-UHFFFAOYSA-N 0.000 description 8
- IPKKHRVROFYTEK-UHFFFAOYSA-N dipentyl phthalate Chemical compound CCCCCOC(=O)C1=CC=CC=C1C(=O)OCCCCC IPKKHRVROFYTEK-UHFFFAOYSA-N 0.000 description 8
- MQHNKCZKNAJROC-UHFFFAOYSA-N dipropyl phthalate Chemical compound CCCOC(=O)C1=CC=CC=C1C(=O)OCCC MQHNKCZKNAJROC-UHFFFAOYSA-N 0.000 description 8
- 230000015556 catabolic process Effects 0.000 description 7
- 238000006731 degradation reaction Methods 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 239000004014 plasticizer Substances 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 239000004927 clay Substances 0.000 description 3
- 238000013016 damping Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 2
- 102000004157 Hydrolases Human genes 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 239000004568 cement Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 206010015287 Erythropenia Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 108020005038 Terminator Codon Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
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- 230000001580 bacterial effect Effects 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
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- 238000001962 electrophoresis Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 238000001952 enzyme assay Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 239000010440 gypsum Substances 0.000 description 1
- 229910052602 gypsum Inorganic materials 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 230000007886 mutagenicity Effects 0.000 description 1
- 231100000299 mutagenicity Toxicity 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- -1 or claim fluidizer Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
Images
Abstract
The invention discloses a hydrolytic enzyme for a diethyl phthalate plasticizer. The amino acid sequence is as shown in SEQ ID NO.2. The invention further discloses genes for coding the hydrolytic enzyme for a diethyl phthalate plasticizer, and applications of the hydrolytic enzyme for the diethyl phthalate plasticizer. The engineering strains constructed by utilizing the genes can efficiently express the hydrolytic enzyme for the diethyl phthalate plasticizer, and the enzymatic preparation produced can be used for removing the diethyl phthalate plasticizer added in the production process of agricultural products and food, and can also be used for restoring environments of soil, water bodies and the like, polluted by the diethyl phthalate plasticizer.
Description
Technical field
The present invention relates to phthalate esters softening agent lytic enzyme, belong to the applied environment microorganism field, be used for the removal of phthalate esters softening agent of the interpolation of agricultural-food, food processing process, can be used for repairing the soil of phthalate esters plasticizer pollution, the environment such as water body simultaneously.
Background technology
Softening agent, or claim fluidizer, plasticizer, be a kind of additive that increases the flexibility of material or make material liquefaction.It adds object and has comprised plastic cement, concrete, wall material, cement, gypsum, food etc.The softening agent kind reaches over one hundred kind, namely is the compound that a group is called phthalate esters but use the most generally.Global fluidizer market in 2004 according to statistics, total amount strides forward about about 5,500,000 tons and towards 6,000,000 tons.The mechanism of action of phthalate esters softening agent is that the Bisphthalate molecule is inserted between the polymer molecular chain, weakened the stress of polymer molecule interchain, the result increased polymer molecular chain movability, reduced the degree of crystallinity of polymer molecular chain, thereby the plasticity of polymkeric substance is increased.Since Bisphthalate not with polymkeric substance generation covalent attachment, so be easy to from polymkeric substance migration out, entered environment and accumulation.The phthalate esters softening agent has larger harm to human body, can cause that the animal dis motility rate reduces, loses weight, hepatic and renal function descends, erythropenia in the blood, has mutagenicity and carinogenicity etc.Therefore Environmental Protection Agency has classified it as preferential environmental pollutant, and its usage quantity has been carried out strict control.But the phthalate esters softening agent of entered environment still exists, carry out many-sided research for the phthalate esters softening agent of removing entered environment in the world at present, proposed to utilize the multiple solution routes such as chemistry, physics, biological method.Wherein biological method has the advantage of nontoxic, noresidue, non-secondary pollution.Microorganism has unique effect in the biology solution route that pollutent is eliminated.The pollutent that utilizes the degrading enzyme that extracts in the microorganism to eliminate in the environment has precedent abroad, and the source of degrading enzyme can be extracted from degradation bacteria, also can adopt genetic engineering means to make up efficient expression strain and obtain.
Summary of the invention
Technical problem to be solved by this invention provides a kind of phthalate esters softening agent lytic enzyme, be used for the removal of phthalate esters softening agent of the interpolation of agricultural-food, food processing process, can be used for repairing simultaneously the environment such as soil, water body of phthalate esters plasticizer pollution.
The technical problem that the present invention also will solve provides the encoding gene of above-mentioned phthalate esters softening agent lytic enzyme.
The technical problem that the present invention will solve at last provides above-mentioned phthalate esters softening agent hydrolysis application of enzymes.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
Phthalate esters softening agent lytic enzyme, its aminoacid sequence contain 315 amino acid altogether shown in SEQ ID NO.2.
The encode gene of phthalate esters softening agent lytic enzyme claimed in claim 1, its nucleotide sequence is shown in SEQ ID NO.1, and this full length gene is 948bp, and G+C content is 62.76%.
A kind of recombinant plasmid, it comprises phthalate esters softening agent hydrolase gene claimed in claim 2.
Wherein, described plasmid is pET29a (+).The pET29a (+) that phthalate esters softening agent hydrolase gene fragment and enzyme cut carries out the enzyme connection, obtains to contain pET29a (+) recombinant plasmid of phthalate esters softening agent hydrolase gene.
A kind of recombinant microorganism, it comprises recombinant plasmid claimed in claim 3.
Wherein, described microorganism is Escherichiacoli BL21 (DE3).Enzyme joins good pET29a (+) the recombinant plasmid transformed host expresses bacterial strain Escherichia coli BL21 (DE3) that contains phthalate esters softening agent hydrolase gene, obtains recombinant microorganism E.coli BL21 (DE3).
The application of above-mentioned phthalate esters softening agent lytic enzyme in removing the phthalate esters softening agent.Above-mentioned phthalate esters softening agent lytic enzyme can be used at the aspects such as phthalate esters plasticizer pollution removal of agricultural-food, food, soil, water body.
The application of above-mentioned phthalate esters softening agent hydrolase gene in removing the phthalate esters softening agent.Above-mentioned phthalate esters softening agent hydrolase gene can be applied at the aspects such as phthalate esters plasticizer pollution removal of agricultural-food, food, soil, water body.
Beneficial effect:
1. from the positive colony of cosmid library, clone phthalate esters softening agent hydrolase gene with shotgun.
2. this full length gene (from the initiator codon to the terminator codon) is 948bp, and G+C content is 62.76%, 315 amino acid of encoding.
3. by the terminal complete phthalate esters softening agent hydrolase gene fragment that contains KpnI and Hind III restriction enzyme site of round pcr amplification, it is connected on the KpnI and Hind III restriction enzyme site of colibacillus high expression vector pET29a (+) (available from Novegen company), transform and express Host Strains E.coli BL21 (DE3) (available from Invitrogen company), carry out the IPTG abduction delivering.
4. the present invention has done enzyme assay, can degrade efficiently dipropyl phthalate, dibutyl phthalate, diamyl phthalate, dihexyl phthalate to the product of phthalate esters softening agent hydrolase gene expression.To the above-mentioned all kinds of Bisphthalate degradation effects of 20ppm all greater than 95%.
5. utilize this gene constructed engineering strain can high efficient expression phthalate esters softening agent lytic enzyme (content of active hydrolase protein account for total protein of cell content 50%), the zymin of production can be used for the removal of phthalate esters plasticizer pollution in agricultural-food, food, soil, the water body.It is more more efficient than other degradation method, economy, safety.
Description of drawings
Fig. 1 phthalate esters softening agent hydrolase gene clone's policy map;
Fig. 2 phthalate esters softening agent hydrolase gene is high efficient expression experimental program figure in BL21 (DE3) (pET29a (+)).
Embodiment
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand, the described content of embodiment only is used for explanation the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1: the clone of phthalate esters softening agent hydrolase gene
1.1 the extraction of positive colony cosmid DNA in the cosmid library
The contained clay of positive colony adopts plasmid extraction test kit (available from Axygen company) to extract in the cosmid library.
1.2pUC118(BamH I) buy in precious biotechnology (Dalian) company limited.
1.3 cutting cosmid DNA, the enzyme of clay adopt Sau3AI partially digested.
1.4DNA the cosmid DNA of recovery enzyme after cutting carry out purifying by electrophoresis (TAE damping fluid), adopt dna fragmentation
Reclaim test kit (available from Axygen company) and reclaim, the DNA of recovery is dissolved in the Tris-HCl(pH8.0 of l0mmol/L) in, place-20 ° of C preservations.
1.5 the enzyme connection is set up following reaction system:
16 ° of C incubations 12 hours.
1.6 the efficient competent cell of preparation bacillus coli DH 5 alpha, concrete grammar is with reference to " the molecular cloning experiment guide " of the volumes such as J. Pehanorm Brooker.
1.7 conversion is got 10 μ l enzyme co-product and transformed 200 μ l competent cells, concrete grammar is with reference to " the molecular cloning experiment guide " of the volumes such as J. Pehanorm Brooker.Be coated with the LB that contains dibutyl phthalate (dibutyl phthalate is the mother liquor of the 50mM that is dissolved into of DMSO) 1.5mM, Amp100mg/L dull and stereotyped, cultivate 24 as a child picking the transformant of hydrolysis is arranged, namely be the positive transformant that contains phthalate esters softening agent hydrolase gene.
1.8 measuring precious biotechnology (Dalian) company limited, gene nucleotide series carries out sequencing, the nucleotides sequence of phthalate esters softening agent hydrolase gene is classified SEQ ID NO.1 as, and 315 coded aminoacid sequences of phthalate esters softening agent hydrolase gene nucleotide sequence are SEQ ID NO.2.
Embodiment 2: the high efficient expression of phthalate esters softening agent hydrolase gene in E.coli BL21 (DE3) (pET29a (+)).
With left end: 5 '-GGGGTACCATGAACGACGGCGCCACTCGTTATACC-3 ' and right-hand member:
5 '-CCCAAGCTTTGCTGCGCCGTTAGCTTCGGCGAC-3 ' is primer, with the PCR phthalate esters softening agent hydrolase gene fragment that increases from clay.
Amplification system:
The pcr amplification program:
A.95 ° C sex change 3min
B.94 ° C sex change 30s, 60 ° of C annealing 30s, 72 ° of C extend 1.0min, carry out 30 circulations
C.72 ° C extends 5min, cool to room temperature
PCR product KpnI and Hind III double digestion
Enzyme is cut system:
In 37 ° of C water-baths, reaction is more than 4 hours
Enzyme is cut product and is carried out 0.75% agarose gel electrophoresis and cut glue and reclaim.
PET29a (+) KpnI and Hind III double digestion (with reference to above-mentioned reaction conditions).
Plasmid after PCR fragment after enzyme is cut and enzyme are cut carries out enzyme connection (with reference to 1.5).
Enzyme joins good pET29a (+) recombinant plasmid transformed that contains phthalate esters softening agent hydrolase gene to expressive host bacterium BL21 (DE3), obtains recombinant microorganism BL21 (DE3).Be coated with the LB that contains dibutyl phthalate 1.5mM, Kan50mg/L, IPTG24mg/L dull and stereotyped.
Picking has the positive transformant of hydrolysis.
Embodiment 3: the enzyme that the checking positive transformant is expressed is to the degradation function of phthalate esters softening agent.
Positive transformant is cultured to OD in the LB substratum
6000.6 between 0.8, add IPTG to concentration 1.0mM, 28 ° of C cultivated 4 hours.100ml bacterium liquid is centrifugal, uses 10ml(50mM, pH7.0) the resuspended thalline of PBS damping fluid, ultrasonication 15 minutes.Get 200 μ l crude enzyme liquids and be added in (10mM, pH9.0) PBS damping fluid that 3ml contains 20ppm dibutyl phthalate (dipropyl phthalate, diamyl phthalate, dihexyl phthalate), the water-bath of 40 ° of C 2 hours.Add 6ml methyl alcohol, abundant mixing is got supernatant liquor after centrifugal and is detected degradation effect with HPLC/MS.Testing conditions is as follows: the temperature of dry gas, 250 ° of C; Dry gas flow velocity (nitrogen), 8L/min; Atomizer gaseous tension (nitrogen), 35psi; Capillary voltage, 4000 volts; The temperature of sheath gas, 300 ° of C; The flow of sheath gas, 10L/min; Spray nozzle voltage, 400 volts.Performance liquid chromatographic column is Eclipse XDB-C
18Post is 4.6 millimeters * 250 millimeters, 5.0 microns granular size (Agilent).Moving phase is 95% methyl alcohol and 5% water (0.1% formic acid), and flow velocity is 0.8ml/min.The degradation effect of dipropyl phthalate, dibutyl phthalate, diamyl phthalate, dihexyl phthalate is all in (such as table 1) more than 95%.
Table 1
| The softening agent title | Degradation effect |
| Dipropyl phthalate | 97.3% |
| Dibutyl phthalate | 100% |
| Diamyl phthalate | 100% |
| Dihexyl phthalate | 98.6% |
Claims (8)
1. phthalate esters softening agent lytic enzyme, its aminoacid sequence is shown in SEQ ID NO.2.
2. the encode gene of phthalate esters softening agent lytic enzyme claimed in claim 1, its nucleotide sequence is shown in SEQ ID NO.1.
3. a recombinant plasmid is characterized in that, it comprises phthalate esters softening agent hydrolase gene claimed in claim 2.
4. recombinant plasmid according to claim 3 is characterized in that, described plasmid is pET29a (+).
5. a recombinant microorganism is characterized in that, it comprises recombinant plasmid claimed in claim 3.
6. recombinant microorganism according to claim 5 is characterized in that, described microorganism is Escherichia coliBL21 (DE3).
7. the application of phthalate esters softening agent lytic enzyme claimed in claim 1 in removing the phthalate esters softening agent.
8. the application of phthalate esters softening agent hydrolase gene claimed in claim 2 in removing the phthalate esters softening agent.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310008487.3A CN103013944B (en) | 2013-01-10 | 2013-01-10 | Hydrolytic enzyme for diethyl phthalate plasticizer |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310008487.3A CN103013944B (en) | 2013-01-10 | 2013-01-10 | Hydrolytic enzyme for diethyl phthalate plasticizer |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109037766A (en) * | 2017-06-09 | 2018-12-18 | 河南师范大学 | A kind of biodegradable composite solid electrolyte film and preparation method thereof |
| CN110038250A (en) * | 2019-04-30 | 2019-07-23 | 江南大学 | A method of the phthalate of degradation metal ion or organic solvent |
| CN110373345A (en) * | 2019-05-08 | 2019-10-25 | 华东理工大学 | DEHP hydrolase and gene and its application in the degradation of phthalate plasticiser |
| CN112125947A (en) * | 2020-08-17 | 2020-12-25 | 赣州禾绿康健生物技术有限公司 | Method and equipment for removing plasticizer from ginsenoside extract |
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2013
- 2013-01-10 CN CN201310008487.3A patent/CN103013944B/en not_active Expired - Fee Related
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| US4133752A (en) * | 1976-06-02 | 1979-01-09 | Agency Of Industrial Science & Technology | Method for decomposition of phthalic acid esters by use of microorganisms |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109037766A (en) * | 2017-06-09 | 2018-12-18 | 河南师范大学 | A kind of biodegradable composite solid electrolyte film and preparation method thereof |
| CN110038250A (en) * | 2019-04-30 | 2019-07-23 | 江南大学 | A method of the phthalate of degradation metal ion or organic solvent |
| CN110373345A (en) * | 2019-05-08 | 2019-10-25 | 华东理工大学 | DEHP hydrolase and gene and its application in the degradation of phthalate plasticiser |
| CN112125947A (en) * | 2020-08-17 | 2020-12-25 | 赣州禾绿康健生物技术有限公司 | Method and equipment for removing plasticizer from ginsenoside extract |
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| CN103013944B (en) | 2014-01-01 |
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