CN102268396A - Amidase gene dimtH for degrading dimethoate, chlorpropham and propanil amide, and encoded protein and application thereof - Google Patents
Amidase gene dimtH for degrading dimethoate, chlorpropham and propanil amide, and encoded protein and application thereof Download PDFInfo
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Abstract
The invention belongs to the fields of environmental microbiology and agriculture, and relates to an amidase gene dimtH for degrading dimethoate, chlorpropham and propanil amide, and an encoded protein and application thereof. The amidase gene dimtH for degrading the dimethoate, the chlorpropham and the propanil amide has the full length of 1,395bp, and the sequence shown as SEQ ID NO.1; and a product encoded by the gene, namely amidase DimtH comprises 465 amino acids and has the sequence shown as SEQ ID NO.2. The DimtH can degrade a pesticide of dimethoate and herbicides of chlorpropham and propanil amide. The amidase gene dimtH can be used for constructing transgenic crops capable of degrading the pesticide of dimethoate and the herbicides of chlorpropham and propanil amide, also can be used for the removal of residues of the pesticide of dimethoate and the herbicides of chlorpropham and propanil amide from soil and water and the bioconversion in medicine synthesis, and has important theoretical and application value.
Description
Technical field
The invention belongs to applied environment microorganism and agriculture field, relate to amidase gene dimtH and the encoded protein matter and the application thereof of degrade Rogor, Y 3 and Stam F-34.
Background technology
China has a large population and a few land, and using chemical pesticide in a large number is the requisite measure that guarantees the high crop yield stable yields, but also brings severe contamination simultaneously.Show according to Chinese Academy of Sciences's data that soil detects, because the refractory organics of main pesticide grown, cause the residual rate of soil and farm crop higher, as reaching more than 90% at the Yangtze River Delta and Pearl River Delta agricultural land soil recall rate at present, be subjected to the agricultural-food of pesticidal contamination all the more so, detecting the result in 2009 as international green peace organization shows, China's 90% agricultural-food detect pesticide residue, the residual agricultural chemicals more than at least 5 kinds of 66% sample, the Qingdao poison leek incident that takes place in April, 2010 is exactly that the organic phosphorous insecticide pollution causes, and has caused baneful influence.The annual herbicide damage area of China reaches 3,000 ten thousand mu in addition, and wherein serious poisoning area reaches 5,000,000 mu, causes tens yuan loss every year.The microorganism recovery technique is a kind of novel biology in situ recovery technique, has effectively, and expense is low, and non-secondary pollution is fit to advantages such as big area pollution of area source reparation, is the main flow and the developing direction of soil organic pollutant recovery technique.
Sterilant Rogor and weedicide Y 3, Stam F-34 etc. are the agricultural chemicals of amide containing key, use extensively in China, the degradation property microorganism strains of sterilant Rogor and weedicide Y 3, these several agricultural chemicals of Stam F-34 of degrading has at present had report, but the degrading genes of these several agricultural chemicals of degrading yet there are no report.The degrading genes that obtains sterilant Rogor and amide containing key agricultural chemicals such as weedicide Y 3, Stam F-34 is being administered pesticide residue, eliminate in its poisoning technical research and have following effect and function, (1) by modern biotechnology degrading genes is imported crop and make up corresponding Herbicid resistant genetically modified crops, (two) make degradation bacterial agent or zymin with chemical residual degradation in the soil by modern microbial fermentation technology with degradation bacteria strains and gene.Degrading genes also can be used for useful Chemicals and the bio-transformation of medicine synthetic in addition.Therefore degrading genes has very important theory and using value in eliminating such herbicide damage and bio-transformation field.
Summary of the invention
The objective of the invention is above-mentioned deficiency, amidase gene dimtH and the encoded protein matter and the application thereof of degraded Rogor, Y 3 and Stam F-34 are provided at prior art.This amidase gene can be used for making up the genetically modified crops of degraded sterilant Rogor and weedicide Y 3, Stam F-34, sterilant Rogor and weedicide Y 3, the residual removal of Stam F-34 in soil, the water body.
Purpose of the present invention can be achieved through the following technical solutions:
Secondary coccus Dimd-2 (Paracoccus sp.) is kept at Chinese typical culture collection center, and deposit number is CCTCC M 2011234, and preservation date is on July 4th, 2011.
A kind of amidase gene dimtH, its nucleotides sequence classify SEQ ID NO.1 as.
The to degrade secondary coccus Dimd-2 (Paracoccus sp.) of Rogor, Y 3 and Stam F-34 of the strain that amidase gene dimtH of the present invention clone self-sizing obtains.Bacterial strain Dimd-2 is kept at Chinese typical culture collection center, deposit number is CCTCC M 2011234, preservation date is on July 4th, 2011, and mass spectrometry results shows that the crude enzyme liquid of bacterial strain Dimd-2 can be the fracture of the amido linkage of sterilant Rogor and weedicide Y 3, Stam F-34 and these target agricultural chemicals of degrading.
The strategy of taking of the amidase gene of clone's degraded Rogor, Y 3 and Stam F-34 is the shotgun (see figure 1), adopt the selection markers of a kind of amide containing key compound paracetamol as the target Ntn hydrolase, paracetamol (colourless) its amido linkage fracture under Ntn hydrolase catalysis generates the p-aminophenol of brown.At first extract total DNA of bacterial strain Dimd-2 (CCTCC M 2011234), the plasmid pUC118 enzyme that total DNA adopts partially digested back of Sau3AI and BamHI to cut with enzyme connects, enzyme connects total DNA library that product transformed into escherichia coli competent cell makes up bacterial strain Dimd-2, the library that builds is transformed in the bacillus coli DH 5 alpha, and the clone's daughter colony that contains the amidase gene of can degrade Rogor, Y 3 and Stam F-34 can produce brown pigment containing on the LB flat board of acetyl aminophenol.Utilize this cloning process to carry out high-throughout screening to the library.
The gene library that makes up with top Policy Filtering shotgun obtains an energy produces brown pigment on the LB flat board that adds the 100ppm acetyl aminophenol clone's, and further degradation experiment shows this positive colony can degrade Rogor, Y 3 and Stam F-34.Sequencing result shows that this positive colony contains 2,585bp wherein includes 12 potential ORF (greater than 150bp), and these potential ORF is carried out subclone and sequence alignment analysis respectively, the size of determining the gene of coding target Ntn hydrolase at last is 1395bp, called after dimtH.This is the amidase gene of being cloned into can degrade Rogor, Y 3 and Stam F-34 first.
The Ntn hydrolase DimtH that described amidase gene dimtH is coded, its aminoacid sequence is: SEQ ID NO.2.
The recombinant expression vector that contains described amidase gene dimtH.
Preferably the described amidase gene dimtH of claim 1 is inserted gained between the NdeI of pET-29a (+) and the EcoRI site.
The genetic engineering bacterium that contains described amidase gene dimtH.
The starting strain of described genetic engineering bacterium is e. coli bl21 (DE3).
The application of described amidase gene dimtH in the genetically modified crops that make up degraded sterilant Rogor, weedicide Y 3 or Stam F-34.
Application during described amidase gene dimtH sterilant Rogor and weedicide Y 3, Stam F-34 in removing water body is residual.
The application of described Ntn hydrolase DimtH in degraded sterilant Rogor, weedicide Y 3 or Stam F-34.
Application during described Ntn hydrolase DimtH sterilant Rogor and weedicide Y 3, Stam F-34 in removing water body is residual.
Beneficial effect
1. the present invention clones amidase gene dimtH with the shotgun success from bacterial strain Dimd-2 (CCTCC M 2011234).Show that at the GenBank comparison result this gene is a new gene, total length (from the initiator codon to the terminator codon) is 1395bp, 465 amino acid of encoding.
2. amidase gene dimtH of the present invention can in 8hr, degrade fully Rogor, Y 3 and the Stam F-34 of 100mg/L.DimtH can be used for making up the genetically modified crops of antiweed Y 3 and Stam F-34, also can be used for the removal and the bio-transformation of medicine synthetic of sterilant Rogor and weedicide Y 3 and Stam F-34 in soil, the water body, has very important theory and using value.
Description of drawings
Fig. 1 amidase gene dimtH clone's policy map.
Fig. 2 amidase gene dimtH is expression strategy figure in BL21 (pET-29a (+)).
Fig. 3 amidase gene dimtH protein electrophoresis collection of illustrative plates;
A: albumen marker; B: the DimtH albumen of purifying; C:IPTG inductive BL21 (DimtH) crude enzyme liquid; D: inductive BL21 not.
The catalytic Rogor degraded product of Fig. 4 DimtH GC/MS collection of illustrative plates;
A: the GC collection of illustrative plates of the enzyme liberating reaction solution dichloromethane extract of Rogor; B: retention time is the material one-level mass spectrum of 7.51min; C: retention time is the material one-level mass spectrum of 6.96min.
The catalytic Stam F-34 degraded product of Fig. 5 DimtH GC/MS collection of illustrative plates;
A: the GC collection of illustrative plates of the enzyme liberating reaction solution dichloromethane extract of Stam F-34; B: retention time is the material one-level mass spectrum of 8.88min; C: retention time is the material one-level mass spectrum of 6.37min.
The catalytic Y 3 degraded product of Fig. 6 DimtH GC/MS collection of illustrative plates;
A: the GC collection of illustrative plates of the enzyme liberating reaction solution dichloromethane extract of Y 3;
B: retention time is the material one-level mass spectrum of 5.99min;
C: retention time is the material one-level mass spectrum of 4.03min.
The biochemical reaction approach of Fig. 7 amine enzyme DimtH degraded Rogor.
The biochemical reaction approach of Fig. 8 amine enzyme DimtH degraded Stam F-34.
The biochemical reaction approach of Fig. 9 amine enzyme DimtH degraded Y 3.
Biomaterial preservation information
Secondary coccus Dimd-2 (Paracoccus sp.) is kept at Chinese typical culture collection center (CCTCC), place China Wuhan, and Wuhan University, deposit number is CCTCC M 2011234, preservation date is on July 4th, 2011.
Embodiment
Embodiment 1
(1) separation of amides pesticide degradation bacteria Dimd-2 (CCTCC M 2011234)
Being used for the enrichment matrix of enrichment amides pesticide degradation bacteria strains takes from the active sludge of Yangzhou insecticide factory agricultural chemicals waste water biochemical treatment tank, gets active sludge 5.0g and adds in the salt culture medium of 100ml basis, adds 50mgL
-1Fluorine bell urea, 30 ℃, 180rmin
-1Cultivated 7d days, the inoculum size with 5% is transferred in the identical substratum, transfers continuously 3 times, and the gradient dilution pregnant solution gets 10
-4～10
-7Each 0.1mL of dilution pregnant solution coats and is added with 100mgL
-1On the solid medium flat board of fluorine bell urea, behind 30 ℃ of cultivation 4d, single colony inoculation that picking grows is in containing 100mgL
-1In the substratum of various amides pesticides (fluorine bell urea, Rogor, Stam F-34 and Y 3), 30 ℃, 180rmin
-1Shaking table is cultivated 5d, verifies its degradation effect.Basis salt culture medium prescription is: 5.0 glucose, 1.0NH
4NO
3, 1.0NaCl, 1.5K
2HPO
4, 0.5KH
2PO
4, 0.2MgSO
47H
2O.pH 7.0. solid medium adds 15.0g agar.
The verification method of degradation effect: in nutrient solution, add isopyknic methylene dichloride and carry out the full dose extraction, standing demix behind the thermal agitation, after getting the methylene dichloride volatilization fully of 1ml lower floor then, add 1mL dissolve with methanol (chromatographically pure), filter with filter membrane (aperture 0.22 μ m).Adopt fluorine bell urea content in the high-performance liquid chromatogram determination extracting solution, liquid phase chromatogram condition: moving phase is methyl alcohol: and water (80: 20, V/V), Zorbax C218 ODS Spherex reversed-phase column (5 μ m, 4.6mm * 250mm, Agilent, USA), column temperature is a room temperature, UV-detector, measure wavelength 230nm, sample size 20 μ L, flow velocity is 0.8mLmin-1.External standard method is pressed peak area quantification.Adopt the content of Rogor, Stam F-34 and Y 3 in the gas chromatographic detection extracting solution, detection method: column type: BD-5MS quartz capillary column (15m * 0.25mm * 0.25 μ m); Sample feeding amount: 2 μ L; Splitting ratio: 30; Carrier gas: helium; Flow rate of carrier gas 1ml/min; Injector temperature is 230 ℃, and column temperature is 200 ℃.
From pregnant solution, be separated to 1 strain amides pesticide degradation bacteria, called after Dimd-2, this bacterium degradation rate to fluorine bell urea, Rogor, Stam F-34 and the Y 3 of 100mgL-1 in 7d all reaches more than 80%.
(2) evaluation of amides pesticide degradation bacteria Dimd-2 and biological characteristics
Strain morphology and physio-biochemical characteristics are measured reference literature common bacteria system identification handbook (eastern elegant pearl chief editor, Beijing, Science Press, 2001) and are carried out.
Bacterial strain Dimd-2 belongs to Gram-negative bacteria, the bacterium colony yellow, and neat in edge, the circular and projection of bacterium colony, moistening smooth, opaque, easy picking; Thalline is spherical, does not observe flagellum.Oxydase, oxydase and the nitrate reduction positive can not hydrolyzed starches, do not produce hydrogen sulfide, gelatine liquefication, VP, methyl red and urease negative.Can utilize L-arabinose, D-glucose, D-seminose and D-fructose to produce acid, can assimilate carbon sources such as D-glucose, D-seminose and D-sorbyl alcohol, but can not assimilate the D-wood sugar.
16S rRNA gene order Phylogenetic Analysis: high salt method is adopted in the extraction of the total DNA of bacterial strain, and with total DNA as template, carry out the amplification of 16S rRNA gene order.The primer that is used for amplified reaction is a pair of universal primer, upstream primer: 5 '-AGAGTTTGATCCTGGCTCAG-3 ' (SEQ ID No.5), downstream primer: 5 '-TACCTTGTTACGACTT-3 ' (SEQ ID No.6).25 μ L PCR reaction systems are: template 1 μ L, dNTP (25mmol/L) 2 μ L, each 1 μ L of primer (1mmol/L), 10 * Taq damping fluid, 2.5 μ L, Mg2+ (25mmol/L) 1.5 μ L, Taq enzyme (5U/ μ L) 0.3 μ L, ultrapure water 15.7 μ L.Polymerase chain reaction condition: 95 ℃ of pre-sex change 5min; 94 ℃ of sex change 0.5min, 52 ℃ of annealing 1min, 72 ℃ are extended 1min, circulate 30 times; 72 ℃ are extended 10min.Adopt PCR to reclaim the gene fragment that test kit (AXYGEN company) reclaims 16S rDNA, agarose gel electrophoresis detects the size (about 1.5kb) of amplified production, checks order behind the TA clone (being finished by BIOAISA company).Sequencing result is by on-line analysis, with the RDP database (https: // type strain 16S rRNA gene order in rdp.cme.msu.edu/) carries out similarity relatively.The result shows that bacterial strain Dimd-2 and paracoccus homology are the highest, reach 99.2% with the homology of Paracoccus aminovoransJCM 7685 (T), reach 97.8% with the homology of paracoccus type species Paracoccus denitrificans DSM 413 (T).Is paracoccus (Paracoccus sp.) in conjunction with physiological and biochemical property with this dientification of bacteria.This bacterial strain delivered be positioned at Chinese Wuhan, the Chinese typical culture collection center of Wuhan University (being called for short CCTCC) preservation, deposit number is CCTCC M 2011234, preservation date is on July 4th, 2011.
The clone of embodiment 2 amidase gene dimtH
(1) extraction of the total DNA of bacterial genomes
After bacterial strain Dimd-2 (CCTCC M 2011234) cultivates in the LB substratum in a large number, adopt the CTAB method to extract the genome DNA of high purity, big segmental Dimd-2, be dissolved in the TE damping fluid (pH8.0), place-20 ℃ of preservations, concrete grammar is with reference to " the fine works molecular biology experiment guide " of volumes such as F Ao Sibai.
(2) pUC118 (BamHI) buys in precious biotechnology (Dalian) company limited.
(3) it is partially digested that the enzyme of total DNA is cut the total DNA employing of Paracoccus sp.Dimd-2 Sau3AI.
(4) recovery of DNA
Total DNA after enzyme is cut carries out purifying by electrophoresis (TAE damping fluid), adopts axygen biosciences (China) to reclaim test kit and reclaims, and the DNA of recovery is dissolved among the TrisHCl of 10mmol/L (pH8.0), places-20 ℃ of preservations.
(5) enzyme connects
Set up following reaction system:
16 ℃ of incubations 12 hours.
(6) transform and screen
Adopt the selection markers of a kind of amide containing key compound paracetamol as the target Ntn hydrolase, paracetamol (colourless) its amido linkage fracture under Ntn hydrolase catalysis generates the p-aminophenol of brown.Get 10 μ l enzymes connect product transform 200 μ l bacillus coli DH 5 alpha competent cells (TaKaRa, Code:D9057), " fine works molecular biology experiment guide " P23 that concrete grammar is compiled with reference to F. Ao Sibai etc.Coating contains the LB flat board of 100mg/kg penbritin and 100mg/kg paracetamol, select clone's that produces brown pigment after cultivating 24h, can the further checking clone Rogor of degrading, clone's be seen the separation of (1) amides pesticide degradation bacteria among the embodiment 1 to the degraded test method of Rogor.
(7) gene nucleotide series is measured
Entrust Shanghai Ying Jun Bioisystech Co., Ltd to carry out sequencing positive colony of can degrade Rogor, Y 3 and the Stam F-34 of acquisition in (6), the nucleotides sequence of amidase gene dimtH is classified SEQ IDNO.1 as, and 465 aminoacid sequences being shifted onto according to amidase gene dimtH nucleotide sequence are SEQ ID NO.2.
(1) pcr amplification of amidase gene dimtH
With forward primer: 5 '-TCTGGA
CATATGATACCGAGACTGACCAACG-3 ' (SEQ ID NO.3) and reverse primer: 5 '-TCTGGA
GAATTCGCCTTCCATAAGAGCGCCGATAGC-3 ' (SEQ ID NO.4) is a primer, amplifies amidase gene dimtH sequence with PCR from the total DNA of Paracoccus sp.Dimd-2.
Amplification system:
The pcr amplification program:
A.95 ℃ sex change 3min;
B.95 ℃ sex change 1.5min, 53 ℃ of annealing 0.5min, 72 ℃ are extended 1.5min, carry out 25 circulations;
C.72 ℃ extend 10min, cool to room temperature.
(2) PCR product NdeI and EcoRI double digestion.
Enzyme is cut system:
In 37 ℃ of water-baths, more than the reaction 3h.Enzyme is cut product and is carried out 2% agarose gel electrophoresis and cut glue and reclaim.
(3) pET-29a (+) NdeI and EcoRI double digestion (with reference to 2.2).
(4) transform
(2) pET-29a (+) that enzyme cuts in the recovery fragment in and (3) carries out enzyme and connects pET-29a (+) recombinant plasmid pET-29a (+)-dimtH that (reference example 1 step (5)) obtains amide containing enzyme gene dimtH.Recombinant plasmid pET-29a (+)-dimtH is transformed into the recombinant microorganism BL21 (DimtH) that expressive host bacterium BL21 (DE3) obtains amide containing enzyme gene dimtH.
(5) expression of DimtH, purifying and functional verification
BL21 (DimtH) is cultured to OD in the LB substratum
600nmBe between 0.6 to 0.8, add IPTG, cultivated 4 hours for 30 ℃ to concentration 1mM.100ml bacterium liquid is centrifugal, with 10ml (50mM, pH 7.0) the resuspended thalline of PBS damping fluid, ultrasonication (Auto Science, UH-650B ultrasonic processor is 30%intensity) 5 minutes, centrifugal, collect supernatant, with the nickel ion affinity chromatograph post supernatant has been carried out purifying and got Ntn hydrolase DimtH (see figure 3).
(6) DimtH vitality test
Enzyme reaction system alive: 50mM phosphoric acid buffer (pH 7.0), 0.2mM target substrate (Rogor, Y 3 and Stam F-34), Ntn hydrolase DimtH 50 μ l, 30 ℃ of reaction 20min.Each reaction picks up counting to add enzyme, adds 3ml methylene dichloride termination reaction and thermal agitation, and organic phase is through anhydrous sodium sulfate dehydration after the layering, and the content of Rogor, Y 3 and Stam F-34 is measured (detection method sees 2.7) with high resolution gas chromatography.An enzyme activity unit (U) is defined as: at pH 7.0, under 30 ℃ of conditions of temperature, per minute catalysis reduces the required enzyme amount of 1 μ mol target substrate.
Degraded test shows the Ntn hydrolase DimtH behind the purifying can degrade Rogor, Y 3 and the Stam F-34 of 100mg/kg in 6hr.Zymetology test shows that DimtH lives to the ratio enzyme of Rogor, Y 3 and Stam F-34 and reaches 82,51 and 55U/mg protein respectively.
(7) meta-bolites determines
The dichloromethane extract of the Rogor in the step (6), Y 3 and Stam F-34 enzyme liberating reaction solution is dissolved in 100 μ L methyl alcohol after nitrogen dries up, utilize the meta-bolites in the GC/MS assaying reaction liquid.Detection method: column type: BD-5MS quartz capillary column (15m * 0.25mm * 0.25 μ m); The intensification condition: 50 ℃ of pillar initial temperatures, keep 1min, be warming up to 210 ℃ with 10 ℃/min, keep 2min again, be warming up to 240 ℃ with 20 ℃ of min-1 again, keep 10min again; Sample feeding amount: 2 μ L; Splitting ratio: 30; Carrier gas: helium; Flow rate of carrier gas 1ml/min; Injector temperature is 250 ℃.The mass spectrum operational condition: ion source is the EI source; 250 ℃ of ion source temperatures; Mass scanning scope (m/z): 30-650.
The GC/MS collection of illustrative plates of the dichloromethane extract of the enzyme liberating reaction solution of Rogor, Stam F-34 and Y 3 is seen Fig. 4, Fig. 5 and Fig. 6, draw Rogor, Stam F-34 and Y 3 by the spectrum data analysis and behind DimtH hydrolytic cleavage amido linkage, generate O respectively, O-dimethyl-S-(N-formic acid methyl) phosphorodithioate, 3,4-dichlorphenamide bulk powder and m-chloro aniline, the biochemical reaction approach is seen Fig. 7-9.
It is microbe-derived as follows that the present invention uses:
PUC118 (BamHI) is available from precious biotechnology (Dalian) company limited,
Intestinal bacteria DH10B is available from Shanghai Ying Jun Bioisystech Co., Ltd,
Colibacillus high expression vector pET-29a (+) is available from Novegen company,
Expressive host bacterium e. coli bl21 (DE3) is available from Shanghai Ying Jun Bioisystech Co., Ltd.
Claims (10)
1. secondary coccus Dimd-2 (Paracoccus sp.) is kept at Chinese typical culture collection center, and deposit number is CCTCC M 2011234, and preservation date is on July 4th, 2011.
2. amidase gene dimtH, its nucleotides sequence is classified SEQ ID NO.1 as.
3. the coded Ntn hydrolase DimtH of the described amidase gene dimtH of claim 2, its aminoacid sequence is: SEQ ID NO.2.
4. the recombinant expression vector that contains the described amidase gene dimtH of claim 2.
5. recombinant expression vector according to claim 3 is characterized in that it being that the described amidase gene dimtH of claim 2 is inserted gained between the NdeI of pET-29a (+) and the EcoRI site.
6. the genetic engineering bacterium that contains the described amidase gene dimtH of claim 2, the preferred e. coli bl21 of the starting strain of described genetic engineering bacterium (DE3).
7. the application of the described amidase gene dimtH of claim 2 in the genetically modified crops that make up degraded sterilant Rogor, weedicide Y 3 or Stam F-34.
8. the application of the described amidase gene dimtH of claim 2 sterilant Rogor and weedicide Y 3, Stam F-34 in removing water body in residual.
9. the application of the described Ntn hydrolase DimtH of claim 3 in degraded sterilant Rogor, weedicide Y 3 or Stam F-34.
10. the application of the described Ntn hydrolase DimtH of claim 3 sterilant Rogor and weedicide Y 3, Stam F-34 in removing water body in residual.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111139255A (en) * | 2018-11-06 | 2020-05-12 | 南京农业大学 | Propanil amidase gene pamD and coding protein and application thereof |
| CN114672438A (en) * | 2022-04-18 | 2022-06-28 | 淮北师范大学 | Carbamate herbicide degrading strain, broad-spectrum amidase gene and application |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2041944C1 (en) * | 1992-12-28 | 1995-08-20 | Акционерное общество закрытого типа "Биотехинвест" | Strain of bacteria paracoccusdenitrificans being destructor of aniline, 3-chloraniline,4-chloraniline and 3,4-dichloraniline |
| CA2107762C (en) * | 1991-04-26 | 1997-04-08 | Richard L. Tyndall | Amoebae/bacteria consortia and uses for degrading wastes and contaminants |
| CN1800355A (en) * | 2005-12-23 | 2006-07-12 | 南京农业大学 | Bacterium for degrading monocron pesticide residue and produced bacterium formulation |
| WO2007093523A1 (en) * | 2006-02-16 | 2007-08-23 | Wacker Chemie Ag | Method for the anaerobic degradation of organosilicon compounds by bacteria of the genus parac0ccus |
| CN101186898A (en) * | 2007-12-24 | 2008-05-28 | 北京大学 | Paracoccus and application thereof in nitrogenous heterocyclic compound degradation |
| US20100184121A1 (en) * | 2007-07-23 | 2010-07-22 | Henkel Ag & Co. Kgaa | Removal of by-products from crosslinkable preparations |
-
2011
- 2011-07-25 CN CN 201110208381 patent/CN102268396B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2107762C (en) * | 1991-04-26 | 1997-04-08 | Richard L. Tyndall | Amoebae/bacteria consortia and uses for degrading wastes and contaminants |
| RU2041944C1 (en) * | 1992-12-28 | 1995-08-20 | Акционерное общество закрытого типа "Биотехинвест" | Strain of bacteria paracoccusdenitrificans being destructor of aniline, 3-chloraniline,4-chloraniline and 3,4-dichloraniline |
| CN1800355A (en) * | 2005-12-23 | 2006-07-12 | 南京农业大学 | Bacterium for degrading monocron pesticide residue and produced bacterium formulation |
| WO2007093523A1 (en) * | 2006-02-16 | 2007-08-23 | Wacker Chemie Ag | Method for the anaerobic degradation of organosilicon compounds by bacteria of the genus parac0ccus |
| US20100184121A1 (en) * | 2007-07-23 | 2010-07-22 | Henkel Ag & Co. Kgaa | Removal of by-products from crosslinkable preparations |
| CN101186898A (en) * | 2007-12-24 | 2008-05-28 | 北京大学 | Paracoccus and application thereof in nitrogenous heterocyclic compound degradation |
Non-Patent Citations (3)
| Title |
|---|
| 《International Biodeterioration & Biodegradation》 20110131 M. Santoshkumar et al. Utilization of aliphatic nitrile by Paracoccus sp. SKG isolated from chemical waste samples 153-159 1-10 第65卷, 第1期 * |
| 《南京农业大学博士学位论文》 20080115 贾开志 久效磷降解菌的分离、坚定及降解酶基因的克隆 1-10 , * |
| 《浙江工商大学硕士学位论文》 20090215 蔡晶晶 链霉菌（Streptomyces sp.）的筛选及其降解水肿含氮杂环化合物的特性研究 1-10 , * |
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| CN111139255A (en) * | 2018-11-06 | 2020-05-12 | 南京农业大学 | Propanil amidase gene pamD and coding protein and application thereof |
| CN111139255B (en) * | 2018-11-06 | 2022-07-01 | 南京农业大学 | Propanil amidase gene pamD and coding protein and application thereof |
| CN114672438A (en) * | 2022-04-18 | 2022-06-28 | 淮北师范大学 | Carbamate herbicide degrading strain, broad-spectrum amidase gene and application |
| CN114672438B (en) * | 2022-04-18 | 2023-06-30 | 淮北师范大学 | Carbamate herbicide degrading strain, broad-spectrum amidase gene and application |
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