CN106244507A - The application of Burkholderia pyrrocinia and the method for degraded phthalic acid 2 Octyl Nitrite thereof - Google Patents
The application of Burkholderia pyrrocinia and the method for degraded phthalic acid 2 Octyl Nitrite thereof Download PDFInfo
- Publication number
- CN106244507A CN106244507A CN201610880270.5A CN201610880270A CN106244507A CN 106244507 A CN106244507 A CN 106244507A CN 201610880270 A CN201610880270 A CN 201610880270A CN 106244507 A CN106244507 A CN 106244507A
- Authority
- CN
- China
- Prior art keywords
- phthalic acid
- octyl nitrite
- burkholderia pyrrocinia
- burkholderia
- pyrrocinia
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Emergency Management (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Business, Economics & Management (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Toxicology (AREA)
- Molecular Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to application and the method for degraded phthalic acid 2 Octyl Nitrite thereof of Burkholderia pyrrocinia, belong to environmental microorganism application.The purposes coughing up bulkholderia cepasea B1213 of the present invention is degraded phthalic acid 2 Octyl Nitrite；The degradation rate of phthalic acid 2 Octyl Nitrite be may be up to 73.49%；Described bulkholderia cepasea B1213 of coughing up is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center；Deposit number is CGMCCNo.12806；The preservation time is on July 21st, 2016.The present invention uses the method coughing up bulkholderia cepasea B1213 degraded phthalic acid 2 Octyl Nitrite, is that Burkholderia pyrrocinia B1213 contacts with phthalic acid 2 Octyl Nitrite under conditions of yeast extract exists.Compared with the method for antibacterial, fungus degrading phthalic acid 2 Octyl Nitrite before the present invention, the method for the present invention, strain source has new meaning, it is higher to the degradation rate of phthalic acid 2 Octyl Nitrite, and degradation cycle is short, simple to operate.
Description
Technical field
The present invention relates to application and the method for degraded phthalic acid 2-Octyl Nitrite thereof of Burkholderia pyrrocinia,
Belong to environmental microorganism application.
Background technology
Phthalic acid 2-Octyl Nitrite (Bison(2-ethylhexyl) ortho-phthalate), vehicle economy HP, be
The environmental hormone class organic synthesis compound that one class is important, is mainly used as polrvinyl chloride (PVC) plastics in the industrial production and increases
Mould agent, improve plasticity and the flexibility of plastics.
In plastics, DEHP is connected with hydrogen bond or Van der Waals force with plastic molecules, it is easy to by drip washing, migrate or evaporation etc.
Mode enters in food, air, drinking water and other material；DEHP can also pass through flue dust in plastics-production or combustion process
Sedimentation is discharged in environment.Many Results of Animal display DEHP have Reproductive and developmental toxicity, immunotoxicity, embryo
The multiple toxicity such as toxicity, hepatotoxicity and carcinogenecity, and there is the quasi-waverider vehicle causing thyroxine metabolism to change.The U.S.
Environmental Protection Agency (EPA), European Union and China National Environmental Monitoring Center have been listed in priority pollutants blacklist the most.
In environment, the decomposition of DEHP, method for transformation and technology exploration have become the important research direction of environmental pollution improvement.
But DEHP hydrolysis in natural environment, photodissociation speed are slowly, belong to hard-degraded substance.There are some researches show, DEHP's
Hydrolysising half-life reaches 2000, and Photolysis Half is also required to 105 years.Compared with hydrolysis and the mechanical degradation such as photodissociation, based on micro-life
The biodegradation process of thing metabolism has the feature such as functional microorganism multiformity, process simple, low cost, environmental friendliness so that
Microbial degradation is considered as the most effective approach of DEHP permineralization in natural environment.Therefore, efficient-decomposition DEHP is screened
Microorganism also illustrates its catabolic pathway, has reality meaning for in-depth about the research and application eliminating DEHP environmental hazard
Justice.
In recent years, the microbial degradation of DEHP is studied the most widely, and the bacterial strain of energy efficient degradation DEHP is in a large number
Through isolated from all kinds of environment, including the activated sludge etc. of Rhizophora apiculata Blume, soil, ocean, river and waste water treatment plant, micro-life
Species does not include a large amount of antibacterial and some fungi.Research shows, separate sources and different types of microorganism are dropped at DEHP
Solution approach aspect has larger difference, and the degradation mechanism presented also is not quite similar.Described degradation mechanism includes playing degraded
Effect key enzyme and enzyme system, degradation pathway in key node and key influence factor, microorganism own metabolism with degraded effect
Relation, microbial degradation broad spectrum activity and specificity and the relation etc. with DEHP structure thereof between Guo.
Burkholderia pyrrocinia (Burkholderia pyrrocinia), its application is mainly promoting plant growing
Biological control with plant disease；Have no the report about utilizing Burkholderia pyrrocinia degraded DEHP at present.
Summary of the invention
It is an object of the invention to provide the new strains of a kind of phthalic acid 2-Octyl Nitrite of can degrading；And this bacterial strain
Application and the method that uses this strains for degrading phthalic acid 2-Octyl Nitrite.
Technical scheme
The present invention screens from soil and has isolated the bacterial strain that a strain is new；It is shaft-like gram negative bacteria, and can not be formed
Spore；Identified, this bacterial strain be a kind of Burkholderia pyrrocinia (Burkholderia pyrrocinia)；Named pyrrole
Cough up bulkholderia cepasea B1213；It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center
(CGMCC)；Deposit number is CGMCCNo.12806；The preservation time is on July 21st, 2016.
The Burkholderia pyrrocinia B1213 of the present invention can degrade phthalic acid 2-Octyl Nitrite；Pyrroles primary gram
Hall moral Salmonella B1213 may be up to 55.66-73.49% to the degradation rate of phthalic acid 2-Octyl Nitrite.
Use the one of which of the Burkholderia pyrrocinia B1213 degraded phthalic acid 2-Octyl Nitrite of the present invention
Method is: under conditions of yeast extract exists, Burkholderia pyrrocinia B1213 with DEHP contacts.Wherein, yeast carries
The amount taking thing can affect degradation rate.So, said method, it is preferred that at the bar that improvement BSM basis salt fluid medium exists
Under part, Burkholderia pyrrocinia B1213 with DEHP contacts；Described improvement BSM basis salt fluid medium, contains in every 1L
There is a following component: yeast extract 2-10g, ammonium sulfate 0.5g, sodium chloride 4.0g, three hypophosphite monohydrate potassium 0.5g, seven hydrated sulfuric acid
Magnesium 0.4g, distilled water surplus；pH7.0.
Concrete, it is that DEHP is added improvement BSM basis salt fluid medium；By Burkholderia pyrrocinia
B1213 seed liquor is inoculated in improvement BSM basis salt fluid medium, constant temperature shake under conditions of 150-200 rpm, 30-35 DEG C
Swing cultivation.
Said method, it is preferred that in the salt fluid medium of improvement BSM basis, the content of yeast extract is 7g/L.
Said method, it is preferred that condition of culture is rotating speed 175rpm, temperature 30 DEG C, under other identical conditions, this
Under part, the degradation rate of bacterium B1213 is the highest.
Said method, Burkholderia pyrrocinia B1213 seed liquor is relative to improvement BSM basis salt fluid medium
The change of inoculum concentration, the DEHP degradation rate in the unit interval is had a significant effect；Under normal circumstances, inoculum concentration is the most, unit
In time, degradation rate is the highest；But to final degradation rate (degradation rate when DEHP content reaches stable in fermentation liquid) the most substantially
Impact.
Said method, described Burkholderia pyrrocinia B1213 seed liquor is by Burkholderia pyrrocinia B1213
Cultivate acquisition.Under conditions of obtaining Burkholderia pyrrocinia B1213, those skilled in the art by routine operation are
Burkholderia pyrrocinia B1213 seed liquor can be obtained.
In the present invention, for the percentage ratio of the content of certain composition, if not otherwise specified, refer both to percentage by weight
(w/w).
Technical term explanation used by the present invention: rpm is Speed unit, and 1 rpm refers to that rotation per minute is gone around.
Beneficial effect:
(1) separation and Culture goes out to degrade the Burkholderia pyrrocinia B1213 of phthalic acid 2-Octyl Nitrite first；
(2) Burkholderia pyrrocinia B1213 is 55.66-73.49% to the degradation rate of phthalic acid 2-Octyl Nitrite；
(3) present invention degrades antibacterial, fungus degrading O-phthalic before the method for phthalic acid 2-Octyl Nitrite, with the present invention
The method of acid 2-Octyl Nitrite is compared, and has new meaning on strain source, and degradation rate is high, and degradation cycle is short, simple to operate.
Preservation information:
Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center；
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica；
Preservation date: on July 21st, 2016；
Deposit number: CGMCCNo.12806；
Classification And Nomenclature: Burkholderia pyrrocinia (Burkholderia pyrrocinia).
Accompanying drawing explanation
Fig. 1 is the Burkholderia pyrrocinia B1213 morphological characteristic under the microscope of the present invention；In Fig. 1, Bi Kabai
Ke Huoerde Salmonella B1213 is shaft-like gram negative bacteria, and can not form spore；
Fig. 2 is the 16SrDNA sequential system developmental analysis of Burkholderia pyrrocinia B1213；
Fig. 3 is that the recA sequential system of Burkholderia pyrrocinia B1213 grows tree analysis.
Detailed description of the invention
In following embodiment, if no special instructions, it is method commonly used in the art.Agents useful for same or the unreceipted production of instrument
Manufacturer person, be can by city available from conventional products.
The qualification of embodiment 1 bacterial strain B1213
Bacterial strain B1213 is isolatable from soil, and its form is as it is shown in figure 1, belong to shaft-like gram negative bacteria, it is impossible to form spore.
Extract its genomic DNA, and utilize 16s rDNA and BCR1 primer to carry out polymerase chain reaction for this genomic DNA
(Polymerase Chain Reaction, PCR) is to breed specific DNA sequence, and utilizes American National Biotechnology Information
Center (National Center for Biotechnology Information is called for short NCBI) data base carries out bacterial strain ratio
Right.
(1) 16S rDNA sequence analysis:
16s rDNA forward primer 27F:5 '-AGA GTT TGA TCC TGG CTC AG-3 ', as shown in SEQ ID NO.1；
16s rDNA reverse primer 1492R:5'-ACG GTT ACC TTG TTA CGA CTT-3', such as SEQ ID NO.2 institute
Show；
Amplification gained 16S rDNA sequence is as shown in SEQ ID NO:3, and sequence is 1411bp.By this amplification gained 16S
RDNA sequence compares with the gene order of the related strain in GenBank data base, carries out sequence ratio with MEGA4.1 software
Right, use and face a connection method phylogenetic tree construction, through 1000 stochastic sampling, calculate Bootstrap value, constructed system
Grow tree such as Fig. 3.Figure is grown tree node and only shows that Bootstrap value is more than 50% numerical value, upper target " T " intermediate scheme bacterium
Strain.Bacterial strain " B1213 " and type strain Burkholderia stabilis LMG 28156, homology can be found from NCBI
Property reaches 99.7%.
(2) recA gene sequencing
BCR1 gene forward primer: 5'-TgA CCg CCg AgA AgA gCA A-3', as shown in SEQ ID NO.4；
BCR2 gene reverse primer: 5'-CTC TTC TTC gTC CAT CgC CTC-3', as shown in SEQ ID NO.5；
Amplification gained recA gene order is as shown in SEQ ID NO:6, and sequence is 974bp.By this amplification gained recA base
Because the gene order of sequence with the related strain in GenBank data base compares, carry out sequence ratio with MEGA4.1 software
Right, use and face a connection method phylogenetic tree construction, through 1000 stochastic sampling, calculate Bootstrap value, constructed system
Grow tree such as Fig. 4.Figure is grown tree node and only shows that Bootstrap value is more than 50% numerical value, upper target " T " intermediate scheme bacterium
Strain.Bacterial strain " B1213 " and type strain Burkholderia pyrrocinia DSM 10685T can be found out from NCBI
(CP011503) homology reaches 97.9%.
Therefore can be determined that B1213 is a kind of brand-new Burkholderia pyrrocinia.Bacterium belonging to it is confirmed through identifying
After Zhong, Burkholderia pyrrocinia B1213 is preserved in China Committee for Culture Collection of Microorganisms on July 21st, 2016
Common micro-organisms center；Deposit number is CGMCCNo.12806；This biomaterial survival test is tested and passes through this examination
Test.
Prepared by embodiment 2 Burkholderia pyrrocinia B1213 liquid submerged culture liquid
(1) slant culture: Burkholderia pyrrocinia B1213 is inoculated on slant medium, cultivates 48 hours at 40 DEG C,
Obtain inclined-plane bacterial strain.Slant medium used, the composition contained in every 1L is: glucose 2.0g, peptone 1.0g, yeast extract
0.5g, agar 1.8g, distilled water surplus, pH is neutral, 115 DEG C of sterilizing 20 min.
(2) inclined-plane inoculation is taken in sterilized seed culture medium, in 175 rpm, constant temperature shake under conditions of 30 DEG C
Swing cultivation 24h, obtain seed culture fluid.Seed culture medium used, the composition contained in every 1L is: ammonium sulfate 0.5g, sodium chloride
4.0g, three hypophosphite monohydrate potassium 0.5g, bitter salt 0.4g, agar powder 15g, yeast extract 5g, distilled water surplus,
PH7.0,121 DEG C of sterilizing 25min.
(3) by the DEHP(technical pure of 10 μ L) add the improvement BSM basis salt liquid culture containing 50mL with sterile working
In the shaking flask of base, the seed culture fluid of Burkholderia pyrrocinia B1213 is inoculated in shaking flask with the inoculum concentration of 1mL,
175r/min, isothermal vibration cultivates 72h under conditions of 30 DEG C；Obtain fermentation liquid.Improvement BSM basis used salt fluid medium, often
The composition contained in 1L is: yeast extract 5g, ammonium sulfate 0.5g, sodium chloride 4.0g, three hypophosphite monohydrate potassium 0.5g, seven hydration sulfur
Acid magnesium 0.4g, distilled water surplus, pH7.0,121 DEG C of sterilizing 15min.
Embodiment 3 DEHP standard curve is formulated and assay
(1) high performance liquid chromatography (HPLC) makes standard curve: the 600 μ L DEHP mark product that take, with methanol as solvent, are prepared as
The DEHP solution of 1mg/mL, takes 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, 1.2mL, 1.4mL after diluting ten times fixed respectively
Hold in 2mL centrifuge tube, i.e. Concentraton gradient (μ g/mL) is 10,20,30,40,50,60,70 to have machine filter by 0.45 μm respectively
Membrane filtration is placed in liquid phase bottle to be measured.Efficient liquid phase testing conditions in the present embodiment 3 is: chromatographic column is Sepax Gp-
C18 post (150mm*4.6mm, 5.0 μm)；Flowing is acetonitrile-water (83:17, V/V) mutually；Sample size 10 μ L；Flow velocity 1.0mL/
min；Column temperature 25C °；UV-detector wavelength 210nm.With the peak area of DEHP as abscissa, with the concentration of DEHP as vertical coordinate,
Make standard curve；And then obtain peak area-concentration equation.
The degradation rate of embodiment 4 high-efficient liquid phase technique detection DEHP
(1) in the fermentation liquid that embodiment 2 obtains, add the acetonitrile with fermentation liquid equivalent DEHP is extracted, ultrasonic
Take after (40KHZ, 300W) assisted extraction 30min centrifugal after 2mL solution acetonitrile is settled to 10mL mixing (12000rpm,
10min), the supernatant after centrifugal is filtered with organic filter membrane (0.45 μm), discard just filtrate, take subsequent filtrate and be placed in liquid phase
In bottle, with the testing conditions of embodiment 3 step (1), high performance liquid chromatograph (HPLC) is used to detect.The operation time is
20 minutes, reading retention time, at the peak area value of about 7 min, according to the peak area-concentration equation of embodiment 3, must continue filter
The concentration of DEHP in liquid, and then calculate the residual concentration of DEHP in fermentation liquid.
(2) computing formula of degradation rate: degradation rate %=(C0-C)/C0*100%, C0 is for using Burkholderia pyrrocinia
DEHP mass concentration (μ g/mL) (i.e. mixed liquor in shaking flask before embodiment 2 step (3) fermentation culture before B1213 degraded
DEHP mass concentration: 0.1713 μ g/mL) in, C is the DEHP residual concentration (μ g/mL) in fermentation liquid.It is computed, implements
The degradation rate of example 2 is 66.73%.
The optimization of embodiment 5 Burkholderia pyrrocinia B1213 degraded DEHP
Seed liquor inoculum concentration in embodiment 2 step (3) is changed into successively 2,3,4,5,6mL, other operations are with embodiment 2.Press
According to the DEHP concentration of the step measurements fermentation liquid of embodiment 4, and then calculate the degradation rate of DEHP.Under the conditions of different vaccination amount,
DEHP degradation rate is as shown in table 1.
The optimization of embodiment 6 Burkholderia pyrrocinia B1213 degraded DEHP
The content of the yeast extract of improvement BSM basis used salt fluid medium in embodiment 2 step (3) is revised successively
Be 2,3,4,6,7,8,10g/L；Other operations are with embodiment 2.According to the DEHP concentration of the step measurements fermentation liquid of embodiment 4,
And then calculate the degradation rate of DEHP.Under the conditions of the content of different yeast extracts, DEHP degradation rate is as shown in table 2.
Table 1
| Seed liquor inoculum concentration (mL) | DEHP degradation rate (%) |
| 0 | 0 |
| 1 | 66.73 |
| 2 | 68.29 |
| 3 | 70.25 |
| 4 | 70.63 |
| 5 | 70.54 |
| 6 | 70.21 |
Table 2
| The content (g/L) of yeast extract | DEHP degradation rate (%) |
| 0 | 0 |
| 2 | 55.66 |
| 3 | 58.35 |
| 4 | 59.18 |
| 5 | 66.73 |
| 6 | 70.38 |
| 7 | 73.49 |
| 8 | 72.15 |
| 10 | 72.08 |
<110>Beijing Technology and Business University
<120>Burkholderia pyrrocinia application and degraded phthalic acid 2-Octyl Nitrite method
<160>6
<210>1
<211>20
<212>DNA
<213>synthetic
<400>1
AGAGTTTGAT CCTGGCTCAG 20
<210>2
<211>21
<212>DNA
<213>synthetic
<400>2
ACGGTTACCT TGTTACGACT T 21
<210>3
<211>1411
<212>DNA
<213>synthetic
<400>3
ACATGCAGTC GAACGGCAGC ACGGGTGCTT GCACCTGGTG GCGAGTGGCG AACGGGTGAG 60
TAATACATCG GAACATGTCC TGTAGTGGGG GATAGCCCGG CGAAAGCCGG ATTAATACCG 120
CATACGATCT ACGGATGAAA GCGGGGGACC TTCGGGCCTC GCGCTATAGG GTTGGCCGAT 180
GGCTGATTAG CTAGTTGGTG GGGTAAAGGC CTACCAAGGC GACGATCAGT AGCTGGTCTG 240
AGAGGACGAC CAGCCACACT GGGACTGAGA CACGGCCCAG ACTCCTACGG GAGGCAGCAG 300
TGGGGAATTT TGGACAATGG GCGAAAGCCT GATCCAGCAA TGCCGCGTGT GTGAAGAAGG 360
CCTTCGGGTT GTAAAGCACT TTTGTCCGGA AAGAAATCCT TGGCTCTAAT ACAGTCGGGG 420
GATGACGGTA CCGGAAGAAT AAGCACCGGC TAACTACGTG CCAGCAGCCG CGGTAATACG 480
TAGGGTGCGA GCGTTAATCG GAATTACTGG GCGTAAAGCG TGCGCAGGCG GTTTGCTAAG 540
ACCGATGTGA AATCCCCGGG CTCAACCTGG GAACTGCATT GGTGACTGGC AGGCTAGAGT 600
ATGGCAGAGG GGGGTAGAAT TCCACGTGTA GCAGTGAAAT GCGTAGAGAT GTGGAGGAAT 660
ACCGATGGCG AAGGCAGCCC CCTGGGCCAA TACTGACGCT CATGCACGAA AGCGTGGGGA 720
GCAAACAGGA TTAGATACCC TGGTAGTCCA CGCCCTAAAC GATGTCAACT AGTTGTTGGG 780
GATTCATTTC CTTAGTAACG TAGCTAACGC GTGAAGTTGA CCGCCTGGGG AGTACGGTCG 840
CAAGATTAAA ACTCAAAGGA ATTGACGGGG ACCCGCACAA GCGGTGGATG ATGTGGATTA 900
ATTCGATGCA ACGCGAAAAA CCTTACCTAC CCTTGACATG GTCGGAATCC CGCTGAGAGG 960
TGGGAGTGCT CGAAAGAGAA CCGATACACA GGTGCTGCAT GGCTGTCGTC AGCTCGTGTC 1020
GTGAGATGTT GGGTTAAGTC CCGCAACGAG CGCAACCCTT GTCCTTAGTT GCTACGCAAG 1080
AGCACTCTAA GGAGACTGCC GGTGACAAAC CGGAGGAAGG TGGGGATGAC GTCAAGTCCT 1140
CATGGCCCTT ATGGGTAGGG CTTCACACGT CATACAATGG TCGGAACAGA GGGTTGCCAA 1200
CCCGCGAGGG GGAGCTAATC CCAGAAAACC GATCGTAGTC CGGATTGCAC TCTGCAACTC 1260
GAGTGCATGA AGCTGGAATC GCTAGTAATC GCGGATCAGC ATGCCGCGGT GAATACGTTC 1320
CCGGGTCTTG TACACACCGC CCGTCACACC ATGGGAGTGG GTTTTACCAG AAGTGGCTAG 1380
TCTAACCGCA AGGAGGACGG TCACCACGGT A 1411
<210>4
<211>19
<212>DNA
<213>synthetic
<400>4
TgACCgCCgA gAAgAgCAA 19
<210>5
<211>21
<212>DNA
<213>synthetic
<400>5
CTCTTCTTCg TCCATCgCCT C 21
<210>6
<211>974
<212>DNA
<213>synthetic
<400>6
TCATGCGCAT GGGCGACGGC GAGGCGGCCG AAGACATCCA GGTCGTCTCC ACGGGTTCGC 60
TGGGGCTCGA CATCGCGCTT GGCGTCGGCG GCTGCCGCGC GGCCGGGTGG TCGAGATCTA 120
CGGCCCGGAA TCGTCCGGTA AAACCACGCT CACGCTGCAG GTCATCGCCG AACTGCAGAA 180
GATCGGCGGC ACGGCCGCGT TCATCGACGC CGAACACGCG CTCGACGTTC AATATGCGTC 240
GAAGCTCGGC GTGAACGTGC CGGAACTGCT GATCTCGCAG CCGGACACCG GCGAACAGGC 300
ACTGGAAATC ACCGATGCGC TGGTGCGCTC GGGCTCGGTC GACATGATCG TCATCGACTC 360
GGTCGCGGCG CTCGTGCCGA AGGCCGAAAT CGAAGGCGAG ATGGGCGATT CGCTGCCGGG 420
CCTGCAGGCT CGCCTGATGT CGCAGGCGCT GCGCAAGCTG ACCGGCACGA TCAAGCGCAC 480
GAACTGCCTG GTGATCTTCA TCAACCAGAT TCGCATGAAG ATCGGCGTGA TGTTCGGCAA 540
CCCGGAAACC ACGACGGGCG GCAACGCGCT GAAGTTCTAT GCGTCGGTGC GTCTCGATAT 600
CCGCCGGATC GGCTCGATCA AGAAGAACGA CGAGGTGATC GGCAACGAAA CCCGCGTGAA 660
GGTCGTCAAG AACAAGGTGT CGCCGCCGTT CCGCGAAGCG ATCTTCGACA TCCTGTATGG 720
CGAGGGCATT TCGCGTCAGG GCGAGATCAT CGATCTCGGC GTGCAGGCAA AGATCGTCGA 780
CAAGGCGGGC GCCTGGTACA GCTACAACGG CGAGAAGATC GGCCAGGGCA AGGACAACGC 840
GCGTGAATTC CTGCGCGAGA ATCCGGAAAT CGCACGCGAG ATCGAAAACC GCATCCGCGA 900
ATCGCTCGGC GTCGTCGCCA AGGCGCTGGC GGCCGCACTC GCGCAGATCG AGAAGCAGTT 960
CGGCAAAGGG TCGA 974
Claims (7)
1. Burkholderia pyrrocinia (Burkholderia pyrrocinia) a kind of purposes of B1213, it is used for adjacent benzene of degrading
Dioctyl phthalate 2-Octyl Nitrite；Described Burkholderia pyrrocinia B1213 is preserved in Chinese microorganism strain preservation conservator
Can common micro-organisms center；Deposit number is CGMCCNo.12806；The preservation time is on July 21st, 2016.
2. one kind uses Burkholderia pyrrocinia B1213 degraded phthalic acid 2-Octyl Nitrite described in claim 1
Method, it is characterised in that: under conditions of yeast extract exists, Burkholderia pyrrocinia B1213 and phthalic acid
2-Octyl Nitrite contacts.
Method the most according to claim 2, it is characterised in that in the condition that improvement BSM basis salt fluid medium exists
Under, Burkholderia pyrrocinia B1213 contacts with phthalic acid 2-Octyl Nitrite；The salt liquid training of described improvement BSM basis
Support base, containing following component in every 1L: yeast extract 2-10g, ammonium sulfate 0.5g, sodium chloride 4.0g, three hypophosphite monohydrate potassium
0.5g, bitter salt 0.4g, distilled water surplus；pH7.0.
Method the most according to claim 3, it is characterised in that yeast extract in the salt fluid medium of improvement BSM basis
Content be 7g/L.
5. according to the method described in claim 3 or 4, it is characterised in that phthalic acid 2-Octyl Nitrite to be added improvement BSM
Basis salt fluid medium；Burkholderia pyrrocinia B1213 seed liquor is inoculated in improvement BSM basis salt liquid culture
Base, cultivates at isothermal vibration under conditions of 150-200 rpm, 30-35 DEG C.
Method the most according to claim 5, it is characterised in that rotating speed 175rpm, temperature 30 DEG C.
Method the most according to claim 6, it is characterised in that described Burkholderia pyrrocinia B1213 seed liquor is
Acquisition is cultivated by Burkholderia pyrrocinia B1213.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610880270.5A CN106244507B (en) | 2016-10-09 | 2016-10-09 | The application of Burkholderia pyrrocinia and its method for the ethylhexyl of phthalic acid 2 of degrading |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610880270.5A CN106244507B (en) | 2016-10-09 | 2016-10-09 | The application of Burkholderia pyrrocinia and its method for the ethylhexyl of phthalic acid 2 of degrading |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106244507A true CN106244507A (en) | 2016-12-21 |
| CN106244507B CN106244507B (en) | 2018-04-10 |
Family
ID=57612434
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610880270.5A Expired - Fee Related CN106244507B (en) | 2016-10-09 | 2016-10-09 | The application of Burkholderia pyrrocinia and its method for the ethylhexyl of phthalic acid 2 of degrading |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106244507B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112980722A (en) * | 2019-09-29 | 2021-06-18 | 北京工商大学 | Burkholderia polyphylla culture method and application thereof in catalytic degradation of harmful esters of white spirit |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002142754A (en) * | 2000-11-08 | 2002-05-21 | I M B Kk | Bacterium capable of scavenging phthalic ester |
| CN105316251A (en) * | 2015-02-15 | 2016-02-10 | 华东理工大学 | Strain with efficient phthalic acid esters (PAEs) degrading function and application of strain |
| CN103589659B (en) * | 2013-09-18 | 2016-08-17 | 中国科学院南京土壤研究所 | One strain spherical Rhodococcus fascians WJ4 and the application in phthalic acid ester contaminated soil remediation thereof |
-
2016
- 2016-10-09 CN CN201610880270.5A patent/CN106244507B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002142754A (en) * | 2000-11-08 | 2002-05-21 | I M B Kk | Bacterium capable of scavenging phthalic ester |
| CN103589659B (en) * | 2013-09-18 | 2016-08-17 | 中国科学院南京土壤研究所 | One strain spherical Rhodococcus fascians WJ4 and the application in phthalic acid ester contaminated soil remediation thereof |
| CN105316251A (en) * | 2015-02-15 | 2016-02-10 | 华东理工大学 | Strain with efficient phthalic acid esters (PAEs) degrading function and application of strain |
Non-Patent Citations (3)
| Title |
|---|
| MIN ZHANG ET AL.: "Isolation, Identification and Biodegradation Characteristics of a Phthalate Ester Degrading Bacterium", 《AGRICULTURAL SCIENCE & TECHNOLOGY》 * |
| 张付海等: "一株邻苯二甲酸酯降解菌降解特性研究", 《农业环境科学学报》 * |
| 徐衍舜等: "开发及分离新型分解邻苯二甲酸二酯(DEHP)细菌之研究", 《台湾大学农业化学研究所学位论文》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112980722A (en) * | 2019-09-29 | 2021-06-18 | 北京工商大学 | Burkholderia polyphylla culture method and application thereof in catalytic degradation of harmful esters of white spirit |
| CN112980722B (en) * | 2019-09-29 | 2022-10-14 | 北京工商大学 | Method for culturing Burkholderia polyphylla and application of Burkholderia polyphylla in catalytic degradation of harmful esters of white spirit |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106244507B (en) | 2018-04-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106244508B (en) | One plant of Burkholderia pyrrocinia and its application | |
| Yang et al. | Community composition specificity and potential role of phosphorus solubilizing bacteria attached on the different bloom-forming cyanobacteria | |
| CN102154173A (en) | Separation and application of phthalate ester high-efficiency degrading bacteria | |
| CN107904188A (en) | One plant of achromobacter with monomethyl amine degradation capability and its application | |
| CN113930365A (en) | Pseudomonas aeruginosa for degrading polycyclic aromatic hydrocarbon and application thereof | |
| CN104830708A (en) | Crude oil degrading bacteria strain and application thereof | |
| CN106635872B (en) | One plant of Mo Haiwei bacillus and its application | |
| CN114107092B (en) | Endophyte Gordonia L191 for degrading phthalate and application thereof | |
| CN103881947A (en) | Brevendimonas diminuta strain for forming biofilm and application thereof in cyanide-containing wastewater treatment | |
| CN106477729A (en) | The preparation method of seawater microbial flocculants DHS | |
| CN103045496A (en) | Preparation method of high-efficient degrading bacterium agent for phthalate ester environmental hormone | |
| CN111154685A (en) | Klebsiella variicola for degrading tetracycline and application thereof | |
| WO2008062557A1 (en) | Novel microorganism capable of microbially decomposing chloro-, methylthio- and methoxy-triazine agrichemicals | |
| CN102787089B (en) | Acinetobacterbeijerinckii and application of acinetobacterbeijerinckii | |
| CN115386520B (en) | Rhodococcus pyridine-philic RL-GZ01 strain and application thereof | |
| CN106244507A (en) | The application of Burkholderia pyrrocinia and the method for degraded phthalic acid 2 Octyl Nitrite thereof | |
| CN114231448B (en) | Burkholderia contaminated with indole degradation capability and application thereof | |
| CN103966127B (en) | The red coccus of one strain katsura tree and screening technique and application | |
| CN110283757A (en) | One plant of grease degrading strain IUMR B53 and its application | |
| CN113755407B (en) | Paenibacillus mucilaginosus, extracellular polysaccharide prepared by same and application of extracellular polysaccharide in preparation of microbial flocculant | |
| CN104388365A (en) | Efficient 2, 4, 6-trichlorophenol degrading bacterial strain and separation method thereof | |
| CN110157637A (en) | Enterobacteria Z1 and klebsiella Z2 composite bacteria agent removal high nitrogen pollutant effluents and application | |
| CN106282069B (en) | A kind of pair coccus and the application in sewage purification | |
| CN105154365A (en) | Mycobacterium YC-RL4 and application thereof | |
| CN106244506B (en) | The method of the application of Mo Haiwei bacillus and its naphthalene of degrading |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180410 Termination date: 20191009 |