CN106282057B - A kind of bacterial strain of degrading polyaromatic hydrocarbon and its application - Google Patents
A kind of bacterial strain of degrading polyaromatic hydrocarbon and its application Download PDFInfo
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- CN106282057B CN106282057B CN201610668012.0A CN201610668012A CN106282057B CN 106282057 B CN106282057 B CN 106282057B CN 201610668012 A CN201610668012 A CN 201610668012A CN 106282057 B CN106282057 B CN 106282057B
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- bacterial strain
- polycyclic aromatic
- hydrocarbon
- degrading
- polyaromatic hydrocarbon
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- 125000005575 polycyclic aromatic hydrocarbon group Chemical group 0.000 title claims abstract description 87
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 61
- 230000000593 degrading effect Effects 0.000 title claims abstract description 38
- 241000588919 Citrobacter freundii Species 0.000 claims abstract description 17
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 8
- 239000002689 soil Substances 0.000 claims abstract description 8
- UFWIBTONFRDIAS-UHFFFAOYSA-N Naphthalene Chemical compound C1=CC=CC2=CC=CC=C21 UFWIBTONFRDIAS-UHFFFAOYSA-N 0.000 claims description 22
- GVEPBJHOBDJJJI-UHFFFAOYSA-N fluoranthene Chemical compound C1=CC(C2=CC=CC=C22)=C3C2=CC=CC3=C1 GVEPBJHOBDJJJI-UHFFFAOYSA-N 0.000 claims description 22
- BBEAQIROQSPTKN-UHFFFAOYSA-N pyrene Chemical compound C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 BBEAQIROQSPTKN-UHFFFAOYSA-N 0.000 claims description 20
- YNPNZTXNASCQKK-UHFFFAOYSA-N phenanthrene Chemical compound C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 claims description 12
- 241000894006 Bacteria Species 0.000 claims description 11
- DXBHBZVCASKNBY-UHFFFAOYSA-N 1,2-Benz(a)anthracene Chemical compound C1=CC=C2C3=CC4=CC=CC=C4C=C3C=CC2=C1 DXBHBZVCASKNBY-UHFFFAOYSA-N 0.000 claims description 8
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims 3
- 241000193830 Bacillus <bacterium> Species 0.000 claims 1
- 230000015556 catabolic process Effects 0.000 abstract description 20
- 238000006731 degradation reaction Methods 0.000 abstract description 20
- 230000000694 effects Effects 0.000 abstract description 10
- 238000001228 spectrum Methods 0.000 abstract description 3
- 239000000758 substrate Substances 0.000 abstract description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 12
- 239000000243 solution Substances 0.000 description 10
- 239000002609 medium Substances 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 7
- 239000007836 KH2PO4 Substances 0.000 description 6
- 239000001888 Peptone Substances 0.000 description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 6
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 6
- 239000003344 environmental pollutant Substances 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 6
- 231100000719 pollutant Toxicity 0.000 description 6
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 6
- 108010080698 Peptones Proteins 0.000 description 5
- 235000019319 peptone Nutrition 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- MWPLVEDNUUSJAV-UHFFFAOYSA-N anthracene Chemical compound C1=CC=CC2=CC3=CC=CC=C3C=C21 MWPLVEDNUUSJAV-UHFFFAOYSA-N 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- 150000004945 aromatic hydrocarbons Chemical class 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 150000002220 fluorenes Chemical class 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 108010082340 Arginine deiminase Proteins 0.000 description 1
- HJKOVIKHMJATAZ-UHFFFAOYSA-N C1=CC=CC2=CC=CC=C21.C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 Chemical compound C1=CC=CC2=CC=CC=C21.C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 HJKOVIKHMJATAZ-UHFFFAOYSA-N 0.000 description 1
- 206010007269 Carcinogenicity Diseases 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FMMWHPNWAFZXNH-UHFFFAOYSA-N ERM-AC051 Natural products C1=C2C3=CC=CC=C3C=C(C=C3)C2=C2C3=CC=CC2=C1 FMMWHPNWAFZXNH-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-L Malonate Chemical compound [O-]C(=O)CC([O-])=O OFOBLEOULBTSOW-UHFFFAOYSA-L 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 108010044588 Phenylalanine decarboxylase Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 208000031320 Teratogenesis Diseases 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 238000013475 authorization Methods 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000007670 carcinogenicity Effects 0.000 description 1
- 231100000260 carcinogenicity Toxicity 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000002856 computational phylogenetic analysis Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000006114 decarboxylation reaction Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 150000002475 indoles Chemical class 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000299 mutagenicity Toxicity 0.000 description 1
- 230000007886 mutagenicity Effects 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 239000002957 persistent organic pollutant Substances 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- -1 polycyclic aromatic hydrocarbons naphthalenes Chemical class 0.000 description 1
- 125000003367 polycyclic group Chemical group 0.000 description 1
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Inorganic materials [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000002910 solid waste Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
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- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
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- Biotechnology (AREA)
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- General Health & Medical Sciences (AREA)
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Abstract
The present invention relates to a kind of bacterial strain of degrading polyaromatic hydrocarbon and its applications, belong to technical field of bioengineering.It solves the problems, such as to be the new strains in order to realize the high ring polycyclic aromatic hydrocarbon that can degrade, bacterial strain and its application of a kind of degrading polyaromatic hydrocarbon are provided, the bacterial strain is Citrobacter freundii L2-14, is CCTCC NO:M2016371 in the deposit number of China typical culture collection center；And the bacterial strain is for repairing polycyclic aromatic hydrocarbons contaminated soil.The bacterial strain of degrading polyaromatic hydrocarbon of the present invention, having using polycyclic aromatic hydrocarbon is ability that carbon source and the energy are grown, and can be to the effect of the quick solution drop of polycyclic aromatic hydrocarbons contaminated realization, and degradation substrate spectrum is wide, degradation rate is high.
Description
Technical field
The present invention relates to a kind of bacterial strain of degrading polyaromatic hydrocarbon and its applications, belong to technical field of bioengineering.
Background technique
Polycyclic aromatic hydrocarbon is a kind of persistence organic pollutant.In soil environment it is polycyclic aromatic hydrocarbons contaminated mostly come from petroleum and
The substance of hydrocarbon-containiproducts such as the production and application and timber of its derived product, organic high molecular compound, paper it is incomplete
Burning.Since Polycyclic aromatic hydrocarbons and Derivatives have carcinogenicity, mutagenicity and teratogenesis, polycyclic aromatic hydrocarbons contaminated reparation is asked at present
Topic has caused the very big concern of people.
Currently, for the pollutant of these multiring aromatic hydrocarbons, certain particular kind of microorganisms can be catalyzed polycyclic aromatic hydrocarbon into
The various chemical reactions such as row redox, decarboxylation, hydrolysis and dehydration, are allowed to final mineralising.It is polycyclic aromatic hydrocarbons contaminated compared with other
Repair process, using microbial degradation can more effective, these pollutants difficult to degrade of degrading to low power consuming, and microbial degradation
It is also the main path of polycyclic aromatic hydrocarbons contaminated reparation now.The sieve of a large amount of degrading polycyclic aromatic hydrocarbons microorganisms is carried out both at home and abroad thus
Select work.Up to the present, separated multiple bacterial strains with degrading polycyclic aromatic hydrocarbons ability out.
However, can only degrade naphthalene (2 ring), fluorenes (3 mostly just in these degrading polycyclic aromatic hydrocarbons microorganisms reported at present
Ring) etc. low ring polycyclic aromatic hydrocarbon, and when the phenyl ring number of polycyclic aromatic hydrocarbon meets or exceeds 4, bio-toxicity is significantly raised, raw
Biodegradable significantly reduces.Although also there is the bacterial strain screened that can degrade the effect of high ring polycyclic aromatic hydrocarbon in the prior art,
If Chinese patent (Authorization Notice No.: CN102943052B) discloses the polycyclic aromatic hydrocarbon-degrading bacteria of one plant of resistance to heavy metal, also can
The ability of enough degrading polycyclic aromatic hydrocarbons naphthalenes, phenanthrene, fluoranthene, pyrene, benzanthracene and BaP.But screening this at present some has
Degrading polycyclic aromatic hydrocarbons can especially degrade high ring polycyclic aromatic hydrocarbon bacterial strain it is still relatively fewer, make its application receive it is certain
Limitation.Therefore, how to screen to obtain more bacterial strains, the microbial strains for allowing to the high ring polycyclic aromatic hydrocarbon of efficient degradation seem outstanding
It is application that is important, keeping wider use biodegrade polycyclic aromatic hydrocarbons contaminated.
Summary of the invention
The present invention for it is above-mentioned the problems of in the prior art, the present invention provides a kind of bacterium of degrading polyaromatic hydrocarbon
Strain and its application solve the problems, such as it is how to realize a kind of new strains of high ring polycyclic aromatic hydrocarbon that can degrade.
An object of the present invention technical scheme is that, a kind of bacterium of degrading polyaromatic hydrocarbon
Strain, which is characterized in that the bacterial strain is Citrobacter freundii L2-14, in the guarantor of China typical culture collection center
Hiding number is CCTCC NO:M2016371.
The bacterial strain of degrading polyaromatic hydrocarbon of the present invention is screen from polycyclic aromatic hydrocarbons contaminated soil one plant new
Bacterial strain belongs to citrobacter freundii (Citrobacter freundii), is named as L2-14, the bacterial strain is in 2016 07
The moon is preserved in China typical culture collection center on the 04th, and deposit number is CCTCC NO:M2016371, during preservation place is
Wuhan Wuhan University, state, address are Luojiashan, Wuchang, Wuhan City, Hubei Province.
The bacterial strain of degrading polyaromatic hydrocarbon of the present invention can be under 15~35 DEG C, 6.0~8.5 condition of culture of pH value more quite
It is long；Strain morphology feature is Gram-negative, rod-shaped, peritrichous, no gemma, not raw pod membrane；Bacterial strain is in LB solid medium
Upper culture can form round translucent bacterium colony, surface wettability, neat in edge for 18 hours；Meanwhile this bacterial strain is to polycyclic aromatic hydrocarbon dirt
Dye especially has fast degradation ability to high ring polycyclic aromatic hydrocarbon, and degradation substrate spectrum is wide, degradation rate is high, to also make it
There is wide application potential in a variety of polycyclic aromatic hydrocarbons contaminated reparations.
In the bacterial strain of above-mentioned degrading polyaromatic hydrocarbon, preferably, the 16S rRNA sequence such as SEQ.NO1 of the bacterial strain
It is shown.Another expression-form of bacterial strain has corresponding base sequence.
In the bacterial strain of above-mentioned degrading polyaromatic hydrocarbon, preferably, the bacterial strain has the ability of degrading polycyclic aromatic hydrocarbons.
Since this bacterial strain can be grown by carbon source of polycyclic aromatic hydrocarbon, also especially there is preferable degradation energy to high cycloaromatics
Power realizes the repair process to contaminated soil to make the ability with degrading polycyclic aromatic hydrocarbons.
In the bacterial strain of above-mentioned degrading polyaromatic hydrocarbon, polycyclic aromatic hydrocarbon therein can be low ring polycyclic aromatic hydrocarbon such as naphthalene, fluorenes
Etc., it is also possible to high cycloaromatics, this bacterial strain all has preferable degradation capability to these polycyclic aromatic hydrocarbons.Preferably, described
Polycyclic aromatic hydrocarbon is with polycyclic aromatic hydrocarbons more than 4 rings.When meeting or exceeding 4 due to the phenyl ring number of polycyclic aromatic hydrocarbon, biology
Toxicity is significantly raised, and biodegradability significantly reduces, and the bacterial strain that the present invention is obtained by screening is to more than 4 rings more
Cycloaromatics has good adaptability, equally has preferable degradation effect to the polycyclic aromatic hydrocarbon of these epipodiums, so as to right
The polycyclic aromatic hydrocarbons contaminated environment of epipodium achievees the effect that reparation.As a further preference, the polycyclic aromatic hydrocarbon is selected from luxuriant and rich with fragrance, glimmering
Anthracene, pyrene, benzanthracene or BaP.
In the bacterial strain of above-mentioned degrading polyaromatic hydrocarbon, preferably, the bacterial strain have using polycyclic aromatic hydrocarbon be carbon source with
The ability that the energy is grown.It can be grown by carbon source and the energy of polycyclic aromatic hydrocarbon just because of the bacterial strain, to realize
To the effect of pollutant degrading polycyclic aromatic hydrocarbons.Certainly, this bacterial strain can also be in for example common beef extract-peptone of nutrient medium, general
It is grown in logical LB, nutrient agar.
The second object of the present invention is to what is be achieved by the following technical programs, a kind of above-mentioned degrading polyaromatic hydrocarbon
Bacterial strain application, which is characterized in that the bacterial strain is for repairing polycyclic aromatic hydrocarbons contaminated soil.
Since degrading polyaromatic hydrocarbon bacterial strain of the invention can be grown by carbon source and the energy of polycyclic aromatic hydrocarbon, thus
Also it is provided with the ability degraded to polycyclic aromatic hydrocarbon pollutant, pollutant is dropped by applying it in contaminated soil
Solution, thus achieve the effect that rehabilitating soil, and since this bacterial strain also has preferable degradation capability for high ring polycyclic aromatic hydrocarbon, from
And also improve the popularity of its application.
In conclusion having the advantage that compared with prior art
1. the bacterial strain of degrading polyaromatic hydrocarbon of the present invention, can be to the polycyclic aromatic hydrocarbons contaminated realization effect that quickly solution drops, especially
It is that have fast degradation ability to high ring polycyclic aromatic hydrocarbon, and degradation substrate spectrum is wide, degradation rate is high.
2. the bacterial strain of degrading polyaromatic hydrocarbon of the present invention, there is the energy grown using polycyclic aromatic hydrocarbon as carbon source and the energy
Power is not necessarily to additional other carbon sources and the energy, it will be able to realize the effect of targetedly solution drop polycyclic aromatic hydrocarbon pollutant.
Detailed description of the invention
The bacterial strain Citrobacter freundii L2-14 of this degrading polyaromatic hydrocarbon of Fig. 1 is bent to the degradation of naphthalene at any time
Line chart.
The bacterial strain Citrobacter freundii L2-14 degradation to pyrene naphthalene at any time of this degrading polyaromatic hydrocarbon of Fig. 2
Curve graph.
The bacterial strain Citrobacter freundii L2-14 of this degrading polyaromatic hydrocarbon of Fig. 3 is at any time to the drop of BaP
Solution curve figure.
Specific embodiment
Below by specific embodiments and the drawings, the technical solutions of the present invention will be further described, but this
Invention is not limited to these examples.
Embodiment 1
The separation and screening of this degrading polyaromatic hydrocarbon bacterial strain Citrobacter freundii L2-14
From certain solid waste dismantling field acquisition away from the topsoil sample from 15cm under earth's surface, 1mL is added by every 1g pedotheque
The ratio (pedotheque: the ratio of domestication liquid is 1g:1mL) of domestication liquid is added domestication liquid and carries out domestication culture, domestication culture item
Part is 25 DEG C of temperature, revolving speed 100rpm, time 72h.Wherein, the formula of above-mentioned domestication liquid are as follows:
(NH4)2SO4: 2g/L；MgSO4: 0.2g/L；K2HPO4: 4g/L；KH2PO4: 6g/L；Polycyclic aromatic hydrocarbon (naphthalene, phenanthrene, fluoranthene,
Pyrene, benzanthracene and BaP) mixture 200mg/L, wherein naphthalene, phenanthrene, fluoranthene, pyrene, benzanthracene and BaP additional amount can be with
It is equivalent, different contents can also be used, these polycyclic aromatic hydrocarbons is mainly added and play preferable domestication effect.Then, will
Above-mentioned domestication mixture is that the ratio of 1:50 is inoculated in inorganic salts Selective agar medium and carries out selection culture in mass ratio, wherein
Inorganic salts Selective agar medium formula are as follows: peptone: 5g/L；(NH4)2SO4: 2g/L；MgSO4: 0.2g/L；K2HPO4: 4g/L；
KH2PO4: 6g/L；Polycyclic aromatic hydrocarbon (naphthalene, phenanthrene, fluoranthene, pyrene, benzanthracene and BaP) mixture 200mg/L, wherein naphthalene, phenanthrene, glimmering
Anthracene, pyrene, benzanthracene and BaP additional amount can be equivalent, different contents can also be used；Select condition of culture for temperature
25 DEG C, revolving speed 100rpm, time 72h of degree, obtains selecting culture accordingly.
Above-mentioned selection culture is subjected to gradient dilution with phosphate buffer solution, chooses the gradient dilution liquid of suitable concentration
In on solid inorganic salt Selective agar medium, selection single colonie is separated dilution spread, that is, isolating one plant of Strain Designation is L2-
14.The bacterial strain is preserved in China typical culture collection center on July 4th, 2016, deposit number CCTCC NO:
M2016371。
Meanwhile the bacterial strain is identified, wherein the 16SrDNA gene PCR amplification of bacterial strain L2-14 is sequenced by Shanghai beauty
Lucky biotech firm completes, and obtains the sequence that length is 1430bp, and specific sequencing result is shown in SEQ NO.1.Meanwhile it being tied according to sequencing
Fruit phylogenetic tree construction selects gene order of the homology greater than 97% to carry out Phylogenetic Analysis, phylogenetic tree analysis
The result shows that the evolutionary distance of bacterial strain L2-14 and Citrobacter freundii is nearest, combining form, colony characteristics and physiology
Biochemical characteristic, identification bacterial strain belong to citrobacter freundii (Citrobacter freundii), i.e. this Citrobacter
Freundii L2-14 bacterial strain.
By this Citrobacter freundii L2-14 strain inoculated in LB solid medium (tryptone 1.0%, ferment
Female extract 0.5%, sodium chloride 1.0%, agar 2%) the round translucent bacterium colony of culture formation in 18 hours, surface wettability, side
Edge is neat.And the physiological and biochemical property of bacterial strain Citrobacter freundii L2-14 is as shown in table 1 below:
Table 1:
| Certified variety | Qualification result |
| Indoles generates | - |
| M.R experiment | + |
| V.P experiment | - |
| Gelatin hydrolysis | - |
| Urinate enzyme experiment | + |
| H2S | + |
| Phenylalanine decarboxylase | - |
| Arginine dihydrolase | - |
| KNO3 | + |
| KCN | + |
| Malonate utilizes | - |
| Citrate utilizes | + |
| Glucose utilization | + |
| Maltose utilizes | + |
| Arabinose utilizes | + |
| Xylose utilization | + |
| Mannitol utilizes | + |
| Sucrose utilizes | + |
Embodiment 2
This Citrobacter freundii L2-14 bacterial strain is used for degrading polycyclic aromatic hydrocarbons
It takes this bacterial strain Citrobacter freundii L2-14 to be seeded in LB solid medium to be activated；It chooses again
The LB liquid medium that single colonie is inoculated in 5mL is cultivated, and is cultivated under conditions of 28 DEG C of temperature and revolving speed 100rpm
To bacterium solution OD600≈1；Then, bacterium solution is inoculated in mineral-salts glucose culture medium according to 2% inoculum concentration, at 28 DEG C of temperature
And it cultivates under conditions of revolving speed 100rpm to bacterium solution OD600≈1；Wherein, mineral-salts glucose culture medium prescription are as follows: peptone:
5g/L；Glucose: 10g/L；(NH4)2SO4: 2g/L；MgSO4: 0.2g/L；K2HPO4: 4g/L；KH2PO4: 6g/L, culture terminate
Afterwards, corresponding bacterium solution is obtained.
Above-mentioned bacterium solution 10000r/min centrifuge is centrifuged 5 minutes, lower layer's bacterial sediment is taken, is 7.0~7.5 with pH
Phosphate buffer washs 2 times, spare.Above-mentioned thallus is taken to be resuspended in inorganic salts-naphthalene, inorganic salts-pyrene, inorganic in equal volume respectively
It is that 100rpm carries out culture 12 days controlled at 25 DEG C, revolving speed in salt-BaP culture medium.Efficient liquid phase is used in the process
Naphthalene in above-mentioned three kinds of systems of chromatography detection different time, pyrene, BaP content test degradation effect；
Wherein, inorganic salts-naphthalene culture medium prescription are as follows: peptone: 2g/L；(NH4)2SO4: 2g/L；MgSO4: 0.2g/L；
K2HPO4: 4g/L；KH2PO4: 6g/L；Naphthalene: 200mg/L；
Inorganic salts-pyrene culture medium prescription are as follows: peptone: 2g/L；(NH4)2SO4: 2g/L；MgSO4: 0.2g/L；K2HPO4:
4g/L；KH2PO4: 6g/L；Pyrene: 200mg/L；
Inorganic salts-BaP culture medium prescription are as follows: peptone: 2g/L；(NH4)2SO4: 2g/L；MgSO4: 0.2g/L；
K2HPO4: 4g/L；KH2PO4: 6g/L；BaP: 200mg/L.
Wherein, testing result is as shown in Figs. 1-3；It can be seen from the figure that after 12 days, naphthalene, pyrene, BaP in above-mentioned system
Degradation rate be respectively 84.6%, 84.6%, 62.6%, thus also illustrate bacterial strain of the invention have preferably degrade it is polycyclic
The ability of aromatic hydrocarbons, while the ability for the high ring polycyclic aromatic hydrocarbon that also can preferably degrade.Equally, by inorganic salts-BaP culture medium
In BaP be replaced by luxuriant and rich with fragrance, fluoranthene or benzanthracene, after 12 days, degradation rate can reach 60% or more.
Specific embodiment described in the present invention only illustrate the spirit of the present invention by way of example.The neck of technology belonging to the present invention
The technical staff in domain can make various modifications or additions to the described embodiments or replace by a similar method
In generation, however, it does not deviate from the spirit of the invention or beyond the scope of the appended claims.
It is skilled to this field although present invention has been described in detail and some specific embodiments have been cited
For technical staff, as long as it is obvious for can making various changes or correct without departing from the spirit and scope of the present invention.
Claims (5)
1. a kind of bacterial strain of degrading polyaromatic hydrocarbon, which is characterized in that the bacterial strain of the degrading polyaromatic hydrocarbon is Freund citric acid
Bacillus (Citrobacter freundii) L2-14 is CCTCC NO in the deposit number of China typical culture collection center:
M2016371；The 16SrDNA sequence of the bacterial strain is as shown in SEQ NO.1.
2. the bacterial strain of degrading polyaromatic hydrocarbon according to claim 1, which is characterized in that the polycyclic aromatic hydrocarbon is with 4 rings
Above polycyclic aromatic hydrocarbon.
3. the bacterial strain of degrading polyaromatic hydrocarbon according to claim 1, which is characterized in that the polycyclic aromatic hydrocarbon be selected from naphthalene, phenanthrene,
Fluoranthene, pyrene, benzanthracene or BaP.
4. the bacterial strain of degrading polyaromatic hydrocarbon according to claim 1 to 3, which is characterized in that the bacterial strain has
The ability grown using polycyclic aromatic hydrocarbon as carbon source and the energy.
5. a kind of bacterial strain application of the degrading polyaromatic hydrocarbon as described in claim 1-4 any one, which is characterized in that the bacterium
Strain is for repairing polycyclic aromatic hydrocarbons contaminated soil.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102943052A (en) * | 2012-11-22 | 2013-02-27 | 台州职业技术学院 | Heavy metal-resistant polycyclic aromatic hydrocarbon (PAHs) degrading bacteria and application thereof in remediation of composite contaminated soil |
| CN102994404A (en) * | 2012-10-12 | 2013-03-27 | 台州职业技术学院 | Bacterial strain for degrading polycyclic aromatic hydrocarbon and application of bacterial strain cooperated with surface active agent in soil remediation |
| CN104673683A (en) * | 2015-02-27 | 2015-06-03 | 台州职业技术学院 | Salt-tolerance polycyclic aromatic hydrocarbon degrading bacterium Tzyjm and application of salt-tolerance polycyclic aromatic hydrocarbon degrading bacterium Tzyjm in marine polluted environment |
| CN104877934A (en) * | 2015-02-27 | 2015-09-02 | 台州职业技术学院 | Salt-tolerant polycyclic aromatic hydrocarbon degradation bacterium strain Tzymx and application thereof in marine pollution environment |
-
2016
- 2016-08-15 CN CN201610668012.0A patent/CN106282057B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102994404A (en) * | 2012-10-12 | 2013-03-27 | 台州职业技术学院 | Bacterial strain for degrading polycyclic aromatic hydrocarbon and application of bacterial strain cooperated with surface active agent in soil remediation |
| CN102943052A (en) * | 2012-11-22 | 2013-02-27 | 台州职业技术学院 | Heavy metal-resistant polycyclic aromatic hydrocarbon (PAHs) degrading bacteria and application thereof in remediation of composite contaminated soil |
| CN104673683A (en) * | 2015-02-27 | 2015-06-03 | 台州职业技术学院 | Salt-tolerance polycyclic aromatic hydrocarbon degrading bacterium Tzyjm and application of salt-tolerance polycyclic aromatic hydrocarbon degrading bacterium Tzyjm in marine polluted environment |
| CN104877934A (en) * | 2015-02-27 | 2015-09-02 | 台州职业技术学院 | Salt-tolerant polycyclic aromatic hydrocarbon degradation bacterium strain Tzymx and application thereof in marine pollution environment |
Non-Patent Citations (3)
| Title |
|---|
| Anaerobic Degradation of Biphenyl by the Facultative Anaerobic Strain Citrobacter freundii BS2211;V. G. Grishchenkov等;《Applied Biochemistry and Microbiology》;20020331;第38卷(第2期);125-128 * |
| Degradation of polycyclic aromatic hydrocarbons by microbial consortia enriched from three soils using two different culture media;Manli Wu等;《Environmental Pollution》;20130731;第178卷;152-158 * |
| 胜利油田土壤中石油降解菌分离和鉴定及代谢产物的研究;姚瑶等;《南京农业大学学报》;20131231;第36卷(第1期);65-71 * |
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