CN109609428A - The screening technique of polycyclic aromatic hydrocarbon-degrading bacteria - Google Patents
The screening technique of polycyclic aromatic hydrocarbon-degrading bacteria Download PDFInfo
- Publication number
- CN109609428A CN109609428A CN201811582315.6A CN201811582315A CN109609428A CN 109609428 A CN109609428 A CN 109609428A CN 201811582315 A CN201811582315 A CN 201811582315A CN 109609428 A CN109609428 A CN 109609428A
- Authority
- CN
- China
- Prior art keywords
- polycyclic aromatic
- aromatic hydrocarbon
- medium
- soil
- screening technique
- 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
- 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
- C12N1/36—Adaptation or attenuation of cells
-
- 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
- C12N1/02—Separating microorganisms from their culture media
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of screening technique of polycyclic aromatic hydrocarbon-degrading bacteria, which includes: that (1) is cultivated to by polycyclic aromatic hydrocarbon is added in polycyclic aromatic hydrocarbons contaminated soil, the soil after being cultivated；(2) soil after culture is repeated into step (1)；(3) it takes step (2) resulting soil to be diluted with sterile water, prepares bacteria suspension；Bacteria suspension is coated on using polycyclic aromatic hydrocarbon to cultivate on the Double-Medium of sole carbon source, the bacterium colony for whether generating transparent circle on Double-Medium is observed；(4) if failing to generate the bacterium colony of transparent circle, repeatedly step (1) to (3) on Double-Medium；(5) bacterium colony is purified to get polycyclic aromatic hydrocarbon-degrading bacteria.Method of the invention is compared with the traditional method, and substantially reduces the screening time of polycyclic aromatic hydrocarbon-degrading bacteria, and screening is carried out after taming 28 days to soil under normal circumstances can be obtained polycyclic aromatic hydrocarbon-degrading bacteria.
Description
Technical field
The invention belongs to microbial degradation polycyclic aromatic hydrocarbon technical fields, more particularly, to a kind of polycyclic aromatic hydrocarbon-degrading bacteria
Screening technique.
Background technique
Polycyclic aromatic hydrocarbon (PAHs) refers to the chemical combination that two or more phenyl ring are rearranged with chain, horn shape or string-like
Object mostlys come from the imperfect combustion of the organic matters such as timber, fossil fuel, coai-tar product or food, and polycyclic aromatic hydrocarbon is in ring
It is widely distributed in border, there is detection in atmosphere, water body, soil, deposit and food.Many types have carcinogenic, cause
Abnormal, mutagenesis, and its toxicity and carcinogenicity increase with the increase of phenyl ring number.
Photochemical breakdown can occur for polycyclic aromatic hydrocarbon in environment and by oxidizing in environment in addition to small part
Except, it largely relies primarily on biodegradationpathway and fades away, therefore microbial degradation is that control is polycyclic aromatic hydrocarbons contaminated more
Cost-effective method.And the base of microbial degradation polycyclic aromatic hydrocarbon ability is exactly studied and is utilized in the acquisition of polycyclic aromatic hydrocarbon-degrading bacteria
Plinth.
Currently used polycyclic aromatic hydrocarbon-degrading bacteria screening technique is liquid submerged culture method, is in minimal medium
The bacterium sources such as soil are added, by the screening for carrying out polycyclic aromatic hydrocarbon-degrading bacteria after enrichment of repeatedly transferring.But this method is due to four
The molecular weight of the polycyclic aromatic hydrocarbon of ring or more is big, hydrophobicity is strong, water-soluble low, and therefore, currently used shaking culture is only fitted
Together in screening tricyclic and the degradation bacteria of the following polycyclic aromatic hydrocarbon of tricyclic, and for the degrading microorganism at Fourth Ring and its above polycyclic aromatic hydrocarbon
Screening, it may appear that the period is long, sifts out the problems such as probability is small, and the degradation bacterium series stability sifted out is poor, degradation rate is not high.Meanwhile
Since polycyclic aromatic hydrocarbon-degrading bacteria growth rate is slower, it is longer to will cause growth cycle in bacterium source later by repeatedly switching
Polycyclic aromatic hydrocarbon-degrading bacteria is lost, and finally resulting in the bacterium that enrichment comes out is the polycyclic aromatic hydrocarbon tolerance for not having degrading polycyclic aromatic hydrocarbons ability
Bacterium.
Summary of the invention
The object of the present invention is to provide a kind of screening techniques of polycyclic aromatic hydrocarbon-degrading bacteria, short using this method screening time,
It is easier to obtain efficient polycyclic aromatic hydrocarbon-degrading bacteria, is especially suitable for the Fourth Ring slower to degradation rate and the above degrading polycyclic aromatic hydrocarbons
The screening of bacterium.
To achieve the goals above, the present invention provides a kind of screening technique of polycyclic aromatic hydrocarbon-degrading bacteria, the screening techniques
Include:
(1) it is cultivated to by polycyclic aromatic hydrocarbon is added in polycyclic aromatic hydrocarbons contaminated soil, the soil after being cultivated；
(2) soil after the culture is repeated into step (1)；
(3) it takes step (2) resulting soil to be diluted with sterile water, prepares bacteria suspension；The bacteria suspension is coated on more
The bacterium that transparent circle whether is generated on the Double-Medium observed to be cultivated on the Double-Medium of sole carbon source for cycloaromatics
It falls；
(4) if failing to generate the bacterium colony of transparent circle, repeatedly step (1) and (3) on the Double-Medium；
(5) bacterium colony is purified to get the polycyclic aromatic hydrocarbon-degrading bacteria.
Technical solution of the present invention has the following beneficial effects:
(1) persistently addition polycyclic aromatic hydrocarbon tames the microbial flora in soil to the present invention in the soil, Neng Goubao
The slower polycyclic aromatic hydrocarbon-degrading bacteria of card growth and breeding can grow in same domestication system, avoid taming in traditional shaking flask
The risk for causing polycyclic aromatic hydrocarbon-degrading bacteria to be lost in switching process；
(2) polycyclic aromatic hydrocarbon of addition in the soil of the invention is to carry out domestication to microbial flora and in traditional flask system
Middle addition polycyclic aromatic hydrocarbon domestication is compared, and uniformity of the PAHs in soil after mixing and stirring is than the uniformity in flask system
It is good, it is more preferable to the domestication effect of microbial flora；
(3) method of the invention overcomes cumbersome feature during conventional screen bacterium acclimation method, has and saves people
Work, easy to operate feature；
(4) method of the invention is compared with the traditional method, and substantially reduces the screening time of polycyclic aromatic hydrocarbon-degrading bacteria, generally
In the case of to soil tame 28 days after carry out screening can be obtained polycyclic aromatic hydrocarbon-degrading bacteria.
Other features and advantages of the present invention will then part of the detailed description can be specified.
Detailed description of the invention
Exemplary embodiment of the invention is described in more detail in conjunction with the accompanying drawings, it is of the invention above-mentioned and its
Its purpose, feature and advantage will be apparent, wherein in exemplary embodiment of the invention, identical reference label
Typically represent same parts.
The selection culture that the screening technique that Fig. 1 shows the polycyclic aromatic hydrocarbon-degrading bacteria of embodiment according to the present invention 1 obtains
The pyrene degradation bacterium colony of transparent circle on base plate.
Specific embodiment
The preferred embodiment of the present invention is described in more detail below.Although the following describe preferred implementations of the invention
Mode, however, it is to be appreciated that may be realized in various forms the present invention without that should be limited by the embodiments set forth herein.Phase
Instead, these embodiments are provided so that the present invention is more thorough and complete, and can be by the scope of the present invention completely
It is communicated to those skilled in the art.
The present invention provides a kind of screening technique of polycyclic aromatic hydrocarbon-degrading bacteria, which includes:
(1) it is cultivated to by polycyclic aromatic hydrocarbon is added in polycyclic aromatic hydrocarbons contaminated soil, the soil after being cultivated；
(2) soil after the culture is repeated into step (1)；
(3) it takes step (2) resulting soil to be diluted with sterile water, prepares bacteria suspension；The bacteria suspension is coated on more
The bacterium that transparent circle whether is generated on the Double-Medium observed to be cultivated on the Double-Medium of sole carbon source for cycloaromatics
It falls；
(4) if failing to generate the bacterium colony of transparent circle, repeatedly step (1) and (3) on the Double-Medium；
(5) bacterium colony is purified to get the polycyclic aromatic hydrocarbon-degrading bacteria.
Method of the invention overcomes heavy workload in conventional method, degradation bacteria is easily lost, degradation bacteria pick-up rate is low etc.
Disadvantage.If can continue the addition polycyclic aromatic hydrocarbon into soil because the addition polycyclic aromatic hydrocarbon domestication time is inadequate and continue to tame, without weight
Newly start to tame, compared with conventional screening methods, avoids the generation of repeated workload and the loss of polycyclic aromatic hydrocarbon-degrading bacteria.
The present invention maintains polycyclic aromatic hydrocarbon by the way that the soil after the culture is repeated step (1) with Soil Microorganism
Toxicity stress under.
Step (4) in method of the invention is by continuing to tame and docile to having already passed through polycyclic aromatic hydrocarbon acclimated microorganism flora
Change, to continue the abundance of polycyclic aromatic hydrocarbon-degrading bacteria in raising microbial flora.
In accordance with the present invention it is preferred that the polycyclic aromatic hydrocarbon is at least one of phenanthrene, pyrene, anthracene, fluoranthene and benzo (a) pyrene.
In accordance with the present invention it is preferred that step (1) includes: to part by adding polycyclic virtue in polycyclic aromatic hydrocarbons contaminated soil
Hydrocarbon solution is uniformly mixed by polycyclic aromatic hydrocarbons contaminated soil with residue, is cultivated after the solvent is volatilized.
It in the present invention, is mixed, is prevented by polycyclic aromatic hydrocarbons contaminated soil with residue again after being evaporated completely after the solvent is volatilized
Only the addition of solvent generates harm to Soil Microorganism flora.
In accordance with the present invention it is preferred that the polycyclic aromatic hydrocarbon solution is the acetone soln of polycyclic aromatic hydrocarbon.
In accordance with the present invention it is preferred that the temperature of the culture is 25-35 DEG C in step (1), the time of culture is 14-
28d is 400-500mg/kg with the addition concentration by the poidometer of polycyclic aromatic hydrocarbons contaminated soil, the polycyclic aromatic hydrocarbon.Soil
Water content is adjusted to the field capacity of soil.
In accordance with the present invention it is preferred that the diluted multiple is 10 in step (3)3-105Times；The temperature of the culture is
25-35 DEG C, the time of culture is 7-14d.
Preferably, step (3) is that step (2) resulting soil is taken to mix, vibrate, stand with sterile water, is taken
Clear liquid；The supernatant is diluted with sterile water, prepares bacteria suspension；The bacteria suspension is coated on using polycyclic aromatic hydrocarbon as sole carbon
It is cultivated on the Double-Medium in source, the bacterium colony for whether generating transparent circle on the Double-Medium is observed.
It by the Double-Medium of sole carbon source of polycyclic aromatic hydrocarbon is solid medium in accordance with the present invention it is preferred that described, on
Layer is the minimal medium of the polycyclic aromatic hydrocarbon containing sole carbon source, and lower layer is the minimal medium of not carbonaceous sources.
Of the invention is the Double-Medium of sole carbon source the preparation method comprises the following steps: the nothing that high-temperature sterilization is crossed using polycyclic aromatic hydrocarbon
Machine salt culture medium solution is poured into sterile petri dish, is formed by curing lower layer's culture medium；By the inorganic salts training through high-temperature sterilization
Support based sols mixed with polycyclic aromatic hydrocarbon solution after draw it is a certain amount of be spread evenly across on cooling lower layer's culture medium, make polycyclic aromatic hydrocarbon
It is evenly distributed on culture medium, forms upper layer culture medium.
In accordance with the present invention it is preferred that the solution includes: (NH in terms of the solution of the minimal medium of 1L4)2SO42-4g, KH2PO40.3-0.7g, Na2HPO40.3-0.7g, MgSO40.2-0.5g, trace element solution 0.5-
It is 6-7 that 1.5mL, agar 10-20g, which adjust pH with the NaOH solution of 5mol/L, and surplus is sterile distilled water；With the described micro of 1L
Element Solution meter, the trace element solution include: FeCl30.1-0.5g, FeSO4·7H2O 0.1-0.5g, MnSO4·H2O
0.1-0.2g, ZnSO40.1-0.2g, CoCl20.1-0.3g, surplus are sterile distilled water.
In accordance with the present invention it is preferred that described using polycyclic aromatic hydrocarbon as in the upper layer culture medium of the Double-Medium of sole carbon source
Polycyclic aromatic hydrocarbon concentration is 400-500mg/L.
In accordance with the present invention it is preferred that the method for the purifying include: by the bacterium colony that can generate transparent circle it is new with
Polycyclic aromatic hydrocarbon is separation of crossing on the Double-Medium of sole carbon source.
The present invention is further illustrated by the following examples:
Embodiment 1
The present embodiment provides a kind of screening technique of polycyclic aromatic hydrocarbon-degrading bacteria, the screening technique is as follows:
(1) it the acquisition of bacterium source: acquires around certain coke-oven plant by polycyclic aromatic hydrocarbons contaminated soil, crosses 2mm sieve, be immediately placed on 4
It is saved backup in DEG C refrigerator；
(2) pyrene degradation bacteria is enriched with: take 50g polycyclic aromatic hydrocarbon pollution to cultivate in 1L sterile beaker, it is living under the conditions of 30 DEG C
Change 24 hours；10g soil, the pyrene acetone soln that addition 4mL concentration is 5000mg/L are taken after soil activating；Acetone is waved after 1 hour
It distributes entirely, the 10g soil for being added to polycyclic aromatic hydrocarbon and residue 40g soil is stirred evenly, the initial concentration of pyrene in soil is made
Soil is transferred in 150mL conical flask by 400mg/kg, is placed in 30 DEG C of incubators stationary culture 14 days；It was cultivated after 14 days
Soil continue add 5000mg/L pyrene acetone soln 4mL, continue stationary culture 14 days in 30 DEG C of incubators；
(3) Selective agar medium preparation method is as follows:
The preparation method of lower layer: by the solution of minimal medium, high-temperature vapour sterilizing is solidified in minimal media based sols
Before pour into culture dish, it is spare after cooling.Wherein, the preparation of the solution of minimal medium are as follows: by (NH4)2SO43g, KH2PO4
0.5g, Na2HPO40.5g, MgSO40.3g, trace element solution 1mL, pH=7.0 and agar 15g are uniformly mixed, and add sterile steaming
Distilled water is settled to 1000mL, with 5M NaOH solution tune medium pH=6.5.The preparation of trace element solution are as follows: by FeCl3
0.3g, FeSO4·7H2O 0.3g, MnSO4·H2O 0.15g, ZnSO40.14g and CoCl20.2g is uniformly mixed, and adds sterile steaming
Distilled water is settled to 1000mL.
The preparation method on upper layer: by minimal medium solution, 5000mg/L pyrene is added in high-temp steam sterilizing after sterilizing
Acetone soln mixes with minimal media based sols, so that the final concentration of minimal media based sols is reached 500mg/L, inhaled with liquid-transfering gun
The not solidified culture medium mixed solution of 5mL is taken to be uniformly coated on lower layer's minimal medium, it is spare after cooling.
(4) pyrene bacterial isolation: taking the soil 5g for having cultivated 28 days and 45g sterile water in 250mL sterile conical flask,
It is vibrated 2 hours in the constant temperature oscillator that revolving speed is 150rpm, then takes out conical flask, 30 minutes are stood, by supernatant sterile
Under the conditions of carry out 10 times of serial dilutions to 10-5, prepare bacteria suspension；10 are taken respectively-3, 10-4With 10-5The bacterium of three concentration is outstanding
100 μ L of liquid is coated on using pyrene on the Double-Medium of sole carbon source, to be placed in 30 DEG C of constant incubators and cultivates 1-2 weeks, observation
The generation situation of degradation bacteria transparent circle on Selective agar medium plate.It after two weeks can be in Selective agar medium (using pyrene as sole carbon source
Double-Medium) on observe the bacterium colony (as shown in Figure 1) that can generate transparent circle.
(6) pyrene degradation bacteria purifies: picking can generate the bacterium colony of transparent circle on Selective agar medium plate, train in new selection
It supports and utilizes trilinear method scribing line purifying on base plate (using pyrene as the Double-Medium of sole carbon source).Above-mentioned scribe step is repeated, directly
The single no miscellaneous bacteria of colonial morphology on to Selective agar medium plate.Finally obtain the different pyrene degradation bacteria of two plant shape states.
Pyrene degrading strain identification and preservation: 16S rRNA gene sequence is carried out respectively to two plants of pyrene degradation bacterias obtained by the above method
Column measurement and Phylogenetic Analysis.Two plants of pyrene degradation bacterias are Mycobacterium (Mycobacterium sp.) respectively as the result is shown
With Pseudonocardia (Pseudonocardia sp.).Two plants of bacterium are saved with desivac.
Embodiment 2
(1) acquisition of bacterium source: the bacterium source of the present embodiment is from Liaoning chemical plant polycyclic aromatic hydrocarbon pollution.
(2) degradation bacteria is enriched with: the present embodiment is by adding pyrene and fluoranthene respectively in contaminated soil come to micro- in soil
Biology is enriched with.The adding method of pyrene and fluoranthene is in the same manner as in Example 1, after adding pyrene and fluoranthene, two domestication system soil
The initial concentration of pyrene and fluoranthene is 500mg/kg in earth.Two domestication systems are placed in 30 DEG C of constant incubators and cultivate 14
It, continues to add pyrene and fluoranthene into soil respectively, makes the concentration of pyrene and fluoranthene increase 500mg/kg, addition in soil after adding
Method is the same as embodiment 1.
(3) preparation method of Selective agar medium is the same as embodiment 1.
(4) screening technique of degradation bacteria is the same as embodiment 1.Selective agar medium is cultivated in the incubator to 4th week and is not observed
To the generation of degradation transparent circle.
(5) step (2) and (4) are repeated, find to produce a large amount of transparent circles on pyrene and fluoranthene Selective agar medium after 14 days,
Illustrate the degradation bacteria that a large amount of pyrenes and fluoranthene have been filtered out on the Selective agar medium containing pyrene and fluoranthene.
(6) purifying of degradation bacteria and identification method are identical as the method in embodiment 1；
The degradation bacteria that discovery pyrene and fluoranthene are identified finally by 16SrRNA gene order is same bacterium, belongs to branch bar
Pseudomonas (Mycobacterium sp.).The degradation capability of pyrene and fluoranthene is found by studying the bacterium, the bacterium only contain pyrene and
It is cultivated 14 days in the inorganic medium of fluoranthene, the pyrene and fluoranthene degradation rate to 100mg/L are respectively 73.2% and 89.5%.
Embodiment 3
The difference of the present embodiment and embodiment 1 are as follows: using certain steel plant's polycyclic aromatic hydrocarbon pollution as degradation bacteria come
Bacterium source is tamed with the phenanthrene that pollutant is 500mg/kg concentration in source, other screening steps and technological parameter with embodiment 1
It is identical.
The bacterium that can generate degradation circle is obtained on the culture medium using phenanthrene as sole carbon source, can confirm as polycyclic aromatic hydrocarbon
Luxuriant and rich with fragrance degradation bacteria can primarily determine that the phenanthrene degradation bacteria of acquisition is by 16S rRNA gene sequencing and Phylogenetic Analysis
Rhod (Rhodochrous sp.).
Various embodiments of the present invention are described above, above description is exemplary, and non-exclusive, and
It is not limited to disclosed each embodiment.Without departing from the scope and spirit of illustrated each embodiment, for this skill
Many modifications and changes are obvious for the those of ordinary skill in art field.
Claims (10)
1. a kind of screening technique of polycyclic aromatic hydrocarbon-degrading bacteria, which is characterized in that the screening technique includes:
(1) it is cultivated to by polycyclic aromatic hydrocarbon is added in polycyclic aromatic hydrocarbons contaminated soil, the soil after being cultivated；
(2) soil after the culture is repeated into step (1)；
(3) it takes step (2) resulting soil to be diluted with sterile water, prepares bacteria suspension；The bacteria suspension is coated on polycyclic virtue
The bacterium colony that transparent circle whether is generated on the Double-Medium observed to be cultivated on the Double-Medium of sole carbon source for hydrocarbon；
(4) if failing to generate the bacterium colony of transparent circle, repeatedly step (1) to (3) on the Double-Medium；
(5) bacterium colony is purified to get the polycyclic aromatic hydrocarbon-degrading bacteria.
2. screening technique according to claim 1, wherein the polycyclic aromatic hydrocarbon is phenanthrene, pyrene, anthracene, fluoranthene and benzo (a) pyrene
At least one of.
3. screening technique according to claim 1, wherein step (1) includes: to part by polycyclic aromatic hydrocarbons contaminated soil
Middle addition polycyclic aromatic hydrocarbon solution is uniformly mixed by polycyclic aromatic hydrocarbons contaminated soil with residue, is cultivated after the solvent is volatilized.
4. screening technique according to claim 3, wherein the polycyclic aromatic hydrocarbon solution is the acetone soln of polycyclic aromatic hydrocarbon.
5. screening technique according to claim 1, wherein in step (1), the temperature of the culture is 25-35 DEG C, culture
Time be 14-28d, with by the poidometer of polycyclic aromatic hydrocarbons contaminated soil, the addition concentration of the polycyclic aromatic hydrocarbon is 400-
500mg/kg。
6. screening technique according to claim 1, wherein in step (3), the diluted multiple is 103-105Times；Institute
The temperature for stating culture is 25-35 DEG C, and the time of culture is 7-14d.
7. screening technique according to claim 1, wherein described to be by the Double-Medium of sole carbon source of polycyclic aromatic hydrocarbon
Solid medium, upper layer are the minimal medium of the polycyclic aromatic hydrocarbon containing sole carbon source, and lower layer is the inorganic salts culture of not carbonaceous sources
Base.
8. screening technique according to claim 7, wherein described molten in terms of the solution of the minimal medium of 1L
Liquid includes: (NH4)2SO42-4g, KH2PO40.3-0.7g, Na2HPO40.3-0.7g, MgSO40.2-0.5g, microelement are molten
It is 6-7 that liquid 0.5-1.5mL, agar 10-20g, which adjust pH with the NaOH solution of 5mol/L, and surplus is sterile distilled water；With the institute of 1L
Trace element solution meter is stated, the trace element solution includes: FeCl30.1-0.5g, FeSO4·7H2O 0.1-0.5g,
MnSO4H2O 0.1-0.2g, ZnSO40.1-0.2g, CoCl20.1-0.3g, surplus are sterile distilled water.
9. screening technique according to claim 7, wherein described using polycyclic aromatic hydrocarbon as the Double-Medium of sole carbon source
Polycyclic aromatic hydrocarbon concentration is 400-500mg/L in the culture medium of upper layer.
10. screening technique according to claim 1, wherein the method for the purifying includes: that can generate transparent circle for described
Bacterium colony cross on the new Double-Medium using polycyclic aromatic hydrocarbon as sole carbon source separation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811582315.6A CN109609428B (en) | 2018-12-24 | 2018-12-24 | Method for screening polycyclic aromatic hydrocarbon degrading bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811582315.6A CN109609428B (en) | 2018-12-24 | 2018-12-24 | Method for screening polycyclic aromatic hydrocarbon degrading bacteria |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109609428A true CN109609428A (en) | 2019-04-12 |
| CN109609428B CN109609428B (en) | 2021-11-30 |
Family
ID=66010519
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811582315.6A Active CN109609428B (en) | 2018-12-24 | 2018-12-24 | Method for screening polycyclic aromatic hydrocarbon degrading bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109609428B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110283772A (en) * | 2019-07-25 | 2019-09-27 | 北京高能时代环境技术股份有限公司 | A kind of preparation method of functional flora that repairing petroleum hydrocarbon contaminated soil and underground water |
| CN111705001A (en) * | 2020-07-23 | 2020-09-25 | 桂林理工大学 | Method for rapidly screening polycyclic aromatic hydrocarbon degrading bacteria |
| CN116689477A (en) * | 2023-06-25 | 2023-09-05 | 江苏大学 | Bioremediation method of degradation flora composite edible fungus residues of polycyclic aromatic hydrocarbon contaminated soil |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101597590A (en) * | 2009-07-24 | 2009-12-09 | 中国科学院南京土壤研究所 | The double-layer plate method of fast simple screening polycyclic aromatic hydrocarbon-degrading and screening thereof are dull and stereotyped |
-
2018
- 2018-12-24 CN CN201811582315.6A patent/CN109609428B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101597590A (en) * | 2009-07-24 | 2009-12-09 | 中国科学院南京土壤研究所 | The double-layer plate method of fast simple screening polycyclic aromatic hydrocarbon-degrading and screening thereof are dull and stereotyped |
Non-Patent Citations (2)
| Title |
|---|
| R. C. JOHN ET AL.: "Polycyclic Aromatic Hydrocarbon-degrading Bacteria from Aviation Fuel Spill Site at Ibeno, Nigeria", 《BULL ENVIRON CONTAM TOXICOL》 * |
| 周宜开 主编: "《中华医学百科全书 公共卫生学 环境卫生学》", 31 March 2017, 中国协和医科大学出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110283772A (en) * | 2019-07-25 | 2019-09-27 | 北京高能时代环境技术股份有限公司 | A kind of preparation method of functional flora that repairing petroleum hydrocarbon contaminated soil and underground water |
| CN111705001A (en) * | 2020-07-23 | 2020-09-25 | 桂林理工大学 | Method for rapidly screening polycyclic aromatic hydrocarbon degrading bacteria |
| CN116689477A (en) * | 2023-06-25 | 2023-09-05 | 江苏大学 | Bioremediation method of degradation flora composite edible fungus residues of polycyclic aromatic hydrocarbon contaminated soil |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109609428B (en) | 2021-11-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sarkar et al. | Microbial Consortium: A New Approach in EffectiveDegradation of Organic Kitchen Wastes | |
| CN104388312B (en) | Method for screening petroleum degrading bacteria, method for preparing petroleum degrading bacteria microbial inoculum from screened bacteria and application of microbial inoculum | |
| CN102533619B (en) | Preparation method of hydrochloric ether efficient aerobic degradation mixed bacteria and application thereof | |
| CN109609428A (en) | The screening technique of polycyclic aromatic hydrocarbon-degrading bacteria | |
| JP5383724B2 (en) | Oil-degrading microorganism, microorganism-immobilized carrier, wastewater treatment method, and wastewater treatment system | |
| CN108753662B (en) | Low-temperature dimethyl phthalate high-efficiency degrading bacterium and fermentation method thereof | |
| CN106434470B (en) | A kind of polycyclic aromatic hydrocarbon-degrading bacteria and its application | |
| JP5630855B2 (en) | Oil-degrading microorganism, microorganism-immobilized carrier, wastewater treatment method, and wastewater treatment system | |
| CN103215204A (en) | Arthrobacter strain for efficiently degrading phenanthrene and application thereof | |
| CN109913387A (en) | Degrade N-Methyl pyrrolidone enterobacteria and application in the treatment of waste water | |
| CN105647838B (en) | Skin spy's acinetobacter calcoaceticus and application thereof | |
| CN110229766A (en) | Aoxidize microbacterium and its application in degradable organic pollutant | |
| CN117603853A (en) | Klebsiella SCSIO 43801 and application thereof | |
| CN102373191A (en) | Dominant consortium flora for degrading urban waste water and preparation method thereof | |
| KR100761339B1 (en) | -2Novel strain Pseudomonas sp. KS-2P useful for degradation of endosulfan and toxic endosulfan sulfate | |
| CN101654656B (en) | Separation and purification method and application of aerobic degradation pure culture of polycyclic aromatic hydrocarbon | |
| CN104845898B (en) | Providence (Providencia sp.) 2D of one high-efficiency degradation dibutyl phthalate | |
| CN110743907A (en) | Microorganism and plant combined in-situ remediation process for petroleum-polluted soil | |
| CN110511975A (en) | A kind of brominated waste-water treatment efficient bacterium screening and culturing experimental method | |
| KR102665552B1 (en) | Novel Novosphingobium sp. strain, and uses thereof | |
| CN110157637A (en) | Enterobacteria Z1 and klebsiella Z2 composite bacteria agent removal high nitrogen pollutant effluents and application | |
| CN114940957A (en) | Paracoccus pantotrophus with facultative denitrification synchronous nitrogen and phosphorus removal performance | |
| CN109370931A (en) | A kind of composite bacteria agent of efficient degradation polycyclic aromatic hydrocarbon and its application | |
| CN107586751A (en) | One plant of dioxanes degradation bacteria D2 and its application | |
| CN108410758B (en) | Triazole degrading bacteria and application thereof in treatment of wastewater containing triazole |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |