CN105950501B - The general bacterium of one plant of degrading polycyclic aromatic hydrocarbons class organic pollutant - Google Patents
The general bacterium of one plant of degrading polycyclic aromatic hydrocarbons class organic pollutant Download PDFInfo
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- CN105950501B CN105950501B CN201610325152.8A CN201610325152A CN105950501B CN 105950501 B CN105950501 B CN 105950501B CN 201610325152 A CN201610325152 A CN 201610325152A CN 105950501 B CN105950501 B CN 105950501B
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- 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
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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
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- 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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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
- C02F2101/327—Polyaromatic Hydrocarbons [PAH's]
Abstract
The invention discloses the general Pseudomonas of one plant of degrading polycyclic aromatic hydrocarbons class organic pollutant (Pantoea sp.) PYR16, belong to microorganism and environmental organic pollutant repairing and treating field, the bacterial strain is deposited in China typical culture collection center, deposit number are as follows: CCTCC NO:M2016227.The a variety of polycyclic aromatic hydrocarbons of bacterial strain energy efficient degradation disclosed in this invention have the adaptation of extensive environment and preventing from heavy metal ability, can be colonized in rugged environment and repair by the soil or water body of heavy metal and polycyclic aromatic hydrocarbon composite pollution.
Description
Technical field
The invention belongs to microorganisms and environmental organic pollutant repairing and treating field, and in particular to one plant of degrading polycyclic aromatic hydrocarbons
The general bacterium (Pantoea sp.) of class organic pollutant, application and a kind of degrading polycyclic aromatic hydrocarbons the invention further relates to the general bacterium
The method of class organic pollutant.
Background technique
Polycyclic aromatic hydrocarbon (abbreviation PAHs) refers to what one kind was arranged by two or more phenyl ring with linear, horn shape or tufted
Condensed ring hydrocarbon, wherein the PAHs with 4 phenyl ring and 4 or more is referred to as polycyclic aromatic hydrocarbon with high molecular weight (High
Molecular weight-PAHs, abbreviation HMW-PAHs).The source of polycyclic aromatic hydrocarbon mainly includes two aspects: being on the one hand certainly
The activity on right boundary itself, such as forest fire, animals and plants remains are rotted, petroleum coal is formed, volcanic eruption；Another aspect
It is mankind's pollution behavior, such as the burning of the imperfect combustion, municipal solid wastes of fossil fuel, oil exploitation, processing transport
And Chemical Manufacture and the discharge of vehicle exhaust etc..Mankind's activity is most of polycyclic aromatic hydrocarbons contaminated to ambient contribution, is
The generally acknowledged main pollution source of environment polycyclic aromatic hydrocarbon.Polycyclic aromatic hydrocarbon is typically found in petroleum chemicals, rubber, plastics, lubricating oil, prevents
In the substances such as rust oil and unburnt organic compound, it can be occurred over long distances by the flowing of atmosphere and water body
Migration, therefore in the various surrounding mediums such as be widely distributed in atmosphere, soil, water body, deposit and plant.Due to polycyclic virtue warp
Water-soluble low, strong adsorption, long half time, allow to be continuously present in environment the several years, usually with aromatic ring number and molecule
The increase of amount, bioavailability is poorer, therefore repairs the polycyclic fragrance of high molecular weight compared to low molecular weight polycyclic aromatic hydrocarbons (PAH)
It is more difficult.The fat-soluble of polycyclic aromatic hydrocarbon allows to be enriched to step by step by food chain in the internal of food chain end people,
And it is distributed to in-vivo tissue, by the catalytic action during protein ketone-dilute alcohol tautomerism, lead to the irreversible of albumen
Become, and then makes cell that canceration occur.Therefore grave danger, the pollution amelioration of polycyclic virtue warp are caused to human health and the ecosystem
It is research hotspot both domestic and external.
Traditional polycyclic aromatic hydrocarbons contaminated reparation rely primarily on physical chemistry reparation (chemical oxidation, soil moved in improve the original or method of digging,
Pollutant in the removal of the methods of stripping, thermal desorption, burning, landfill or fixing soil), these methods often processing cost it is high,
Reparation is not thorough, is easy to produce secondary pollution and is not suitable for large-scale application.In addition, single pollutant is constituted in nature
Though environmental pollution happen occasionally, in fact the Single Pollution in absolute sense is not present, pollute more tool associations and
The combined pollution that comprehensive i.e. multiple pollutant is formed, since heavy metal can be continuously present in soil ring as polycyclic aromatic hydrocarbon
In border, therefore heavy metal-polycyclic aromatic hydrocarbon composite pollution is the principal mode of soil pollution.
Biological prosthetic is to repair pollutant using microorganism, it not only overcome physical chemistry repair process it is at high cost,
The defects of reparation is not thorough, and secondary pollution will not be generated, there is the microbial bacterial agent of repairing effect to be also suitble to pollutant
Large-scale application.The essence of microorganism remediation is using the organic pollutant in edaphon specificity metabolism soil, by it
It is converted into the process of cell other carbon-based macromoleculars and carbon dioxide.Therefore the core of organic pollutant microorganism remediation technology is
Accretion rate is fast, the screening of the strong degradation bacteria of environmental suitability.And the enrichment isolation indigenous microorganism directly from environment, not only have
Effect avoids the biosafety issues that genetic engineering bacterium occurs in practical applications, but also is a kind of simple and convenient ideal
Technological means.Screening obtains a collection of polycyclic aromatic hydrocarbon-degrading bacteria, including pseudomonad to people from all kinds of ecological environments at present
(Pseudomonas), arthrobacterium (Arthrobacter), Rhodococcus sp (Rhodococcus), mycobacteria
(Mycobacterium), bacillus (Bacillus), Sphingol single-cell (Sphingomonas), Aeromonas
(Aeromonas), Bai Yelin kirschner bacterium (Beijerinckia), nocardia (Nocardioides), bacterium of unlinking
(Cycloclasticus), Burkholderia (Burkholderia), comamonas (Polaromonas) and some thermophilic detest
For example thermophilic oily bacterium (Neptunomonas) of the Pseudomonas of oxygen, Janus bacterium (Janibacter), Nocard's bacillus (Nocardia), denaturation bacterium
(Deltaproteobacteria) and Alcaligenes (Alcaligenes) etc..
Summary of the invention
The object of the present invention is to provide a kind of general bacterium of degrading polycyclic aromatic hydrocarbons class organic pollutant.
Applicant separates from the soil of oil pollution obtains one plant of bacterium for capableing of fast degradation polyaromatic hydrocarbon pollutant
Strain is accredited as general Pseudomonas (Pantoea sp.) by Physiology and biochemistry identification and ribosomes 16S rDNA gene sequencing,
It is named as Pantoea sp.PYR16.The bacterial strain is delivered in the Wuhan University of Wuhan City, Hubei Province on April 26th, 2016
China typical culture collection center (CCTCC) preservation, deposit number are as follows: CCTCC NO:M 2016227.
Test result shows:
1) bacterial strain has the efficient degradation ability to pyrene, and the pyrene for the high concentration that can degrade.
2) bacterial strain is within the scope of 6-10 in pH, best to the degradation effect of pyrene, therefore is suitble to faintly acid to alkaline environment
Under the conditions of polycyclic aromatic hydrocarbon pollutant removal.
3) bacterial strain has extensive NaCl concentration adaptability.
4) bacterial strain has extensive Acclimation temperature ability, is more suitable for the polycyclic aromatic hydrocarbon organic pollutant under cryogenic conditions
It removes, is conducive to carry out soil in-situ reparation with northern area throughout the year in China.
5) bacterial strain is capable of the toxic action of heavy metal tolerance in a big way, maintains to polyaromatic hydrocarbon pollutant
Efficient degradation has heavy metal-polycyclic aromatic hydrocarbon composite pollution environment remediation ability.
6) bacterial strain has good degradation function to a variety of PAHs such as phenanthrene, fluorenes, fluoranthene, pyrene, especially for 4 rings and 4 rings
Polycyclic aromatic hydrocarbons (PAH) degradation effect below is significant, this is conducive to the bacterium area seriously polluted applied to combined pollutant.
7) bacterial strain have extensive preventing from heavy metal ability, this be conducive to the bacterium be colonized in rugged environment repair weighed
The soil or water body of metal and polycyclic aromatic hydrocarbon composite pollution.
More detailed technical solution is shown in that " specific embodiment " is described.
Detailed description of the invention
Fig. 1 is growth and degradation curve of the bacterial strain PYR16 under 200mg/L pyrene concentration.
Fig. 2 is degradation curve of the bacterial strain PYR16 to different starting pyrene concentration.
Fig. 3 be bacterial strain PYR16 under condition of different pH to the degradation capability of pyrene.
Fig. 4 be bacterial strain PYR16 under the conditions of different Na ion concentrations to the degradation capability of pyrene.
Fig. 5 be bacterial strain PYR16 under condition of different temperatures to the degradation capability of pyrene.
Fig. 6 be bacterial strain PYR16 under different heavy metal ion existence conditions to the degradation capability of pyrene.
Fig. 7 is degradation capability of the bacterial strain PYR16 to variety classes polycyclic aromatic hydrocarbons (PAH).
Specific embodiment
The present invention is described in detail with reference to embodiments.It should be noted that the embodiment of the present invention only limits
It is illustrated in for the present invention, without restriction effect.Involved in embodiment in relation to screening technique, buffer and
Common culture medium formula etc. can refer to Zhao Bin, " Microbiology Experiment " and " Molecular Cloning:A Laboratory guide " institute of He Shaojiang chief editor
Description content (referring to J. Pehanorm Brooker etc., 2002, Molecular Cloning:A Laboratory guide, the third edition, golden winter wild goose etc. (translating), science
Publishing house, Beijing).Other various experimental implementations, are the ordinary skill in the art involved in the present invention, without spy in text
Do not mentionlet alone bright part, those skilled in the art be referred to the present patent application day before various common tool books, section
Skill document or relevant specification, handbook etc. are practiced.
The separation of embodiment 1PYR16 bacterial strain is identified
It uses from the long-term petroleum-polluted soil of Changqing oilfields to screen soil, specific screening scheme is as follows:
1) take 10g soil, be added in the minimal medium MSM containing 200mg/L pyrene, 28 DEG C shake culture 15 days,
It is connected in the above-mentioned minimal medium MSM culture medium containing 200mg/L pyrene with 10% inoculum concentration again；Continuous switching 5 times.
2) the obtained enrichment culture liquid of 0.1ml step 1) is taken, is applied on the MSM solid plate containing pyrene, 28 DEG C static
After culture 15 days, the bacterium colony to grow on plate is transferred on the MSM solid plate containing pyrene, continues culture growth, Zhi Daoping
Occurs single bacterium colony on plate.
3) single colonie to grow out on MSM solid plate of the picking containing pyrene, is inoculated into the MSM liquid containing 200mg/L pyrene
In body culture medium, 28 DEG C shake culture 15 days, obtain bacterial strain of the invention.
Morphology and molecular biology identification further have been carried out to bacterial strain.
Firstly, the morphological feature to bacterial strain has carried out preliminary observation, the results showed that, which is Gram-negative
Bacterium, rod-short.
Meanwhile inventor separates the genomic DNA for obtaining bacterial strain, then with the universal primer of bacterial 16 S rRNA gene
27F and 1492R primer pair DNA carries out PCR amplification；The pcr amplification product of acquisition is sequenced, sequencing result such as sequence table
Shown in SEQ ID NO:1.Then, using NCBI nucleic acid database, BLAST points have been carried out according to bacterial strain 16S rRNA gene order
Analysis, the results showed that the 16S rRNA gene order of bacterial strain and the homology of general Pseudomonas are up to 99%, in conjunction with Gram's staining and shape
The Strain Designation is Pantoea sp.PYR16 by state observation, inventor.
Degradation feature of the embodiment 2PYR16 bacterial strain to pyrene
It is basic culture medium, degradation of the inventor to PYR16 bacterial strain to pyrene using MSM culture medium using pyrene as sole carbon source
Ability is tested.Firstly, taking out the bacterial strain of preservation from -80 DEG C, it is then seeded into R2It is activated 1 day in A solid medium；It chooses
Single colonie is taken to be inoculated into 5 milliliters of R2It cultivates in A fluid nutrient medium to OD600=0.5；Then it takes 0.5ml bacterium solution to be inoculated into 25ml to contain
Have in the MSM culture medium of the pyrene of 200mg/L, is placed in 28 DEG C of shake cultures.In above-mentioned degradation system, the access amount of PYR16 bacterial strain
About 105-106Left and right.Every the one bottle of culture of taking-up in 24 hours, OD is measured respectively600Value, and utilize acetone/n-hexane (1:1)
All remaining pyrenes in triangular flask are extracted, then carry out pyrene content analysis using GC-FID.Three bottles of progress parallel analysis are taken every time.
As shown in Figure 1, the pyrene in culture medium is degraded 70% or more the 15th day after inoculation PYR16 bacterial strain, by the 20th day,
82% or more pyrene is all completely removed in degradation system.This absolutely proves that bacterial strain provided by the present invention has quickly, thoroughly
Remove the ability of pyrene.
Further, PYR16 bacterial strain is inventors tested a larged to the degradation rate and degradation effect of the pyrene of different initial concentrations.
As shown in Fig. 2, the pyrene (be less than 200mg/L) of low concentration, PYR16 bacterial strain 20 days i.e. degradable pyrene is up to 80% or more；And when starting
When pyrene concentration is increased to 800mg/L, about 80% or more removal rate can be completed within 20 days after inoculation PYR16 bacterial strain.This table
Bright, PYR16 bacterial strain provided by the present invention has the ability of quickly removal high concentration pyrene.
The influence of embodiment 3pH, sodium ion and temperature to PYR16 bacterial strain pyrene degradation capability
It is respectively 4,5,6,7,8,9,10 that MSM culture medium hydrochloric acid or sodium hydroxide solution, which are adjusted to pH, first, then
10 are inoculated with according to the operating method of embodiment 25-106The PYR16 bacterial strain of left and right, 28 DEG C of shake cultures, 10 days post analysis cultivating systems
The concentration of middle residual pyrene.As shown in figure 3, being within the scope of 6-10 in pH, PYR16 bacterial strain is able to maintain that the degradation effect to pyrene,
But when pH is lower than 6, degradation capability will be significantly inhibited.Therefore, the conjunction of pyrene degradation bacteria provided by the present invention
The pH of suitable processing environment should be adjusted between 6-10, also, when the pH for handling sample is adjusted to 6-10, PYR16 bacterium
Strain can maintain 34% or more the degradation efficiency to pyrene substance.Therefore bacterial strain provided by the present invention is suitble to faintly acid to alkaline ring
Polycyclic aromatic hydrocarbon pollutant removal under the conditions of border.
Similar, inventor adds NaCl in above-mentioned MSM culture medium, make its concentration be respectively 0%, 1%, 2%, 3%,
4%, 5%, 6%, 7%, 8%, then 10 are inoculated with according to the operating method of embodiment 25-106The PYR16 bacterial strain of left and right, 28 DEG C of shakes
Swing the concentration that pyrene is remained in 10 days post analysis cultivating systems of culture.As shown in figure 4, the NaCl concentration in cultivating system is less than
When 3%, degradation rate can maintain 35% or more；When NaCl concentration is increased to 8% or so, PYR16 is to pyrene
Degradation rate remains within 30% or so.
In addition, inventor is inoculated with 10 according to the operating method of embodiment 25-106The PYR16 bacterial strain of left and right, then sets respectively
In 4 DEG C, 16 DEG C, 28 DEG C, 37 DEG C, 42 DEG C shake culture 10 days, then analyze cultivating system in remain pyrene concentration.Such as Fig. 5 institute
Show, under 28 DEG C, 37 DEG C of cultivation temperatures, PYR16 bacterial strain is able to maintain that 65% or more the degradation rate of pyrene, wherein 37 DEG C
Under cultivation temperature, above-mentioned strain is able to maintain that best degradation effect；Once and temperature promoted to 42 DEG C, degradation efficiency will decline
To 60% or so.This shows that PYR16 strain growth adaptive temperature provided by the present invention is relatively wide and is more suitable under cryogenic conditions
The removing of environment pyrene pollutant, this is beneficial to carry out soil pyrene pollution in-situ immobilization with northern area throughout the year in China.
Influence of 4 heavy metal ion of embodiment to the pyrene degradation effect of PYR16
According to inoculation method described in embodiment 2, PYR16 is inoculated into the MSM culture medium containing 200mg/L pyrene, simultaneously
It is separately added into 150mg/L manganese sulfate, 75mg/L manganese sulfate, 250mg/L copper sulphate, 125mg/L copper sulphate, 28 DEG C of shake cultures 21
After it, the concentration of remaining pyrene in degradation system is analyzed respectively.As shown in Figure 6: the manganese sulfate of 75mg/L is for PYR16 bacterial strain
For pyrene degradation capability almost without inhibition, and under the manganese sulfate existence condition of 150mg/L, PYR16 connects the degradation rate of 200mg/L pyrene
Nearly 40% or so；And PYR16 degradation capability influenced by copper sulphate it is bigger, under 125mg/L copper sulphate existence condition, bacterial strain
PYR16 bacterial strain still is able to maintain 25% or so degradation rate, and when concentration of copper sulfate is increased to 250mg/L, drop
Solution rate drops to 16% or so.This shows that PYR16 bacterial strain provided by the present invention being capable of heavy metal tolerance in a big way
Toxic action maintains the efficient degradation ability to polyaromatic hydrocarbon pollutant, has heavy metal-polycyclic aromatic hydrocarbon composite pollution environment
The ability of reparation.
Degradation effect of the embodiment 5PYR16 bacterial strain to variety classes PAHs
According to inoculation method described in embodiment 2, PYR16 is inoculated into respectively containing 200mg/L phenanthrene, 200mg/L pyrene,
In the MSM culture medium of 200mg/L fluorenes and 200mg/L fluoranthene, 28 DEG C of shake cultures after a week, are analyzed remain in degradation system respectively
Pyrene concentration.As shown in Figure 7: PYR16 has reached 98% or more to luxuriant and rich with fragrance degradation rate under identical condition of culture, almost will
Completely, also relatively good to the degradation effect of fluorenes and pyrene, degradation rate has respectively reached 60% and 35%, and can for biology for phenanthrene degradation
The extremely low fluoranthene of usability also degrades 23%, this also further illustrate polycyclic aromatic hydrocarbons (PAH) phenyl ring number increase its to biology
Toxicity is bigger, and degradation process can be more complicated, and degradation cycle also can be relatively long.Bacterial strain PYR16 is to a variety of PAHs on the whole
There is good degradation function, significant especially for 4 rings and 4 rings polycyclic aromatic hydrocarbons (PAH) degradation effect below, this is conducive to the bacterium
The area seriously polluted applied to combined pollutant.
Embodiment 6PYR16 bacterial strain is to variety classes heavy metal ion resistance
The bacterial strain that preservation is taken out from -80 DEG C, is then seeded into R2It is activated 1 day in A solid medium；The inoculation of picking single colonie
To 5 milliliters of R2It cultivates in A fluid nutrient medium to OD600=0.5.Be respectively configured in 2 milliliters of sterile centrifugation tubes containing 50mg/L,
Heavy metal solution (the configuration liquid are as follows: 10*10 of five concentration gradients of 100mg/L, 150mg/L, 200mg/L, 250mg/L6A huge sum of money
Belong to mother liquor and R2A culture medium), cultured PYR16 bacterium solution is then accessed into the heavy metal species solution according to 2% inoculum concentration
In, 28 DEG C of shaking table cultures are placed in, observation in every two days is primary and records its upgrowth situation.PYR16 is to MnCl known to the following table 12With
CuSO4There is larger tolerance, is containing 150mg/LMnCl respectively2And 250mg/LCuSO4Culture medium in still can grow,
It is to K2CrO4With Cd (NO3)2Maximum suppression growth concentration also have 100mg/L, this can be from different heavy metals pair and PYR16 generation
The mechanism of action difference for thanking to organic matter related enzyme systems is related.Show that the bacterial strain has extensive preventing from heavy metal ability, this is conducive to
The bacterium is colonized in rugged environment and repairs by heavy metal and polycyclic aromatic hydrocarbons contaminated soil or water body.
Growing state of the 1 bacterial strain PYR16 of table under different heavy metal concentrations
Note :+can grow；-- it cannot grow.
Claims (4)
1. general Pseudomonas (Pantoea sp.) PYR16 bacterial strain of one plant of degrading polycyclic aromatic hydrocarbons class organic pollutant, it is characterised in that:
It is deposited in China typical culture collection center, deposit number are as follows: CCTCC NO:M 2016227.
2. application of the general Pseudomonas PYR16 bacterial strain described in claim 1 in degrading polycyclic aromatic hydrocarbons class organic pollutant, described more
Cyclophane hydro carbons organic pollutant is phenanthrene, fluorenes, fluoranthene, pyrene.
3. a kind of method of degrading polycyclic aromatic hydrocarbons class organic pollutant, including claim 1 institute is inoculated with into contaminated sample
The step of general Pseudomonas PYR16 bacterial strain stated, the multiring aromatic hydrocarbon organic pollutant are phenanthrene, fluorenes, fluoranthene, pyrene.
4. a kind of microbial bacterial agent for degrading polycyclic aromatic hydrocarbons class organic pollutant, active constituent is described in claim 1
General Pseudomonas PYR16 bacterial strain, the multiring aromatic hydrocarbon organic pollutant be phenanthrene, fluorenes, fluoranthene, pyrene.
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| CN107306532B (en) * | 2017-06-13 | 2021-09-07 | 南京农业大学 | Method for simultaneously removing USEPA PAHs in plant body by using composite PAHs degrading bacteria |
| CN110408562B (en) * | 2019-07-12 | 2021-05-04 | 南京农业大学 | Preparation method and application of compound microbial agent for repairing cadmium-polluted soil and promoting plant growth |
| CN110773562B (en) * | 2019-11-05 | 2021-10-22 | 北京高能时代环境技术股份有限公司 | Microbial remediation method for polycyclic aromatic hydrocarbon in heavy metal-polycyclic aromatic hydrocarbon combined contaminated soil |
| CN113528371B (en) * | 2021-04-25 | 2023-06-02 | 中国石油大学(华东) | Composite microbial agent and restoration of pyrene-heavy metal composite polluted environment by same |
| CN115820494B (en) * | 2022-12-02 | 2024-03-12 | 华南理工大学 | Pantoea dispersa capable of efficiently degrading polycyclic aromatic hydrocarbon and application thereof |
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