Pseudomonas nicosulfurensis DY-8 strain and application thereof in degradation of phenanthrene and benzopyrene

Abstract

The invention discloses a Pseudomonas nicosulfurones bacterial strain and an application of the Pseudomonas nicosulfurones bacterial strain in degradation of phenanthrene and benzopyrene. According to the invention, a degradation strain Pseudomonas nicosulfurensis DY-8, which takes phenanthrene and benzopyrene as carbon sources, is obtained by domestication, separation and identification from petroleum-polluted soil in Ningbo. The invention also relates to a method for preparing the degradation strain Pseudomonas nicosulfurensis DY-8 with phenanthrene and benzopyrene as carbon sources. The DY-8 can utilize phenanthrene and benzopyrene as carbon sources, and after the DY-8 is cultured in an inorganic salt culture solution with the initial concentration of phenanthrene and benzopyrene being 50 mg.L <-1 > and 25 mg.L <-1 > respectively for 7 days, the degradation rates of phenanthrene and benzopyrene can reach 70% or above. Therefore, the strain has good application potential in the aspect of bioremediation of polycyclic aromatic hydrocarbon.

Description

Technical Field:

[0001] The present invention belongs to the field of degradation of organic pollutants, and specifically relates to the strain Pseudomonas nicosulfuronedens DY-8 and its application in the degradation of phenanthrene and benzo[a]pyrene. Background Art:

[0002] The acceleration of the modern industrial process is accompanied by the increasingly serious industrial pollution, especially persistent organic pollutants such as polycyclic aromatic hydrocarbons (PAHs). PAHs have potential carcinogenic, teratogenic, mutagenic and bioaccumulative properties, posing a major threat to the ecological environment and human health. Therefore, the problem of PAHs pollution has attracted much attention. Most of the main pollutants in important organic pollution sites such as chemical industrial parks and their surrounding soils, oil production areas, mining areas, sewage irrigation areas, etc. contain high concentrations of polycyclic aromatic hydrocarbons. Phenanthrene is a tricyclic aromatic hydrocarbon, and benzo[a]pyrene is a polycyclic aromatic hydrocarbon containing benzene rings. They both have potential carcinogenic, teratogenic, mutagenic and bioaccumulative properties, and will cause great harm to the ecological environment and human health.

[0003] The natural attenuation of toxic and harmful organic pollutants in the environment mainly depends on the metabolic action of related microorganisms. Bioremediation technology has the advantages of low cost, good effect, no secondary pollution, etc., and is the most potential remediation method for PAHs pollution repair at present. Currently, there are few reported strains for the degradation of phenanthrene and benzo[a]pyrene, mainly including Pseudomonas, Mycobacterium and Rhodococcus, etc. Since most microorganisms in the environment are unculturable, many microorganisms, especially those with specific functions, cannot be isolated and obtained by pure culture methods. Therefore, screening out strains that can effectively degrade high concentrations of phenanthrene and benzo[a]pyrene has important application value and practical significance. In this experiment, phenanthrene and benzo[a]pyrene with mass concentrations of 50 mg·L -1 and 25 mg·L -1 were used as the substrates for strain degradation, in order to provide data support for the biological treatment of polycyclic aromatic hydrocarbons. Summary of the Invention:

[0004] The first object of the present invention is to provide a strain Pseudomonas nicosulfuronedens DY-8 with the ability to degrade phenanthrene and / or benzo[a]pyrene. It was deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC) on February 26, 2025. Address: 5th Floor, Building 59, No. 100 Yard, Xianlie Middle Road, Guangzhou, Guangdong Province, Zip Code: 510070, Deposit Number: GDMCC No: 65987.

[0005] This study reports that the strain is Pseudomonas nicosulfuronedens, which can be isolated from various terrestrial and aquatic habitats. There are relatively few reports on the pollutant degradation of Pseudomonas nicosulfuronedens at present, and there have been no studies on the degradation of phenanthrene and benzo[a]pyrene by Pseudomonas nicosulfuronedens at home and abroad. In this study, a strain DY-8 that uses high-concentration phenanthrene and benzo[a]pyrene as carbon sources was domesticated and isolated from an oil-polluted soil in Ningbo, identified, and its growth characteristics were studied. At the same time, a microbial agent was prepared to explore its degradation characteristics of phenanthrene and benzo[a]pyrene, providing a reference for the bioremediation of PAHs-polluted environments.

[0006] The second object of the present invention is to provide the use of the above-mentioned Pseudomonas nicosulfuronedens DY-8 in the degradation of phenanthrene and / or benzo[a]pyrene.

[0007] Preferably, the degradation of phenanthrene and / or benzo[a]pyrene is the degradation of phenanthrene and / or benzo[a]pyrene in oil-polluted soil.

[0008] Preferably, Pseudomonas nicosulfuronedens DY-8 is applied to an environment polluted by phenanthrene and / or benzo[a]pyrene for the degradation of phenanthrene and / or benzo[a]pyrene.

[0009] The third object of the present invention is to provide a phenanthrene and / or benzo[a]pyrene-degrading microbial agent, which contains the above-mentioned Pseudomonas nicosulfuronedens DY-8 as an active ingredient.

[0010] The fourth object of the present invention is to provide a method for degrading phenanthrene and / or benzo[a]pyrene, which is to sprinkle the above-mentioned Pseudomonas nicosulfuronedens DY-8 into an environment containing phenanthrene and / or benzo[a]pyrene for the degradation of phenanthrene and / or benzo[a]pyrene.

[0011] Preferably, Pseudomonas nicosulfuronedens DY-8 is sprinkled into an environment polluted by phenanthrene and / or benzo[a]pyrene for the degradation of phenanthrene and / or benzo[a]pyrene.

[0012] Preferably, Pseudomonas nicosulfuronedens DY-8 is sprinkled into oil-polluted soil for the degradation of phenanthrene and / or benzo[a]pyrene.

[0013] The present invention domesticates and separates a degradation strain DY-8 using phenanthrene and benzopyrene as carbon sources from a petroleum-contaminated soil in Ningbo. According to the strain morphology, 16S rDNA gene sequencing analysis and phylogenetic analysis, the strain is identified as Pseudomonas nicosulfuronedens DY-8. The optimal environmental conditions for the growth of the strain are: temperature of 28°C, pH value of 7, and no addition of sodium chloride; the 16S rDNA gene sequencing analysis results of DY-8 show that the strain most similar to it is Pseudomonas nicosulfuronedens strain LAM1902 (99.86%). DY-8 can use phenanthrene and benzopyrene as carbon sources, and the initial concentrations of phenanthrene and benzopyrene are 50 mg·L -1 and 25 mg·L -1 After culturing in inorganic salt culture medium for 7 days, the degradation rate of phenanthrene and benzopyrene can reach more than 70%. Therefore, this strain has good application potential in the bioremediation of polycyclic aromatic hydrocarbons.

[0014] Pseudomonas nicosulfuronedens DY-8, which was deposited in Guangdong Microbiological Culture Collection Center (GDMCC) on February 26, 2025, address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province, Postal Code: 510070, and the deposit number is: GDMCC No: 65987. Description of the drawings:

[0015] Figure 1 1 is the morphological characteristics of strain DY-8 in Example 1, wherein a is the front and back of DY-8 grown on a solid medium with LB as the carbon source for 12 hours; b is a scanning electron microscope image of DY-8: no flagella, scale bar is 2 μm.

[0016] Figure 2 It is the phylogenetic relationship of strain DY-8 and its related bacteria based on 16s rRNA gene sequence in Example 1. The construction method is the neighbor-joining method. The bootstrap value is set to repeat 1000 times. Only the results with a bootstrap value greater than 50% are shown in the figure. The scale 0.005 represents the substitution rate of each nucleotide.

[0017] Figure 3 The strain DY-8 in Example 2 was grown under different culture temperatures, salinities and pH values.

[0018] Figure 4 is the degradation efficiency of strain DY-8 in Example 3 in an inorganic salt medium containing high concentrations of phenanthrene and benzopyrene (the initial concentrations of phenanthrene and benzopyrene were 50 mg·L -1 and 25 mg·L -1 ). Detailed implementation manners:

[0019] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. In addition, the technical features involved in the various implementation manners of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0020] Example 1 Isolation and identification of Pseudomonas nicosulfuronedens DY-8

[0021] 1 Materials and methods

[0022] 1.1 Sample source

[0023] Soil samples were collected near a certain petroleum polluted factory in Ningbo and were acclimated for a long time with high-concentration phenanthrene and benzo[a]pyrene as carbon sources respectively. Through multiple screening and isolation and purification, efficient phenanthrene and benzo[a]pyrene degrading bacteria were obtained.

[0024] 1.2 Culture media

[0025] 1.2.1 Inorganic salt medium

[0026] The inorganic salt medium is used for the enrichment culture of microorganisms in the samples and the degradation experiments of phenanthrene and benzo[a]pyrene under pure bacteria conditions. The formula of this medium is shown in Table 1 (containing phenanthrene and benzo[a]pyrene solutions). Its preparation method is to add each component to the solvent water, mix evenly, and sterilize to obtain.

[0027] Table 1 Formula of inorganic salt medium

[0028]

[0029] 1.2.2 Nutrient medium

[0030] The nutrient medium is used for the conventional cultivation of microorganisms such as the isolation, purification, preservation, and activation of bacteria. The types and components of the liquid nutrient medium used in this experiment are shown in Table 2. If a solid medium needs to be prepared for the experiment, only 1.5 - 2% agar powder needs to be added on the basis of the original medium formula. If there is no special instruction for the cultivation conditions of the strain, the pH of the medium is adjusted to 7. The preparation method of the nutrient medium is to add each component to the solvent water, mix evenly, and sterilize to obtain.

[0031] Table 2 Components of Luria-Bertani medium (LB)

[0032]

[0033] 1.3 Domestication, screening and isolation of strains

[0034] The collected contaminated soil was added to the enrichment medium (the above inorganic salt medium), and phenanthrene and benzo[a]pyrene with concentrations of 50 mg·L -1 and 25 mg·L -1 were used as the degradation substrates respectively, and placed in a 28 °C incubator for dark shaking culture. The inorganic salt medium with phenanthrene and benzo[a]pyrene as the carbon source was used for strain domestication, and 7 days was one domestication cycle. A 10% inoculum was transferred to a fresh enrichment medium with the same culture system and the above enrichment process was repeated. This was repeated three times.

[0035] The obtained fourth-generation enrichment culture samples were spread and separated by the dilution plate method, and the samples were separated with the nutrient medium. The spread samples were placed in the original culture temperature condition for cultivation. After about 48 hours, obvious single colonies formed on the surface of the medium. According to the characteristics such as the morphological size, color, transparency, etc. of the colonies, several different single colonies were picked and streaked and purified on the nutrient medium plate for cultivation. If different characteristic single colonies could still be observed on the plate after streaking and purification, they were streaked and separated again until only single colonies with the same characteristics could be observed on the same plate. 1 strain of strain DY-8 with high-efficiency degradation performance for both phenanthrene and benzo[a]pyrene was screened in the experiment. The purified single colonies were picked into the corresponding liquid nutrient medium and cultured to the logarithmic phase, and the bacterial liquid and sterile glycerol were mixed and aliquoted into sterile 2 ml cryotubes (glycerol concentration was 15%), and placed at -80 °C for long-term storage.

[0036] 1.4 Identification of strains

[0037] Strain DY-8 was identified according to its morphological characteristics and molecular biological characteristics.

[0038] 1.4.1 Morphological characteristics

[0039] DY-8 is a bacterium isolated from an oil-contaminated soil in Ningbo. After activation, it can form colonies with a diameter of about 1.0 mm, white, smooth surface, slightly convex upward, opaque, without spores, without flagella, and the strain individuals are short rod-shaped on the plate made of the inorganic salt medium under aerobic conditions at 28 °C after growing for 12 h. The cell size is approximately 0.4 - 0.6 × 1.2 - 1.8 μm( Figure 1 ).

[0040] 1.4.2 Molecular biological characteristics

[0041] The identification of molecular biological characteristics mainly includes sequencing and the construction of phylogenetic trees. Before sequencing and constructing phylogenetic trees, it is necessary to extract the DNA of bacteria (the bacterial genomic DNA rapid extraction kit used in the experiment is from Beijing Aidlab Biotechnologies Co., Ltd.). In order to study the taxonomy of bacteria, it is usually necessary to amplify the 16S rRNA gene and construct a phylogenetic tree. The amplified gene is a section of DNA that constitutes a part of the rRNA encoded in prokaryotes. Because of its high conservation, specificity, and appropriate sequence length, it is usually used to detect and identify bacteria.

[0042] Polymerase chain reaction (PCR) is mainly used to amplify different gene fragments. PCR requires different primers (27F and 1492R); the system of PCR amplification reaction: 10×buffer 2.5 μl, Mg 2+ (25 mmol / l) 1.5 μl, dNTP (25 mmol / l) 0.3 μl, forward primer (10 mmol / l) 0.5 μl, reverse primer (10 mmol / l) 0.5 μl, Taq enzyme: 0.25 μl, DNA group template 0.1 μl, deionized water 19.35 μl. The conditions of PCR amplification reaction: denaturation at 95°C, annealing at 55°C, extension at 72°C. This process is cycled 30 times, and extension is carried out at 72°C for 10 min. After the PCR reaction is completed, it is stored at 4°C. After amplifying the required gene, a gel block is prepared with 0.75 - 1% agarose and added with the nucleic acid stain GelRed. The PCR product and the DNA marker (maker) containing various length fragments are added to the gel block and placed in an electrophoresis apparatus. The electrophoresis apparatus is filled with TBE (Tris-borate) buffer, and the electrophoresis apparatus is operated at a certain voltage for 20 min and then taken out and observed under a 300 nm ultraviolet lamp to determine the success of the PCR product amplification reaction. Then the successfully amplified PCR product is sent to BGI Tech Solutions Co., Ltd. for sequencing, and the sequencing primers are the same as the amplification primers.

[0043] The bacterial 16S rRNA gene sequences obtained by sequencing were uploaded to EzTaxon-e (http: / / eztaxon-e.ezbiocloud.net / ). This website will compare the submitted sequences with the 16S rRNA gene sequences of the type strains of recognized species to obtain the similarity information between the sequences. According to the results of sequence alignment analysis, the corresponding type strain can be selected as the reference strain for the isolated strains in this experiment. At the same time, the 16S rRNA gene sequence of the reference strain can be obtained, and a phylogenetic analysis can be constructed to prove the differences between the reference strain and the isolated strains in the experiment, so as to identify the isolated strains. The phylogenetic tree was constructed using the MEGA 5.05 program. Usually, the neighbor-joining method, the minimum evolution method, and the maximum parsimony method are used to construct the phylogenetic tree. Among them, the neighbor-joining method is the most commonly used, and the bootstrap value is usually set to be calculated 1000 times repeatedly.

[0044] A 16S rRNA gene sequence with a length of 1393 bp was obtained by PCR and gene sequencing. Through 16S rRNA gene alignment, it was found that the gene similarity of this strain with Pseudomonas nicosulfuronedens strain LAM1902 (accession number MN007089 in GenBank) was 99.86%. From the above results, it can be concluded that the isolated bacterium DY-8 in this experiment is Pseudomonas nicosulfuronedens.

[0045] A phylogenetic tree was constructed using the 16S rRNA gene sequence of DY-8 and the 16S rRNA gene sequences with relatively high similarity to it, so as to obtain the homology results between the 16S rRNA gene of DY-8 and the 16S rRNA genes with relatively high similarity to it. The phylogenetic tree constructed by the neighbor-joining method is shown in Figure 2 . At present, there are few reports on the application of this strain in the environmental field. Therefore, obtaining highly efficient phenanthrene- and benzo[a]pyrene-degrading bacteria has important theoretical and practical significance for the treatment and in-depth remediation of soil PAHs pollution containing phenanthrene and benzo[a]pyrene.

[0046] The 16S rRNA gene sequence of DY-8 is shown in SEQ ID NO.1, specifically:

[0047]

[0048] From the above results, it can be concluded that the strain DY-8 isolated in this experiment is the species of Pseudomonas nicosulfuronedens. It was named Pseudomonas nicosulfuronedens DY-8, which was deposited in Guangdong Microbiological Culture Collection Center (GDMCC) on February 26, 2025, address: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City, Guangdong Province, Postal Code: 510070, and the deposit number is: GDMCC No: 65987.

[0049] Example 2 Growth conditions of Pseudomonas nicosulfuronedens DY-8

[0050] Determination of growth temperature:

[0051] Prepare the liquid nutrient medium required for the growth of the strain (Example 1), and sterilize it in an autoclave after preparation. Inoculate the activated strain Pseudomonas nicosulfuronedens DY-8 into the culture medium (experimental group), use a culture medium without inoculation of bacteria as a control (control group), and culture the culture medium at different temperatures for 12 hours. The control group and the experimental group corresponding to each temperature have three replicates, and observe the growth of bacteria. When the results are difficult to distinguish with the naked eye, use a visible-ultraviolet spectrophotometer to measure the absorbance of the culture medium at a wavelength of λ=600nm, and finally obtain the growth temperature and the optimal growth temperature range of the new bacteria. The test temperatures are as follows: 13°C, 18°C, 23°C, 28°C, 33°C, and 38°C.

[0052] Determination of growth pH:

[0053] Prepare the liquid nutrient medium required for the growth of the strain (Example 1), and adjust the pH of the culture medium with the following buffer systems: pH 4.0 - 5.0, 0.1 mol / l sodium citrate and 0.1 mol / l citric acid; pH 6.0 - 8.0, 0.1 mol / l NaOH and 0.1 mol / l KH₂PO₄; pH 9.0 - 10.0, 0.1 mol / l NaHCO₃ and 0.1 mol / l Na₂CO₃; pH 11.0, 0.1 mol / l NaOH and 0.05 mol / l Na₂HPO₄. Inoculate Pseudomonas nicosulfuronedens DY-8 into the medium, with three replicates for each pH, and use the medium without inoculated bacteria as a control. Place the medium in the optimal temperature for the growth of the new bacteria and culture for 12 h, and observe the growth of the bacteria. When the results are difficult to distinguish by the naked eye, measure the absorbance value of the medium at a wavelength of λ = 600 nm with a visible-ultraviolet spectrophotometer. Finally, obtain the pH range for the growth of the new bacteria and the optimal growth pH range. The tested pH values are as follows: 4.0, 5.0, 6.0, 7.0, 8.0, 9.0.

[0054] Salt concentration tolerance:

[0055] Prepare the liquid nutrient medium required for the growth of the strain (Example 1), and adjust the salt concentration of the medium. Inoculate the activated new bacteria Pseudomonas nicosulfuronedens DY-8 into the sterilized medium, with three replicates for each salt concentration, and use the medium without inoculated bacteria as a control. Place the medium under the optimal conditions for the growth of the new bacteria and culture for 12 h, and observe the growth of the bacteria. When the situation is difficult to distinguish by the naked eye, measure the absorbance value of the medium at a wavelength of λ = 600 nm with a visible-ultraviolet spectrophotometer. Finally, obtain the salt concentration range that the new bacteria can tolerate. The tested salt concentrations are as follows: mass fraction 0%, 1%, 2%, 3%, 4%, 5%.

[0056] The carbon source utilization, acid production and other tests are all tested using API ID 32GN and API 20NE microbial identification kits (BioMérieux).

[0057] The results are as Figure 3 shown. In the nutrient broth medium, DY-8 can grow under the temperature conditions of 13 - 38 °C, and the optimal growth temperature is 28 °C, the enrichment temperature of this bacterium; this bacterium can grow under the pH conditions of 4.0 - 9.0, and the optimal growth pH is 7.0; the salt tolerance ability of this bacterium is weak, and it can grow under the conditions of a salt concentration of 0% to 4%, and grows best under salt-free conditions.

[0058] Example 3: Degradation Experiment of Phenanthrene and Benzo[a]pyrene by Pseudomonas nicosulfuronedens DY-8

[0059] The activated strain Pseudomonas nicosulfuronedens DY-8 was inoculated into the inorganic salt culture medium (Example 1) containing an initial phenanthrene concentration of 50 mg / L or an initial benzo[a]pyrene concentration of 25 mg / L at an inoculation amount of 10% by mass fraction, and cultured under dark shaking for 7 days at a temperature of 28 °C, pH 7.0, and without sodium chloride addition. The treatment without adding strain DY-8 was used as the control group.

[0060] Samples from each treatment were taken for chemical analysis, and the specific steps were as follows: (1) Sample pretreatment: Dichloromethane was added to each culture sample for extraction, and at the same time, 5 μL of a recovery indicator with a concentration of 200 mg / L was added (for phenanthrene and benzo[a]pyrene treatment samples, deuterated polycyclic aromatic hydrocarbons were added). After sufficient shaking, it was transferred to a separatory funnel and left to stand. After stratification, the organic phase was collected, the lower layer liquid was put back into the flask and extracted again with an equal volume of dichloromethane. The extraction solutions were combined and transferred to a flat-bottomed flask containing an appropriate amount of activated copper chips for rotary evaporation until concentrated to about 2 mL. A small amount of n-hexane (about 5 mL) was added, and rotary evaporation was carried out until 2 mL remained. This was repeated three times to replace the organic solvent with n-hexane. The concentrated solution after replacement was purified using a glass-packed column (with a diameter of about 9 mm). The column packing from bottom to top was 3 cm of 3% deactivated neutral alumina, 3 cm of 3% deactivated silica gel, and 1 cm of anhydrous sodium sulfate. The column was activated with an appropriate amount of n-hexane, and a 15 mL n-hexane / dichloromethane (volume ratio 1:1) mixed reagent was used to wash the packed column, and the eluate was collected in a brown reagent bottle at about 15 mL and concentrated to about 0.5 mL by nitrogen blowing. Finally, it was transferred to a 1.5 mL vial and stored frozen. Before measurement on the instrument, 5 μL of the internal standard hexamethylbenzene with a concentration of 200 mg / L was added. (2) Instrumental analysis: The content of PAHs in each treatment sample was determined by using an Agilent 7890 gas chromatograph - 5975 mass spectrometer in combination. The chromatographic column used was an Agilent DB 5-MS capillary chromatographic column (column length 30 m, inner diameter 0.25 mm, film thickness 0.25 μm). The obtained data was processed using an Agilent chromatographic workstation, and the quantification of phenanthrene and benzo[a]pyrene was carried out using a 6-point calibration curve and the internal standard method. The determination of the microbial cell concentration was carried out by the photoelectric turbidimetry method, expressed as OD, that is, the optical density value of the ultraviolet light transmitted through the measured bacterial liquid sample at a wavelength of 600 nm.

[0061] According to the determination and analysis by GC-MS, it was found that strain DY-8 could degrade phenanthrene and benzo[a]pyrene, and after culturing for 7 days in the inorganic salt culture medium containing 50 mg / L and 25 mg / L of phenanthrene and benzo[a]pyrene, the degradation rate could reach over 70%( Figure 4) It is illustrated that DY-8 is a strain that can degrade both phenanthrene and benzo[a]pyrene and has a strong tolerance to these two compounds.

[0062] Conclusion:

[0063] 1) A phenanthrene- and benzo[a]pyrene-degrading bacterium DY-8 that can grow using phenanthrene and benzo[a]pyrene as carbon sources respectively was enriched and isolated from oil-polluted soil in Ningbo.

[0064] 2) The strain DY-8 can form colonies that are about 1.0 mm in diameter, white, smooth on the surface, slightly convex upward, opaque, without spores, without flagella, and the individual strain is short rod-shaped. According to molecular biology analysis, it can be concluded that the bacterium DY-8 isolated in this experiment is the strain Pseudomonas nicosulfuronedens, and its phylogenetic tree was drawn. There are few reports on the application of this strain at present, especially the research on using it to degrade phenanthrene and benzo[a]pyrene has not been reported.

[0065] 3) The optimal growth conditions for the strain DY-8 are a temperature of 28 °C, a pH of 7.0, and no NaCl addition. DY-8 can use phenanthrene and benzo[a]pyrene as carbon sources respectively to degrade them. After culturing in an inorganic salt culture solution with initial concentrations of phenanthrene and benzo[a]pyrene of 50 mg·L -1 and 25 mg·L -1 for 7 days, the degradation rate can reach over 70%. In summary, DY-8 is a strain that can degrade phenanthrene and benzo[a]pyrene and has a strong tolerance to phenanthrene and benzo[a]pyrene, has a strong adaptability to polycyclic aromatic hydrocarbons, and has good application potential in bioremediation.

Claims

1. Pseudomonas nicosulfuronedens DY-8, with the deposit number: GDMCC No: 65987.

2. Use of Pseudomonas nicosulfuronedens DY-8 according to claim 1 in degrading phenanthrene and / or benzo[a]pyrene.

3. The application according to claim 2, wherein The degradation of phenanthrene and / or benzo[a]pyrene is the degradation of phenanthrene and / or benzo[a]pyrene in petroleum-polluted soil.

4. The application according to claim 2, characterized in that It is to apply Pseudomonas nicosulfuronedens DY-8 in an environment polluted by phenanthrene and / or benzo[a]pyrene to degrade phenanthrene and / or benzo[a]pyrene.

5. A phenanthrene and / or benzo[a]pyrene degrading bacterial agent, characterized in that, Contains Pseudomonas nicosulfuronedens DY-8 according to claim 1 as an active ingredient.

6. A method for degrading phenanthrene and / or benzo[a]pyrene, characterized in that, Sprinkle Pseudomonas nicosulfuronedens DY-8 according to claim 1 into an environment containing phenanthrene and / or benzo[a]pyrene to degrade phenanthrene and / or benzo[a]pyrene.

7. The method according to claim 6, wherein It is to sprinkle Pseudomonas nicosulfuronedens DY-8 into an environment polluted by phenanthrene and / or benzo[a]pyrene to degrade phenanthrene and / or benzo[a]pyrene.

8. The method according to claim 6, characterized in that, It is to sprinkle Pseudomonas nicosulfuronedens DY-8 into petroleum-polluted soil to degrade phenanthrene and / or benzo[a]pyrene.

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